Withholding methotrexate after vaccination with ChAdOx1 nCov19 in patients with rheumatoid or psoriatic arthritis in India (MIVAC I and II): results of two, parallel, assessor-masked, randomised controlled trials.
There is a necessity for an optimal COVID-19 vaccination strategy for vulnerable population groups, including people with autoimmune inflammatory arthritis on immunosuppressants such as methotrexate, which inhibit vaccine-induced immunity against SARS-CoV-2. Thus, we aimed to assess the effects of withholding methotrexate for 2 weeks after each dose of ChAdOx1 nCov-19 (Oxford-AstraZeneca) vaccine (MIVAC I) or only after the second dose of vaccine (MIVAC II) compared with continuation of methotrexate, in terms of post-vaccination antibody titres and disease flare rates. MIVAC I and II were two parallel, independent, assessor-masked, randomised trials. The trials were done at a single centre (Dr Shenoy's Centre for Arthritis and Rheumatism Excellence; Kochi, India) in people with either rheumatoid arthritis or psoriatic arthritis with stable disease activity, who had been on a fixed dose of methotrexate for the preceding 6 weeks. Those with previous COVID-19 or who were positive for anti-SARS-CoV-2 nucleocapsid antibodies were excluded from the trials. People on high-dose corticosteroids and rituximab were also excluded, whereas other disease-modifying antirheumatic drugs were allowed. In MIVAC I, participants were randomly assigned (1:1) to stop methotrexate treatment for 2 weeks after each vaccine dose or to continue methotrexate treatment. In MIVAC II, participants who had continued methotrexate during the first dose of vaccine were randomly assigned (1:1) to withhold methotrexate for 2 weeks after the second dose of vaccine or to continue to take methotrexate. The treating physician was masked to the group assignments. The primary outcome for both MIVAC I and MIVAC II was the titre (absolute value) of anti-receptor binding domain (RBD) antibody measured 4 weeks after the second dose of vaccine. All analyses were done per protocol. The trials were registered with the Clinical Trials Registry- India, number CTRI/2021/07/034639 (MIVAC I) and CTRI/2021/07/035307 (MIVAC II). Between July 6 and Dec 15, 2021, participants were recruited to the trials. In MIVAC I, 250 participants were randomly assigned and 158 completed the study as per the protocol (80 in the methotrexate hold group and 78 in the control group; 148 [94%] were women and 10 [6%] were men). The median post-vaccination antibody titres in the methotrexate hold group were significantly higher compared with the control group (2484·0 IU/mL, IQR 1050·0-4388·8 vs 1147·5 IU/mL, 433·5-2360·3; p=0·0014). In MIVAC II, 178 participants were randomly assigned and 157 completed the study per protocol (76 in the methotrexate hold group and 81 in the control group; 135 [86%] were women and 22 [14%] were men). The methotrexate hold group had higher post-vaccination antibody titres compared with the control group (2553·5 IU/ml, IQR 1792·5-4823·8 vs 990·5, 356·1-2252·5; p<0·0001). There were no reports of any serious adverse events during the trial period. Withholding methotrexate after both ChAdOx1 nCov-19 vaccine doses and after only the second dose led to higher anti-RBD antibody titres compared with continuation of methotrexate. However, withholding methotrexate only after the second vaccine dose resulted in a similar humoral response to holding methotrexate after both vaccine doses, without an increased risk of arthritis flares. Hence, interruption of methotrexate during the second dose of ChAdOx1 nCov-19 vaccine appears to be a safe and effective strategy to improve the antibody response in patients with rheumatoid or psoriatic arthritis. Indian Rheumatology Association.


Biochemical metabolism in the anterior cingulate cortex and cognitive function in major depressive disorder with or without insomnia syndrome.
Major depressive disorder (MDD) and insomnia have been linked to deficiencies in cognitive performance. However, the underlying mechanism of cognitive impairment in MDD patients with insomnia symptoms (IS) remains unclear. This study aimed to explore the effects of IS in patients with MDD by comparing cognitive function indices among those with IS, those without insomnia symptoms (NIS), and healthy controls (HCs). In addition, we assessed whether the dysfunction of central nervous system (CNS) is one of the important pathophysiologic mechanisms of IS in patients with MDD by comparing the biochemical metabolism ratios in the anterior cingulate cortex (ACC). Fifty-five MDD with IS, 39 MDD without IS, and 47 demographically matched HCs underwent the MATRICS Consensus Cognitive Battery (MCCB) assessment and proton magnetic resonance spectroscopy (1H-MRS). MCCB cognitive scores and biochemical metabolism in ACC were assessed and compared between groups. Compared to the HCs group, IS and NIS groups scored significantly lower in seven MCCB cognitive domains (speed of processing, attention/vigilance, working memory, verbal learning, visual learning, reasoning problem solving and social cognition). IS group showed a lower speed of processing and lower Cho/Cr ratio in the left ACC vs. NIS group and HCs. Also, in IS group, the Cho/Cr ratio in the left ACC was positively correlated with the composite T-score. Patients with comorbidity of MDD with IS may exhibit more common MCCB cognitive impairments than those without IS, particularly speed of processing. Also, dysfunction of ACC may underlie the neural substrate of cognitive impairment in MDD with IS.


Vagal control of lower oesophageal sphincter motility in the cat.
1. The effects of vagal efferent fibre stimulation on the smooth muscle of the lower oesophageal sphincter have been studied on the anaesthetized animal and on the isolated and perfused organ.2. In both muscle layers (longitudinal and circular) vagal stimulation elicits two types of electromyographic (e.m.g.) potentials: (a) excitatory junction potentials (e.j.p.s) where there is a depolarization of the smooth muscle fibres. E.j.p.s can give rise to spike potentials inducing a contraction of the sphincter; (b) inhibitory junction potentials (i.j.p.s) where there is hyperpolarization of the smooth muscle fibres, often followed by a transient depolarization which may initiate spikes (post-inhibitory rebound).3. Pure i.j.p.s are observed after atropine treatment which suppresses e.j.p.s. Under these conditions, a long lasting vagal stimulation induces a long duration hyperpolarization concomitant with an opening of the lower oesophageal sphincter followed after the cessation of stimulation by a powerful rebound leading to a strong contraction which closes the sphincter.4. Several arguments, pharmacological (action of acetylcholine (ACh), atropine and hexamethonium) and physiological (threshold and latency of responses) lead to the following conclusions. Preganglionic vagal fibres are cholinergic and they activate (a) intramural excitatory cholinergic neurones; (b) intramural non-adrenergic inhibitory neurones (purinergic neurones). Preganglionic fibres leading to inhibition have a higher threshold than those leading to excitation. Both excitatory and inhibitory pathways are interconnected inside the intramural network. In particular, activation of intramural inhibitory neurones, by relaxing the oesophagus orally to the lower oesophageal sphincter, inhibits intramural excitatory neurones and subsequently blocks vagal excitatory responses.5. Two functions may be attributed to the vagal extrinsic innervation: (a) closure of the lower oesophageal sphincter by maintaining the basal tone of the sphincter; this would imply that at rest the inhibitory control is supplanted by the excitatory one; (b) sphincter opening during swallowing by suppressing the excitatory stimulus and reinforcing the inhibitory one (it may be recalled that after bilateral vagotomy, swallowing is no longer followed by a relaxation of the sphincter).


Predicting ICU admission and death in the Emergency Department: A comparison of six early warning scores.
To compare the ability of the most used Early Warning Scores (EWS) to identify adult patients at risk of poor outcomes in the emergency department (ED). Single-center, retrospective observational study. We evaluated the digital records of consecutive ED admissions in patients ≥ 18 years from 2010 to 2019 and calculated NEWS, NEWS2, MEWS, RAPS, REMS, and SEWS based on parameters measured on ED arrival. We assessed the discrimination and calibration performance of each EWS in predicting death/ICU admission within 24 hours using ROC analysis and visual calibration. We also measured the relative weight of clinical and physiological derangements that identified patients missed by EWS risk stratification using neural network analysis. Among 225,369 patients assessed in the ED during the study period, 1941 (0.9%) were admitted to ICU or died within 24 hours. NEWS was the most accurate predictor (area under the receiver operating characteristic [AUROC] curve 0.904 [95% CI 0.805-0.913]), followed by NEWS2 (AUROC 0.901). NEWS was also well calibrated. In patients judged at low risk (NEWS < 2), 359 events occurred (18.5% of the total). Neural network analysis revealed that age, systolic BP, and temperature had the highest relative weight for these NEWS-unpredicted events. NEWS is the most accurate EWS for predicting the risk of death/ICU admission within 24 h from ED arrival. The score also had a fair calibration with few events occurring in patients classified at low risk. Neural network analysis suggests the need for further improvements by focusing on the prompt diagnosis of sepsis and the development of practical tools for the measurement of the respiratory rate.


Association Between Physical Activity and Sedentary Behavior With Depressive Symptoms Among US High School Students, 2019.
The prevalence of depression among US adolescents has increased during the past decade. Previous studies found relationships among physical activity, sedentary behavior, and depression, but more recent information is needed to inform research and practice. We used national surveillance data to assess the association of physical activity and sedentary behavior with depressive symptoms among US high school students. This study included 13,526 high school students from the 2019 Youth Risk Behavior Survey. The dependent variable was the presence of depressive symptoms in the past year that lasted almost every day for at least 2 weeks in a row and interfered with usual activities. The independent variables were physical activity (overall activity, muscle-strengthening exercises, participation on sports teams) and sedentary behavior (watching television, using a computer or digital device). We used weighted multivariable logistic regression to evaluate the association of physical activity and sedentary behavior variables with depressive symptoms, while controlling for demographic characteristics and other health behaviors. The prevalence of depressive symptoms was 36.7%. Participating in physical activity 5 or more days in the past 7 days (adjusted odds ratio [aOR], 0.81; 95% CI, 0.68-0.97) and participating on 1 or more sports teams in the past year (aOR, 0.66; 95% CI, 0.55-0.78) were associated with reduced odds of depressive symptoms. Using a computer or digital device for 3 or more hours per school night was associated with higher odds of reporting depressive symptoms (aOR, 1.61; 95% CI, 1.41-1.85). Inadequate physical activity and excessive sedentary behavior are associated with depressive symptoms among US high school students. Interventions targeting physical activity and sedentary behavior may be a public health strategy to reduce depressive symptoms in this population.


