Bacterial mutagenicity test data: collection by the task force of the Japan pharmaceutical manufacturers association.
Ames test is used worldwide for detecting the bacterial mutagenicity of chemicals. In silico analyses of bacterial mutagenicity have recently gained acceptance by regulatory agencies; however, current in silico models for prediction remain to be improved. The Japan Pharmaceutical Manufacturers Association (JPMA) organized a task force in 2017 in which eight Japanese pharmaceutical companies had participated. The purpose of this task force was to disclose a piece of pharmaceutical companies' proprietary Ames test data. Ames test data for 99 chemicals of various chemical classes were collected for disclosure in this study. These chemicals are related to the manufacturing process of pharmaceutical drugs, including reagents, synthetic intermediates, and drug substances. The structure-activity (mutagenicity) relationships are discussed in relation to structural alerts for each chemical class. In addition, in silico analyses of these chemicals were conducted using a knowledge-based model of Derek Nexus (Derek) and a statistics-based model (GT1_BMUT module) of CASE Ultra. To calculate the effectiveness of these models, 89 chemicals for Derek and 54 chemicals for CASE Ultra were selected; major exclusions were the salt form of four chemicals that were tested both in the salt and free forms for both models, and 35 chemicals called "known" positives or negatives for CASE Ultra. For Derek, the sensitivity, specificity, and accuracy were 65% (15/23), 71% (47/66), and 70% (62/89), respectively. The sensitivity, specificity, and accuracy were 50% (6/12), 60% (25/42), and 57% (31/54) for CASE Ultra, respectively. The ratio of overall disagreement between the CASE Ultra "known" positives/negatives and the actual test results was 11% (4/35). In this study, 19 out of 28 mutagens (68%) were detected with TA100 and/or TA98, and 9 out of 28 mutagens (32%) were detected with either TA1535, TA1537, WP2uvrA, or their combination. The Ames test data presented here will help avoid duplicated Ames testing in some cases, support duplicate testing in other cases, improve in silico models, and enhance our understanding of the mechanisms of mutagenesis.


Behavioral and emotional outcomes at preschool age in children born very preterm: The role of breast milk feeding practices.
Breast milk feeding (BMF) improved neurodevelopment in children born very preterm (VPT, <32 weeks of gestation), but knowledge about its effect on other mental health outcomes remains limited. To estimate the association of BMF practices with behavioral and emotional problems at preschool age in children born VPT. We studied 263 children born VPT during 2011-12 and enrolled in the Portuguese EPICE cohort. At the age of 3, information on BMF initiation and duration was collected and behavioral and emotional problems were assessed using the parents' completed Child Behavior Checklist 1.5-5 years (CBCL/1½-5). Children were categorized for all CBCL/1½-5 sub-scales and for Diagnostic and Statistical Manual of Mental Disorders (DSM5)-oriented scales. Risk ratios were estimated to assess the association of BMF with subclinical/clinical problems, fitting a Poisson regression. Behavioral or emotional subclinical/clinical problems were found in almost 20% of children (11.8% in the clinical range). BMF was consistently associated with lower adverse behavioral and emotional outcomes, particularly risks of externalizing problems, somatic complaints, aggressive behavior, as well as autism spectrum and attention deficit/hyperactivity symptoms, although the magnitude of the unadjusted risks was attenuated by adjustment for relevant confounders and wider confidence intervals included the null. Lower exposure to BMF seemed to increase the risk of adverse behavioral and emotional outcomes at preschool age in children born VPT. These results raise questions about explanatory pathways and strengthen evidence underpinning BMF promotion for VPT children.


The Impact of Prenatal Diagnosis on Clinical Outcomes of Isolated Vascular Rings From a Statewide Paediatric Cardiology Tertiary Service.
Vascular rings, including right aortic arch with aberrant left subclavian artery (RAA-ALSCA), double aortic arch (DAA) and pulmonary artery sling (PAS), are congenital anomalies that may cause airway and oesophageal compression. As prenatal detection has improved, literature comparing clinical outcomes of antenatally versus postnatally diagnosed cases continues to emerge. The aim is to define a statewide tertiary paediatric institution's clinical profile and outcomes of prenatal versus postnatally diagnosed isolated vascular rings. A retrospective single-centre review of isolated RAA-ALSCA, DAA and PAS between 1 January 1999 and 31 December 2020 was conducted. Clinical characteristics, surgical and follow-up information were collected. Antenatal and postnatally diagnosed groups were compared. Out of 123 cases diagnosed with isolated vascular rings, 98 (79.7%) cases had RAA-ALSCA, 21 (17.1%) with DAA, 4 (3.3%) with PAS. The antenatal detection rate was 73.6% in the past decade; 20.3% had a genetic disorder, of which 48% had 22q11.21 microdeletion. Of prenatally diagnosed cases, 31.3% developed symptoms, commonly stridor and dysphagia, at a median age of 2.0 months (IQR 0.0-3.0), compared to a median age of diagnosis for the postnatal cohort of 9 months (IQR 1.0-40.7). Postnatally diagnosed cases were more likely to present with symptoms, primarily respiratory distress, than prenatally diagnosed cases (p=0.006). Fifty-nine (59) cases (50% antenatally diagnosed) required vascular ring division; 6.8% had residual symptoms following surgery. Antenatal diagnosis has improved and leads to better parental awareness and more timely, appropriate intervention. Postnatally diagnosed patients were older, more likely to be symptomatic, underwent more investigations and were commenced on more medications for symptom management prior to diagnosis. One in five cases of isolated vascular ring anomalies carried a genetic diagnosis, which has important implications on prenatal counselling and genetic testing.


Microbiological Studies on the Influence of Essential Oils from Several Origanum Species on Respiratory Pathogens.
Essential oils (EOs) with established and well-known activities against human pathogens might become new therapeutics in multidrug-resistant bacterial infections, including respiratory tract infections. The aim of this study was to evaluate the antimicrobial activity of EOs obtained from several samples of Origanum vulgare, O. syriacum, and O. majorana cultivated in Poland. EOs were analyzed by GC-MS and tested against four bacterial strains: Staphylococcus aureus (MRSA), Haemophilus influenzae, Haemophilus parainfluenzae, and Pseudomonas aeruginosa. Chemical analyses showed that the Eos were characterized by a high diversity in composition. Based on the chemical data, four chemotypes of Origanum EOs were confirmed. These were carvacrol, terpineol/sabinene hydrate, caryophyllene oxide, and thymol chemotypes. Thin-layer chromatography-bioautography confirmed the presence of biologically active antibacterial components in all tested EOs. The highest number of active spots were found among EOs with cis-sabinene hydrate as the major compound. On the other hand, the largest spots of inhibition were characteristic to EOs of the carvacrol chemotype. Minimal inhibitory concentrations (MICs) were evaluated for the most active EOs: O. vulgare 'Hirtum', O. vulgare 'Margarita', O. vulgare 'Hot & Spicy', O. majorana, and O. syriacum (I) and (II); it was shown that both Haemophilus strains were the most sensitive with an MIC value of 0.15 mg/mL for all EOs. O. majorana EO was also the most active in the MIC assay and had the highest inhibitory rate in the anti-biofilm assay against all strains. The most characteristic components present in this EO were the trans-sabinene hydrate and terpinen-4-ol. The strain with the least sensitivity was the MRSA with an MIC of 0.6 mg/mL for all EOs except for O. majorana, where the MIC value reached 0.3 mg/mL. Scanning electron microscopy performed on the Haemophilus influenzae and Haemophilus parainfluenzae biofilms showed a visible decrease in the appearance of bacterial clusters under the influence of O. majorana EO.


