Secondary health conditions in people with spinal cord injury in South Africa: Prevalence and associated factors.
People with spinal cord injury (SCI) experience preventable secondary health conditions (SHCs) that worsen the disability, reduce the quality of life and affect health and wellbeing. There is limited information on the prevalence of SHCs and the associated factors to inform planning and practice in South Africa (SA). To identify the prevalence of SHCs and the associated factors in people with SCI. We conducted a retrospective review of patients' medical records at a tertiary academic hospital and a rehabilitation hospital in Gauteng Province, SA. Data collected included demographic data, injury profile, SHCs and associated factors. Data were summarised using descriptive statistics of frequency and percentages. Fisher's exact test was used to determine the association between SHCs and sociodemographic and clinical variables. The Mann-Whitney U-test was used to determine the associated risk factors for SHCs. Multinomial regression was used to determine the predictors of the frequency of SHCs. A total of 425 records were reviewed, 68.0% of patients were male, and the median (interquartile range) age was 45 (35 - 56) years. The majority (93.7%) of the patients had SHCs, and 78% had ≥2 SHCs. Significant predictors of having >3 SHCs were the duration of SCI (p=0.01), site of injury in the upper (p=0.03) and lower (p=0.01) thoracic spine, being unemployed (p=0.04), and public hospital income classification (H0 = social grant/unemployed, H1 = earning ZAR0 - 70 000 per annum single income/ZAR0 - 100 000 per annum household income, and Road Accident Fund; p=0.03, p=0.03 and p=0.01, respectively). Secondary health conditions were prevalent among patients with SCI. People with SCI experience multimorbidity that requires multiple management strategies. These findings point to the need for prevention strategies to minimise the occurrence of SHCs.


The interpretation of shadows on the adult chest radiograph.
In this article I consider that part of the interpretation of a chest radiograph that is concerned with abnormal opacities. The recognition of various shadow patterns and their interpretation in terms of gross pathology is central to the approach.


High-performance and -efficiency cardiomyocyte-based potential biosensor for temporal-specific detection of ion channel marine toxins.
Paralytic shellfish toxins (e.g., saxitoxin, STX; gonyautoxin-2, GTX-2) and tetrodotoxin (TTX) are highly toxic and widely distributed ion channel marine toxins which specifically block the voltage-dependent sodium channels (VDSCs), causing great harm to human health. It is urgent to exploit new detection methods with high specificity and high efficiency. Here, a portable high-throughput cardiomyocyte-based potential biosensor was established with cardiomyocytes, a 16-well microelectrodes (MEs) sensor and a robust 32-channel recording system, which presented high-quality and high-consistency extracellular field potential (EFP) signals in each well with a long duration of 80 h. The feature parameters, including firing rate (FR), spike amplitude (SA), spike slope (SS), spike duration (SD) and field potential duration (FPD), were extracted from EFP to quantitatively assess the toxic effects of these ion channel toxins. Importantly, the biosensor showed temporal specificity and parametric selectivity under toxin treatments, and FR, SS and SD were the optimal parameters to STX, TTX and GTX-2, respectively. This biosensor can rapidly detect 0.29 ng/mL STX, 0.30 ng/mL TTX and 0.16 ng/mL GTX-2 within 5 min, 10 min and 15 min, respectively. Thus, our novel multi-well cardiomyocyte-based biosensor will be a promising tool for high-effective detection of ion channel toxins.


The impact of therapeutic massage on adult residents living with complex and high level disabilities: A brief report.
Research into the effects of touch in disabled adults in residential care remains largely unexplored in the current literature. Evidence suggests however, that massage therapy may improve mood state, including anxiety and stress, reduce pain and improve sleep/wake behavior and fatigue. These benefits are of importance as they have substantial impact on quality of life. This pilot study evaluated the effect of therapeutic massage on the quality of life of adults with complex care needs living in residential care. Participants were recruited from three residential homes (Queensland, Australia) for 18-65 year olds with severe disability. 25 participants were recruited and received a massage program consisting of five weeks of twice weekly massages. Structured interviews were conducted pre-post intervention. Additionally, mood was ascertained preceding and following each massage session. Mood of participants improved markedly immediately following massage session (p < 0.05) and pre-massage mood was observed to increase over the study period. However, pre- and post-intervention measures indicated massage did not improve pain, sleepiness, depression or stress levels or sustain positive mood three days post-intervention. Participants' satisfaction with their current health significantly improved (Z = -2.51, p = 0.012), as did their satisfaction with their current happiness (Z = -2.06, p = 0.04), suggesting that massage therapy offered some improvement in quality of life. The results of this pilot indicates that massage may be of benefit to people living with high care needs and represents a practical innovation providing tactile stimulation that may be integrated into care.


On the benefit of aberration-corrected HAADF-STEM for strain determination and its application to tailoring ferroelectric domain patterns.
Revealing strains on the unit-cell level is essential for understanding the particular performance of materials. Large-scale strain variations with a unit-cell resolution are important for studying ferroelectric materials since the spontaneous polarizations of such materials are strongly coupled with strains. Aberration-corrected high-angle-annular-dark-field scanning transmission electron microscopy (AC-HAADF-STEM) is not so sensitive to the sample thickness and therefore thickness gradients. Consequently it is extremely useful for large-scale strain determination, which can be readily extracted by geometrical phase analysis (GPA). Such a combination has various advantages: it is straightforward, accurate on the unit-cell scale, relatively insensitive to crystal orientation and therefore helpful for large-scale. We take a tetragonal ferroelectric PbTiO3 film as an example in which large-scale strains are determined. Furthermore, based on the specific relationship between lattice rotation and spontaneous polarization (Ps) at 180° domain-walls, the Ps directions are identified, which makes the investigation of ferroelectric domain structures accurate and straightforward. This method is proposed to be suitable for investigating strain-related phenomena in other ferroelectric materials.


Design, synthesis, molecular docking, molecular dynamic simulation, and MMGBSA analysis of 7-O-substituted 5-hydroxy flavone derivatives.
A series of chrysin derivatives were designed, synthesized, and evaluated for their antibacterial activity against four different bacterial strains. We have synthesized new propyl-substituted and butyl-substituted chrysin-piperazine derivatives, which show marvellous inhibition against E. coli and S. aureus. The free hydroxyl group at the C-5 position of chrysin improved therapeutic efficacy in vivo and was a beneficial formulation for chemotherapy. All synthesized compounds were confirmed by various spectroscopic techniques such as IR, NMR, HPLC, and mass spectrometry. The compounds exhibited moderate to good inhibition, and their structure-activity relationship (SAR) has also been illustrated. Among the synthesised compounds, compounds 4 and 10 were the most active against S. pyogenes and E. coli, with 12.5 g/mL MICs; additionally, compound 12 exhibits significant activity on both the S. aureus and E. coli stains. Based on the promising activity profile and docking score of compound 12, it was selected for 100 ns MD simulation and post-dynamic binding free energy analysis within the active sites of S. aureus TyrRS (PDB ID: 1JIJ) and E. coli DNA GyrB (PDB ID: 6YD9) to investigate the stability of molecular contacts and to establish how the newly synthesized inhibitors fit together in the most stable conformations.Communicated by Ramaswamy H. Sarma.


Incidental discovery of subtle-variant septooptic dysplasia in an adult with headaches: a novel case with pathophysiologic implications.
Septooptic dysplasia (SOD) is a congenital central nervous system malformation syndrome classically associated with the triad of agenesis of the septum pellucidum, optic nerve hypoplasia, and pituitary abnormalities. It has been suggested that SOD may result from in utero vascular insults. We present the case of an adult male with personal and family histories of intracranial vascular pathology in whom SOD was incidentally discovered, and we describe how the specific abnormalities in this case could be related to vascular pathology.


Ecotoxicity of parathion during its dissipation mirrored by soil enzyme activity, microbial biomass and basal respiration.
The application of parathion (PTH) in agriculture can result in its entry into the soil and threaten the soil environment. Monitoring the PTH residues and assessing toxicity on soil health are of paramount importance to the public. Herein, the dissipation of PTH and concomitant influence on microbial activities [FDA hydrolase (FDA‒H), microbial biomass carbon (MBC) and basal respiration (BR)] in coastal solonchaks were investigated. Results showed that the dissipation of PTH in tested soil declined linearly, and the half-lives varied from 5.6 to 56.8 days, depending on pollutant concentrations. The FDA‒H activity and MBC were negatively affected by PTH pollution and exhibited a significantly positive correlation. Two‒way ANOVA analysis demonstrated that microbial activities were affected not only by PTH dose and incubation time but also by their interactions. The integrated biomarker response (IBR/n) index values on day 120 were between 1.02 and 2.89, larger than those on day 1 during PTH dissipation. This implied that the soil quality did not recover though there was no PTH residue in the soil at the end of the experiment. These findings suggested that microbial activities integrated with IBR/n index could elucidate the hazardous impacts of PTH dissipation on biochemical cycling and microorganisms in soil.


Put on notice. New York report could have hospital execs, boards looking over their shoulders.
A New York state commission wants to give state officials the power to remove management and boards at poorly performing hospitals in Brooklyn and possibly recognition that the borough's healthcare system is dysfunctional. "There's some competition that really doesn't benefit anybody" and is inefficient, says Richard Becker, left, of Brooklyn Hospital Center.


Heart Transplant in Asia.
The prevalence of heart failure has increased in Asia. A significant proportion of patients with heart failure and left ventricular dysfunction end up with advanced heart failure or end-stage heart disease. These patients may be placed on the waiting list for heart transplant. There are more than 10 countries in Asia that have an active heart transplant program. The number of heart transplants performed is limited despite an increase in the number of patients with end-stage heart failure mainly because of donor shortage, which may be related to religious belief and inefficient allocation policy.


Hydrogen peroxide-mediated SERCA cysteine 674 oxidation contributes to impaired cardiac myocyte relaxation in senescent mouse heart.
A hallmark of aging of the cardiac myocyte is impaired sarcoplasmic reticulum (SR) calcium uptake and relaxation due to decreased SR calcium ATPase (SERCA) activity. We tested the hypothesis that H2O2-mediated oxidation of SERCA contributes to impaired myocyte relaxation in aging. Young (5-month-old) and senescent (21-month-old) FVB wild-type (WT) or transgenic mice with myocyte-specific overexpression of catalase were studied. In senescent mice, myocyte-specific overexpression of catalase (1) prevented oxidative modification of SERCA as evidenced by sulfonation at Cys674, (2) preserved SERCA activity, (3) corrected impaired calcium handling and relaxation in isolated cardiac myocytes, and (4) prevented impaired left ventricular relaxation and diastolic dysfunction. Nitroxyl, which activates SERCA via S-glutathiolation at Cys674, failed to activate SERCA in freshly isolated ventricular myocytes from senescent mice. Finally, in adult rat ventricular myocytes in primary culture, adenoviral overexpression of SERCA in which Cys674 is mutated to serine partially preserved SERCA activity during exposure to H2O2. Oxidative modification of SERCA at Cys674 contributes to decreased SERCA activity and impaired myocyte relaxation in the senescent heart. Strategies to decrease oxidant levels and/or protect target proteins such as SERCA may be of value to preserve diastolic function in the aging heart.