Ocular symptoms of moyamoya disease.
We examined four patients with moyamoya disease who had ocular symptoms. The diagnosis of the disease was based on an abnormal vascular network in the cerebral basilar region, observed by carotid angiography. One patient had amaurosis fugax. A second patient demonstrated bilaterally decreased visual acuity and visual spatial agnosia. The other two patients had homonymous hemianopsia; one also had transient diplopia and the other had temporal pallor of the optic disks. We found that patients with moyamoya disease exhibited various ocular symptoms but had few abnormal intraocular findings.


ENT pathology and diplopia.
ENT pathology causes diplopia in a minority of patients. Because the various ENT conditions that can produce diplopia are relatively unusual they often escape early detection. We describe the various ENT disorders that can produce diplopia and illustrate this with our experience between 1992 and 1998. We wish to emphasise the benefit which can be obtained from undertaking an ENT history and examination in arriving at the correct diagnosis.


[Histological evidences of using human external jugular vein to reconstruct carotid artery: a comparative study with greater saphenous vein].
To investigate the histological architecture of human external jugular vein and greater saphenous vein and to determine the feasibility of using autogenous external jugular vein for reconstruction of carotid artery defects. Specimens of 3 external jugular veins and greater saphenous vein and 1 segment of common carotid artery resected due to involvement of malignant tumor were available. The specimens were subjected to HE staining and examined under light microscopy. In addition, elastic fiber stain and Smactin stain were conducted to detect the amount and location of the elastic fibers and smooth muscles in the venous specimens. The smooth muscle cells in external jugular vein and greater saphenous vein were different both in amount and orientation of arrangement. There were more smooth muscle cells in the tunica media of the external jugular vein arranged circumferentially, while in greater saphenous vein, the smooth muscle cells were mainly located in the adventitia and arranged longitudinally. There were fewer elastic fibers in the venous walls. Histologically, it seemed more reasonable to reconstruct the carotid artery with external jugular vein.


Psychiatric morbidity among West Africans and West Indians living in London.
All west Africans and West Indians living in Camberwell who made a psychiatric contact between 1 January 1965 and 31 December 1968 were selected using the Camberwell Psychiatric Register as a sampling frame. Inception rates of psychiatric illness were then calculated using the 1966 10% census figures for West Africans and West Indians in Camberwell. The rates found were compared to those among the British living in Camberwell. For each major diagnostic caterogy, with the exception of reactive depression and paranoid states, the inception rates are significantly higher among West Africans than West Indians. They are also significantly higher among West Indians than British. However, the differences in inception rates are generally greater between West Africans and West Indians than between the latter and the British.


Ring pessary for all pelvic organ prolapse.
To evaluate the risk factors of unsuccessful fitting of ring pessary in pelvic organ prolapse (POP), continuation rate, and adverse events. The ring pessary was offered to 100 symptomatic POP patients. POP was staged according to Pelvic Organ Prolapse Quantification (POP-Q) system. A successful fitting was defined as when patients continued pessary use for more than 2 weeks. Adverse events were evaluated in patients who had a successful fitting. Most patients (71%) were in POP-Q stage III-IV and 77 patients were fitted at the initial visit. Successful fitting was reported in 62 patients (62%). However, 52 patients (52%) continued to use the pessary at 13 months median follow-up. The most common reason for discontinuation was frequent pessary expulsion. Short vaginal length ≤ 6 cm and wide introitus > 4 fingerbreadths were the significant risk factors for unsuccessful fitting. New onset of stress urinary incontinence (SUI) developed in 28.2% of patients. One third and 21% of patients had vaginal discharge and vaginal erosion, respectively. Only half of the patients could manage the pessary either by themselves or with their caregivers. The ring pessary fitting can be attempted in all POP cases irrespective of stage. Short vaginal length <6 cm and wide introitus >4 fingerbreadths were the risk factors for unsuccessful fitting. It has an acceptable continuation rate and manageable adverse events. The self-care of pessary is an important strategy to minimize adverse events.


TEAD4-YAP interaction regulates tumoral growth by controlling cell-cycle arrest at the G1 phase.
TEA domain transcription factor 4 (TEAD4), which has critical functions in the process of embryonic development, is expressed in various cancers. However, the important role of TEAD4 in human oral squamous cell carcinomas (OSCCs) remain unclear. Here we investigated the TEAD4 expression level and the functional mechanism in OSCC using quantitative reverse transcriptase-polymerase chain reaction, Western blot analysis, and immunohistochemistry. Furthermore, TEAD4 knockdown model was used to evaluate cellular proliferation, cell-cycle analysis, and the interaction between TEAD4 and Yes-associated protein (YAP) which was reported to be a transcription coactivator of cellular proliferation. In the current study, we found that TEAD4 expression increased significantly in vitro and in vivo and correlated with tumoral size in OSCC patients. TEAD4 knockdown OSCC cells showed decreased cellular proliferation resulting from cell-cycle arrest in the G1 phase by down-regulation of cyclins, cyclin-dependent kinases (CDKs), and up-regulation of CDK inhibitors. We also found that the TEAD4-YAP complex in the nuclei may be related closely to transcriptions of G1 arrest-related genes. Taken together, we concluded that TEAD4 might play an important role in tumoral growth and have potential to be a therapeutic target in OSCCs.


The genus Alterosa Blahnik, 2005 (Trichoptera, Philopotamidae, Philopotaminae) in northeastern Brazil, including the description of three new species and an identification key for the genus.
Alterosa Blahnik, 2005 contains 35 described species distributed in southern and southeastern Brazil. Three new species of Alterosa from northeastern Brazil are described and illustrated, Alterosa amadoi sp. n., Alterosa castroalvesi sp. n. and Alterosa caymmii sp. n., the first records of the genus from northeastern Brazil. An identification key for all known species of the genus is also presented.


Localization of dipole by boundary element method in three dimensional reconstructed monkey brain.
The boundary element method was applied to 3-dimensionally reconstructed monkey brain to localize a current source generator (dipole) of somatosensory evoked potentials (SEPs) recorded from epidural electrodes under assumption that a brain is electrically homogeneous and is bounded by a high resistant substance (skull). Absolute mean deviation of estimated dipoles from actual dipoles artificially generated by electrical stimulation at known coordinates in the brain were within 1-3 mm. The estimated dipole computed from SEPs by right median nerve stimulation at latency of P10-N10 waves was located in area 3b of the somatosensory hand area in monkey. Our data suggest that this technique of locating dipoles by the boundary element method applied to a reconstructed brain allows accurate determination of the 3-dimensional coordinates of a dipole.


A quantitative study on the tail flick test in the rat.
Tail flick latency and response temperature were measured in awake rats. Both of these parameters varied with the change in rate of increase of radiant heat stimulus and the area of skin irradiated. The caloric factor applied to the tail before the response occurred was calculated approximately from the time-temperature curves monitored over the tail and the response latencies. The magnitude of the caloric factor over a tentative threshold temperature was the crucial factor for the induction of tail flick response, because it was similar for the various irradiating conditions. It was proposed that analgesic effects in the tail flick test may be expressed using this caloric factor which allows direct comparison of studies in which different base-line latencies are used.


Systematic integration of protein-affecting mutations, gene fusions, and copy number alterations into a comprehensive somatic mutational profile.
Somatic mutations occur as random genetic changes in genes through protein-affecting mutations (PAMs), gene fusions, or copy number alterations (CNAs). Mutations of different types can have a similar phenotypic effect (i.e., allelic heterogeneity) and should be integrated into a unified gene mutation profile. We developed OncoMerge to fill this niche of integrating somatic mutations to capture allelic heterogeneity, assign a function to mutations, and overcome known obstacles in cancer genetics. Application of OncoMerge to TCGA Pan-Cancer Atlas increased detection of somatically mutated genes and improved the prediction of the somatic mutation role as either activating or loss of function. Using integrated somatic mutation matrices increased the power to infer gene regulatory networks and uncovered the enrichment of switch-like feedback motifs and delay-inducing feedforward loops. These studies demonstrate that OncoMerge efficiently integrates PAMs, fusions, and CNAs and strengthens downstream analyses linking somatic mutations to cancer phenotypes.


Efficacy of Multicomponent Tympanometry in Identifying Middle Ear Disorder.
Multi component tympanometry is not widely used in clinics due lack of evidence supporting its usefulness. The present study was undertaken to investigate the efficacy of multicomponent tympanometry for 1000 Hz probe tone in detecting middle ear pathology. Data were collected from 20 ears of 10 participants with no history of middle ear pathology (Group I) and 39 ears of 26 participants (Group II) with history of middle ear infection or ear-pain or blocking sensation. Single component tympanometry for 226 Hz probe tone and Susceptancec and Conductance tympanograms for 1000 Hz probe tone was carried out on all the participants. Results revealed normal tympanogram for both the probe tones for all the participants of Group I. It was observed that 53.84% of the ears could be identified as having middle ear pathology using 226 Hz probe tone whereas the percentage of correct identification increased to 94.87% when 1000 Hz probe tone was used. They are in accordance with the concept that subtle changes in the middle ear which cannot be detected by 226 Hz tympanogram can be identified using 1000 Hz multicomponent tympanogram. These findings highlight the efficiency of 1000 Hz probe tone in detecting mass dominant middle ear pathology.


Antigen cocktails: valid hypothesis or unsubstantiated hope?
After more than three decades of research into the development of vaccines against parasites, a substantial number of antigens have been identified that, as purified native proteins or recombinant proteins, induce some protection against the target parasite. Very few achieve a degree of efficacy likely to make them candidates for single-antigen vaccines. Therefore, multi-antigen or 'cocktail' vaccines are proposed based on the assumption that such cocktails will show enhanced efficacy. This assumption, although often poorly acknowledged, has become central to much vaccine research. The experimental evidence for it, however, is extremely scarce and contradictory. The efficacy of multicomponent vaccines deserves greater experimental attention than it has received.


Human leukemic K562 cells: differential effects of 5-azacytidine on DNA methylation of epsilon-, gamma-globin and 7SL RNA genes.
5 Azacytidine ribonucleoside (5 Aza CR), greatly enhances erythroid differentiation of the K562(h) cell line, with a sharp increase of embryonic and fetal globin gene expression. This phenomenon is correlated with the undermethylation of gamma-globin but not of epsilon-globin, as the epsilon-globin gene is already extensively undermethylated before 5AzaCR induction. By contrast no variations in both DNA methylation and expression are observed in 7SL RNA genes.