Contraction-induced enhancement of relaxation during high force contractions of mouse lumbrical muscle at 37°C.
Repeated stimulation of unfatigued rodent fast-twitch skeletal muscle accelerates the kinetics of tension relaxation through an unknown mechanism. This effect varies with muscle type and stimulation parameters, and has been observed at physiological temperatures for submaximal but not maximal contractions. The purpose of this study was to compare relaxation kinetics of C57BL/6 mouse lumbrical muscles ex vivo from maximal isometric force (500 Hz for 20 ms) when evoked before (pre) and after (post) an intervening tetanic contraction at 37°C. During post contractions, we noted significant increases in the rate of tension decline during both the slow linear phase and the fast exponential phase of relaxation, as well as a reduced duration of the slow phase of relaxation compared with pre contractions (all P<0.05). This is the first demonstration of enhanced slow and fast relaxation phases from maximal isometric tension induced by prior stimulation in intact muscle at a physiological temperature.


Non-coding RNAs: new players in eukaryotic biology.
The completion of the human, mouse and other eukaryotic genomes were important scientific milestones, but they were just small steps towards the understanding of eukaryotic biology. Recent transcriptome analysis and different experimental approaches have identified a surprisingly large number of non-coding RNAs (ncRNAs) in eukaryotic cells. ncRNAs comprise microRNAs, anti-sense transcripts and other Transcriptional Units containing a high density of stop codons and lacking any extensive "Open Reading Frame". They have been shown to regulate gene expression by novel mechanisms such as RNA interference, gene co-suppression, gene silencing, imprinting and DNA demethylation. It is becoming clear that these novel RNAs perform critical functions during development and cell differentiation. There is also mounting evidence of their involvement in cancer and neurological diseases. Together, all this information indicates that ncRNAs are emerging as a new class of functional transcripts in eukaryotes. Therefore, great challenges lie in the years ahead: understanding the molecular biology of higher organisms will require revealing all proteins (Proteome), all ncRNAs (RNome) and their interactions (Interactome) in the complex molecular scenario within eukaryotic cells.


Clinical relevance of thyroid-stimulating immunoglobulins in graves' ophthalmopathy.
Thyroid-stimulating immunoglobulins (TSIs) likely mediate Graves' ophthalmopathy (GO). The clinical relevance of these functional autoantibodies was assessed in GO. Cross-sectional trial. A total of 108 untreated patients with GO. Thyroid-stimulating immunoglobulins, assessed with a novel bioassay, bind to the thyrotropin receptor (TSHR) and transmit signals for cyclic adenosine monophosphate (cAMP)-dependent activation of luciferase gene expression. The cAMP/cAMP response element-binding protein/cAMP-regulatory element complex induces luciferase that is quantified after cell lysis. The TSI levels were correlated with activity and severity of GO and compared with a TSHR binding inhibitory immunoglobulin (TBII) assay. Thyroid-stimulating immunoglobulins, activity and severity of GO, diplopia, and TBII. Thyroid-stimulating immunoglobulins were detected in 106 of 108 patients (98%) with GO. All 53 hyperthyroid patients were TSI positive versus 47 patients (89%) who were TBII positive. All 69 patients with active GO were TSI positive, whereas only 58 of 69 patients (84%) were TBII positive. Thyroid-stimulating immunoglobulins correlated with the activity (r=0.83, P < 0.001) and severity (r=0.81, P < 0.001) of GO. All 59 patients with GO with diplopia were TSI positive, and 50 of 59 patients (85%) were TBII positive. Among patients with moderate-to-severe and mild GO, 75 of 75 (100%) and 31 of 33 (94%) were TSI positive compared with TBII positivity in 63 of 75 (84%) and 24 of 33 (73%), respectively. The TSI levels were higher in moderate-to-severe versus mild GO (489%±137% vs. 251%±100%, P < 0.001). Chemosis and GO activity predicted TSI levels alone (P < 0.001, multivariable analysis). The TSI levels were higher in patients with chemosis (527%±131%) than in patients without chemosis (313%±127%, P < 0.001). Thyroid-stimulating immunoglobulins show more significant association with clinical features of GO than TBII and may be regarded as functional biomarkers for GO. Proprietary or commercial disclosure may be found after the references.


Hypothalamic modulation of energy expenditure.
The acute effects of electrical stimulation of the hypothalamus on energy expenditure as measured by indirect calorimetry were investigated in 20 unanaesthetized rats. Thirty sec of stimulation increased both O2 consumption and respiratory quotient (R.Q.). The largest magnitude hypermetabolic response (39% mean peak increase in O2 consumption) was produced by stimulation of the ventromedial hypothalamic nucleus. Stimulation of the lateral hypothalamus produced hypermetabolic effects similar to but smaller than those produced by medial stimulation. A number of considerations suggest that the hypermetabolism is not secondary to changes in motor activity, carbohydrate utilization or blood glucose levels. Consequently, these data suggest that the hypothalamus modulates energy expenditure through changes in non-shivering thermogenesis. These metabolic changes may modulate the effects of various hypothalamic manipulations on body weight.


Compliance and its role in successful treatment of an advanced periodontal case: review of the literature and a case report.
Periodontal disease is a chronic disease that is perceived by many patients to be nonthreatening. Periodontal therapy has been shown to be less effective if a regular periodontal maintenance schedule is not followed after completion of active therapy. Periodontal maintenance is an integral part of successful periodontal therapy.


Vedolizumab for children with intestinal graft-versus-host disease: a case report and literature review.
Acute graft-versus-host disease (aGVHD) is a challenging complication of allogeneic hematopoietic stem cell transplantation, and alternative therapies for patients showing inadequate response to steroids are limited. Vedolizumab, an anti-α4β7 integrin antibody widely used for treating inflammatory bowel diseases, has recently been studied in adult patients with steroid-refractory intestinal aGVHD. However, few studies have examined its safety and effectiveness in pediatric patients with intestinal aGVHD. We report the case of a male patient with intestinal late-onset aGVHD treated with vedolizumab. He underwent allogeneic cord blood transplantation for warts, hypogammaglobulinemia, infections, and myelokathexis (WHIM) syndrome and developed intestinal late-onset aGVHD 31 months after transplantation. The patient was refractory to steroids; however, vedolizumab was initiated 43 months after transplantation (at the age of 7 years) and the symptoms of intestinal aGVHD were alleviated. Additionally, favorable endoscopic findings were observed, such as reduction of erosion and regenerative epithelial growth. We also evaluated the efficacy of vedolizumab in 10 patients with intestinal aGVHD (9 from the literature review and the present case). Six patients (60%) showed an objective response to vedolizumab. No serious adverse events were observed in any patients. Vedolizumab is a potential treatment option for steroid-refractory intestinal aGVHD in pediatric patients.


Photo-activated affinity-site cross-linking of antibodies using tryptophan containing peptides.
Affinity-based conjugation methods for antibodies can produce defined and reproducible conjugates. This requires that the target antibody has an affinity site for the ligand and that the ligand has a reactive site. These requirements are critical for the conjugation of antibodies designed for diagnostic and therapeutic application. Our laboratory has discovered a novel affinity of antibodies for the amino acid tryptophan using an azido derivative of tryptophan. Here we show that tryptophan without the azido group can be photo-cross-linked to antibodies. Biotinylated tryptophan peptides are photolysed into monoclonal and polyclonal antibodies and such biotinylated antibodies are used in avidin-based ELISA. With the simple and gentle tryptophan-affinity photo-conjugation of peptides, antibodies can be conjugated with peptides to enhance their potency and expand their targeting range.