Optimal sampling sites for repeat prostate biopsy: a recursive partitioning analysis of three-dimensional 26-core systematic biopsy.
To explore an optimal combination of sampling sites to detect prostate cancer in a repeat biopsy setting. A transrectal ultrasound-guided systematic three-dimensional 26-core biopsy (3D26PBx), a combination of transrectal 12 and transperineal 14 core biopsies, was performed in 235 Japanese men with prior negative biopsy. Using recursive partitioning, we evaluated cancer detection of all possible combinations of sampling sites and selected the combination that provides the highest cancer detection rate at a given number of biopsy cores. Prostate cancer was detected in 87 of the 235 (37%) men. The 3D26PBx improved cancer detection by 89% relative to the conventional transrectal sextant biopsy. Neither Gleason score nor percentage of Gleason 4/5 cancers differed between cancers with and without positive cores within the transrectal sextant-sampling sites. A three-dimensional combination of transrectal and transperineal approaches outperformed either transrectal or transperineal approach alone. Recursive partitioning revealed that a three-dimensional 16-core (transrectal eight cores plus transperineal eight cores) biopsy could detect all the cancers with the minimum number of cores. We propose a three-dimensional combination of transrectal eight cores taken from the far lateral peripheral zone and the parasagittal base, and transperineal eight cores taken from the anterior and posterior apex and the transition zone as an optimal set of sampling sites for repeat biopsy.


Characterization of the RNA dependent DNA polymerase of a new human T-lymphotropic retrovirus (lymphadenopathy associated virus).
We described here the characteristics of the Reverse Transcriptase activity associated with the Lymphadenopathy Associated Virus ( LAV ). A critical concentration of non ionic detergent, all four deoxyribonucleosides triphosphates and the divalent cation Mg2+ are required for optimal endogenous enzyme activity. The endogenous reaction product is digested by DNase and not by RNase and its synthesis is only slightly inhibited by actinomycin D. Exogenous reactions are optimal using poly A oligo dT12 -18 or poly Cm oligo dG12 -18 as template primer and Mg2+ as divalent cation. This enzyme can be distinguished from other cellular DNA polymerases activities and from Terminal deoxynucleotidyl Transferase (TdT) by purification from LAV infected T lymphocytes using phosphocellulose column.


Immune-mediated peripheral neuropathies and voltage-gated sodiums channels.
Antibodies to GM1 ganglioside are found in some patients with the Guillain-Barré syndrome and multifocal motor neuropathy, and may alter neuronal excitability. We measured voltage-gated sodium channel (VGSC) function by 22Na+ influx in a motor neuronal cell line (NSC19) in which we demonstrated GM1 ganglioside and tetrodotoxin-sensitive VGSC function. We were unable to detect any effect of peripheral neuropathy plasmas, with or without complement, on VGSC function in NSC19 cells.


Bromelain, a cysteine protease from pineapple (Ananas comosus) stem, is an inhibitor of fungal plant pathogens.
This study aimed to evaluate the effect of bromelain, a cysteine protease isolated from pineapple (Ananas comosus), on growth of several agronomically important fungal pathogens. Purification of bromelain from pineapple stems was carried out by chromatography techniques, and its antimicrobial activity was tested against the fungal pathogens Fusarium verticillioides, Fusarium oxysporum and Fusarium proliferatum by broth microdilution assay. A concentration of 0.3 μmol l(-1) of bromelain was sufficient for 90% growth inhibition of F. verticillioides. The capability of bromelain to inhibit fungal growth is related to its proteolytic activity. The study demonstrates that stem bromelain exhibits a potent antifungal activity against phytopathogens and suggests its potential use as an effective agent for crop protection. The results support the use of a natural protease that accumulates at high levels in pineapple stems as alternative to the use of chemical fungicides for crop protection.


Cytostatic activity of organic compounds and their average quasi-valence number.
It is found that the average quasi-valence numbers of alkylating cytostatics lie in the region of potential carcinogens, while the average quasi-valence numbers of antimetabolites predominantly cover the region of noncarcinogens. Implications of this finding on the design of new drugs are discussed.


Development and validation according to the SANTE guidelines of a QuEChERS-UHPLC-QTOF-MS method for the screening of 204 pesticides in bivalves.
A qualitative screening high resolution mass spectrometry method was developed and validated according to the EU SANTE/12682/2019 guidelines for the analysis of 204 pesticides in seven commercial bivalve species spiked at three concentrations (0.01, 0.05 and, 0.1 mg.kg-1). Samples were extracted using QuEChERS and analysed using ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry. The QuEChERS method was optimised by the Taguchi Orthogonal Array approach. The best conditions were obtained with pure ACN, MgSO4/NaCl as extraction salts, MgSO4/PSA/C18 as clean-up, and the non-dilution of extracts. The impact of different HRMS acquisition modes on detection and identification rates were also evaluated. The screening detection limits were determined to be 0.01 mg.kg-1 and 0.1 for 66% and 87% of pesticides, respectively. These screening procedure was finally applied to different bivalve samples using target and suspect analysis. This allowed the identification of diuron and its metabolite 1-(3,4-dichlorophenyl)-3-methylurea in the investigated samples.


The SLC22 drug transporter family.
The SLC22 family comprises organic cation transporters (OCTs), zwitterion/cation transporters (OCTNs), and organic anion transporters (OATs). These transporters contain 12 predicted alpha-helical transmembrane domains (TMDs) and one large extracellular loop between TMDs 1 and 2. Transporters of the SLC22 family function in different ways: (1) as uniporters that mediate facilitated diffusion in either direction (OCTs), (2) as anion exchangers (OAT1, OAT3 and URAT1), and (3) as Na(+)/ l-carnitine cotransporter (OCTN2). They participate in the absorption and/or excretion of drugs, xenobiotics, and endogenous compounds in intestine, liver and/or kidney, and perform homeostatic functions in brain and heart. The endogenous substrates include monoamine neurotransmitters, choline, l-carnitine, alpha-ketoglutarate, cAMP, cGMP, prostaglandins, and urate. Defect mutations of transporters of the SLC22 family may cause specific diseases such as "primary systemic carnitine deficiency" or "idiopathic renal hypouricemia" or change drug absorption or excretion.


Anesthetic management of external cephalic version.
Breech presentation is common at term and its reduction through external cephalic version represents a noninvasive opportunity to avoid cesarean delivery and the associated maternal morbidity. In addition to uterine relaxants, neuraxial anesthesia is associated with increased success of version procedures when surgical anesthetic dosing is used. The intervention is likely cost effective given the effect size and the avoided high costs of cesarean delivery.


Quality of life, pain of prepectoral and subpectoral implant-based breast reconstruction with a discussion on cost: A systematic review and meta-analysis.
Prepectoral implant-based breast reconstruction (PIBR) has regained popularity, despite decades-long preference for subpectoral implant placement. This paper aims to compare patient-reported outcomes (PRO) between prepectoral and subpectoral approaches to implant-based breast reconstruction (IBBR). The primary PRO was with the BREAST-Q, and postoperative pain scores, while the secondary outcomes were complication rates. A comprehensive literature search of the PubMed library was performed. All studies on patients undergoing IBBR after mastectomy that compared prepectoral to subpectoral placement and PROM or postoperative pain were included. A total of 3789 unique studies of which 7 publications with 216 and 332 patients who received prepectoral and subpectoral implants, respectively, were included for meta-analysis. Patients with prepectoral implant placement had significantly higher satisfaction with the outcome (p = 0.03) and psychosocial well-being (p = 0.03) module scores. The pain was lower in patients with prepectoral implants on postoperative day 1 (p<0.01) and day 7 (p<0.01). The subgroup analysis of prepectoral breast implants showed that complete acellular dermal matrix coverage had lower rates of wound dehiscence (p<0.0001), but there were no significant differences in complications between one-stage and two-stage procedures. Overall, patients with prepectoral implants reported higher BREAST-Q scores and lower postoperative pain and lower complications rates than patients with subpectoral implants. In appropriately selected patients, prepectoral implant placement with ADM coverage, be it the primary placement of an implant or placement of a tissue expander before definitive implant placement, should be the modality of choice in patients who choose IBBR. Further research should focus on patient selection, strategies to reduce cost and cost-benefit analysis of PIBR.


Ethyl-eicosapentaenoate (E-EPA) attenuates motor impairments and inflammation in the MPTP-probenecid mouse model of Parkinson's disease.
Parkinson's disease (PD) is a neurodegenerative disorder, characterized by hypokinesia, but also mood and cognitive disorders. Neuropathologically, PD involves loss of nigrostriatal dopamine (DA) and secondary non-dopaminergic abnormalities. Inflammation may contribute to PD pathogenesis, evident by increased production of pro-inflammatory cytokines. PD onset has been positively associated with dietary intake of omega-(n)-6 polyunsaturated fatty acids (PUFA). On the other hand, omega-(n)-3 PUFA may benefit PD. One of these n-3 PUFA, eicosapentaenoic acid (EPA), is a neuroprotective lipid with anti-inflammatory properties, but its neuroprotective effects in PD are unknown. Thus, we presently tested the hypothesis that EPA can protect against behavioral impairments, neurodegeneration and inflammation in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-probenecid (MPTP-P) mouse model of PD. MPTP-P injections caused hypokinesia in the rotorod and pole test, hyperactivity in the open field, and impaired mice on the cued version (procedural memory) of the Morris water maze. MPTP-P caused a loss of nigrostriatal DA and altered neurochemistry in the frontal cortex and hippocampus. Furthermore, striatal levels of pro-inflammatory cytokines were increased, while the brain n-3/n-6 lipid profile remained unaltered. Feeding mice a 0.8% ethyl-eicosapentaenoate (E-EPA) diet prior to MPTP-P injections increased brain EPA and docosapentaenoic acid (DPA) but not docosahexaenoic acid (DHA) or n-6 PUFA. The diet attenuated the hypokinesia induced by MPTP-P and ameliorated the procedural memory deficit. E-EPA also suppressed the production of pro-inflammatory cytokines. However, E-EPA did not prevent nigrostriatal DA loss. Based on this partial protective effect of E-EPA, further testing may be warranted.