Association between vomiting in the first trimester and preterm birth: a retrospective birth cohort study in Wuhan, China.
Although vomiting in the first trimester has been reported to be associated with preterm birth (PTB), findings supporting this association remain inconsistent. Our aim was to assess the association between vomiting and PTB, as well as evaluate if the association is modified by pre-pregnancy body mass index (BMI). A retrospective cohort study. Wuhan, a central city of China. A total of 317 463 pregnant women who had a live, singleton newborn from 1 January 2010 to 23 May 2016 were enrolled in our study. PTB was defined as gestational age <37 gestational weeks. Gestational age was calculated using reports from mothers based on the first day of their last menstrual period. An ultrasound was routinely used to determine gestational age before 12 gestational weeks. Of the 317 463 pregnant women, 29.88% (94 857) experienced vomiting in the first trimester and 5.00% (15 889) experienced a PTB. Vomiting in the first trimester increased the risk for PTB and the multivariable adjusted OR was 1.05 (95% CI 1.02 to 1.09). In the stratified analyses, the association of vomiting in the first trimester was significant among underweight women (adjusted OR=1.08, 95% CI 1.04 to 1.17) and normal pre-pregnancy BMI women (adjusted OR=1.06, 95% CI 1.02 to 1.11), but not in overweight women (adjusted OR=1.01, 95% CI 0.90 to 1.14) and obese women (adjusted OR=0.93, 95% CI 0.73 to 1.19). Our study indicates that vomiting in the first trimester was associated with PTB. Additionally, women with underweight and normal pre-pregnancy BMI who experienced vomiting are more likely to have a PTB.


Corrosion of pure titanium sternal wire.
Titanium and its alloys are considered reliable materials commonly used in heart surgeries. A patient had severe inflammation at the median sternotomy wound develop after open heart surgery. Sternal wires were removed and were found to be seriously corroded on most of the surface. Energy dispersive spectroscopy identified titanium as the only elemental component of the explanted wire with grade 2 purity, and the polarization curve showed satisfactory corrosion resistance similar to commercial pure titanium. Prolonged inflammation might have caused extensive corrosion of titanium in the present case.


Predictors of successful emergency shock wave lithotripsy for acute renal colic.
The role of emergency shockwave lithotripsy (SWL) in persistent pain control in patients with ureteral stones is not well established. The aim of this study is to evaluate efficacy as well as the predicting variables for successful early SWL patients with symptomatic ureteral stones. Eighty-six patients with a persistent renal colic secondary to single ureteral stone (6-12 mm) were prospectively enrolled in this study. SWL was performed within 24 h of the onset of flank pain. Pain control and stone-free rate after emergency SWL session were 58.1% and 44.2%, respectively. Seven patients required post-SWL ureteroscopy and ureteral stent placement for uncontrolled pain. The overall 3-month stone-free rate after SWL monotherapy was 83.7%. On multivariate analysis, predictors for pain relief after emergency SWL were lower Hounsfield (HU) stone density, mild hydronephrosis (HN) at presentation and presentation during the first colic episode. Lower HU stone density was the single predictor of successful stone clearance after single emergency SWL session on multivariate analysis. In conclusion, early SWL is feasible and effective in management of ureteral stones presented by renal colic with low HU.


Gene expression networks involved in multiple cellular programs coexist in individual hepatocellular cancer cells.
The gene expression networks of a single cell can be used to reveal cell type- and condition-specific patterns that account for cell states, cell identity, and its responses to environmental changes. We applied single cell sequencing datasets to define mRNA patterns and visualized potential cellular capacities among hepatocellular cancer cells. The expressing numbers and levels of genes were highly heterogenous among the cancer cells. The cellular characteristics were dependent strongly on the expressing numbers and levels of genes, especially oncogenes and anti-oncogenes, in an individual cancer cell. The transcriptional activations of oncogenes and anti-oncogenes were strongly linked to inherent multiple cellular programs, some of which oppose and contend against other processes, in a cancer cell. The gene expression networks of multiple cellular programs proliferation, differentiation, apoptosis, autophagy, epithelial-mesenchymal transition, ATP production, and neurogenesis coexisted in an individual cancer cell. The findings give rise a hypothesis that a cancer cell expresses balanced combinations of genes and undergoes a given biological process by rapidly transmuting gene expressing networks.


Chronic kidney disease mineral bone disorder in childhood and young adulthood: a 'growing' understanding.
Chronic kidney disease (CKD) mineral and bone disorder (MBD) comprises a triad of biochemical abnormalities (of calcium, phosphate, parathyroid hormone and vitamin D), bone abnormalities (turnover, mineralization and growth) and extra-skeletal calcification. Mineral dysregulation leads to bone demineralization causing bone pain and an increased fracture risk compared to healthy peers. Vascular calcification, with hydroxyapatite deposition in the vessel wall, is a part of the CKD-MBD spectrum and, in turn, leads to vascular stiffness, left ventricular hypertrophy and a very high cardiovascular mortality risk. While the growing bone requires calcium, excess calcium can deposit in the vessels, such that the intake of calcium, calcium- containing medications and high calcium dialysate need to be carefully regulated. Normal physiological bone mineralization continues into the third decade of life, many years beyond the rapid growth in childhood and adolescence, implying that skeletal calcium requirements are much higher in younger people compared to the elderly. Much of the research into the link between bone (de)mineralization and vascular calcification in CKD has been performed in older adults and these data must not be extrapolated to children or younger adults. In this article, we explore the physiological changes in bone turnover and mineralization in children and young adults, the pathophysiology of mineral bone disease in CKD and a potential link between bone demineralization and vascular calcification. A higher resolution version of the Graphical abstract is available as Supplementary information.


Expanded trinucleotide repeats in the TBP/SCA17 gene mapped to chromosome 6q27 are associated with schizophrenia.
Schizophrenia has a complex and non-Mendelian mode of inheritance. Recently, trinucleotide repeat (TNR)-containing genes have been considered as the candidate genes predisposing to schizophrenia. The purpose of this study was to determine whether a genetic association could be observed between schizophrenia and the TNR polymorphisms within the KLHL1AS/SCA8, PPP2R2B/SCA12, and TBP/SCA17 genes. We studied 100 unrelated schizophrenia patients and 124 controls without evident neurodegenerative or psychiatric disorders. The overall allele frequency distributions of the KLHL1AS/SCA8 and PPP2R2B/SCA12 genes were not significantly different between the schizophrenic patients and the control subjects (P>0.05). The allele frequency distribution in the schizophrenic patients was significantly different from that in the controls at the TBP/SCA17 gene (P=0.0149), with an increased frequency of 36 repeats in the patients and two patients carrying 45 TNR expansions were identified. TBP/SCA17 is the TATA box binding protein gene mapped to chromosome 6q27. The study suggests that TNR expansions of the TBP/SCA17 gene may contribute to the genetic risk of schizophrenia in rare cases.


Cryopreservation of human platelets. General protocols, development of freezing techniques and clinical applications.
A review is presented of the recent advances in cryopreservation of human platelets with particular reference to the technical protocol reported and its development, using either DMSO, glycerol or HES as additives. The methods for assaying viability of frozen-thawed platelets are described and the results are reported according to the cryoprotector used. The clinical applications of frozen platelets are discussed with reference to the clinical experience with DMSO and glycerol. An organization scheme for a frozen platelet bank is proposed.


A review on recent developments in mass spectrometry instrumentation and quantitative tools advancing bacterial proteomics.
Proteomics has evolved substantially since its early days, some 20 years ago. In this mini-review, we aim to provide an overview of general methodologies and more recent developments in mass spectrometric approaches used for relative and absolute quantitation of proteins. Enhancement of sensitivity of the mass spectrometers as well as improved sample preparation and protein fractionation methods are resulting in a more comprehensive analysis of proteomes. We also document some upcoming trends for quantitative proteomics such as the use of label-free quantification methods. Hopefully, microbiologists will continue to explore proteomics as a tool in their research to understand the adaptation of microorganisms to their ever changing environment. We encourage them to incorporate some of the described new developments in mass spectrometry to facilitate their analyses and improve the general knowledge of the fascinating world of microorganisms.


The effect of phenytoin on the action potential of a vertebrate spinal neuron.
The effect of phenytoin (PTN) 20 microgram/ml was tested on the passive membrane properties and the action potential of the dorsal cells in the spinal cord of the river lamprey. Dorsal cells are primary sensory neurons with no synaptic input, and thus allow examination of membrane properties in the absence of contaminating synaptic currents. PTN did not affect the resting membrane potential and slightly raised the input resistance. However, it greatly raised the threshold voltage and current for activation of action potentials by intracellularly injected current. It also reduced the maximum rate of rise of the action potential, the spike overshoot and the spike undershoot, while increasing the spike duration. In contrast to findings in other vertebrate sensory neurons, dorsal cell action potentials were blocked in zero sodium or tetrodotoxin, and not affected by zero calcium or 1 mM manganese. Thus they PTN on dorsal cells is that it partially blocks the activated sodium conductance increase of the action potential. Because a long delay was observed for maximal effect of PTN and for washout, it is postulated that the drug may require partitioning into the lipid membrane, or entry into the cell for its pharmacological action.


Immunology of multisystem ocular disease.
In this review based on Prof. James' 1983 George Nelms Wise Lecture, the authors summarize proven and speculated relationships involved in the immunology of multisystem ocular disease. Extensive tables elucidate the mechanisms underlying such conditions as, systemic lupus erythematosus, polyarteritis nodosa, Sjögren's syndrome, Wegener's granulomatosis, scleritis, sarcoidosis, Behcet's disease, idiopathic retinal vasculitis, sympathetic ophthalmia, Vogt-Koyanagi-Harada syndrome, and dysthyroid eye disease.


Effects of HDV infection and pegylated interferon α treatment on the natural killer cell compartment in chronically infected individuals.
Although hepatitis delta is considered an immune-mediated disease, adaptive immune responses to hepatitis delta virus (HDV) are hardly detectable. Thus, the role of other immune responses, including those mediated by natural killer (NK) cells, must be considered in HDV pathogenesis and in treatments with immune-stimulating agents such as interferon (IFN)α. However, the phenotype and function of NK cells in chronic HDV infection, or in HDV-infected individuals undergoing IFNα treatment, have not been extensively studied. We performed an extensive analysis of NK cells in chronically HDV-infected patients before and during treatment with IFNα, and compared the results with those for patients with HBV mono-infection as well as healthy controls. In untreated HDV-infected patients, a higher than normal frequency of NK cells was observed in peripheral blood with unaltered phenotypic NK cell differentiation status. In contrast, long-term IFNα treatment of HDV-infected patients caused a significant change in NK cell differentiation status, with selective loss of terminally differentiated NK cells and, in parallel, a relative enrichment in immature NK cell subsets. Treatment was associated with marked functional impairment of the NK cells, which was independent of the changes in NK cell differentiation status. Furthermore, treatment polarised NK cell IFN signalling from STAT4 towards STAT1 dependency. Strikingly, a high frequency of CD56(dim) NK cells at baseline was positively associated with IFNα treatment outcome in the patients. We describe in detail how HDV infection, and IFNα treatment of this infection, affects the NK cell compartment and what consequences this has for the functional capacity of NK cells.