Comprehensive Analysis to Uncover Determinants of Patient Appointment Compliance in Ophthalmology at the Kresge Eye Institute, USA.
Appointment compliance (AC) has a significant impact on patient care; however, determinants of AC in Ophthalmology and its subspecialties remains elusive. We performed a five-year retrospective analysis across Kresge Eye Institute (KEI) and its affiliated Michigan locations. A total of 597,364 appointments across >13 subspecialties were included. AC was the primary outcome of interest. Compliant (CO) and non-compliant (NC) groups were compared to the following variables: patient characteristics (gender, race, age, insurance), appointment rank (relative to patient history), scheduling location, month, and ophthalmic specialty, in regard to arrival and no-show. Among all appointments, 59.77% were associated with a female patient and 79.16% of the total number of appointments depicted patient compliance. AC differed concerning specialty, with retina representing the highest compliance across all appointments. Among 200+ insurance providers, Medicare was most frequently used and represented the highest share of CO appointments. African Americans were the primary ethnicity served by KEI and had the highest number of NC appointments. Our study demonstrates the impact of patient demographics, appointment characteristics, and ophthalmic subspecialty on AC. A better understanding of these determinants could allow for an increased CO for Ophthalmology practices.


Modeling of malachite green adsorption onto novel polyurethane/SrFe12O19/clinoptilolite nanocomposite using response surface methodology and biogeography-based optimization-assisted multilayer neural network.
This research studied the modeling of malachite green (MG) adsorption onto novel polyurethane/SrFe12O19/clinoptilolite (PU/SrM/CLP) nanocomposite from aqueous solutions by the application of biogeography-based optimization (BBO) algorithm-assisted multilayer neural networks (MNN-BBO) as a new evolutionary algorithm in environmental science. The PU/SrM/CLP nanocomposite was successfully fabricated and characterized by some spectroscopic analyses. Four variables influencing the removal efficiency were modeled by MNN-BBO and response surface methodology (RSM). The MNN-BBO model gave higher percentage removal (99.6%) about 7.6% compared to the RSM technique. Under optimal conditions obtained by MNN-BBO, the four independent variables including pH, shaking rate, initial concentration, and adsorbent dosage were 6.5, 255 rpm, 50 mg.L-1, and 0.08 g, respectively. Under these conditions, the results were fitted well to the Langmuir isotherm with a monolayer maximum amount of sorbate uptake (qmax) of 68.49 mg.g-1 and the pseudo-first-order kinetic pattern with the rate constant (K1) of 0.01 min-1 with the R2 values of 0.9248 and 0.9980, respectively. The results of thermodynamics demonstrated that the MG uptake was not spontaneous due to the positive value of the adsorption ΔG. In addition, the positive values of ΔS (0.079 kJ/mol K) and ΔH (30.816 kJ/mol) indicated the feasible operation and endothermic approach, respectively. Besides, the wastewater investigations showed that the nanocomposite could be used as a new promising sorbent for efficient removal of MG (R% > 72) and magnetically separable from the real samples.


Post-match recovery profile of leukocyte cell subsets among professional soccer players.
This study assessed the impact of cumulative match time on the distribution of CD45+ cell subtests in the capillary blood of professional soccer players. Twenty-two males (aged 18-30 years) took part in the 36-week study. Participants playing up to 540 in cumulative match time and less than 30 min in each single match during the observation period formed the control group. White blood cell (WBC) phenotyping and creatine kinase (CK) plasma activity analyses were performed. Also, counts for WBC subsets were determined. No significant differences in the hematological parameters or lymphocyte and NK cell percentages were observed between the control and study groups. Changes in the T cell percentage were significant during weeks 11 and 30 and in Th and Tc cell percentages during weeks 2 and 26. Significant correlations were found between the cumulative match time and Th, NK, and B cell percentages; monocyte counts; and CK activity in the control group. However, for the study group, correlations were found between cumulative match time and Th, Tc, and B cell percentages; CK activity; and the CK ratio. Our study suggests that the distribution of CD45+ cells might be a useful tool for monitoring the immune status of professional soccer players.


Miniaturized cardiopulmonary bypass in coronary artery bypass surgery: marginal impact on inflammation and coagulation but loss of safety margins.
Inflammation and coagulation disturbances are common consequences of cardiopulmonary bypass (CPB). Recently, miniaturized closed CPB circuits without cardiotomy suction and venous reservoir have been proposed to reduce complication rates. We compared outcomes with conventional (CCPB) and miniaturized cardiopulmonary bypass (MCPB) after coronary artery bypass operations (CABG) with respect to inflammation and coagulation. Thirty patients (23% female; aged 67.9 +/- 9.0 years) were prospectively randomly assigned to undergo isolated CABG with CCPB or MCPB. Conventional CPB had a pump prime of 1, 600 mL. Miniaturized CPB consisted of a centrifugal pump, arterial filter, heparinized tubing, and oxygenator with a priming volume of 800 mL. Shed blood was removed by a cell-saving device and reinfused. Measurements included interleukin (IL)-2 receptor, IL-6, IL-10, tumor necrosis factor receptor 55 and 75, C reactive protein, leukocyte differentiation, d-dimers, fibrinogen, and thrombocytes at six time points. In both groups no major complication occurred. However, two dangerous air leaks occurred in the closed MCPB circuit, demonstrating the narrow safety margins. Operative handling was also more difficult owing to limitations in venting and fluid management. International normalized ratio (p = 0.03) and antithrombin III (p = 0.04) levels were elevated during CPB in the CCPB group, most likely owing to differences of the intraoperative anticoagulation management. Repeated measures analysis revealed that not a single parameter of inflammation or clinical outcome showed significant differences among groups. Use of a MCPB affected inflammation and coagulation variables only marginally and did not lead to clinical relevant changes as assessed by blood loss, need for blood products, and intensive care unit and clinical stays. However, safety margins for volume loss, air emboli, and weaning from CPB decrease, because of the closed MCPB circuit.


Eigenvalue and field equations of three-layered uniaxial fibers and their applications to the characteristics of long-period fiber gratings with applied axial strain.
By using the Debye potentials, the exact eigenvalue equations and the corresponding field distributions of the core and cladding modes for three-layered, radially stratified, and dielectric uniaxial optical fibers are derived completely; the modes include TE, TM, and hybrid HE/EH modes. The strain characteristics of long-period fiber gratings' (LPFGs') with applied axial strain are investigated theoretically by studying three-layered uniaxial optical fibers. When uniform axial strain is applied to fiber, the core, and inner and outer cladding become uniaxial crystal optically; that is, the optical axes are parallel to the fiber's axis. Analytic expressions of the strain sensitivities of LPFGs are derived. The analysis reveals that the strain sensitivities of LPFGs based on various cladding modes, including the shift value and direction of the resonance wavelength, differ greatly.


Do non-insulin-dependent diabetes mellitus and cardiovascular disease share common antecedents?
Recent evidence suggests that non-insulin-dependent diabetes mellitus (NIDDM) and cardiovascular disease, rather than being related as underlying disease and complication, share common genetic and environmental antecedents, that is, they "spring from the same soil." Fetal and early-life nutritional deficiencies appear to predispose persons to both NIDDM and cardiovascular disease in later life. The insulin resistance syndrome, including abdominal obesity, may constitute the intermediate link between fetal and early-life nutritional deficiency and later disease. The insulin resistance syndrome includes insulin resistance, hyperinsulinemia, abdominal obesity, dyslipidemia with high triglyceride and low high-density lipoprotein cholesterol levels, and hypertension. Each element of the insulin resistance syndrome has been firmly established as a risk factor for development of diabetes. In addition, most of these elements are also well-recognized cardiovascular risk factors, although the weight of evidence now suggests that hyperinsulinemia itself is not. This last point is significant because of concern that aggressive insulinization of diabetic patients, which has been proved to reduce microvascular complications, might paradoxically increase the risk for large-vessel atherosclerosis. Available clinical trials suggest that this fear is unwarranted, but definitive trials are needed to resolve this important clinical question.