Diet quality in an ethnically diverse sample of children and adolescents with autism spectrum disorder compared with nationally representative data.
Children with developmental disabilities are at an increased risk of unhealthy eating habits, which may contribute to compromised growth and development. Children with autism spectrum disorder (ASD) exhibit unique risk factors for unhealthy dietary patterns, including sensory issues and cognitive rigidity at mealtimes. This cross-sectional study examined diet quality in a sample of children with ASD in Florida compared to nationally representative National Health and Nutrition Examination Survey (NHANES) 2009-2014/2013-2014 data using the Healthy Eating Index-2015 (HEI-2015). A 24-h food record was completed by 41 parents of children with ASD aged 2-17 years, and food and beverage items consumed by each child were reported per standardized 24-h recall protocol. Two models were used to compare mean total and component HEI-2015 scores to NHANES means: (1) comparing means for our full sample to published NHANES means for children aged 2-18 years and (2) a matched model with subjects matched 1:1 by age, gender, race/ethnicity, and parent education level. HEI component scores were significantly lower (poorer) in children with ASD for whole fruit, total vegetables, dairy, total protein foods, and seafood and plant protein. Whole grains, fatty acids, added sugars, and refined grains scores were higher (better) in our sample. However, total HEI scores and HEI scores for all 13 components were similar among children with ASD and the matched cases from the NHANES data. There are potential discrepancies in diet quality between children with ASD and general population. Further research with a larger sample size, reporting both total and component HEI scores, is needed.


Relationships between neural crest cells and catecholamine in suckling mice.
In suckling mice injected i.p. with a 27 mg/kg dose of L-hydrochloric acid isoproterenol, multiple neural crest tumors developed and cell death of neural crest cells occurred. It is speculated that neural crest derivatives may be beta-receptor cells and contain regulatory units of neurotransmission mediated through cyclic AMP.


Insights into the inhibitory mechanism of skullcapflavone II against α-synuclein aggregation and its mediated cytotoxicity.
The dangerous self-assembled and infectious seeds of α-synuclein (αSN) play primary roles in Parkinson's disease. Accordingly, the inhibition of αSN fibrillation and elimination of toxic aggregates are the main therapeutic strategies. Skullcapflavone II (S.FII), a compound isolated from S. pinnatifida, has shown multiple neuroprotective features. Herein, we demonstrated that S.FII inhibited αSN aggregation with IC50 of 7.2 μM. It increased nucleation time and decreased fibril elongation rate and the species formed in the presence of S.FII were unable to act as seeds. Additionally, S.FII inhibited both secondary nucleation and seeding of αSN and disaggregated the mature preformed fibrils as well. The species formed in the presence of S.FII showed less toxicity. It also preserved neurite length and dopamine content of SH-SY5Y cells and attenuated the inflammatory responses in mixed glial cells. The Localized Surface Plasmon Resonance (LSPR) analysis indicated that S.FII interacts with αSN. Docking simulation studies on αSN fibrils revealed that S.FII could interact with the key residues of the salt bridges and glutamine ladder, which might lead to the destruction of fibril's structures. We also showed that S.FII passes through the blood-brain barrier in vitro and in vivo. Overall, these findings elucidate the neuroprotective roles of S.FII in reducing αSN pathogenicity.


Prevalence of overweight/obesity, hypertension and its associated factors among women from Northeast India.
To assess prevalence of overweight/obesity, hypertension and its risk factors among women from North-eastern States of India. A community based cross-sectional study was carried out in two north-eastern States. Information was collected on household's socio-economic & demographic particulars. Height (cm), weight (kg), waist and hip circumferences and blood pressure were measured on all the selected women. Association between overweight/obesity, hypertension with socio-demographic variables was tested using Chi-square and logistic regression was done. Total 1047 women were covered from both the states. The prevalence of chronic energy deficiency was 19% & 10%, overweight/obesity (BMI≥23) was 17.5% & 26% and hypertension was 15% & 17% (age adjusted 19.6% & 17%) respectively among women from Meghalaya & Nagaland. The prevalence of pre-hypertension was observed more among women from Nagaland (36.5%) as compared to Meghalaya (18.3%). Only 31% women were aware of hypertension and 6% had history of hypertension and 82% of them were on treatment. The prevalence of hypertension was observed high among 36-49 years and among overweight/obese women. On logistic regression, only age, BMI and use of additional salt was observed to be significantly associated with hypertension, while living in pucca house was associated with overweight/obesity. The prevalence of hypertension was similar in both the states (15-17%) and pre-hypertension was high among women from Nagaland. This is warning sign for women in Nagaland. There is an urgent need to undertake more health & nutrition education sessions along with regular check-up for early diagnosis and treatment of hypertension.


Control of action as mediated by the human frontal lobe.
Conscious control of action involves the voluntary initiation and the continuous adjustment of motor activity. Neuroimaging data provide evidence that the plan for a movement is developed with respect to the behavioral context in prefrontal cortex, while the synergies of a motor program are coded by premotor cortex and the specific movement parameters by the motor cortex. It is suggested that the initiational aspects of conscious motor activity are implemented in a medial system of information flow and the integrative aspects in a lateral system of the human frontal lobe.


A two-tiered mechanism for stabilization and immobilization of E-cadherin.
Epithelial tissues maintain a robust architecture which is important for their barrier function, but they are also remodelled through the reorganization of cell-cell contacts. Tissue stability requires intercellular adhesion mediated by E-cadherin, in particular its trans-association in homophilic complexes supported by actin filaments through beta- and alpha-catenin. How alpha-catenin dynamic interactions between E-cadherin/beta-catenin and cortical actin control both stability and remodelling of adhesion is unclear. Here we focus on Drosophila homophilic E-cadherin complexes rather than total E-cadherin, including diffusing 'free' E-cadherin, because these complexes are a better proxy for adhesion. We find that E-cadherin complexes partition in very stable microdomains (that is, bona fide adhesive foci which are more stable than remodelling contacts). Furthermore, we find that stability and mobility of these microdomains depend on two actin populations: small, stable actin patches concentrate at homophilic E-cadherin clusters, whereas a rapidly turning over, contractile network constrains their lateral movement by a tethering mechanism. alpha-Catenin controls epithelial architecture mainly through regulation of the mobility of homophilic clusters and it is largely dispensable for their stability. Uncoupling stability and mobility of E-cadherin complexes suggests that stable epithelia may remodel through the regulated mobility of very stable adhesive foci.


Assisted reproductive treatment outcomes of women with endometriomas: Either with or without previous ovarian surgery.
The present study aimed to evaluate and compare the assisted reproductive technique (ART) outcomes of women with endometriomas either with or without prior endometrioma surgery. A total of 122 women with endometriomas underwent intracytoplasmic sperm injection-embryo transfer (ICSI-ET) at a tertiary IVF Center, between 2014 and 2019, were included in this retrospective study. Of this group, 38 patients had recurrent endometriomas and 84 patients had primary endometrioma without a previous endometrioma surgery. The outcomes of ART treatment including cancellation rates before ET, numbers of oocytes obtained, implantation, clinical pregnancy and live birth rates were compared between the groups. A logistic regression model including potential confounders as age and presence of male factor infertility was used to evaluate the possible effect of recurrent endometriomas on the live birth. The baseline characteristics of the groups were similar. The poor ovarian response rate, defined as the harvest of fewer than four oocytes, was identified in 35.7% and 42.1% of primary and recurrent endometrioma groups, respectively. The implantation (27.2 ± 42.7% vs 24.1 ± 41.4%, P = .74), clinical pregnancy/ET (30.9% vs 27.6%, P = .93 ) and live birth rates/ET (22.1% vs 17.2%, P = .79) CONCLUSION: The ART outcomes of patients with primary and recurrent endometriomas do not seem different in terms of response to ovarian stimulation and live birth rates after ICSI. These results may indicate that the recurrence of the endometrioma might not have a further detrimental effect on ART outcome than the disease itself.


Evaluating the autoinduction expression system and one-step purification for high-level expression and purification of gallbladder-derived rhIL-1Ra.
Recent advancement in fermentation technologies resulted in the increased yields of recombinant proteins of biopharmaceutical and medicinal importance. Consequently, there is an important task to develop simple and easily scalable methods that can facilitate the production of high-quality recombinant protein. Most of the recent reports described the expression of recombinant human IL-1 receptor antagonist (rhIL-1Ra) in Escherichia coli using isopropyl-β-d-thiogalacto pyranoside (IPTG), a nonmetabolizable and expensive compound, as an expression inducer. In this study, we describe the expression and one-step purification of gallbladder-derived rhIL-1Ra by autoinduction in E. coli. This method includes special media that automatically induce the target protein expression from T7 promoter and allow the production of the target protein in high yield than the conventional IPTG induction method. In addition to fermentation process improvements, one-step purification strategy is essential to make the process economical. We developed a single-step cation exchange chromatography and obtained 300 mg/L of rhIL-1Ra with 98% purity. Purified protein was characterized by SDS-PAGE and Ion exchange HPLC (IEX-HPLC). The described method can be used to scale up the production of rhIL-1Ra and other recombinant proteins.


Thrombosis of second-generation drug-eluting stents in real practice results from the multicenter Spanish registry ESTROFA-2 (Estudio Español Sobre Trombosis de Stents Farmacoactivos de Segunda Generacion-2).
This study sought to evaluate second-generation drug-eluting stent (DES) thrombosis in clinical practice. First-generation DES are associated with a significant incidence of late thrombosis. There is paucity of data regarding real practice late thrombosis incidence and predictors with second-generation DES, zotarolimus-eluting stent (ZES), and everolimus-eluting stents (EES). A prospective, large-scale, non-industry-linked multicenter registry was designed. Complete clinical-procedural data and systematic follow-up of all patients treated with these stents was reported in a dedicated registry supported by the Spanish Working Group on Interventional Cardiology. From 2005 to 2008, 4,768 patients were included in 34 centers: 2,549 treated with ZES, and 2,219 with EES. The cumulative incidence of definite/probable thrombosis for ZES was 1.3% at 1 year and 1.7% at 2 years and for EES 1.4% at 1 year and 1.7% at 2 years (p = 0.8). The increment of definite thrombosis between the first and second year was 0.2% and 0.25%, respectively. In a propensity score analysis, the incidence remained very similar. Ejection fraction (adjusted hazard ratio [HR]: 0.97; 95% confidence interval [CI]: 0.95 to -0.99; p = 0.008), stent diameter (adjusted HR: 0.37; 95% CI: 0.17to 0.81; p = 0.01) and bifurcations (adjusted HR: 2.1; 95% CI: 1.14 to 3.7; p = 0.02) emerged as independent predictors of thrombosis. In the subgroup of patients with bifurcations, the use of ZES was independently associated with a higher thrombosis rate (adjusted HR: 4; 95% CI: 1.1 to 13; p = 0.03). In a real practice setting, the incidence of thrombosis at 2 years with ZES and EES was low and quite similar. The incidence of very late thrombosis resulted lower than was reported in registries of first-generation DES. In the subset of bifurcations, the use of ZES significantly increased the risk of thrombosis.