Blocking human protein C anticoagulant activity improves clotting defects of hemophilia mice expressing human protein C.
Hemophilia A and B are hereditary coagulation defects resulting in unstable blood clotting and recurrent bleeding. Current factor replacement therapies have major limitations such as the short half-life of the factors and development of inhibitors. Alternative approaches to rebalance the hemostasis by inhibiting the anticoagulant pathways have recently gained considerable interest. In this study, we tested the therapeutic potential of a monoclonal antibody, HAPC1573, that selectively blocks the anticoagulant activity of human activated protein C (APC). We generated F8-/- or F9-/- hemophilia mice expressing human protein C by genetically replacing the murine Proc gene with the human PROC. The resulting PROC+/+;F8-/- or PROC+/+;F9-/- mice had bleeding characteristics similar to their corresponding F8-/- or F9-/- mice. Pretreating the PROC+/+;F8-/- mice with HAPC1573 shortened the tail bleeding time. HAPC1573 pretreatment significantly reduced mortality and alleviated joint swelling, similar to those treated with either FVIII or FIX, of either PROC+/+;F8-/- or PROC+/+;F9-/- mice in a needle puncture-induced knee-joint bleeding model. Additionally, we found that HAPC1573 significantly improved the thrombin generation of PROC+/+;F8-/- mice but not F8-/- mice, indicating that HAPC1573 enhanced the coagulant activity of hemophilia mice by modulating human APC in vivo. We further documented that HAPC1573 inhibited the APC anticoagulant activity to improve the clotting time of human plasma deficient of FVIII, FIX, FXI, FVII, VWF, FV, or FX. These results demonstrate that selectively blocking the anticoagulant activity of human APC may be an effective therapeutic and/or prophylactic approach for bleeding disorders lacking FVIII, FIX, or other clotting factors.


Characteristics of neuropathic pain and its relationship with quality of life in 72 patients with spinal cord injury.
A cross-sectional study. Neuropathic pain (NP) after spinal cord injury (SCI) tends to be hard to treat, and its heterogeneous properties make it difficult to identify and characterize. This study was conducted to assess the characteristics of SCI-related NP in detail. A single hospital for SCI rehabilitation. This study included 72 patients who were seen at our hospital in 2012 and 2013 and who had sustained SCI at least 3 months before enrollment. The patients completed the Neuropathic Pain Symptom Inventory (NPSI) and the Short Form (SF)-36 Health Inventory. The NPSI score was analyzed for correlations with clinical presentations of SCI and SF-36 subitems. Paresthesia/dysesthesia was the most common subtype of NP after SCI. With regard to location, below-level superficial NP was significantly more intense than at-level pain. Patients who underwent surgery showed significantly less evoked pain compared with patients with non-surgery. Patients reported significantly more severe pain if >1 year had elapsed after the SCI. Patients with an American Spinal Injury Association Impairment Scale grade of B for completeness of injury reported more intense NP than those with other grades. Among the SF-36 subitems, NP correlated significantly with bodily pain, general health and mental health. NP in SCI patients was significantly associated with the location of pain, the time period since the injury, surgery and quality-of-life factors. A more detailed understanding of the characteristics of NP may contribute to better strategies for relieving the pain associated with SCI.


A new predictor of the optimal pulmonary artery reconstruction method during the arterial switch operation.
The aim of this study was to more accurately predict the optimal pulmonary artery (PA) reconstruction procedure (Lecompte manoeuvre or original Jatene procedure) during the arterial switch operation, we focused on the horizontal sectioning (HS) angle between the left hilum PA and the great arteries using preoperative computed tomography imaging. We defined the HS angle α (β) as the angle between the tangential line from the posterior (anterior) wall of the left PA at the hilum to the left anterior (right posterior) surface of the main PA and the tangential line from the left surface of the ascending aorta to the left anterior (right posterior) surface of the main PA. We identified 14 consecutive patients diagnosed with transposition of the great arteries or transposition of the great artery-type double-outlet right ventricle who underwent preoperative computed tomography imaging. The original Jatene or Lecompte procedure was used for 9 (OJ group) and 5 (L group) patients. Relationships of the great arteries of the OJ and L groups were side by side in 8 and 2 patients, oblique in 1 and 1 patient and anteroposterior in 0 and 2 patients, respectively. In the OJ group, β was greater than α in all patients. The median α/β value was 0.618. In group L, α was greater than β in all patients. The median α/β was 1.307. Left PA stenosis caused by stretching was not observed in the L group. Coronary obstruction was not identified in the OJ group. Left PA stenosis behind the neo-ascending aorta was observed in 1 patient in the OJ group and required reoperation. The HS angle may be a useful predictor of optimal intraoperative PA reconstruction during arterial switch operation, especially for side-by-side or oblique relationships.


Targeting of RUNX3 by miR-130a and miR-495 cooperatively increases cell proliferation and tumor angiogenesis in gastric cancer cells.
Mature microRNAs (miRNAs) are 21 to 23 nucleotide noncoding RNA molecules that can downregulate multiple gene expression by mRNA degradation or translational repression. miRNAs are considered to play important roles in cell proliferation, apoptosis, and differentiation during mammalian development. The Runt-related transcription factor 3 (RUNX3) expression and activity are frequently downregulated by various mechanisms in gastric cancer. We have reported that RUNX3 inactivation is crucial for early tumorigenesis. In this study, we investigated the role of miRNAs targeting RUNX3 in early tumorigenesis. miR-130a and miR-495 upregulated under hypoxic conditions that bind to the RUNX3 3'-untranslated region (3'-UTR) were identified in gastric cancer cells by using microarray analysis and bioinformatics programs. Combination of miR-130a and miR-495 inhibited RUNX3 expression at the protein level, but not at the mRNA level. miR-130a and miR-495 significantly inhibited the RUNX3-3'UTR-luciferase activity. Combination of miR-130a and miR-495 significantly decreased apoptosis determined by Annexin V-FITC/propidium iodide staining and flow cytometric analysis, and the expression of Bim in SNU484 gastric cancer cells. In addition, p21 and Bim, RUNX3 target genes, were completely downregulated by the combination of miR-130a and miR-495. Using matrigel plug assay, we found that antagomiRs specific for miR-130a and miR-495 significantly reduced angiogenesis in vivo. In conclusion, targeting miR-130a and miR-495 could be a potential therapeutics to recover RUNX3 expression under hypoxic conditions and in early tumorigenic progression.


Establishment of comorbidity target pools and prediction of drugs candidate for multiple sclerosis and autoimmune thyroid diseases based on GWAS and transcriptome data.
Clinical observation has revealed that multiple sclerosis (MS) and autoimmune thyroid disease (AITD) are strongly correlated. The aim of this study was to explore the shared molecular causes of MS and AITD, and to conduct drug rearrangement on this basis, search for comorbidity drugs and feasible drugs for mutual reference between the two diseases. Based on genome-wide association study (GWAS) data and transcriptome data, susceptibility genes and differentially expressed genes related to MS and AITD were identified by bioinformatics analysis. Pathway enrichment, gene ontology (GO), protein-protein interaction analysis, and gene-pathway network analysis of the above genes were performed to identify a common target pool, including common genes, common hub genes, and common pathways, and to explore the specific pathogenesis of the two diseases, respectively. Drugs that target the common pathways/genes were identified through the Comparative Toxicogenomics Database (CTD), DrugBank database, and Drug-Gene Interaction (DGI) Database. Common hub genes were compared with the target genes of drugs approved for treating MS/AITD and drugs under investigation identified by DrugBank and ClinicalTrials, respectively. We identified a pool of shared targets containing genes and pathways, including 46 common genetic susceptibility pathways and 9 common differentially expressed pathways, including JAK-STAT signaling pathway, Th17 cell differentiation, Th1 and Th2 cell differentiation, PD-L1 expression and PD-1 checkpoint pathway in cancer, etc. In addition, a total of 29 hub genes, including TYK2, JAK1, STAT3, IL2RA, HLA-DRB1, and TLR3, were identified. Drugs approved for treating MS or AITD, such as methylprednisolone, cyclophosphamide, glatiramer, natalizumab, and methimazole, can target the shared genes and pathways, among which methylprednisolone and cyclophosphamide have been shown to be beneficial for the treatment of the two diseases, indicating that these drugs have the potential to become a priority in the treatment of comorbidities. Moreover, drugs targeting multiple common genes and pathways, including tacrolimus, deucravacitinib, and nivolumab, were identified as potential drugs for the treatment of MS, AITD, and their comorbidities. We observed that T-cell activation-related genes and pathways play a major role in the pathogenesis of both MS and AITD, which may be the molecular basis of the comorbidity. Moreover, we identified a variety of drugs which may be used as priority or potential treatments for comorbidities.


Circulating tumor DNA as a Predictive and Prognostic Biomarker in the Perioperative Treatment of Muscle-invasive Bladder Cancer: A Systematic Review.
Predictive and prognostic biomarkers in the perioperative treatment of muscle-invasive bladder cancer (MIBC) are an unmet need. Circulating tumor DNA (ctDNA) holds promise as a biomarker in this setting. To review the evidence of ctDNA as a prognostic and predictive biomarker in the perioperative treatment of MIBC. We systematically reviewed the literature using PubMed, MEDLINE, and Embase databases according to the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) statement. We included prospective studies investigating neoadjuvant and/or adjuvant chemotherapy and/or immunotherapy for MIBC (T2-T4a, any N, and M0) treated with radical cystectomy. We reported ctDNA results to monitor and/or predict disease status, relapse, and progression. The research retrieved 223 records. Six papers were considered for this review based on prespecified inclusion criteria. Our review confirms the prognostic role of ctDNA after cystectomy and shows a potential predictive benefit in using neoadjuvant chemotherapy and preoperative immunotherapy. Circulating tumor DNA was used to monitor recurrence, and changes in ctDNA status anticipated radiological progression with a median difference of time from 101 to 932 d. A subgroup analysis of the phase 3 Imvigor010 trial showed that only ctDNA-positive patients treated with atezolizumab had an improvement in disease-free survival (DFS; hazard ratio [HR] = 3.36, 95% confidence interval [CI]: 2.44-4.62). Clearance of ctDNA after two cycles of adjuvant atezolizumab was associated with improved outcomes (DFS HR = 0.26, 95% CI: 0.12-0.56, p = 0.0014; overall survival HR = 0.14, 95% CI: 0.03-0.59). Circulating tumor DNA is a prognostic factor after cystectomy and may be used to monitor recurrence. In the adjuvant immunotherapy setting, ctDNA might select patients who benefit the most from this strategy. In the perioperative treatment of muscle-invasive bladder cancer, circulating tumor DNA (ctDNA) positivity correlates with the outcomes after cystectomy and might select patients who may benefit from neoadjuvant chemotherapy and/or immunotherapy. Changes in ctDNA status anticipated radiological progression.