Primary structure of bovine chromogranin B deduced from cDNA sequence.
The cDNA sequence of bovine chromogranin B has been determined. Bovine chromogranin B consists of 626 amino acids and contains 16 pairs of basic amino acids which might be used for generation of biological active peptides. A comparison of the amino acid sequences of chromogranin B from various species identifies regions of high homology at the N- and C-termini as well as stretches in the middle of the chromogranin B sequence.


RT-qPCR work-flow for single-cell data analysis.
Individual cells represent the basic unit in tissues and organisms and are in many aspects unique in their properties. The introduction of new and sensitive techniques to study single-cells opens up new avenues to understand fundamental biological processes. Well established statistical tools and recommendations exist for gene expression data based on traditional cell population measurements. However, these workflows are not suitable, and some steps are even inappropriate, to apply on single-cell data. Here, we present a simple and practical workflow for preprocessing of single-cell data generated by reverse transcription quantitative real-time PCR. The approach is demonstrated on a data set based on profiling of 41 genes in 303 single-cells. For some pre-processing steps we present options and also recommendations. In particular, we demonstrate and discuss different strategies for handling missing data and scaling data for downstream multivariate analysis. The aim of this workflow is provide guide to the rapidly growing community studying single-cells by means of reverse transcription quantitative real-time PCR profiling.


Mesoporous platinum nanoparticles as a peroxidase mimic for the highly sensitive determination of C-reactive protein.
The generation of a mesoporous structure in platinum nanoparticles can effectively enhance physical and chemical properties. In this study, mesoporous platinum nanoparticles (MPNs) were synthesized by a soft template-mediated one-pot chemical method. To develop a mesoporous structure, Pluronic F-127 was employed. The Pluronic F-127 surfactant forms self-assembled micelles, and the micelles act as the pore-directing agents in the synthesis of nanoparticles. Scanning electron microscopy results revealed that the MPN had a uniform size of 70 nm on average and a distinct mesoporous structure. The development of a concave mesoporous structure on the surface of the MPNs can increase the surface area and facilitate the efficient transport of reactants. The synthesized MPNs exhibited peroxidase-like activity. Furthermore, the MPNs showed excellent catalytic efficiency compared to HRP, due to the high surface area derived from the presence of the mesoporous structure. The peroxidase-like MPNs were applied to the enzyme-linked immunosorbent assay (ELISA) of C-reactive protein (CRP). The MPN-based ELISA exhibited sensitive CRP detection in the range from 0.24 to 7.8 ng/mL with a detection limit of 0.13 ng/mL. Moreover, the recoveries of the CRP concentrations in spiked human serum were 98.6% and 102%. These results demonstrate that as a peroxidase mimic, the MPNs can replace the natural enzymes in conventional ELISA for sensitive CRP detection.


Assessment of the number of lamellae in the central region of the normal human corneal stroma at the resolution of the transmission electron microscope.
To determine the total number of lamellae within the central region of the human corneal stroma by using a continuous section through the corneal stroma and at the ultrastructural resolution of the electron microscope. Six human eye bank corneas, from individuals aged between 42 and 82 years, were received in corneal storage medium (Optisol GS) and then processed for transmission electron microscopy with buffered 2% glutaraldehyde fixation. Thin sections were mounted on parallel bar copper grids for an uninterrupted full corneal thickness observation. A sequence of overlapping micrographs were taken at 2,600x magnification, printed at a final magnification of 3,640x, and assembled as a montage. The number of lamellae were counted across the corneal stromal strip by three observers, independently, by using a preagreed set of criteria for identifying individual lamella. The average number of lamellae per stroma was determined to be 242+/-4 (range, 234-247). In the more anterior region of the stroma, the density of the lamellae was 50% greater than in the posterior stroma (mean, 57+/-12 per 100 microm anteriorly vs. 38+/-5 per 100 microm posteriorly). Interobserver differences were modest and generally less than the intersample variance. When assessed at the resolution of the transmission electron microscope, the number of stromal lamellae in the central region of the human cornea are higher (at approximately 240 per cornea) than most previous estimates reported from light microscopy studies.


Vhc1, a novel transporter belonging to the family of electroneutral cation-Cl(-) cotransporters, participates in the regulation of cation content and morphology of Saccharomyces cerevisiae vacuoles.
Cation-Cl(-) cotransporters (CCCs) are integral membrane proteins which catalyze the coordinated symport of Cl(-) with Na(+) and/or K(+) ions in plant and mammalian cells. Here we describe the first Saccharomyces cerevisiae CCC protein, encoded by the YBR235w open reading frame. Subcellular localization studies showed that this yeast CCC is targeted to the vacuolar membrane. Deletion of the YBR235w gene in a salt-sensitive strain (lacking the plasma-membrane cation exporters) resulted in an increased sensitivity to high KCl, altered vacuolar morphology control and decreased survival upon hyperosmotic shock. In addition, deletion of the YBR235w gene in a mutant strain deficient in K(+) uptake produced a significant growth advantage over the parental strain under K(+)-limiting conditions, and a hypersensitivity to the exogenous K(+)/H(+) exchanger nigericin. These results strongly suggest that we have identified a novel yeast vacuolar ion transporter mediating a K(+)-Cl(-) cotransport and playing a role in vacuolar osmoregulation. Considering its identified function, we propose to refer to the yeast YBR235w gene as VHC1 (vacuolar protein homologous to CCC family 1).


Nutritional status associates with immunotherapy clinical outcomes in recurrent or metastatic head and neck squamous cell carcinoma patients.
Beyond programmed death-ligand 1 (PD-L1) assessed by the combined positive score (CPS) and tumor mutational burden (TMB), no other biomarkers are approved for immunotherapy interventions. Here, we investigated whether additional clinical and pathological variables may impact on immunotherapy outcomes in recurrent or metastatic (R/M) head and neck squamous cell carcinoma (HNSCC) patients. R/M HNSCC patients treated with immunotherapy were reviewed. Analyzed variables at baseline included: clinicopathological, laboratory, and variables reflecting the host nutritional status such as the prognostic nutritional index (PNI) and albumin. The primary endpoint was progression free survival (PFS). The secondary endpoints were overall survival (OS) and objective response rate (ORR). Univariable and multivariable Cox models were fitted and random forest algorithm was used to estimate the importance of each prognostic variable. A total of 100 patients were treated with immunotherapy; 50% with single agent and 50% with experimental immunotherapy combinations. In the multivariable analysis, both ECOG performance status (HR: 1.73; 95%CI 1.07-2.82; p = 0.03) and PNI levels (10-point increments, HR: 0.66; 0.46-0.95; p = 0.03) were significantly associated with PFS. However, the derived neutrophil to lymphocyte ratio (dNLR) and lactate dehydrogenase (LDH) were not significantly associated with PFS (p-values > 0.15). In the OS analysis, albumin and PNI were the only statistically significant factors in the multivariable model (p < 0.001). In our cohort, PNI and ECOG performance status were most strongly associated with PFS in R/M HNSCC patients treated with immunotherapy. These results suggest that parameters informative of nutritional status should be considered before immunotherapy.