Platelet activating factor (PAF) and activity of its biosynthetic and catabolic enzymes in blood and leukocytes of male patients with newly diagnosed heart failure.
To evaluate platelet activating factor (PAF) levels, its metabolic enzymes activity and its associations with other inflammatory markers in heart failure (HF) patients. PAF, and two of its key biosynthetic enzymes [lyso-PAF acetyltransferase (lyso-PAF-AT) and DTT-insensitive CDP-choline:1-alkyl-2-acetyl-sn-glycerol cholinephosphotransferase (PAF-CPT)] along with its catabolic enzymes [PAF-acetylhydrolase (PAF-AH) and lipoprotein-associated phospholipase-A(2) (Lp-PLA(2))] were measured in serum and leukocytes of twelve newly diagnosed male HF patients. Serum CRP, TNF-alpha, IL-6, sCD14 and CD40L were also determined. PAF ranged from 0.03 to 5.6 pmol/mL. Median lyso-PAF-AT, PAF-CPT, PAF-AH and Lp-PLA(2) activities were 4.1, 68.42, 644.44 pmol/min/mg protein and 51.42 pmol/min/microL correspondingly. Lyso-PAF-AT and PAF-CPT activities positively correlated with CRP, IL-6 and with each other, whereas PAF-CPT activity correlated with sCD14 and CD40L (P<0.05). PAF's biosynthetic enzyme activities correlated with inflammatory and immunologic molecules, which are activated in HF. Our study indicates a potential role of PAF in HF patients.


Recent studies on the enterotropic strain of avian infectious bronchitis virus.
Avian infectious bronchitis (AIB) is an economically important disease of chickens. Recent studies have revealed enterotropism by at least one strain of AIB virus with pathological lesions in parts of the gut. This review highlights the findings of the studies so far made on this enterotropic strain of AIB virus.


The interaction of SIRT4 and Calreticulin during ER stress in glia cells.
Endoplasmic Reticulum (ER) stress is the response that occurs after the dysfunction of ER and its structure. Activated UPR triggers a stress response using ER membrane proteins such as PERK, IRE-1, GRP78, ATF5 ve ATF6. Sirtuins are enzymes that carry out post-translational modifications such as deacetylation and ADP-ribosylation. In our previous study, we identified Calreticulin as a SIRT4-interacting protein via mass spectrometry. Calreticulin binds to misfolded proteins, prevents them from leaving ER, which results in the reduction of ER stress. In this study, we aimed to investigate the interaction between SIRT4 and Calreticulin during ER stress in glia cells (IHA-immortalized human astrocytes). To trigger ER stress in glia cells, we first optimized the dose and the duration of the Tunicamycin which is 2.5 μg/ml concentration for 16 h. SIRT4 gene was silenced with lentiviral particles using 4 MOI (Multiplicity of Infection). In SIRT4-silenced cells, when treated with 2.5 μg/ml Tunicamycin for 16 h, the increase in the expressions of ATF6, GRP78 and the ratio of spliced/unspliced XBP1 mRNA were reduced. This shows that silencing SIRT4 may decrease ER stress. SIRT4-Calreticulin interaction was shown both in control and ER-stress induced glia cells. Additionally, this interaction did not change with the ER stress. SIRT4 only ADP-ribosylates Calreticulin during ER stress. Normally, SIRT4 ADP-ribosylates and deactivates Calreticulin during ER stress condition. When SIRT4 is silenced, the ADP-ribosylation level of Calreticulin decreases resulting in the activation of Calreticulin and the reduction of ER stress. In summary, SIRT4 inhibitors may be investigated as protective agents or drug candidates in neurodegenerative diseases where ER stress mostly underlies as one of the molecular mechanisms.


Acetyl-CoA carboxylase 2-/- mutant mice are protected against fatty liver under high-fat, high-carbohydrate dietary and de novo lipogenic conditions.
Hepatic fat accumulation resulting from increased de novo fatty acid synthesis leads to hepatic steatosis and hepatic insulin resistance. We have shown previously that acetyl-CoA carboxylase 2 (Acc2(-/-)) mutant mice, when fed a high-fat (HF) or high-fat, high-carbohydrate (HFHC) diet, are protected against diet-induced obesity and maintained whole body and hepatic insulin sensitivity. To determine the effect of an ACC2 deletion on hepatic fat metabolism, we studied the regulation of the enzymes involved in the lipogenic pathway under Western HFHC dietary and de novo lipogenic conditions. After completing the HFHC regimen, Acc2(-/-) mutant mice were found to have lower body weight, smaller epididymal fat pads, lower blood levels of nonesterified fatty acids and triglycerides, and higher hepatic cholesterol than wild-type mice. Significant up-regulation of lipogenic enzymes and an elevation in hepatic peroxisome proliferator-activated receptor-γ (PPAR-γ) protein were found in Acc2(-/-) mutant mice under de novo lipogenic conditions. The increase in lipogenic enzyme levels was accompanied by up-regulation of the transcription factors, sterol regulatory element-binding proteins 1 and 2, and carbohydrate response element-binding protein. In contrast, hepatic levels of the PPAR-γ and PPAR-α proteins were significantly lower in the Acc2(-/-) mutant mice fed an HFHC diet. When compared with wild-type mice fed the same diet, Acc2(-/-) mutant mice exhibited a similar level of AKT but with a significant increase in pAKT. Hence, deleting ACC2 ameliorates the metabolic syndrome and protects against fatty liver despite increased de novo lipogenesis and dietary conditions known to induce obesity and diabetes.


The temperate marine phage PhiHAP-1 of Halomonas aquamarina possesses a linear plasmid-like prophage genome.
A myovirus-like temperate phage, PhiHAP-1, was induced with mitomycin C from a Halomonas aquamarina strain isolated from surface waters in the Gulf of Mexico. The induced cultures produced significantly more virus-like particles (VLPs) (3.73 x 10(10) VLP ml(-1)) than control cultures (3.83 x 10(7) VLP ml(-1)) when observed with epifluorescence microscopy. The induced phage was sequenced by using linker-amplified shotgun libraries and contained a genome 39,245 nucleotides in length with a G+C content of 59%. The PhiHAP-1 genome contained 46 putative open reading frames (ORFs), with 76% sharing significant similarity (E value of <10(-3)) at the protein level with other sequences in GenBank. Putative functional gene assignments included small and large terminase subunits, capsid and tail genes, an N6-DNA adenine methyltransferase, and lysogeny-related genes. Although no integrase was found, the PhiHAP-1 genome contained ORFs similar to protelomerase and parA genes found in linear plasmid-like phages with telomeric ends. Southern probing and PCR analysis of host genomic, plasmid, and PhiHAP-1 DNA indicated a lack of integration of the prophage with the host chromosome and a difference in genome arrangement between the prophage and virion forms. The linear plasmid prophage form of PhiHAP-1 begins with the protelomerase gene, presumably due to the activity of the protelomerase, while the induced phage particle has a circularly permuted genome that begins with the terminase genes. The PhiHAP-1 genome shares synteny and gene similarity with coliphage N15 and vibriophages VP882 and VHML, suggesting an evolutionary heritage from an N15-like linear plasmid prophage ancestor.


Antidepressant drug prescription and incidence of COVID-19 in mental health outpatients: a retrospective cohort study.
Currently, the main pharmaceutical intervention for COVID-19 is vaccination. While antidepressant (AD) drugs have shown some efficacy in treatment of symptomatic COVID-19, their preventative potential remains largely unexplored. Analysis of association between prescription of ADs and COVID-19 incidence in the population would be beneficial for assessing the utility of ADs in COVID-19 prevention. Retrospective study of association between AD prescription and COVID-19 diagnosis was performed in a cohort of community-dwelling adult mental health outpatients during the 1st wave of COVID-19 pandemic in the UK. Clinical record interactive search (CRIS) was performed for mentions of ADs within 3 months preceding admission to inpatient care of the South London and Maudsley (SLaM) NHS Foundation Trust. Incidence of positive COVID-19 tests upon admission and during inpatient treatment was the primary outcome measure. AD mention was associated with approximately 40% lower incidence of positive COVID-19 test results when adjusted for socioeconomic parameters and physical health. This association was also observed for prescription of ADs of the selective serotonin reuptake inhibitor (SSRI) class. This preliminary study suggests that ADs, and SSRIs in particular, may be of benefit for preventing COVID-19 infection spread in the community. The key limitations of the study are its retrospective nature and the focus on a mental health patient cohort. A more definitive assessment of AD and SSRI preventative potential warrants prospective studies in the wider demographic.


Acetate mediates a microbiome-brain-β-cell axis to promote metabolic syndrome.
Obesity, insulin resistance and the metabolic syndrome are associated with changes to the gut microbiota; however, the mechanism by which modifications to the gut microbiota might lead to these conditions is unknown. Here we show that increased production of acetate by an altered gut microbiota in rodents leads to activation of the parasympathetic nervous system, which, in turn, promotes increased glucose-stimulated insulin secretion, increased ghrelin secretion, hyperphagia, obesity and related sequelae. Together, these findings identify increased acetate production resulting from a nutrient-gut microbiota interaction and subsequent parasympathetic activation as possible therapeutic targets for obesity.


Emission characteristics and light absorption apportionment of carbonaceous aerosols: A tunnel test conducted in an urban with fully enclosed use of E10 petrol.
To reduce the heavy dependence on petroleum, bioethanol has been increasingly employed as an alternative and sustainable transportation fuel. However, the characteristics of black carbon (BC) emissions from E10 petrol vehicles (i.e., ethanol-gasoline containing 10% ethanol) are still unclear, especially under real driving conditions. Here, a tunnel test was conducted during a cold winter. This tunnel was characterized by heavy traffic comprising more than 98% E10-fueled gasoline vehicles (GVs). Real-time BC concentrations, traffic parameters and meteorological conditions were recorded during the sampling campaign. The average BC concentration inside the tunnel (10.94 ± 5.02 μg m-3) was almost twice the background concentration. Based on aethalometer AE33 in situ measurements and the minimum R-squared (MRS) method, real-time aerosol light absorption was apportioned. The light absorption proportions of BC, primary brown carbon (BrC1) and secondary brown carbon (BrC2) were 79.86%, 2.78% and 17.36%, respectively, at 370 nm. The BC emission factor (EFBC) of the E10-fueled vehicles was 1.09 ± 0.49 mg km-1·veh-1 and 15.24 ± 6.85 mg·(kg fuel)-1, lower than those of traditional gasoline fueled vehicles in previous studies. This study can support the compilation of vehicular BC emission inventories, provide recommendations for biofuel policies and contribute to comprehensively understanding the climatic impact of E10 petrol.