Promise of Real-World Evidence for Patient Centricity in Gulf Cooperation Council Countries: Call to Action.
Presently, Gulf Cooperation Council countries are lagging in the generation of real-world data and use of real-world evidence for patient-centered care compared with the global average. In a collaborative effort, experts from multiple domains of the healthcare environment from the Gulf Cooperation Council countries came together to present their views and recommended key action points for the generation of robust real-world data and leveraging real-world evidence in the countries. The opinions of the experts are presented, along with existing barriers to the effective generation of real-world evidence in the countries. The Gulf Cooperation Council countries are undergoing transformative changes paving the way for improved healthcare measures; however, the challenges in generating reliable, robust, accessible, and secure real-world evidence are persistent. Hence, ongoing public-private engagements, as well as collaborations between regulators, policymakers, healthcare professionals, insurance and pharmaceutical companies, and patients, are warranted. A few notable examples of real-world evidence studies highlighting the benefits of real-world evidence for gaining valuable insights into patient-centric decision making are also discussed. The actionable steps identified for successful real-world evidence generation would provide long-term, real-world evidence-based patient-centric benefits for the countries.


Viral and bacterial infections associated with camel (Camelus dromedarius) calf diarrhea in North Province, Saudi Arabia.
Diarrhea and deaths in new-born camel calves were noticed by veterinary investigators and pastoralist in Saudi Arabia to be very high. Hence, it is thought to be necessary to investigate this problem from the virological and bacteriological point of view. The role of pathogenic bacteria and viruses in six different towns of North Province (Al-Assafia, Arar, Domat Aljandal, Hail, Skaka and Khoa) in Saudi Arabia was studied. Survey was conducted in diarrheic camel calves aged 12 months or younger. In our study calf diarrhea was reported in 184 out of 2308 camels examined clinically during one year, the prevalence of diarrhea was found to be 8.0% in calves ranging from one month to one year. In the present study group A rotavirus and Brucella abortus were detected in 14.7% and 8.98%, respectively, using ELISA technique. Escherichia coli was isolated from diarrheic calf camel (58.2%) 99/170 samples during dry and wet season. Salmonella spp. and Enterococcus spp. were detected in 12% and 8.8% of the specimens, respectively. In this study enterotoxogenic E. coli (ET E. coli) was isolated from 7% of diarrheic camel, which indicates the strong correlation between the camel calf diarrhea and the detection of enterotoxogenic E. coli. This study represented the first report for the detection of group A rotavirus and B. abortus antigen and antibodies in calf camels in Saudi Arabia. It is recommended that the disease should be controlled by vaccination in calf camels.


Ideal sphere-forming culture conditions to maintain pluripotency in a hepatocellular carcinoma cell lines.
Cancer stem cells (CSCs) constitute 1-2 % of cancer tissue and are a major cause of tumor metastasis and recurrence. The culture environment is important in maintaining CSCs in vitro. Sphere formation is one method of culturing CSCs. In this study, we identified and validated optimal culture conditions for sphere formation in hepatocellular carcinoma cells. Huh7 and HepG2 cells were plated in three different media types and were allowed to form spheres. To confirm the pluripotency of sphere cells, the expression of stem cell markers was evaluated. EpCAM, Connexin 32, and Connexin 43 expression was evaluated using reverse transcription-polymerase chain reaction (RT-PCR). The expression of E-cadherin, β-catenin, CD90, and CD133 was evaluated using immunocytochemistry. Flow cytometry was performed to confirm the presence of stem cell markers CD133 and Connexin 32. Cells maintained in medium containing growth factors ((DMEM(+))GF) showed greater cell proliferation than cells in media with fetal bovine serum (FBS) (DMEM(+)FBS) or without FBS (DMEM(-)FBS). Cells cultured in DMEM(+)FBS medium exhibited greater proliferation after 20 days in culture than cells cultured under the other two conditions. Cells cultured in DMEM(-)FBS medium did not proliferate; therefore, this condition was removed from further analysis. RT-PCR and flow cytometry showed that sphere-forming cells cultured in DMEM(+)GF and DMEM(+)FBS media had similar expression of stem cell markers. Therefore, growth factor-free medium is an adaptable, efficient, and cost-effective tool for in vitro cultivation of CSCs.


Incidental paraduodenal hernia found during laparoscopic colectomy.
This paper describes a rare right paraduodenal hernia discovered during an elective laparoscopic colon resection. Our patient was a 60-year-old Asian man with a history of multiple bouts of diverticulitis and a lifelong history of mild constipation and postprandial abdominal pain. Prior CT scans and preoperative barium enema confirmed the diagnosis of diverticular disease, and no other abnormalities were appreciated. At laparoscopic exploration, a right paraduodenal hernia was found with complete herniation of the small intestine under the ascending colon and hepatic flexure. The unclear anatomy prompted conversion to an open laparotomy. This allowed safe reduction of the hernia and sac excision. Adhesions were lysed to relieve a partial duodenal obstruction, and a Ladd's procedure was performed to correct the incomplete rotation. Additionally, a sigmoid colectomy was performed. After prolonged ileus, the patient was discharged on postoperative day 14. At 6-month follow-up, the patient was asymptomatic and doing well.


Roles of Arg-97 and Phe-113 in regulation of distal ligand binding to heme in the sensor domain of Ec DOS protein. Resonance Raman and mutation study.
The direct oxygen sensor protein isolated from Escherichia coli (Ec DOS) is a heme-based signal transducer protein responsible for phosphodiesterase (PDE) activity. Binding of O(2), CO, or NO to a reduced heme significantly enhances the PDE activity toward 3',5'-cyclic diguanylic acid. We report stationary and time-resolved resonance Raman spectra of the wild-type and several mutants (Glu-93 --> Ile, Met-95 --> Ala, Arg-97 --> Ile, Arg-97 --> Ala, Arg-97 --> Glu, Phe-113 --> Leu, and Phe-113 --> Thr) of the heme-containing PAS domain of Ec DOS. For the CO- and NO-bound forms, both the hydrogen-bonded and non-hydrogen-bonded conformations were found, and in the former Arg-97 forms a hydrogen bond with the heme-bound external ligand. The resonance Raman results revealed significant interactions of Arg-97 and Phe-113 with a ligand bound to the sixth coordination site of the heme and profound structural changes in the heme propionates upon dissociation of CO. Mutation of Phe-113 perturbed the PDE activities, and the mutation of Arg-97 and Phe-113 significantly influenced the transient binding of Met-95 to the heme upon photodissociation of CO. This suggests that the electrostatic interaction of Arg-97 and steric interaction of Phe-113 are crucial for regulating the competitive recombination of Met-95 and CO to the heme. On the basis of these results, we propose a model for the role of the heme propionates in communicating the heme structural changes to the protein moiety.


An educational intervention based on family-centered empowerment model to modify high-risk behaviors of brucellosis via mother education.
The relative contribution of a theory-based intervention to modify high-risk behaviors in training programs is a major priority that remains an open question. Here, we tested whether the family-centered empowerment model used in the educational intervention was effective to modify high-risk behaviors of brucellosis via mother education. A quasi experimental study was conducted on 200 women presenting to healthcare practices in rural areas of Torbat-e Jam, Iran, from April 2020 to February 2021. Four rural areas were randomly assigned to the control and intervention groups. The intervention group received the training program, which included four 2-h sessions and consulting support via social network and messaging service. The control group did not receive any training. SPSS16 was implemented to test multiple statistical analyses. Our finding showed in the intervention group compared with the control group, knowledge, attitude, self-efficacy, self-esteem, and behavior outcomes were significantly changed (P < 0.001) across time during baseline through follow-up. Likewise, there are no differences (P > 0.05) in the change in construct of the family-centered empowerment model and risk behaviors in the control group from baseline to follow-up. Intervention based on a family-centered empowerment model is possible and very acceptable to modify high-risk behaviors of brucellosis by increasing an individual's knowledge, changing attitude, and promoting self-efficacy and self-esteem.Trial registration: Iranian Registry of Clinical Trials (IRCT), IRCT20160619028529N12. Registration date: 24/03/2020.


Construction of adipose tissue using a silica expander capsule and cell sheet-assembled of decellularized adipose tissue.
Delayed neovascularization and unstable adipose formation are major confounding factors in adipose tissue engineering. A system using decellularized adipose tissue (DAT), adipose-derived stem cells (ADSCs), and human umbilical vein endothelial cells (HUVECs) has been preliminarily studied, but it requires optimization, as adipogenic and angiogenic capabilities for maintaining a stable construct shape are limited. The current study aimed to address these limitations. Our initial modification involved the addition of exogenous chemokine (C-C motif) ligand 2 (CCL2), which resulted in enhanced adipogenesis and angiogenesis. However, further improvement was required due to delayed blood recanalization. To further optimize the system, a vascularized fibrous capsule derived from an implanted silica expander was utilized as a second modification. We hypothesized this would function as both a microbioreactor to fix the seed cells and exogenous CCL2 locally and as a vascular bed to promote neovascularization. Compared with that of the CCL2 loaded ADSC-HUVECs cell sheet assembled DAT system, adding the silica expander capsule resulted in significantly increased construct stability, new vessel intensity, a greater number of Oil Red O-positive lipid droplets, more enhanced tissue remodeling, and upregulated peroxisome proliferator-activated receptor gamma (PPARγ) & leptin expression. Thus, these two modifications helped optimize the currently available ADSC-HUVEC cell sheet assembled DAT system, providing an adipose tissue construction strategy with enhanced adipogenesis and angiogenesis to reconstruct soft tissue defects. Moreover, close-to-normal leptin expression provided the engineered adipose tissue with a glucometabolic function, in addition to remodeling capabilities. STATEMENT OF SIGNIFICANCE: Delayed neovascularization and unstable adipose formation are the two major problems in tissue engineering adipose. Here, we introduced an adipose tissue engineering construction strategy using a silica expander capsule along with hADSCs-HUVECs cell sheet-assembled DAT in a CCL2-rich microenvironment. Our data suggested that CCL2 could improve angiogenesis and adipogenesis in vitro and in vivo. The addition of tissue expander capsule could further improve the stability of construction and fabricated adipose tissue with increased new vessel intensity, greater numbers of Oil Red O-positive lipid droplets, more enhanced tissue remodeling, and upregulated leptin expression. CCL2 and expander capsule can have clinical utility for soft tissue defects repair, and these two factors can be useful in other tissue engineering.