Circulating cell free DNA testing: are some test failures informative?
The proportion of circulating cell free DNA derived from the feto-placental unit (fetal fraction or FF) correlates with test success and interpretation reliability. Some fetal disorders are associated with systematically lower FF, sometimes resulting in noninformative results. We analyzed results from pregnancies tested in a nested case/control study derived from a cohort of 4664 high-risk pregnancies. Low FF was defined before and after adjusting for maternal weight and gestational age. Compared with euploid pregnancies, the median FF was significantly higher in Down syndrome pregnancies (ratio 1.17) and significantly lower in trisomy 18 and triploid pregnancies (ratios 0.71 and 0.19, respectively). Among 2157 pregnancies tested, 13 (0.6%) had FF <3.0% (all noninformative), including three trisomy 18 and three triploidy fetuses. After adjustment, 16 pregnancies (0.7%) had FF <0.3 multiples of the median (six informative), including one trisomy 18 and three triploidy fetuses. Modeled positive predictive values for low and high-risk populations were 7% and 30%, respectively. Among women with noninformative results attributable to low FF, trisomy 18 and/or triploidy risk are sufficiently high to warrant offering additional assessments (e.g. ultrasound). If the testing indication is ultrasound abnormality, amniocentesis and karyotype/microarray should be considered. © 2014 John Wiley & Sons, Ltd.


Kinematic and Temporospatial Changes in Children with Cerebral Palsy during the Initial Stages of Gait Development.
To identify changes in the gait kinematics and temporospatial parameters of children with bilateral Cerebral Palsy (CP) at 8 months after the onset of independent walking and identify differences to Typical Development (TD) children at the onset of independent walking and at 8 months follow up. Sixteen children with bilateral CP, GMFCS levels I and II, and 15 TD children were recruited. Gait kinematics and temporospatial parameters were recorded using a 3-D gait analysis system; the sagittal plane of the lower limb joints was analyzed. Baseline measurements were recorded at the individual's onset of independent walking and follow up was after 8 months. Compared to baseline, children with bilateral CP demonstrated increased (mean difference ± SE) plantar flexion (11.79 ± 2.96), single support (0.04 ± 0.01), step length (0.2 ± 0.05) and stride length (0.4 ± 0.09), at follow up; all p < .05. Compared to TD children, they also had lower gait speed (0.16 ± 0.05), higher single support (0.02 ± 0.01) and lower maximum knee extension (9.14 ± 4.49) during the swing phase, at baseline and follow up (0.1 ± 0.04, 0.05 ± 0.01, 23.04 ± 4.17, respectively); all p < .05. There are changes in the sagittal plane kinematics and temporospatial parameters of the gait during the first 8 months of independent walking. These indicate gait maturation changes and highlight the impact of walking experience on the gait characteristics of children with bilateral CP.


Bibliometric Analysis and Visualization of Catholic Health Care Research: 1973-2019.
A multi-dimensional construct of Catholic health care is examined using a bibliometric analysis of 181 scientific studies from the Web of Science database. Medical ethics, religion, and health services research by 418 authors from 26 countries reveals developments in the Catholic health care domain since 1973. Using VOSviewer, we map keyword clusters to reveal the structure of research on Catholic health care that transcends religious and secular literatures. The clusters mostly reflect clinical and ethical issues of reproductive health and the end of life, as well as the essence and the future of the Catholic identity from the theological and empirical perspectives.


Effects of PF1022A on adult Angiostrongylus cantonensis in the pulmonary arteries and larvae migrating into the central nervous system of rats.
We examined the effects of PF1022A, newly developing in Japan, on adult Angiostrongylus cantonensis in the pulmonary arteries of rats. Following five and ten successive oral doses at 10 mg/kg per day, the first-stage larvae in rat faeces disappeared completely at 2 weeks after treatment. The treatment completely killed the female worms, but not the male worms. However, numbers of male worms were also decreased after the administration of either five successive oral doses at 10 mg/kg per day for four courses or five successive intraperitoneal doses at 0.5 mg/kg per day. Next, we examined the effects of PF1022A on larval A. cantonensis migrating into the central nervous system (CNS) of rats. Following five successive oral doses at 5 or 10 mg/kg per day and five successive intraperitoneal doses at 0.5 mg/kg per day, lesser killing effects were observed on male as well as female worms. On the basis of these results it is apparent that PF1022A will become a promising anthelmintic available as treatment for tissue-dwelling as well as intestinal nematodes.


Acetylcholine from the nucleus basalis magnocellularis facilitates the retrieval of well-established memory.
Retrieval deficit of long-term memory is a cardinal symptom of dementia and has been proposed to associate with abnormalities in the central cholinergic system. Difficulty in the retrieval of memory is experienced by healthy individuals and not limited to patients with neurological disorders that result in forgetfulness. The difficulty of retrieving memories is associated with various factors, such as how often the event was experienced or remembered, but it is unclear how the cholinergic system plays a role in the retrieval of memory formed by a daily routine (accumulated experience). To investigate this point, we trained rats moderately (for a week) or extensively (for a month) to detect a visual cue in a two-alternative forced-choice task. First, we confirmed the well-established memory in the extensively trained group was more resistant to the retrieval problem than recently acquired memory in the moderately trained group. Next, we tested the effect of a cholinesterase inhibitor, donepezil, on the retrieval of memory after a long no-task period in extensively trained rats. Pre-administration of donepezil improved performance and reduced the latency of task initiation compared to the saline-treated group. Finally, we lesioned cholinergic neurons of the nucleus basalis magnocellularis (NBM), which project to the entire neocortex, by injecting the cholinergic toxin 192 IgG-saporin. NBM-lesioned rats showed severely impaired task initiation and performance. These abilities recovered as the trials progressed, though they never reached the level observed in rats with intact NBM. These results suggest that acetylcholine released from the NBM contributes to the retrieval of well-established memory developed by a daily routine.


Utility of 'back-up' arterio-venous fistulae in patients on peritoneal dialysis and use of haemodialysis catheters.
Patients undergoing peritoneal dialysis may require unanticipated transfer to haemodialysis. Back up fistula are often created in selected patients. These may help reduce the infective burden of haemodialysis (HD) catheter. To study the utility of back-up arterio-venous fistulae (AVF) in patients initiated on peritoneal dialysis (PD) and to determine the rates of HD catheter use in patients requiring conversion to HD. Data on HD transfer and HD catheter usage were retrospectively analysed in all patients initiating PD between January 2010 and December 2014 at Royal Adelaide Hospital (RAH; universal back-up AVF creation at PD commencement) and Princess Alexandra Hospital (PAH; selective back-up AVF creation in 'high risk' patients). A total of 374 patients initiated PD during the study period: 142 in RAH group and 232 in PAH group. The groups were reasonably comparable, except that RAH patients were more likely to be older, Caucasian and diabetic. Transfer to HD occurred in 33 (23%) patients in RAH group and 99 (43%) in the PAH group with respective median times to HD transfer of 289 and 295 days. HD catheter usage was required at the time of HD transfer in 11 (33%) patients at RAH and 64 (65%) in patients at PAH (P < 0.001). AVF complications occurred in 13 (9%) patients in RAH group (fistuloplasty n = 8, transposition n = 2, ligation due to ischaemia n = 2 and construction of new AVF n = 1). Patients undergoing PD frequently require urgent unanticipated transfer to HD and back-up AVF can be successfully utilised in this setting in the majority of cases, which in turn can reduce the infective burden of HD catheter exposure.


Epidemiological study about the mental state of patients after a pulmonary embolism or deep venous thrombosis event.
Venous thromboembolism (VTE) occurs frequently and represents a serious threat to patient health. However, its effects on mental health have not been studied sufficiently. The objective of this study was to investigate whether VTE alters the patients' mental state. We gathered questionnaire data on 100 patients (59 men, 41 women; age ranging from 24 to 85 years) concerning psychological symptoms and body image. Our results show that after a VTE, patients develop psychological symptoms, some of which persist for more than 2 years. Examples of those symptoms include depression, intrusion, and increased scrutiny of the body. Intrusion (flashbacks, nightmares, and other traumatic sensations of reliving the thrombotic event) affected 69% of patients and may lead to social isolation, occasionally including loss of employment. Depression affected more than 50% of patients. Productivity typically decreases, and body functionality and feeling of health changed in 34-76% of patients. However, anxiety, disordered impulse control, and maladjustment were less frequent (40% or less), and patients' emotional attitude to their bodies (liking their bodies or being angry with their bodies, feelings of shame and attractiveness) tended not to change over time. Nevertheless, we feel it may be advisable to identify patients with relevant psychological changes after VTE by means of a short evidence-based questionnaire and to offer them psychological treatment in order to improve management and quality of life of these patients. The goal is, therefore, to develop diagnostic and therapeutic recommendations.