Prolonged systemic delivery of streptokinase using liposome.
To prolong the biological half-life of streptokinase, a thrombolytic agent, streptokinase-bearing liposome with and distearolyphosphatidyl ethanolamine-N-poly (ethylene glycol) 2000 (DSPE-PEG 2000) was prepared and evaluated. Streptokinase-bearing liposomes composed of distearolyphosphatidylcholine (DSPC), cholesterol and cholesterol-3-sulfate with DSPE-PEG 2000 was prepared by the freeze-thawing method and administered via femoral vein to rats (15,000 IU/kg). The activity of streptokinase in plasma was determined by the method based on the amidolytic activity of streptokinase-plasminogen complex. Pharmacokinetic parameters of streptokinase incorporated in liposomes were compared with those of streptokinase alone. The T1/2 and AUCinfinity of streptokinase incorporated in DSPC-PEG liposome increased 16.3- and 6.1-fold, respectively, compared with those of streptokinase alone. Streptokinase-bearing long-circulating liposome could increase the circulation time of streptokinase in blood and expect longer thrombolytic activity compared with streptokinase alone.


Pregnancy in mother with Glanzmann's thrombasthenia and isoantibody against GPIIb-IIIa: Is there a foetal risk?
Glanzmann's thrombasthenia (GT) is a rare autosomal recessive platelet disorder caused by qualitative or quantitative abnormalities of a platelet glycoprotein complex (GPIIb-IIIa) leading to excessive bleeding. Platelet transfusions are the first-line therapy for severe or persistent bleeding and surgery. Isoantibody against GPIIb-IIIa complexes present on normal platelets can be observed in Glanzmann's thrombasthenia type I patients after platelet transfusion possibly leading to platelet transfusion refractoriness. Pregnancy in Glanzmann's thrombasthenia type I women is rare, and severe bleeding can be observed in the peripartum or late postpartum period. Moreover, pregnancy can contribute to the maternal isoimmunization by the passage of the foetal cells into the maternal circulation. The transplacental passage of the maternal isoantibodies can induce moderate to severe foetal thrombocytopenia. We discuss here the case of in utero death at 31 weeks of gestation due to intracranial haemorrhage in an immunized mother and review the literature. Presence of isoantibody prior to gestation or detected during the index pregnancy must be taken into account in evaluating risk for the mother and the foetus.


The influence of body mass index on in vitro fertilization outcome.
To examine whether body mass index (BMI) influences the outcome of in vitro fertilization (IVF). We studied 516 IVF cycles, 438 undergone by nonobese (BMI < or = 30) and 78 by obese (BMI > 30) women who all had an a priori favorable prognosis (age < 40 years and first, second, or third IVF cycle). Pregnancy was achieved in 122 (27.9%) nonobese and 12 (15.4%) obese women. The obese women required significantly longer stimulation and more gonadotropin ampoules, and had lower peak estradiol levels and a significantly lower fertilization rate; obese poor responders had a significantly lower pregnancy rate than nonobese poor responders; and the prevalence of poor responders was significantly higher among obese than nonobese women (28.2% vs 16.9%, P < 0.04). While the likelihood of poor responders was increased among obese women, reasonable conception rates were achieved in nonobese poor responders, and were comparable to the rates in nonobese and obese normal responders.


Application of the Mandible Injury Severity Score to Pediatric Mandibular Fractures.
The Mandible Injury Severity Score (MISS) has been used to evaluate adult mandibular fractures. The purpose of this study was to evaluate the MISS in a cohort of pediatric patients. This was a retrospective study of pediatric patients treated for mandibular fractures over a 20-year period. Patients were included if they had computed tomographic imaging available for review and had at least 1 post-treatment visit. The primary predictor variable was the MISS. Secondary predictors were demographic and injury-associated factors. The outcome was treatment-associated complications. Descriptive, bivariate, and multiple logistic regression statistics were computed. One hundred sixteen patients with mandibular fractures were identified; 73 (62.9%) met the inclusion criteria. The sample's mean age was 8.5 ± 4.1 years; 44% were girls. Motor vehicle collisions (60%) and falls (15.1%) were the most common mechanisms. More than 50% of patients had an extra-mandibular injury. The mean MISS was 13.5 ± 7.8. Forty-five percent of the sample underwent open reduction and internal fixation. Complications were noted in 20.5% of patients, of which malocclusion was the most common (8.2%). Increasing MISS was associated with complications (P < .001). After controlling for the effects of age, mechanism, cervical spine and skull base injuries, and treatment, patients with an MISS of at least 14 were significantly more likely to have a complication (odds ratio = 4.0; 95% confidence interval, 1.05-15.0; P = .04). In pediatric patients with mandibular fractures, increased severity of injury is associated with complications, even after controlling for the effects of multiple confounders, including open treatment.


Rgl-exomiR-7972, a novel plant exosomal microRNA derived from fresh Rehmanniae Radix, ameliorated lipopolysaccharide-induced acute lung injury and gut dysbiosis.
Plant-derived exosome-like nanoparticles (ELNs) have been proposed as a novel therapeutic tool for preventing human diseases. However, the number of well-verified plant ELNs remains limited. In this study, the microRNAs in ELNs derived from fresh Rehmanniae Radix, a well-known traditional Chinese herb for treating inflammatory and metabolic diseases, were determined by using microRNA sequencing to investigate the active components in the ELNs and the protection against lipopolysaccharide (LPS)-induced acute lung inflammation in vivo and in vitro. The results showed that rgl-miR-7972 (miR-7972) was the main ingredient in ELNs. It exerted stronger protective activities against LPS-induced acute lung inflammation than catalpol and acteoside, which are two well-known chemical markers in this herb. Moreover, miR-7972 decreased the production of pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α), reactive oxygen species (ROS) and nitric oxide (NO) in LPS-exposed RAW264.7 cells, thereby facilitating M2 macrophage polarization. Mechanically, miR-7972 downregulated the expression of G protein-coupled receptor 161 (GPR161), activating the Hedgehog pathway, and inhibited the biofilm form of Escherichia coli via targeting virulence gene sxt2. Therefore, miR-7972 derived from fresh R. Radix alleviated LPS-induced lung inflammation by targeting the GPR161-mediated Hedgehog pathway, recovering gut microbiota dysbiosis. It also provided a new direction for gaining novel bioactivity nucleic acid drugs and broadening the knowledge on cross-kingdom physiological regulation through miRNAs.


Characterization of a Novel Protease Inhibitor from the Edible Mushroom Agaricus bisporus.
Protease inhibitors inhibit the activity of protease enzymes; hence, they are essentially involved in the regulation of the metabolic processes involving protease enzymes and the protection of the host organism against external damage due to proteases. These inhibitors are abundantly present in all living organisms but have not been much reported in mushrooms. Mushrooms are one of the major food components of humans, with delicious taste and high nutritional value. Mushrooms also have therapeutic and economic significance. The edible mushrooms with medicinal properties are much in commercial demand. To date, the presence of protease inhibitors has not been reported much in edible mushrooms. The present study reports the characterization of a protease inhibitor isolated from the common white button mushroom Agaricus bisporus. The objective of the present study is to characterize the novel protease inhibitor from Agaricus bisporus in order to determine its nature and activity under varying environmental conditions. The protease inhibitor was characterized through SDS PAGE, gel filtration chromatography, and de novo sequencing in order to determine its molecular mass and sequence, respectively. The optimum pH, temperature, and thermal stability were studied to determine the optimum working range of the protease inhibitor. The protease inhibitory activity (%) was determined in the presence of metal ions, surfactants, oxidizing agents, and reducing agents. The kinetic parameters and the type of inhibition exhibited by the protease inhibitor were determined using casein and trypsin protease enzyme. The protease inhibitor was found to be a low molecular mass compound of 25 kDa. The de novo sequencing matched the inhibitor against a 227 amino acid containing peptide molecular mass of 24.6 kDa molecular mass. The protease inhibitory activity (%) was found highest at pH 7.0 and temperature 50 °C, and the inhibitor was stable from pH 4.0-9.0 and temperature 30-80 °C. In the presence of metal ions, the residual protease inhibitory activity (%) enhanced in the presence of Na+, Mg2+, and Fe3+. The residual activity increased in the presence of the surfactant SDS slightly in comparison to control, while it decreased in the case of Triton-X and Tween 20. The presence of oxidizing agents, hydrogen peroxide and dimethyl sulfoxide decreased the residual inhibitory activity. The protease inhibitor's activity was unaffected by the reducing agents, dithiothreitol and β-mercaptoethanol, at up to 2mM concentration, but it decreased at higher concentrations. The inhibitor exhibited uncompetitive inhibition against trypsin with an inhibitory constant of 166 nM, indicating a strong affinity towards the protease, with a half-life of 93.90 minutes at 37 °C. Protease inhibitors isolated from mushrooms are generally small in size, more stable, and tolerant towards varying external conditions. The protease inhibitor isolated from Agaricus bisporus also exhibited similar characteristics.


Three-Dimensional Crystal Vue Imaging technology assessment of Sylvian fissures at 20-32+6 weeks' normal gestation.
Assess developmental pattern of Sylvian fissures (SF) with Three-Dimensional Crystal Vue Imaging (3D-CVI) at 20-32+6 weeks of gestation. This was a prospective cross-sectional study. Assess 20-32+6 weeks' gestation normal development of fetal brain SF with 3D-CVI imaging. Measure the uncovered area and perimeter of the insula on the Three-Dimensional (3D) image and establish reference ranges for the uncovered area and perimeters of the insula during normal pregnancy 20-32+6 weeks' gestation. Examine intra- and interobserver repeatability of measurements of the uncovered area and perimeter of the insula. A total of 286 normal fetuses from 20 to 32+6 weeks of gestation were studied. The SF first was trapezoidal in the 25 weeks of gestation, gradually becoming triangular as gestational age (GA) increased, and then closing from posterior up to anterior down. The uncovered area and dimension of the insula showed a parabolic curve that first increased and then decreased as GA and head circumference (HC) increased. Reference ranges for measurements of the uncovered area and perimeters of the insula during normal pregnancy 20-32+6 weeks' gestation were established. The intra- and interobserver agreements were reproducible (all ICC > 0.850); there were more than 95% dots in the Bland-Altman plots (95 limits of agreement (LOA)) scale in every figure. 3D-CVI can be used to observe the morphological changes of SF during middle and late pregnancy, which is an intuitive supplementary means for prenatal evaluation of cerebral cortex development, guiding subsequent follow-up and referral for assessment by expert neurosonologists. • A new imaging technique was found to visualize the SF of fetal brain surface. • This technique has the advantages of good consistency and repeatability, simple operation, short time-consuming, and low cost. • Its 3D visualization images can be used to the development and changes of the sulci on the brain surface, it provides a new method to evaluate the development of cerebral cortex.