Electron microscopic investigations of synaptonemal complexes in an infertile human male carrier of a pericentric inversion inv(1)(p32q42). Regular loop formation but defective synapsis including a possible interchromosomal effect.
Electron microscopic investigations of surface spread synaptonemal complexes in spermatocytes from a 37-year-old man ascertained for infertility detected a pericentric inv(1), and subsequent lymphocyte analysis placed the breakpoints at p32 and q42. Most spermatocytes showed a maturation arrest at mid-pachytene explaining the azoospermia. As in two other comparatively large loop-forming pericentric inversions, initiation of synapsis took place in the middle of the inverted segment. Thus there is no indication of interstitial synaptic initiation being restricted to special pairing sites along the length of the chromosome. All spermatocytes investigated at mid-pachytene showed inversion loops, none of which was fully synapsed with a specific delay in pairing of the heterochromatic block 1qh and adjacent segments. The loops were of similar size in all the cells examined and synaptic adjustment had not taken place. There was no indication of a preferential association between the inv(1) bivalent and the XY configuration, and a functional disturbance of the X seems an unlikely reason for the meiotic maturation arrest. The most likely cause may be the failure of adequate synapsis of the inverted segment and the possibly associated pairing abnormalities of other homologues, including asynapsis and/or precocious desynapsis.


Structure of a shark IgNAR antibody variable domain and modeling of an early-developmental isotype.
The new antigen receptor (IgNAR) antibodies from sharks are disulphide bonded dimers of two protein chains, each containing one variable and five constant domains. Three types of IgNAR variable domains have been discovered, with Type 3 appearing early in shark development and being overtaken by the antigen-driven affinity-matured Type 1 and 2 response. Here, we have determined the first structure of a naturally occurring Type 2 IgNAR variable domain, and identified the disulphide bond that links and stabilizes the CDR1 and CDR3 loops. This disulphide bridge locks the CDR3 loop in an "upright" conformation in contrast to other shark antibody structures, where a more lateral configuration is observed. Further, we sought to model the Type 3 isotype based on the crystallographic structure reported here. This modeling indicates (1) that internal Type 3-specific residues combine to pack into a compact immunoglobulin core that supports the CDR loop regions, and (2) that despite apparent low-sequence variability, there is sufficient plasticity in the CDR3 loop to form a conformationally diverse antigen-binding surface.


Effect of intravitreal triamcinolone injection during vitrectomy for idiopathic epiretinal membrane.
To investigate the effect of intravitreal triamcinolone injections as an adjuvant of vitrectomy for idiopathic epiretinal membrane. This study included 58 eyes of 58 patients who were diagnosed as having idiopathic epiretinal membrane and who underwent vitrectomy and membrane peeling. They were divided into 2 groups: 27 eyes in the injected group that received triamcinolone injection during the surgical procedure and 31 eyes in the control group that did not receive injection. The best-corrected visual acuity, foveal thickness, and complications were compared between the two groups. Average visual acuity of the injected group improved from 0.70 logarithm of the minimal angle of resolution (logMAR) (Snellen equivalent, 20/100; range, 20/25-20/200) at baseline to 0.59 logMAR (20/80; range, 20/40-20/200) at 1 month and 0.55 logMAR (20/70; range, 20/30-20/200) at 3 months. Foveal thickness reduced from 456.9 μm (range, 307-740 μm) to 387.9 μm (range, 238-584 μm) at 1 month and 336.4 μm (range, 225-529 μm) at 3 months, respectively. In the control group, average visual acuity improved from 0.63 logMAR (20/80; range, 20/40-20/300) to 0.48 logMAR (20/60; range, 20/25-20/125) and 0.43 logMAR (20/50; range, 20/25-20/160) at 1 month and 3 months, respectively. Foveal thickness reduced from 467.1 μm (range, 328-621 μm) to 376.1 μm (range, 271-429 μm) and 375.1 μm (range, 236-475 μm) at 1 month and 3 months, respectively. There were no significant differences in the visual acuity, foveal thickness, and any complications between the two groups during follow-up. Intravitreal injection of triamcinolone combined with vitrectomy for idiopathic epiretinal membrane did not affect postoperative foveal thickness and functional recovery.


[Quality indicators in hip fracture care after the implementation of an orthogeriatrics team].
The incidence of hip fracture will increase dramatically in the coming decades. It has been associated with high mortality and morbidity. Orthogeriatric programs have improved outcomes in fragility hip fracture worldwide. There is little evidence in Mexico on the impact of orthogeriatrics and adherence to quality indicators. To compare adherence to quality indicators in hip fracture before and after the implementation of an orthogeriatrics team compared to the traditional model. Quasi-experimental study conducted in León, Mexico. Adherence to quality indicators before and after an interconsultant orthogeriatrics team was compared. The indicators measured were: surgical delay, treatment for osteoporosis, early mobilization, type of implant and delirium. We included 83 patients. The surgical delay was reduced from 144 to 116 hours (28 hours), the percentage of patients with early surgery was increased from 7.9 to 18%, in addition the percentage of patients with treatment for osteoporosis was increased and the incidence of delirium was reduced. Adherence to indicators is low, however, it was possible to discreetly increase adherence to these indicators. Internal policies must be generated in public hospitals to improve this adherence and see its impact on long-term outcomes.


Hybrid learning method based on feature clustering and scoring for enhanced COVID-19 breath analysis by an electronic nose.
Breath pattern analysis based on an electronic nose (e-nose), which is a noninvasive, fast, and low-cost method, has been continuously used for detecting human diseases, including the coronavirus disease 2019 (COVID-19). Nevertheless, having big data with several available features is not always beneficial because only a few of them will be relevant and useful to distinguish different breath samples (i.e., positive and negative COVID-19 samples). In this study, we develop a hybrid machine learning-based algorithm combining hierarchical agglomerative clustering analysis and permutation feature importance method to improve the data analysis of a portable e-nose for COVID-19 detection (GeNose C19). Utilizing this learning approach, we can obtain an effective and optimum feature combination, enabling the reduction by half of the number of employed sensors without downgrading the classification model performance. Based on the cross-validation test results on the training data, the hybrid algorithm can result in accuracy, sensitivity, and specificity values of (86 ± 3)%, (88 ± 6)%, and (84 ± 6)%, respectively. Meanwhile, for the testing data, a value of 87% is obtained for all the three metrics. These results exhibit the feasibility of using this hybrid filter-wrapper feature-selection method to pave the way for optimizing the GeNose C19 performance.


L-arginine methylester reduces Ca2+/Cl(-)-dependent L-[3H]glutamate binding and Ca2+-activated neutral protease activity in rat hippocampal membranes.
Specific binding of L-[3H]glutamate was measured in Tris-HCl buffer in rat hippocampal membranes. In these experimental conditions 1 mM CaCl2 induced an increase in binding due to an increase in Bmax. L-Arginine methylester did not modify the Cl(-)-dependent binding of L-[3H]glutamate, but it decreased Ca2+/Cl(-)-stimulated binding in a dose-dependent manner, decreasing Bmax without changing KD. L-Arginine methylester reduced calcium-activated neutral protease activity in a dose-dependent manner. Serine protease inhibitors (aprotinin and di-isopropylfluorophosphate) did not affect L-[3H]glutamate binding, whereas leupeptin reduced it in a dose-dependent manner. L-Arginine did not mimic the effect of L-arginine methylester in either model.


Effects of triclosan on the early life stages and reproduction of medaka Oryzias latipes and induction of hepatic vitellogenin.
Triclosan (2,4,4'-trichloro-2'-hydroxydiphenyl ether) is widely used as antibacterial agent in various industrial products, such as textile goods, soap, shampoo, liquid toothpaste and cosmetics, and often detected in wastewater effluent. In this study, the effects of TCS on the early life stages and reproduction of medaka (Oryzias latipes) were investigated. The 96-h median lethal concentration value of TCS for 24-h-old larvae was 602 microg/l. The hatchability and time to hatching in fertilized eggs exposed to 313 microg/l TCS for 14 days were significantly decreased and delayed, respectively. An assessment of the effects of a TCS 21-day exposure period on the reproduction of paired medaka showed no significant differences in the number of eggs produced and fertility among the control and 20, 100 and 200 microg/l TCS treatment groups. However, concentrations of hepatic vitellogenin were increased significantly in males treated with TCS at 20 and 100 microg/l. In the F(1) generations, although the hatching of embryos in the 20 microg/l treatment showed adverse effects, there was no dose-response relationship between hatchability and TCS treatment levels. These results suggest that TCS has high toxicity on the early life stages of medaka, and that the metabolite of TCS may be a weak estrogenic compound with the potential to induce vitellogenin in male medaka but with no adverse effect on reproductive success and offspring.


Structure and dynamics of warm dense aluminum: a molecular dynamics study with density functional theory and deep potential.
We perform a systematic study on the structure and dynamics of warm dense aluminum (Al) at temperatures ranging from 0.5 to 5.0 eV with molecular dynamics utilizing both density functional theory (DFT) and the deep potential (DP) method. On one hand, unlike the Thomas-Fermi kinetic energy density functional (KEDF), we find that the orbital-free DFT method with the Wang-Teter non-local KEDF yields properties of warm dense Al that agree well with the Kohn-Sham DFT method, enabling accurate orbital-free DFT simulations of warm dense Al at relatively low temperatures. On the other hand, the DP method constructs a deep neural network that has a high accuracy in reproducing short- and long-ranged properties of warm dense Al when compared to the DFT methods. The DP method is orders of magnitudes faster than DFT and is well-suited for simulating large systems and long trajectories to yield accurate properties of warm dense Al. Our results suggest that the combination of DFT methods and the DP model is a powerful tool for accurately and efficiently simulating warm dense matter.