A 20-week randomized controlled trial of estradiol replacement therapy for women aged 70 years and older: effect on mood, cognition and quality of life.
The results of several observational studies suggest that the use of estrogen replacement is associated with better mood, cognitive function and quality of life. Such findings are consistent with those of laboratory-based research showing that estrogen promotes neuronal sprouting, enhances cholinergic activity in the brain, decreases brain and plasma levels of beta-amyloid, increases serotonin postsynaptic responsivity and the turnover of noradrenaline, and inhibits monoamine oxidase activity. However, the findings from the Women's Health Initiative controlled trial showed that hormone replacement (estrogen plus progestin) not only failed to improve mood, cognition and quality of life but also increased the risk of dementia. At present, there is limited information about the effect of unopposed estradiol replacement therapy (ERT) on the mental health outcomes of women at increased risk of cognitive decline (aged 70 years and over). We designed the present randomized, double-blind, placebo-controlled trial to clarify this issue. One hundred and fifteen women were randomized to treatment with estradiol (n=58; 2mg per day) or placebo for a total period of 20 weeks. The outcomes of interest in this study included changes in the Beck Depression Inventory (BDI) scores between baseline and week 20, as well as changes in quality of life scores (as measured by the SF-36) and cognitive function (CAMCOG, Block Design, Memory for Faces, California Verbal Learning Test (CVLT) and verbal fluency (VF)). Nineteen women treated with estradiol and 10 of those treated with placebo discontinued the use of the medication during trial, most frequently due to adverse reactions (OR=4.11, 95% CI=1.29-15.37). Intention-to-treat analysis showed that the active and placebo groups did not differ in their response to treatment in any of the outcome measures (p>0.05). A separate analysis restricted to women who completed the 20-week-trial produced similar negative results. The results of this trial indicate that the use of a relatively high dosage of unopposed estrogen replacement for 20 weeks is not associated with significant changes in cognitive function, mood and quality of life. Other more efficacious and safer interventions need to be devised with the aim of improving the mental state and quality of life of older women.


Primary or idiopathic intestinal pneumatosis: a rare casual endoscopic finding.
A 66-year-old male patient with no specific prior medical history consulted due to constipation without other associated symptoms. A colonoscopy was performed and multiple pseudonodular lesions were detected from the sigmoid to splenic angle in the form of air bubbles with normal-appearing mucosa and a preserved vascular pattern. Gas was released when the biopsies were being taken. An abdominal computed tomography (CT) scan was performed due to the suspicion of an idiopathic or primary intestinal pneumatosis, in which the findings confirmed the diagnosis.


Cortical hyperexcitability: Diagnostic and pathogenic biomarker of ALS.
Cortical hyperexcitability is an early and intrinsic feature of both sporadic and familial forms of amyotrophic lateral sclerosis (ALS).. Importantly, cortical hyperexcitability appears to be associated with motor neuron degeneration, possibly via an anterograde glutamate-mediated excitotoxic process, thereby forming a pathogenic basis for ALS. The presence of cortical hyperexcitability in ALS patients may be readily determined by transcranial magnetic stimulation (TMS), a neurophysiological tool that provides a non-invasive and painless method for assessing cortical function. Utilising the threshold tracking TMS technique, cortical hyperexcitability has been established as a robust diagnostic biomarker that distinguished ALS from mimicking disorders at early stages of the disease process. The present review discusses the pathophysiological and diagnostic utility of cortical hyperexcitability in ALS.


Corneal anesthesia in a case with Vogt-Koyanagi-Harada syndrome.
To add clinical features to the description of the Vogt-Koyanagi-Harada syndrome. Case report. The case presented with a typical medical history of Vogt-Koyanagi-Harada syndrome, including headaches, low-grade fever, nuchal rigidity, and from the eyes bilateral visual loss, a reaction from the anterior chambers, bilateral uveities with localized exudative retinal detachment from the left. In addition there were tonic pupils, anesthesia of the corneas, and an accommodative deficit. Corneal anesthesia, tonic pupils and accommodative impairment can be features of the Vogt-Koyanagi-Harada syndrome.


X-linked retinoschisis maculopathy treated with topical dorzolamide, and relationship to genotype.
To correlate the response of topical dorzolamide (Trusopt; Merck) in patients with X-linked retinoschisis (XLRS) with genotype. We carried out a retrospective evaluation of four patients (seven eyes) with XLRS, treated with topical dorzolamide. The change in best-corrected visual acuity (VA) and central macular thickness (CMT; central 1 mm subfield thickness) from optical coherence tomography (OCT) was analysed over the follow-up period, using Student's t-test. Each patient also had genetic analysis for mutations in the retinoschisisgene (RS1). The mean age at the start of treatment was 14.7±11 years, and mean follow-up duration was 21.7±7.7 months. Mean CMT at the final follow-up was significantly better than at baseline (291±123 vs 352±119 μm, P=0.007); however, mean VA was worse (0.38±0.25 vs 0.31±0.24 logMAR score, P=0.041). All four patients had a mutation in the RS1gene; there was no apparent association between the type of mutation and the response to topical dorzolamide. Topical dorzolamide may have some effect in reducing central macular thickness in patients with XLRS, but this does not necessarily correlate with improvement in VA. In our case series, genotypic information did not predict the response to this treatment.


Mandibular condylar cartilage response to moving 2 molars in rats.
The purpose of this study was to investigate the responses of mandibular condylar cartilage to moving 2 molars in different combinations. Rats were assigned to male and female control and experimental groups (each, n = 5). Elastic rubber bands were used to move medially the maxillary left and the mandibular right first molars in experimental group I. The same method was used to distally move the maxillary left and the mandibular right third molars, 2 mandibular third molars, and 2 maxillary third molars in experimental groups II, III, and IV, respectively. At the end of the eighth week, all condyles were examined histologically. The areas of histologic change as a percentage of total cartilage area were compared by using the Mann-Whitney U test. Cartilage degenerative remodeling was observed in experimental groups II, III, and IV. The percentage areas of degenerative remodeling were higher in female experimental groups II and III than in the female control group, and in female experimental group II than in female experimental group IV and male experimental group II (all, P <0.05). The mandibular condylar cartilage of female rats responded variously to different combinations of molar movement; the most obvious remodeling was observed in groups in which the maxillary left and mandibular right third molars were moved.


Pre-morbid predictors of psychological adjustment to cancer.
This study examined the potential role of social support, neuroticism, and self-efficacy as predictors of the short-term and long-term adaptation to the diagnosis of cancer. Psychological adjustment was defined in terms of psychological distress. It is argued that these factors may provide insights that might be helpful in the provision of medical care to patients. We hypothesized that they would be especially predictive of long-term psychological adjustment. The study was conducted in a truly prospective design with one pre-morbid assessment and two post-morbid assessments used in the present analyses. Participants of a baseline assessment among 5279 subjects (aged 57 years and older) in 1993 were monitored for cancer incidence by their general practitioners. The study sample included 99 cancer patients who had completed all pre-morbid and post-morbid assessments. Multivariate analyses revealed that high neuroticism was associated with higher levels of distress in the short- and long-term. Higher levels of social support were associated with higher levels of distress in the long-term. The direction of this association was opposite to what might be expected on the basis of the literature. Implications of the findings for health care as well as explanations for the social support findings are discussed.