Failure of complete suppression of endogenous glucose production by euglycaemic hyperinsulinaemia in normal humans.
In normal subjects, endogenous glucose production (EGP) is usually assumed to be completely suppressed during euglycaemic clamp studies performed at high insulin levels (> 100 mU L-1). However, this assumption is based on non-steady-state tracer measurements of EGP which are prone to negative errors. We have used purified [6-(3)H]glucose in an optimal tracer infusion protocol to assess the suppression of EGP during 4 h euglycaemic clamps in eight normal men. An insulin infusion rate of 5 mU kg-1 min-1 was chosen to achieve supraphysiological (> 500 mU L-1) plasma insulin concentrations. Using a labelled exogenous glucose infusion, plasma glucose (mean +/- SEM 5.3 +/- 0.1 mmol L-1) and glucose specific activities (mean 100 +/- 3% of basal) were maintained constant from 80 to 240 min. During hyperinsulinaemia, isotopically determined glucose appearance rates (Ra) were greater than glucose infusion rates (GIR) throughout the euglycaemic clamp period (P < 0.001) and EGP (Ra-GIR) was always greater than zero. In seven of the eight subjects studied EGP was partly suppressed but showed a wide variation (EGP 5 to 91% of basal at 80-120 min and 12 to 87% of basal at 200-240 min) while in one subject EGP rose above basal (by 72% at 80-120 min and 49% at 200-240 min). We conclude that EGP is not completely suppressed during euglycaemic clamps at high insulin levels.


Lockdown measures for COVID-19 outbreak and variation in physical activity in patients with heart failure and cardiac implantable devices.
The present study analysed the patterns of physical activity pre-, during and post-lockdown measures for COVID-19 pandemic in patients with chronic heart failure (CHF) and cardiac implantable electronic devices (CIED) under remote monitoring (RM), and assessed the physical activity patterns during these periods. The raw data from 95 patients with CHF (age 67,7 ± 15,1 years, 71,5% male) corresponding to 2238 RM transmissions of the Medtronic Carelink™ network platform was obtained. The physical exercise profiles and the impact of the lockdown measures on the physical behaviour during and after the measures were analysed. According to the level of activity duration in the pre-lockdown, lockdown and post-lockdown periods, the patterns of behaviour were identified (non-recoverees, incomplete recoverees, recoverees and full-recoverees). RM of CHF patients with CIED using the Carelink™ network is useful for close follow-up and identification of heart failure risk status variations. After relieving the confinement measures there were two groups of patients that did not recover the previous physical activity levels. Physical inactivity in patients with CHF can have a significant impact on outcomes.


Electron microscopy of two-dimensional crystals of mitochondrial ATP synthase.
Two-dimensional crystals of the mitochondrial ATP synthase up to 0.4 microns in size were obtained from the detergent-lipid-protein micelles by detergent dialysis. A projected map of the negatively stained crystal was calculated from electron microscopical images by the Fourier-filtering procedure at about 2.8 nm resolution. The unit cell (with not more than two ATP synthase molecules) has the following parameters: a = 13.0 nm, b = 25.6 nm and gamma = 86 degrees. Two alternative models for the crystal structural organization were suggested, viz. with one or two protein molecules per unit cell.


Relationship between neural responses to stress and mental health symptoms in psychogenic nonepileptic seizures after traumatic brain injury.
To utilize traumatic brain injury (TBI) as a model for investigating functioning during acute stress experiences in psychogenic nonepileptic seizures (PNES) and to identify neural mechanisms underlying the link between changes in processing of stressful experiences and mental health symptoms in PNES. We recruited 94 participants: 50 with TBI only (TBI-only) and 44 with TBI and PNES (TBI + PNES). Participants completed mood (Beck Depression Inventory-II), anxiety (Beck Anxiety Inventory), and posttraumatic stress disorder (PTSD) symptom (PTSD Checklist-Specific Event) assessments before undergoing functional magnetic resonance imaging during an acute psychosocial stress task. Linear mixed-effects analyses identified clusters of significant interactions between group and neural responses to stressful math performance and stressful auditory feedback conditions within limbic brain regions (volume-corrected α = .05). Spearman rank correlation tests compared mean cluster signals to symptom assessments (false discovery rate-corrected α = .05). Demographic and TBI-related measures were similar between groups; TBI + PNES demonstrated worse clinical symptom severity compared to TBI-only. Stressful math performance induced relatively greater reactivity within dorsomedial prefrontal cortex (PFC) and right hippocampal regions and relatively reduced reactivity within left hippocampal and dorsolateral PFC regions for TBI + PNES compared to TBI-only. Stressful auditory feedback induced relatively reduced reactivity within ventral PFC, cingulate, hippocampal, and amygdala regions for TBI + PNES compared to TBI-only. Changes in responses to stressful math within hippocampal and dorsal PFC regions were correlated with increased mood, anxiety, and PTSD symptom severity. Corticolimbic functions underlying processing of stressful experiences differ between patients with TBI + PNES and those with TBI-only. Relationships between these neural responses and symptom assessments suggest potential pathophysiologic mechanisms in PNES.


Alpha interferon in the treatment of symptomatic myelofibrosis with myeloid metaplasia.
2 patients with myelofibrosis and myeloid metaplasia had symptomatic splenomegaly and were treated with interferon alpha-2c (IFN alpha-2c). The splenic pain and pressure symptoms disappeared, accompanied by a decrease in the size of the spleen. However, the peripheral blood count worsened and no improvement in the bone marrow fibrosis could be observed.


Task-evoked pupil responses reflect internal belief states.
Perceptual decisions about the state of the environment are often made in the face of uncertain evidence. Internal uncertainty signals are considered important regulators of learning and decision-making. A growing body of work has implicated the brain's arousal systems in uncertainty signaling. Here, we found that two specific computational variables, postulated by recent theoretical work, evoke boosts of arousal at different times during a perceptual decision: decision confidence (the observer's internally estimated probability that a choice was correct given the evidence) before feedback, and prediction errors (deviations from expected reward) after feedback. We monitored pupil diameter, a peripheral marker of central arousal state, while subjects performed a challenging perceptual choice task with a delayed monetary reward. We quantified evoked pupil responses during decision formation and after reward-linked feedback. During both intervals, decision difficulty and accuracy had interacting effects on pupil responses. Pupil responses negatively scaled with decision confidence prior to feedback and scaled with uncertainty-dependent prediction errors after feedback. This pattern of pupil responses during both intervals was in line with a model using the observer's graded belief about choice accuracy to anticipate rewards and compute prediction errors. We conclude that pupil-linked arousal systems are modulated by internal belief states.


Arterial ketone body ratio as a parameter of hepatic mitochondrial redox state during and after hemorrhagic shock.
This study was aimed at clarifying the usefulness of the arterial ketone body ratio (AKBR), which reflects hepatic mitochondrial redox state and closely correlates with hepatic energy production, for understanding the degree of hepatic mitochondrial damage and the extent of the deterioration in hepatic energy metabolism during or shortly after hemorrhagic shock. Changes in the AKBR of 33 trauma victims who were admitted to our institute in hemorrhagic shock with a systolic pressure lower than 70 mmHg were measured until the patient recovered with the restoration of AKBR to the normal range (> or = 1.0) or until the patient died. During hemorrhagic shock the AKBRs were highly decreased, indicating deteriorated hepatic function. With successive fluid resuscitation the AKBR quickly recovered in 15 surviving patients from an initial value of 0.26 +/- 0.03 toward normal within hours, indicating that hepatic mitochondria are functioning normally. The AKBR recovered to a normal value of 1.10 +/- 0.06 on day 2 (p < 0.001). In 18 expired patients, AKBR did not recover to normal range, even though some of the patients recovered from the shock state. On the other hand, AST, ALT, LDH, and prothrombin time on day 2 were not significantly different from the values on admission, and the changes during the interval were not unidirectional even in the surviving patients, providing no information on the current functional state of the liver. Measurement of AKBR during and shortly after hemorrhagic shock provides timely and accurate information about liver function.


MiR-17-92 and miR-221/222 cluster members target KIT and ETV1 in human gastrointestinal stromal tumours.
Gastrointestinal stromal tumours (GIST) are characterised by high expression of KIT and ETV1, which cooperate in GIST oncogenesis. Our aim was to identify microRNAs that are deregulated in GIST, have a role in GIST pathogenesis, and could potentially be used as therapeutic tool. Differentially expressed microRNAs between primary GIST (n=50) and gastrointestinal leiomyosarcomas (GI-LMS, n=10) were determined using microarrays. Selected microRNA mimics were transfected into GIST-882 and GIST-T1 cell lines to study the effects of microRNA overexpression on GIST cells. Luciferase reporter assays were used to establish regulation of target genes by selected microRNAs. MiR-17-92 and miR-221/222 cluster members were significantly (P<0.01) lower expressed in GIST vs GI-LMS and normal gastrointestinal control tissues. MiR-17/20a/222 overexpression in GIST cell lines severely inhibited cell proliferation, affected cell cycle progression, induced apoptosis and strongly downregulated protein and--to a lesser extent--mRNA levels of their predicted target genes KIT and ETV1. Luciferase reporter assays confirmed direct regulation of KIT and ETV1 by miR-222 and miR-17/20a, respectively. MicroRNAs that may have an essential role in GIST pathogenesis were identified, in particular miR-17/20a/222 that target KIT and ETV1. Delivering these microRNAs therapeutically could hold great potential for GIST management, especially in imatinib-resistant disease.


In patients with severe sepsis, does a single dose of etomidate to facilitate intubation increase mortality?
Currently, single-dose etomidate has not been shown to cause increased mortality in septic patients requiring intubation; however, sufficiently powered randomized trials are required before definitive conclusions can be drawn.


ZLL/AGO10 maintains shoot meristem stem cells during Arabidopsis embryogenesis by down-regulating ARF2-mediated auxin response.
The shoot meristem gives rise to new organs throughout a plant's life by the activity of pluripotent stem cells in the meristem center. Organ initiation at the periphery of the shoot meristem is triggered by the accumulation of the phytohormone auxin at the initiation site. Loss-of-function mutants of the ZWILLE/ARGONAUTE10/PINHEAD (ZLL/AGO10/PNH) gene terminate shoot meristem stem cells late in embryogenesis and can form a leaf or a leaf-like structure instead, indicating that AGO10 activity is required to maintain shoot meristem stem cells undifferentiated. Here, we addressed whether stem cell maintenance by AGO10 involves regulation of auxin. We found that in zll-1 mutants, auxin accumulation and expression of the response reporter DR5:GFP are elevated, and transcription of the Auxin Response Factor 2 (ARF2) gene is upregulated. Downregulation of ARF2 significantly restores stem cells in zll-1 mutants, whereas increased expression of ARF2 enhances differentiation of stem cells in zll-1 mutants. We further found that upregulation of the AGO10 effector gene REVOLUTA restores ARF2 expression and stem cell maintenance in zll-1 embryos. Our results indicate that maintenance of shoot meristem stem cells by AGO10 involves negative regulation of auxin signaling and, via REV-mediated downregulation of ARF2 expression, auxin response.