Attack Site Density of a Highly-efficient PET Hydrolases.
Poly (ethylene terephthalate) (PET) is one of the most abundant polyester materials used in daily life and it is also one of the main culprits of environmental pollution. ICCG (F243I/D238C/S283C/Y127G) is an enzyme that performs four modifications on the leaf branch compost keratase (LCC). It shows excellent performance in the hydrolysis of PET and has a great potential in further applications. Here, we used ICCG to degrade PET particles of various sizes and use the density of attack sites (Γattack) and kinetic parameters to evaluate the effect of particle size on enzyme degradation efficiency. We are surprised to observe that there is a certain relationship between Km and Γattack. In order to further confirm the relationship, we obtained three different enzymes (Y95K, M166S and H218S) by site-directed mutagenesis on the basis of ICCG. The results confirmed that there was a negative correlation between Km and Γattack. In addition, we also found that increasing the affinity between the enzyme and the substrate does not necessarily lead to the increase of degradation rate. These findings show that the granulation of PET and the selection of appropriate particle size are helpful to improve its industrial application value. At the same time, additional protein engineering to increase ICCG performance is realistic, but it can't be limited to enhance the affinity between enzyme and substrate.


Redox imbalance in cystine/glutamate transporter-deficient mice.
Cystine/glutamate transporter, designated as system x(-)(c), mediates cystine entry in exchange for intracellular glutamate in mammalian cells. This transporter consists of two protein components, xCT and 4F2 heavy chain, and the former is predicted to mediate the transport activity. This transporter plays a pivotal role for maintaining the intracellular GSH levels and extracellular cystine/cysteine redox balance in cultured cells. To clarify the physiological roles of this transporter in vivo, we generated and characterized mice lacking xCT. The xCT(-/-) mice were healthy in appearance and fertile. However, cystine concentration in plasma was significantly higher in these mice, compared with that in the littermate xCT(-/-) mice, while there was no significant difference in plasma cysteine concentration. Plasma GSH level in xCT(-/-) mice was lower than that in the xCT(-/-) mice. The embryonic fibroblasts derived from xCT(-/-) mice failed to survive in routine culture medium, and 2-mercaptoethanol was required for survival and growth. When 2-mercaptoethanol was removed from the culture medium, cysteine and GSH in these cells dramatically decreased, and cells started to die within 24 h. N-Acetyl cysteine also rescued xCT(-/-)-derived cells and permitted growth. These results demonstrate that system x(-)(c) contributes to maintaining the plasma redox balance in vivo but is dispensable in mammalian development, although it is vitally important to cells in vitro.


Live Confocal Imaging as a Novel Tool to Assess Liver Quality: Insights From a Murine Model.
In an experimental murine liver clamping model, we aimed to investigate the efficacy of real-time confocal microscopy (RCM) in assessing viability of steatotic livers in comparison to standard assessment tools, including histopathological evaluation. C57Bl/6 mice were subjected to a methionine-choline-deficient diet causing nonalcoholic fatty liver disease or to Lieber DeCarli diet causing ethanol-induced liver injury. Untreated animals served as controls. Liver biopsies were analyzed following challenge with 45 min of warm ischemia time and either 4 h of reperfusion or 24 h of cold storage. Organ quality assessment was performed at defined time points by RCM, histological staining, measurement of serum alanine aminotransferase activity, and expression analyses of proinflammatory cytokines. Additionally, survival analysis was performed. Cold as well as warm ischemia time resulted in a significant decrease in cell viability when compared with naive livers as well as nonischemic-challenged steatotic livers (P < 0.05) as assessed by RCM. Furthermore, RCM revealed the actual cellular damage at early time points, while established methods including H&E-staining and serum transaminase profile failed. In a translational attempt, we demonstrate that RCM is a suitable diagnostic tool to obtain information about functional damage of the liver apart from standard approaches.


Complete Genome Sequence of Salmonella enterica Serovar Enteritidis Bacteriophage f18SE, Isolated in Chile.
Bacteriophage f18SE was isolated from poultry sewage in Olmue, Chile, and lytic activity was demonstrated against Salmonella enterica serovar Enteritidis and serovar Pullorum strains. This bacteriophage has a 41,868-bp double-stranded DNA (ds-DNA) genome encoding 53 coding sequences (CDSs) and belongs to the family Siphoviridae, subfamily Jerseyvirinae.


An investigation of pain experience and management following gynecological day surgery: differences between open and closed surgery.
This study aimed to investigate the pain and activity levels of patients for a 5-day period following gynecological day surgery. Patients recorded their degree of pain and activity on visual analogue scales in a diary; completed diaries were returned by 54.9% of the patients approached (106 of 193). The median age of the women was 28.5 years (range, 15-55 years). Each procedure was classified as either open or closed surgery, depending upon whether an incision was necessary. Most (63.2%) of the patients required analgesics on the first morning after their surgery. Open surgery patients experienced significantly more pain for at least 3 postoperative days and were less active until the fifth day (P < 0.05 by Mann-Whitney tests). Drowsiness (47.2%), nausea (30.2%), and headache (30.2%) were the most common adverse effects experienced in the first postoperative morning. The incidence of adverse effects declined over the five days, but a minority of patients still suffered with headache (14.2%), drowsiness (9.4%), sore throat (7.5%), and nausea (5.3%) on the fifth day following surgery. The percentage of patients who experienced constipation peaked at 19.8% on the second day, most likely reflecting the consumption of codeine-containing analgesics on the first and second postoperative days. While day surgery has an important role in many forms of gynecological surgery, the potential discomfort and recovery time following day surgery should not be underestimated.


Endoscopic Versus Laparoscopic Drainage of Pancreatic Pseudocysts: a Cost-effectiveness Analysis.
Both endoscopic and laparoscopic interventions have a high therapeutic success rate in the management of symptomatic pancreatic pseudocysts; however, neither has been established as the gold standard. A decision tree analysis was performed to examine the costs and outcomes of intervening on pancreatic pseudocysts endoscopically versus laparoscopically. Within the model, a theoretical patient cohort was separated into two treatment arms: endoscopic drainage and laparoscopic drainage. Variables within the model were selected from the published literature. Medicare reimbursements rates (US$) were used to represent costs accumulated during a 3-month perioperative period. Effectiveness was characterized by quality-adjusted life-years (QALYs). A willingness-to-pay of $100,000 per 1 year of perfect health (1 QALY) gained was used as the cost-effectiveness threshold. The model was validated using one-way, two-way, and probabilistic sensitivity analysis. Endoscopic management of symptomatic pancreatic pseudocysts was the dominant strategy, producing 0.22 QALYs more while saving $23,976.37 in comparison to laparoscopic management. This result was further validated by one-way, two-way, and probabilistic sensitivity analysis. For patients presenting with symptomatic pancreatic pseudocysts amenable to either endoscopic or laparoscopic management, endoscopic drainage should be considered first-line therapy.


Adipose tissue expression of UCP1 and PRDM16 genes and their association with postprandial triglyceride metabolism and glucose intolerance.
UCP1 and PRDM16 genes, primarily involved in browning of adipose tissue that can affect lipid metabolism are also associated with diabetes risk. Therefore, we planned to study the adipose tissue expression of UCP1 and PRDM 16 genes in subjects with glucose intolerance to find out its association with postprandial triglyceride (PPTg) measures and T2DM. A total of 30 subjects were recruited in three groups i.e., NGT, prediabetes and T2DM (NDDM + known T2DM) who were matched for age, sex and BMI. An 8-hour standardized fat challenge test was performed to study lipemic responses. UCP1 and PRDM16 genes quantification in adipose tissue was performed by real-time PCR followed by SDS PAGE. UCP1 gene expression in SAT was significantly lower in T2DM and prediabetes as compared to NGT group while PRDM16 gene expression was significantly lower in T2DM group as compared to NGT group. UCP1 gene expression correlated with PPTg measures as well as with glycaemic measures while PRDM16 gene expression correlated with glycaemic measures only. This study found downregulation of PRDM16 and UCP1 gene expression in SAT in subjects with glucose intolerance. The association of UCP1 gene expression with PPTg dysmetabolism may contribute to greater predisposition to T2DM.


SNP calling using genotype model selection on high-throughput sequencing data.
A review of the available single nucleotide polymorphism (SNP) calling procedures for Illumina high-throughput sequencing (HTS) platform data reveals that most rely mainly on base-calling and mapping qualities as sources of error when calling SNPs. Thus, errors not involved in base-calling or alignment, such as those in genomic sample preparation, are not accounted for. A novel method of consensus and SNP calling, Genotype Model Selection (GeMS), is given which accounts for the errors that occur during the preparation of the genomic sample. Simulations and real data analyses indicate that GeMS has the best performance balance of sensitivity and positive predictive value among the tested SNP callers. The GeMS package can be downloaded from https://sites.google.com/a/bioinformatics.ucr.edu/xinping-cui/home/software or http://computationalbioenergy.org/software.html. Supplementary data are available at Bioinformatics online.


The telomeric length and heterogeneity decrease with age in normal human oral keratinocytes.
We have examined the telomere length in NHOK explanted from 28 donors between the ages of 21 and 84 years. Genomic DNA was isolated from exponentially replicating NHOK and digested with HinFI to yield terminal restriction fragments (TRF). The TRF length ranged from 4.1 to 7.0 kbp with a mean of 5.3 +/- 0.8 kbp, which was significantly shorter than that (8.9 +/- 1.0 kbp) of normal human oral fibroblasts (NHOF). The TRF length was inversely correlated to the increase of donor age in NHOK (m=-23 bp per year; r=-0.60; P<0.001). Also, the heterogeneity of TRF length in cultured NHOK decreased with increased donor age (r=-0.38, P<0.05). These data indicated that clonogenic NHOK cells had replicated in situ and showed a progressive shortening of TRF length. The short telomere length and decreased telomeric length heterogeneity in immortalized cells suggested that there is a critical minimum for cell survival.


Use of carbon and nitrogen stable isotope ratios to assess the effects of environmental contaminants on aquatic food webs.
In this study, the value of carbon (delta13C) and nitrogen (delta15N) stable isotope ratios were determined in nymphs of a top-predator, the common backswimmer (Notonecta glauca L.), collected in 18 m3 outdoor freshwater mesocosms used to assess the fate and ecotoxicological effects of a diphenyl ether herbicide, fomesafen, applied alone or in combination with Agral 90 (mixture of polyethoxylated derivatives of nonylphenol). Both treatments had a negative effect on delta13C values which may reflect changes in carbon fluxes across food webs in the treated ponds associated with a shift in phytoplankton structure. A decrease in delta15N values was observed in the nymphs collected in mixture-treated ponds, which was presumably due to an increase in the abundance of rotifers and Chironominae larvae in these ponds. These preliminary results indicate that stable isotope ratios may be used as shortcuts to detect qualitative or quantitative shifts in the structure of aquatic food webs caused by pollutants.