Current investigations on clinical pharmacology and therapeutics of Glucose-6-phosphate dehydrogenase deficiency.
Glucose-6-phospate dehydrogenase (G6PD) deficiency is estimated to affect more than 400 million people world-wide. This X-linked genetic deficiency puts stress on red blood cells (RBC), which may be further augmented under certain pathophysiological conditions and drug treatments. These conditions can cause hemolytic anemia and eventually lead to multi-organ failure and mortality. G6PD is involved in the rate-limiting step of the pentose phosphate pathway, which generates reduced nicotinamide adenine dinucleotide phosphate (NADPH). In RBCs, the NADPH/G6PD pathway is the only source for recycling reduced glutathione and provides protection from oxidative stress. Susceptibility of G6PD deficient populations to certain drug treatments and potential risks of hemolysis are important public health issues. A number of clinical trials are currently in progress investigating clinical factors associated with G6PD deficiency, validation of new diagnostic kits for G6PD deficiency, and evaluating drug safety, efficacy, and pathophysiology. More than 25 clinical studies in G6PD populations are currently in progress or have just been completed that have been examined for clinical pharmacology and potential therapeutic implications of G6PD deficiency. The information on clinical conditions, interventions, purpose, outcome, and status of these clinical trials has been studied. A critical review of ongoing clinical investigations on pharmacology and therapeutics of G6PD deficiency should be highly important for researchers, clinical pharmacologists, pharmaceutical companies, and global public health agencies. The information may be useful for developing strategies for treatment and control of hemolytic crisis and potential drug toxicities in G6PD deficient patients.


Effects of starting powder and thermal treatment on the aerosol deposited BaTiO3 thin films toward less leakage currents.
To prepare high-density integrated capacitors with low leakage currents, 0.2-μm-thick BaTiO3 thin films were successfully deposited on integrated semiconductor substrates at room temperature by the aerosol deposition (AD) method. In this study, the effects of starting powder size were considered in an effort to remove macroscopic defects. A surface morphology of 25.3 nm and an interface roughness of less than 50 nm were obtained using BT-03B starting powder. The nano-crystalline thin films achieved after deposition were annealed at various temperatures to promote crystallization and densification. Moreover, the influence of rapid thermal annealing process on the surface morphology and crystal growth was evaluated. As the annealing temperature increased from room temperature to 650°C, the root mean square (RMS) roughness decreased from 25.3 to 14.3 nm. However, the surface was transformed into rough performance at 750°C, which agreed well with the surface microstructure trend. Moreover, the crystal growth also reveals the changes in surface morphology via surface energy analysis.


Study on potential antitumor mechanism of a novel Schiff base copper(II) complex: synthesis, crystal structure, DNA binding, cytotoxicity and apoptosis induction activity.
A new cytotoxic copper(II) complex with Schiff base ligand [Cu(II)(5-Cl-pap)(OAc)(H(2)O)]·2H(2)O (1) (5-Cl-pap=N-2-pyridiylmethylidene-2-hydroxy-5-chloro-phenylamine), was synthesized and structurally characterized by X-ray diffraction. Single-crystal analysis revealed that the copper atom shows a 4+1 pyramidal coordination, a water oxygen appears in the apical position, and three of the basal positions are occupied by the NNO tridentate ligand and the fourth by an acetate oxygen. The interaction of Schiff base copper(II) complex 1 with DNA was investigated by UV-visible spectra, fluorescence spectra and agarose gel electrophoresis. The apparent binding constant (K(app)) value of 6.40×10(5) M(-1) for 1 with DNA suggests moderate intercalative binding mode. This copper(II) complex displayed efficient oxidative cleavage of supercoiled DNA, which might indicate that the underlying mechanism involve hydroxyl radical, singlet oxygen-like species, and hydrogen peroxide as reactive oxygen species. In addition, our present work showed the antitumor effect of 1 on cell cycle and apoptosis. Flow cytometric analysis revealed that HeLa cells were arrested in the S phase after treatment with 1. Fluorescence microscopic observation indicated that complex 1 can induce apoptosis of HeLa cells, whose process was mediated by intrinsic mitochondrial apoptotic pathway owing to the activation of caspase-9 and caspase-3.


Intra-individual comparison of p-[123I]-iodo-L-phenylalanine and L-3-[123I]-iodo-alpha-methyl-tyrosine for SPECT imaging of gliomas.
Radioactive amino-acids accumulate in gliomas even with an intact blood-brain-barrier. L-3-[(123)I]-iodo-alpha-methyl-tyrosine (IMT) is well established for SPECT imaging of gliomas. Recently, we introduced p-[(123)I]-iodo-L-phenylalanine (IPA) for the characterisation of brain lesions. This study compares both tracers in glioma patients. Eleven patients with gliomas (1 WHO grade 1, 5 grade 2, 1 grade 3, 2 grade 4 gliomas, 1 unconfirmed upgrading and 1 post-therapeutic non-neoplastic lesion) underwent SPECT imaging with IPA (early and delayed acquisitions at 30 min and 3 h) and IMT (early only). Maximum tumour-to-brain ratios (TBR) were calculated using region-of-interest analysis to assess uptake of IMT and IPA. Imaging results were compared to histopathological findings. Early TBRs of IMT and IPA were strongly correlated (r = 0.828, p = 0.002). TBRs were higher for IMT than IPA (1.95+/-0.50 versus 1.79+/-0.42; p < 0.05), but independent from tumour cell density (p > 0.1). Visual interpretation by different observers was more concordant for IMT-SPECT than IPA-SPECT (kappa 1.0 versus 0.774). No differences in early TBRs were observed between low-grade and high-grade gliomas for IMT (1.97+/-0.53 versus 2.21+/-0.44, p > 0.5) or IPA (1.70+/-0.23 versus 2.21+/-0.56, p = 0.167) with a trend to higher TBRs in low-grade tumours for IMT (p = 0.093). In contrast to the known wash-out of IMT, we observed persistent accumulation of IPA in gliomas. IPA shows lower TBRs than IMT, especially in low-grade tumours, so IMT should be preferred for the delineation of low-grade gliomas by SPECT imaging. Due to its prolonged retention, however, IPA remains promising for therapeutic use in gliomas after labelling with I-131.


Diagnostic stability and long-term symptomatic and functional outcomes in first-episode antipsychotic-naïve patients with schizophrenia.
In a prospective cohort design, we investigated: i) diagnostic stability of initially antipsychotic-naïve schizophrenia patients, ii) symptom severity including symptomatic remission, and iii) functional remission including full recovery. We included 143 antipsychotic-naïve patients with first-episode schizophrenia or schizoaffective disorder. After 4-18 years, we clinically re-evaluated diagnosis, symptom severity and functioning for 70 patients. From the nationwide Danish registers, we extracted pragmatic outcome measures for 142 patients. We examined associations between baseline variables (age at diagnosis, sex, and premorbid intelligence) and long-term outcome status (symptomatic and functional remission). At 4-18 years follow-up, 80% met the criteria for schizophrenia or schizoaffective disorder, however, despite the high diagnostic stability 53% met the criteria of symptomatic and/or functional remission. Symptomatic remission characterized 34% of the patients and was associated with female sex, better premorbid intelligence, and a younger age at schizophrenia diagnosis. Functional remission characterized 41% of the patients and 17% of patients met criteria for full recovery both of which were associated with female sex. The clinically re-evaluated patients did not differ from the drop-outs on key register-based variables. We confirm the emerging evidence of a decreasing long-term diagnostic stability of schizophrenia, and a protective role of female sex. The association between premorbid intelligence and symptomatic remission underscores the pertinence of including cognitive deficits in the diagnostic category of schizophrenia. The association between younger age at diagnosis and symptomatic remission may reflect positive effects of early detection or a drift in the interpretation of the diagnostic classification system.