Identification of a Functional Susceptibility Variant for Adolescent Idiopathic Scoliosis that Upregulates Early Growth Response 1 (EGR1)-Mediated UNCX Expression.
Adolescent idiopathic scoliosis (AIS) is a serious health problem affecting 3% of live births all over the world. Many loci associated with AIS have been identified by previous genome wide association studies, but their biological implication remains mostly unclear. In this study, we evaluated the AIS-associated variants in the 7p22.3 locus by combining in silico, in vitro, and in vivo analyses. rs78148157 was located in an enhancer of UNCX, a homeobox gene and its risk allele upregulated the UNCX expression. A transcription factor, early growth response 1 (EGR1), transactivated the rs78148157-located enhancer and showed a higher binding affinity for the risk allele of rs78148157. Furthermore, zebrafish larvae with UNCX messenger RNA (mRNA) injection developed body curvature and defective neurogenesis in a dose-dependent manner. rs78148157 confers the genetic susceptibility to AIS by enhancing the EGR1-regulated UNCX expression. © 2022 American Society for Bone and Mineral Research (ASBMR).


Evaluating the effect of a proposed logistics fee cap on pharmaceuticals in South Africa--a pre and post analysis.
South Africa has proposed the implementation of a maximum logistics fee paid by pharmaceutical manufacturers to wholesalers and distributors. However very little knowledge exists of the effects, unintended or otherwise, of the implementation of these proposed regulations, which are required to guide further policy development and implementation. The objectives of this study was to therefore evaluate the effects of the proposed logistics fee cap on different pharmaceuticals and different dosage forms, as well as to observe the logistics fee contribution to the Single Exit Price. Private sector medicine prices were sourced from the South African Medicine Price Registry as at 20 December 2013. For each medicine the maximum logistics fee was calculated based on the 2012 proposed government guidelines. The logistics fee as a percentage of the final Single Exit Price was calculated, as part of the analysis of results. Out of the 47 medicines in the overall sample from the current study, only 16 medicines showed a decrease in the Single Exit Price with the application of the maximum logistics fee cap. This study reveals the need for greater transparency of the mark ups along the distribution chain as well as further research with regards to the costing of logistics fees of similar pharmaceuticals.


Antinociceptive profiles of non-peptidergic neurokinin1 and neurokinin2 receptor antagonists: a comparison to other classes of antinociceptive agent.
This study compared the antinociceptive properties of systemic administration of selective, non-peptidergic antagonists at neurokinin (NK1 and NK2) receptors to those of other classes of antinociceptive agent. (All doses are in mg/kg.) In mice, the NK1 antagonist, CP 99,994, preferentially (inhibitory dose50 (ID50) = 4.4) inhibited the late phase (LP) as compared to the early phase (EP) (16.1) of formalin-induced licking (FIL). A high dose (17.6) elicited ataxia in the rotarod test. Acetic acid-induced writhing was reduced at intermediate doses (10.0) whereas the tail-flick (TF) response to thermal and mechanical stimuli was inhibited only at high doses (22.7 and 17.7, respectively). Modulation of stimulus intensity did not modify the influence of CP 99,994 upon the response to heat. A similar pattern of data was acquired with RP 67,580, although this NK1 antagonist more potently inhibited writhing (2.8). In contrast, RP 68,651, the inactive isomer of RP 67,580, neither reduced the LP of FIL nor modified writhing indicating that these actions of RP 67,580 were stereospecific. Three further NK1 antagonists, SR 140,333, WIN 51,708 and WIN 62,577, likewise inhibited the LP of FIL and failed to modify the TF response at non-ataxic doses. Further, SR 140,333 (0.5) and WIN 51,708 (1.4) were potent ligands in the writhing procedure. The NK2 antagonist, SR 48,966, mimicked NK1 antagonists in preferentially inhibiting the LP (7.7) as compared to the EP (26.9) of FIL. Further, only at doses higher than those evoking ataxia (20.9) did SR 48,968 modify the TF response (36.5 and 32.0 for heat and pressure, respectively). However, it differed to NK1 antagonists in being inactive in the writhing test (> 40.0). In comparison to these NK1 and NK2 antagonists, the mu-opioid agonists (morphine and fentanyl) and kappa-opioid agonists (enadoline and U 69,593) equipotently inhibited all nociceptive responses at doses not provoking ataxia. While the glycine B receptor partial agonist, (+)-HA 966, selectively blocked the LP of FIL and did not evoke ataxia, the NMDA receptor channel blocker, (+)-MK 801, elicited antinociception only at doses close to those provoking ataxia. Finally, the NSAIDs, indomethacin and ibuprofen, the BK2 antagonist, Hoe 140 and the nitric oxide synthase (NOS) inhibitors, L-NAME and 7 nitroindazole, inhibited the LP (but not the EP) of FIL and (except for L-NAME) also reduced writhing: in contrast, they did not evoke ataxia and were inactive in the TF procedures.(ABSTRACT TRUNCATED AT 400 WORDS)


Development of a platform to investigate long-term potentiation in human iPSC-derived neuronal networks.
Impairment of long-term potentiation (LTP) is a common feature of many pre-clinical models of neurological disorders; however, studies in humans are limited by the inaccessibility of the brain. Human induced pluripotent stem cells (hiPSCs) provide a unique opportunity to study LTP in disease-specific genetic backgrounds. Here we describe a multi-electrode array (MEA)-based assay to investigate chemically induced LTP (cLTP) across entire networks of hiPSC-derived midbrain dopaminergic (DA) and cortical neuronal populations that lasts for days, allowing studies of the late phases of LTP and enabling detection of associated molecular changes. We show that cLTP on midbrain DA neuronal networks is largely independent of the N-methyl-D-aspartate receptor (NMDAR) and partially dependent on brain-derived neurotrophic factor (BDNF). Finally, we describe activity-regulated gene expression induced by cLTP. This cLTP-MEA assay platform will enable phenotype discovery and higher-throughput analyses of synaptic plasticity on hiPSC-derived neurons.


Patient and nurse content preferences for a communication board to facilitate dialogue in the intensive care unit.
The main aim of this study was to identify patient and nurse content preferences for a communication board to facilitate effective communication (dialogue) in the critical care unit. A qualitative research design focusing on explorative and descriptive components was used to address the aim of the research study. The study employed two participant groups. Semi-structured interviews and focus group discussions were conducted with critically ill patients (N = 10) and critical care nurses (N = 30). The study was conducted in four different private hospitals of the same hospital group in Gauteng, South Africa. Four distinct themes were identified in the research regarding the vocabulary items that participants would like to have included. The findings suggest that patients perceive a communication board to be valuable in enhancing communication in the critical care unit; that cultural and linguistic diversity should be considered; and that patients and nurses have varying opinions on topics of priority during communication.


Teleworking from home experiences during the COVID-19 pandemic among public health workers (TelEx COVID-19 study).
When working from home (WFH) became temporarily necessary for staff as a result of the COVID-19 pandemic in 2020, it had to be implemented without significant organisational experience or understanding of WFH and its complexities. This study aims to determine the impacts experienced by staff who have undertaken WFH during the COVID-19 pandemic. This was an observational cross-sectional study using survey with a purposive sampling strategy for staff from corporate and non-clinical departments. These staff undertook WFH during COVID-19 pandemic in 2020. None of these staff had any direct operational roles in a hospital facility and clinical service. Participants' self-reports of their mood while working in their normal workplace and while WFH were collected via the Scale of Positive and Negative Experience (SPANE), a validated affect balance questionnaire. The responses from the open-ended question were analysed using thematic analysis approach. A total of 143 participants completed the survey responses. Majority (61%) WFH for four or more months as a result of the COVID-19 pandemic. Participants rated their skills very highly on the technologies with an average rating of 9 (out of 10) for computer skills, smartphones and videoconferencing/teleconferencing applications. Participants felt WFH was an improvement on normal working, in particular in relation to their ability to concentrate and be productive. The "SPANE" relating to affect balance while WFH was completed by 124 participants (85.7%), resulting in a mean score of 5.45 (S.D. 2.98). The SPANE relating to normal working conditions was completed by 127 participant (88.8%) resulting in a mean score of 2.70 (S.D 3.69). This indicated that while participants' positive emotions typically predominated in both situations, they felt slightly more positive on average with WFH. Over 90% participants reported that they would take the opportunity to WFH again if it were offered. Data obtained from the open-ended questions had complimented the findings of the structure close-ended questions in the benefits of remote working and support for their health and wellbeing. The open-ended questions had provided additional information on challenges which the participants encountered during the WFH experience and their suggested preference to sustain this workplace practice. This study highlighted factors that impacted workers' work processes, productivity, physical and mental health well-being while WFH and provided a foundation for considering how to best support a positive WFH experience.


Determination of propranolol in peritoneal dialysis fluid by high-performance liquid chromatography without extraction.
A rapid, sensitive method for the high-performance liquid chromatographic determination of propranolol in peritoneal dialysis fluid is described. An extraction step is replaced by the use of a C18 Sep-Pak cartridge for sample preparation. The procedure offers an acceptable alternative to sample extraction and will allow for pharmacokinetic studies of propranolol in patients undergoing peritoneal dialysis for chronic renal failure.


Applying the biopsychosocial model to unpack a psychosocial support intervention designed to improve antiretroviral treatment outcomes for adolescents in South Africa.
Adolescents (10 to 19 years) living with HIV (ALHIV) experience disproportionately poor adherence to antiretroviral treatment (ART) compared to other age groups. Several barriers, including psychosocial challenges, contribute to this observation. Psychosocial support (PSS) interventions show promising results as a strategy to deal with the biological and psychosocial challenges faced by ALHIV. However, there is dearth of information on how psychosocial support interventions designed to improve treatment adherence and retention in care among ALHIV are effective. In this commentary, we used the biopsychosocial model to formulate hypotheses on how the components of a PSS intervention could improve adherence and retention in ART care. Psychological wellbeing, coping strategies, social support, self-efficacy, and disclosure are key components in the intervention designed to improve ART adherence and retention in care. The management of ALHIV for improved ART adherence and retention requires recognising and addressing the complex biological, psychological and social issues peculiar to them.