Biomimetic processing of calcium phosphate coating.
Most ceramics used in surgical implantation are better tolerated by the body environment than other biomaterials, particularly metals. However, the broader application of ceramics is limited because they are brittle and prone to catastrophic failure. By contrast. naturally produced ceramic organic biocomposites can possess mechanical properties which make them suitable for use as biomaterials. An example would be the mollusc shell, where an inorganic phase grows onto a charged and organised organic template. This natural biomineralization provides a calcium carbonate phase with satisfactory mechanical properties appropriate for surgical implantation. It is reported how Langmuir Blodgett films have been used as organic templates for calcium phosphate crystallization. Three different Langmuir Blodgett Films: omega tricosenoic acid, stearic acid and octadecylamine have been investigated. The Langmuir Blodgett Films produced had methyl (CH3) and calcium carboxylate (CaCOO) as functional groups. Atomic force microscopy and scanning electron microscopy shows that calcium phosphate crystallization occurs in a similar manner to natural biomineralization. The calcium carboxylate head group appears to have a physical influence on the morphology and distribution of the calcium phosphate crystals. Under the experimental conditions tested, the calcium carboxylate functional head group controls calcium phosphate crystallization by a biomimetic approach. This study strongly suggests that this approach has the potential for producing high performance ceramics comparable with those made by mollusc shells.


Anatomically and physiologically based guidelines for use of the sphenopalatine ganglion block versus the stellate ganglion block to reduce atypical facial pain.
This literature review is designed to develop guidelines needed for the use of a sphenopalatine ganglion block versus a stellate ganglion block to reduce atypical facial pain. We have reviewed the basic anatomy of both ganglia and the physiological responses usually associated with each, and have given an opinion on appropriate use of these therapeutic modalities.


Treatment of gingival recession defects with a coronally advanced flap and a xenogeneic collagen matrix: a multicenter randomized clinical trial.
To evaluate the clinical outcomes of the use of a xenogeneic collagen matrix (CM) in combination with the coronally advanced flap (CAF) in the treatment of localized recession defects. In a multicentre single-blinded, randomized, controlled, split-mouth trial, 90 recessions (Miller I, II) in 45 patients received either CAF + CM or CAF alone. At 6 months, root coverage (primary outcome) was 75.29% for test and 72.66% for control defects (p = 0.169), with 36% of test and 31% of control defects exhibiting complete coverage. The increase in mean width of keratinized tissue (KT) was higher in test (from 1.97 to 2.90 mm) than in control defects (from 2.00 to 2.57 mm) (p = 0.036). Likewise, test sites had more gain in gingival thickness (GT) (0.59 mm) than control sites (0.34 mm) (p = 0.003). Larger (≥3 mm) recessions (n = 35 patients) treated with CM showed higher root coverage (72.03% versus 66.16%, p = 0.043), as well as more gain in KT and GT. CAF + CM was not superior with regard to root coverage, but enhanced gingival thickness and width of keratinized tissue when compared with CAF alone. For the coverage of larger defects, CAF + CM was more effective.


Data gaps in toxicity testing of chemicals allowed in food in the United States.
In the United States, chemical additives cannot be used in food without an affirmative determination that their use is safe by FDA or additive manufacturer. Feeding toxicology studies designed to estimate the amount of a chemical additive that can be eaten safely provide the most relevant information. We analyze how many chemical additives allowed in human food have feeding toxicology studies in three toxicological information sources including the U.S. Food and Drug Administration's (FDA) database. Less than 38% of FDA-regulated additives have a published feeding study. For chemicals directly added to food, 21.6% have feeding studies necessary to estimate a safe level of exposure and 6.7% have reproductive or developmental toxicity data in FDA's database. A program is needed to fill these significant knowledge gaps by using in vitro and in silico methods complemented with targeted in vivo studies to ensure public health is protected.


Gas chromatographic head-space determination of residual acrylonitrile in acrylonitrile-butadiene-styrene resins and migration into a simulated fatty foodstuffs liquid.
Head-space methods are described for the determination of residual acrylonitrile in acrylonitrile-butadiene-styrene resins and in olive oil, which simulates fatty foodstuffs. Dimethylformamide was used as solvent for the resin, with flame ionization detection. The injection of water into the resin dispersion prior to head-space analysis greatly enhances the detection capabilities. The use of a nitrogen-selective detector required dimethyl sulphoxide as the solvent. The determination of acrylonitrile in olive oil was carried out employing both types of detector. The detection sensitivity was much greater with the nitrogen-selective detector.


Effects of dietary levels of brown seaweeds and plant polyphenols on growth and meat quality parameters in growing rabbit.
Growth performances, carcass characteristics and meat quality parameters from growing rabbit fed with two levels of dietary brown seaweed (Laminaria spp) and plant polyphenols were investigated. One hundred and forty-four New Zealand White rabbits were allotted into three dietary treatments containing 0 (C), 0.3% (T1), and 0.6% (T2) of brown seaweed and plant polyphenols mixture for 42 days. Growth performances and carcass weight were improved in T1 group. Vitamin A and E content in Longissimus thoracis and lumborum (LTL) and Semimembranosus (SM) muscle were enhanced in the treated groups. In the SM muscle, the oxidative stability was improved in rabbit fed with both dosages of dietary supplement, and the cholesterol content tended to be lower in T1 than in T2 and C groups. The LTL and SM muscle sensory characteristics were improved. In conclusion, dietary integration with a low dosage of brown seaweed and plant polyphenols is a valid strategy for enhance growth performance and produce healthier rabbit meat.


Temporal and spatial changes of microbial community in an industrial effluent receiving area in Hangzhou Bay.
Anthropogenic activities usually contaminate water environments, and have led to the eutrophication of many estuaries and shifts in microbial communities. In this study, the temporal and spatial changes of the microbial community in an industrial effluent receiving area in Hangzhou Bay were investigated by 454 pyrosequencing. The bacterial community showed higher richness and biodiversity than the archaeal community in all sediments. Proteobacteria dominated in the bacterial communities of all the samples; Marine_Group_I and Methanomicrobia were the two dominant archaeal classes in the effluent receiving area. PCoA and AMOVA revealed strong seasonal but minor spatial changes in both bacterial and archaeal communities in the sediments. The seasonal changes of the bacterial community were less significant than those of the archaeal community, which mainly consisted of fluctuations in abundance of a large proportion of longstanding species rather than the appearance and disappearance of major archaeal species. Temperature was found to positively correlate with the dominant bacteria, Betaproteobacteria, and negatively correlate with the dominant archaea, Marine_Group_I; and might be the primary driving force for the seasonal variation of the microbial community.


A Novel 3-Part Approach to Tackle the Problem of Health Inequities in Early Childhood.
The first 5 years of a child's life are crucial in laying the foundation for their health and developmental trajectory into adulthood. These early years are especially influenced by the surrounding environments in which children live and grow. A large international body of evidence demonstrates that children who experience disadvantage tend to fall increasingly behind over time. At the societal level, these inequities can cause substantial social burdens and significant costs across health, education, and welfare budgets. A contributing factor is that children experiencing adversity are less likely to have access to the environmental conditions that support them to thrive. Many of these factors are modifiable at the community or place level. We argue for three key-though not exhaustive-ideas that collectively could achieve more equitable outcomes for children facing disadvantage and experiencing adversity:We conclude that if adopted, these 3 ideas could contribute to the ability of local communities and networks to identify and respond to factors that address early childhood inequalities.


Transduction of binding energy into hemoglobin cooperativity.
Hemoglobin is a tetrameric molecule consisting of two identical alpha beta dimers which assemble into either of two quaternary structures, T or R. Recent studies on mutant and partially ligated hemoglobins have revealed that cooperativity exists between the alpha and the beta hemes of each dimeric half-molecule and have led to a symmetry rule for quaternary T-->R switching: the quaternary R structure is energetically favored over the T structure when each dimeric half-molecule contains at least one ligated subunit.


PET measurement of adenosine stimulated absolute myocardial blood flow for physiological assessment of the coronary circulation.
Considerable awareness has been raised of late of the need to reduce radiation exposure and control costs of x-ray and radionuclide imaging procedures. PET/CT cameras are now widely available and in conjunction with appropriate radionuclides and commercially available software make quantitative measurement of absolute MBF feasible for routine clinical practice. Quantitative measurement of absolute MBF under condition of coronary vasodilation permits independent assessment of the functional status of each of the three major coronary perfusion zones and so obviates the need for rest MBF determination in the great majority of cases. Coronary microvascular function also may be assessed in this same way. Thus, the stress-only protocol with quantitative PET measurement of MBF provides essential information required for clinical decision making related to need for catheterization and intervention for patients with known or suspected ischemic heart disease. Moreover, the single PET determination of maximal MBF in contrast to the usual rest/stress procedure addresses both safety and cost concerns. The present review focuses on: (1) quantitative PET measurements of myocardial blood flow for physiological assessment of the coronary circulation and (2) the value and potential limitations of performing stress only imaging in the clinical context.


The fertilization process in Lithocarpus dealbatus (Fagaceae) and its implication on the sexual reproduction evolution of Fagales.
The pistillate flowers of Lithocarpus dealbatus show two pollen tube (PT) arresting sites (the style-joining and micropyle) within the pistil during the postpollination-prezygotic stage. The PT, arrested at the pre-ovule stage, enhanced PT competition allowing the most compatible PTs to enter the ovary to ensure the highest fertilization success. During the shift from animal pollination to wind pollination, plants require a series of changes in reproductive traits. The mode of pollination is striking labile in Fagaceae. Lithocarpus is insect pollinated and is closely related to wind-pollinated Quercus. Little is known about the sexual reproduction of Lithocarpus. This study aimed to reveal the sexual reproduction of Lithocarpus dealbatus and to explore the evolutionary pattern of the key sexual reproduction traits to better understand their possible role in labile pollination. We found that after pollination, L. dealbatus PTs grew slowly in the style reaching style-joining in mid-January of the second year; then PT growth was arrested at style-joining for four months. Only two to three PTs resumed growth in mid-May to reach the micropyle, where PT growth ceased for one month before one PT resumed growth and passed through the micropyle to the embryo sac. Fagaceae showed a generalized mating system. Vast pollen production, small-sized pollen grains, long stigmatic receptive time, and reduced perianth were compatible with beetle pollination syndrome, representing the plesiomorphic status in Fagaceae. A large stigmatic surface and dry pollen grains linked to wind pollination might be independently derived several times in fagaceous lineages. Beetle pollination syndrome can cope with the uncertainty of pollinators to ensure conspecific pollen capture, which represents pre-adaptation status and has a selective advantage when conditions change, favouring wind pollination. The arrest of the PT at style-joining is a unique mechanism in later derived fagaceous lineages to enhance PT competition and promote outcrossing.