Morphological and molecular characterization of a species of Tetratrichomonas present in feces of Brazilian sheep (Ovis aries) and goats (Capra hircus).
The trichomonads form part of the phylum Parabasalia, a complex assemblage of diverse species of flagellated protists, with some members recognized as pathogens of men and/or animals. Associations, probably as commensals, between the species Tetratrichomonas ovis and sheep were reported in North America during the 1960s based on morphological and cultural characteristics. Intriguingly, no subsequent studies of this topic have been published. Feces, collected from sheep (n = 55) and goats (n = 14), reared on small-scale, production facilities in Southeastern Brazil, were examined for parabasalids. Protozoa, demonstrating morphologies and motility characteristic of trichomonads, were detected by direct microscopy in 64% of sheep and 43% of goat samples. In contrast to T. ovis, none of the samples could be cultured in Diamond's medium; however, cultures were obtained for three goat and seventeen sheep samples in peptonized broth. Based on morphological analyses, all isolates were classified as members of the genus Tetratrichomonas. Sequencing of the ITS1-5.8S rRNA gene-ITS2 region revealed three highly similar genotypes that were essentially identical to sequences reported for Tetratrichomonas spp. isolated from the preputial cavity of cattle in the USA and Southern Brazil. The findings of this study extend and enhance our knowledge of parasitism in small ruminants by parabasalids.


Patient perspectives on community pharmacy administered and dispensing of methadone treatment for opioid use disorder: a qualitative study in the U.S.
Pharmacy administration and dispensing of methadone treatment for opioid use disorder (PADMOUD) may address inadequate capability of opioid treatment programs (OTPs) in the US by expanding access to methadone at community pharmacies nationally. PADMOUD is vastly underutilized in the US. There is no published US study on OUD patients' perspectives on PADMOUD. Data are timely and needed to inform the implementation of PADMOUD in the US to address its serious opioid overdose crisis. Patient participants of the first completed US trial on PADMOUD through electronic prescribing for methadone (parent study) were interviewed to explore implementation-related factors for PADMOUD. All 20 participants of the parent study were invited to participate in this interview study. Each interview was recorded and transcribed verbatim. Thematic analysis was conducted to identify emergent themes. Seventeen participants completed the interview. Patients' perspectives on PADMOUD were grouped into five areas. Participants reported feasibility of taking the tablet formulation of methadone at the pharmacy and identified benefits from PADMOUD (e.g., better access, efficiency, convenience) compared with usual care at the OTP. Participants perceived support for PADMOUD from their family/friends, OTP staff, and pharmacy staff. PADMOUD was perceived to be a great option for stable patients with take-home doses and those with transportation barriers. The distance (convenience), office hours, and the cost were considered factors most influencing their decision to receive methadone from a pharmacy. Nonjudgmental communication, pharmacists' training on methadone treatment, selection of patients (stable status), workflow of PADMOUD, and protection of privacy were considered key factors for improving operations of PADMOUD. This study presents the first findings on patient perspectives on PADMOUD. Participants considered pharmacies more accessible than OTPs, which could encourage more people to receive methadone treatment earlier and help transition stable patients from an OTP into a local pharmacy. The findings have timely implications for informing implementation strategies of PADMOUD that consider patients' views and needs.


Utilization and outcomes with low dose tissue plasminogen activator as intravenous thrombolytic therapy for ischaemic stroke at Aga Khan University Hospital, Karachi: a retrospective analysis.
To determine the utilisation of intravenous tissue plasminogen activator at a certain dose for ischaemic stroke thrombolysis and to compare the outcomes with those of a different dosage mentioned in literature. The retrospective study was conducted at the Aga Khan University Hospital, Karachi, and comprised medical records from January, 2007, to October, 2016, of all patients having received intravenous tissue plasminogen activator for ischaemic stroke thrombolysis. Primary safety outcome variables included symptomatic intracerebral haemorrhage after the start of treatment (0.6mg/kg) and death within three months as per the modified Rankin scale 6. Secondary efficacy outcome variable was functional independence as per modified Rankin scale 0-2 at three months. The outcomes were compared with those mentioned in literature with a dose of 0. 9mg/kg. Of the 79 patients, 52 (66%) were male and 27 (34 %) were female. Median pre-treatment tissue plasminogen activator score was 12 (interquartile range: 8-15). Overall utilisation of t-PA remained at 1.7%. Symptomatic intracerebral haemorrhage was not seen in our cohort while it was seen in 107 (1.7%) patients at the higher dose. Using another definition, it was seen in 3 (3.8%) patients versus 468 (7.3%) patients at the higher dose. Functional independence was seen in 40 (50.6%) patients at three months compared to 3362 (54.8%) patients at the higher dose. Low-dose intravenous thrombolytic therapy for ischaemic stroke patients was found to be safe and efficacious, and yielded comparable results with those obtained at a higher dose..


Causal assessment in evidence synthesis: A methodological review of reviews.
In fields (such as population health) where randomised trials are often lacking, systematic reviews (SRs) can harness diversity in study design, settings and populations to assess the evidence for a putative causal relationship. SRs may incorporate causal assessment approaches (CAAs), sometimes called 'causal reviews', but there is currently no consensus on how these should be conducted. We conducted a methodological review of self-identifying 'causal reviews' within the field of population health to establish: (1) which CAAs are used; (2) differences in how CAAs are implemented; (3) how methods were modified to incorporate causal assessment in SRs. Three databases were searched and two independent reviewers selected reviews for inclusion. Data were extracted using a standardised form and summarised using tabulation and narratively. Fifty-three reviews incorporated CAAs: 46/53 applied Bradford Hill (BH) viewpoints/criteria, with the remainder taking alternative approaches: Medical Research Council guidance on natural experiments (2/53, 3.8%); realist reviews (2/53, 3.8%); horizontal SRs (1/53, 1.9%); 'sign test' of causal mechanisms (1/53, 1.9%); and a causal cascade model (1/53, 1.9%). Though most SRs incorporated BH, there was variation in application and transparency. There was considerable overlap across the CAAs, with a trade-off between breadth (BH viewpoints considered a greater range of causal characteristics) and depth (many alternative CAAs focused on one viewpoint). Improved transparency in the implementation of CAA in SRs in needed to ensure their validity and allow robust assessments of causality within evidence synthesis.


[Social representations of the vaccine].
Vaccination has made it possible to combat many infectious diseases by reducing their impact in terms of mortality as well as morbidity. However, reservations about this primary prevention measure, as demonstrated by the difficulties in introducing vaccination against COVID-19. Through the analysis of social representations, we try to understand the reasons for these obstacles.


Correlates of hepatitis B among patients with mental illness in Brazil.
To assess correlates of hepatitis B among adults with mental illness under care in Brazil. Cross-sectional national multicenter study of 2206 patients with mental illnesses randomly selected from 26 public mental health services. Sociodemographic and behavioral data were obtained from face-to-face interviews and psychiatric diagnoses from medical charts. Serology testing was conducted, and prevalence rate ratios were estimated by log-binomial regression. The weighted prevalence rates of current hepatitis B virus (HBV) infection (HBsAg+) and previous HBV exposure (anti-HBc+) were 2.0% [95% confidence interval (CI): 1.5%-2.7%] and 17.1% (95% CI: 16.0%-19.0%), respectively. Correlates of HBsAg+ included male gender, younger age (18-29 years), unstable place of residence, intellectual disability, main psychiatric diagnosis of dementia, presence of other medical comorbitidy, use of alcohol/drugs during sex, more than one sexual partner and use of cocaine. Correlates of anti-HBc+ included male gender, older age (≥30 years), black skin color, lower education, unstable place of residence, currently hospitalized, intellectual disability, history of any sexually transmitted disease or syphilis, poor HIV knowledge, history of imprisonment and sexual violence. Hepatitis B is an important comorbidity among psychiatric patients in Brazil. Screening for HBV, effective prevention and intervention strategies, including universal HBV immunization, should be routine practices in these mental health services.


Combination of common mtDNA variants results in mitochondrial dysfunction and a connective tissue dysregulation.
Mitochondrial dysfunction can be associated with a range of clinical manifestations. Here, we report a family with a complex phenotype including combinations of connective tissue, neurological, and metabolic symptoms that were passed on to all surviving children. Analysis of the maternally inherited mtDNA revealed a novel genotype encompassing the haplogroup J - defining mitochondrial DNA (mtDNA) ND5 m.13708G>A (A458T) variant arising on the mtDNA haplogroup H7A background, an extremely rare combination. Analysis of transmitochondrial cybrids with the 13708A-H7 mtDNA revealed a lower mitochondrial respiration, increased reactive oxygen species production (mROS), and dysregulation of connective tissue gene expression. The mitochondrial dysfunction was exacerbated by histamine, explaining why all eight surviving children inherited the dysfunctional histidine decarboxylase allele (W327X) from the father. Thus, certain combinations of common mtDNA variants can cause mitochondrial dysfunction, mitochondrial dysfunction can affect extracellular matrix gene expression, and histamine-activated mROS production can augment the severity of mitochondrial dysfunction. Most important, we have identified a previously unreported genetic cause of mitochondrial disorder arising from the incompatibility of common, nonpathogenic mtDNA variants.


A model for bacterial colonization of sinking aggregates.
Sinking aggregates provide important nutrient-rich environments for marine bacteria. Quantifying the rate at which motile bacteria colonize such aggregations is important in understanding the microbial loop in the pelagic food web. In this paper, a simple analytical model is presented to predict the rate at which bacteria undergoing a random walk encounter a sinking aggregate. The model incorporates the flow field generated by the sinking aggregate, the swimming behavior of the bacteria, and the interaction of the flow with the swimming behavior. An expression for the encounter rate is computed in the limit of large Péclet number when the random walk can be approximated by a diffusion process. Comparison with an individual-based numerical simulation is also given.


Use of computational fluid dynamics studies in predicting aneurysmal degeneration of acute type B aortic dissections.
Whereas uncomplicated acute type B aortic dissections are often medically managed with good outcomes, a subset develop subacute or chronic aneurysmal dilation. We hypothesized that computational fluid dynamics (CFD) simulations may be useful in identifying patients at risk for this complication. Patients with acute type B dissection complicated by rapidly expanding aortic aneurysms (N = 7) were compared with patients with stable aortic diameters (N = 7). Three-dimensional patient-specific dissection geometries were generated from computed tomography angiography and used in CFD simulations of pulsatile blood flow. Hemodynamic parameters including false lumen flow and wall shear stress were compared. Patients with rapid aneurysmal degeneration had a growth rate of 5.3 ± 2.7 mm/mo compared with those with stable aortic diameters, who had rates of 0.2 ± 0.02 mm/mo. Groups did not differ in initial aortic diameter (36.1 ± 2.9 vs 34.4 ± 3.6 mm; P = .122) or false lumen size (22.6 ± 2.9 vs 20.2 ± 4.5 mm; P = .224). In patients with rapidly expanding aneurysms, a greater percentage of total flow passed through the false lumen (78.3% ± 9.3% vs 56.3% ± 11.8%; P = .016). The time-averaged wall shear stress on the aortic wall was also significantly higher (12.6 ± 3.7 vs 7.4 ± 2.8 Pa; P = .028). Hemodynamic parameters derived from CFD simulations of acute type B aortic dissections were significantly different in dissections complicated by aneurysm formation. Thus, CFD may assist in predicting which patients may benefit from early stent grafting.