The developmental context of cell-cycle control in plants.
Plant growth is characterised both by continued growth and organogenesis throughout development, as well as by environmental influences on the rate and pattern of these processes. This necessitates a close relationship between cell cycle control, differentiation and development that can be readily observed and studied. The sequencing of the Arabidopsis genome has revealed the full complexity of cell cycle regulators in plants, creating a challenge to understand how these genes control plant growth and differentiation, and how they are integrated with intrinsic and external signals. Here, we review the control of the cell cycle and examine how it is integrated with proliferative activity within meristems, and during the differentiation processes leading to leaf and lateral root formation.


The effect of duration of exposure on sulfuric acid-induced pulmonary function changes in asthmatic adolescent subjects: a dose-response study.
To evaluate the pulmonary effects of varying doses of sulfuric acid, adolescent subjects with asthma were exposed to 35 or 70 micrograms/m3 sulfuric acid for 45 or 90 min. Exposure was carried out during intermittent moderate exercise. The pulmonary functions measured before and after exposure were FEV1, FVC, and total respiratory resistance. The 45 min exposures were associated with larger decreases in FEV1 (-6% or -3%) than the 90 min exposures (-1% or +2%). Analysis of variance of the change in FEV1 among the exposures revealed that the 45 min exposure to 35 micrograms/m3 was significant (p = 0.03). The p value for 45 min exposure to 70 micrograms/m3 was not significant (p = 0.08). Using analysis of variance, neither of the 90 min exposures was associated with a significant decrease in FEV1 compared to air exposure. Also, none of the changes in FVC or RT was significant. When baseline to post-exposure changes were compared for each of the five test atmospheres using paired t tests, both of the 45 min exposures were associated with statistical significance (p < 0.001 for 35 micrograms/m3 and p < 0.005 for 70 micrograms/m3). This baseline to post exposure change was not statistically significant for the 90 min exposures. The reason for the lesser effect on pulmonary function at increased exposure duration is not known; it may be due to changes in either varying deposition patterns or changes in buffering capacity of the cells lining the airways. With respect to individual sensitivities to H2SO4, the data showed a significant consistency across test atmospheres.


Phyto-genotoxicity of arsenic contaminated soil from Lakhimpur Kheri, India on Vicia faba L.
The present experiment was designed to evaluate physico-chemical characteristics and phyto-genotoxicity of arsenic (As) contaminated soil collected from different sites of Lakhimpur, Uttar Pradesh (UP), India by employing Vicia faba L. The analyses included various biochemical as well as cyto-genotoxicity assays. The results showed that soil pH was slightly acidic to neutral in nature. The bulk density (1.18-1.23 gcm-3), particle density (2.51-2.59 gcm-3) and porosity (44-53%) varied in different places. The level of available nutrients, nitrogen, phosphorus and potassium was found to vary as 124-165 mgkg-1, 173-186 mgkg-1 and 48-98 mgkg-1, respectively. The maximum As levels were found in soil of Fulvareya (27.13 mgkg-1) and Atareya (24.12 mgkg-1), the level of As in water samples of these sites were 0.19 mgl-1and 0.21 mgl-1, respectively. Phytotoxicity of the As present in soils was evident through significant increases in stress metabolites, hydrogen peroxide (H2O2), malondialdehyde (MDA) and carbonyl groups in root and shoot of V. faba. Cyto-genotoxic effects were also seen through reduced mitotic index (MI) and increased mitotic depression (MD), relative abnormality rate (RAR) as well as other chromosomal abnormalities along with micronuclei in root meristematic cells of V. faba. The phytotoxicity and cyto-genotoxicity assessment suggests the harmful soil properties that might affect biota.


Translational model of melphalan-induced gut toxicity reveals drug-host-microbe interactions that drive tissue injury and fever.
Conditioning therapy with high-dose melphalan (HDM) is associated with a high risk of gut toxicity, fever and infections in haematopoietic stem cell transplant (HSCT) recipients. However, validated preclinical models that adequately reflect clinical features of melphalan-induced toxicity are not available. We therefore aimed to develop a novel preclinical model of melphalan-induced toxicity that reflected well-defined clinical dynamics, as well as to identify targetable mechanisms that drive intestinal injury. Male Wistar rats were treated with 4-8 mg/kg melphalan intravenously. The primary endpoint was plasma citrulline. Secondary endpoints included survival, weight loss, diarrhea, food/water intake, histopathology, body temperature, microbiota composition (16S sequencing) and bacterial translocation. Melphalan 5 mg/kg caused self-limiting intestinal injury, severe neutropenia and fever while impairing the microbial metabolome, prompting expansion of enteric pathogens. Intestinal inflammation was characterized by infiltration of polymorphic nuclear cells in the acute phases of mucosal injury, driving derangement of intestinal architecture. Ileal atrophy prevented bile acid reabsorption, exacerbating colonic injury via microbiota-dependent mechanisms. We developed a novel translational model of melphalan-induced toxicity, which has excellent homology with the well-known clinical features of HDM transplantation. Application of this model will accelerate fundamental and translational study of melphalan-induced toxicity, with the clinical parallels of this model ensuring a greater likelihood of clinical success.


Characterising salivary peptidome across diurnal dynamics and variations induced by sampling procedures.
This study aimed to characterise diurnal dynamics of salivary peptidome and variations induced by sampling procedures. A supervised short-term longitudinal study was conducted amongst ten healthy participants. Saliva samples were collected by different procedures (stimulated/unstimulated conditions, forepart/midstream segments) on three consecutive days. The peptidome compositions of saliva samples were analysed using matrix-assisted laser desorption/ionisation-time of flight-mass spectrometry (MALDI-TOF MS). The salivary peptidome exhibited a stable trend generally, even though some diurnal dynamics happened in aspects of both overall structure and certain single peptides. The results indicated saliva samples collected under unstimulated and stimulated conditions have significantly different structures of peptidome, whilst the peptidome profile of stimulated saliva was more abundant than that of unstimulated saliva. It was also indicated that the midstream segment effect might exist in the segmented process of saliva sampling. In summary, salivary peptidome was able to maintain stability though some dynamic changes might happen within a short-term period. Stimulated and unstimulated saliva samples had significantly different peptidome profiles, whilst the stimulated whole saliva was a larger pool of low molecular weight peptides. The stability of the salivary peptidome highlights the reliability of salivary peptidome as a source of diagnostic biomarker. We recommend keeping one collection condition (stimulated/unstimulated) consistently within one study on salivary peptidome. Stimulated whole saliva would be preferred if more abundant peptidome profile is needed.


Risk of Soil-Transmitted Helminthiasis among Agrarian Communities of Kogi State, Nigeria.
Soil-transmitted helminths (STH) have remained a major threat to humans, especially children in developing countries, including Nigeria. Interventions have always been geared towards school-aged children, neglecting preschool-aged children and occupational risk adults. The Soil-Transmitted Helminthiasis Advisory Committee (STHAC) recently suggested incorporating other at-risk groups. This study assessed the associated risk of STH infection among agrarian communities of Kogi State, Nigeria. A total of 310 individuals of all ages participated in the cross-sectional survey. Stool samples were analyzed using standard Kato-Katz method. A total of 106 (34.2%) individuals were infected with at least one STH. Hookworm was the most prevalent (18.1%); followed by Ascaris lumbricoides (16.8%). Worm intensity was generally light. Prevalence of infection was similar between four age groups considered (preschool, school, 'women of reproductive age' and older at-risk group). Poor socio-economic status (SES) was a major risk for STH infection. Using a 20-asset based criteria, 68 (23.1%) and 73 (24.7%) of 295 questionnaire respondents were classified into first (poorest) and fifth (richest) wealth quintiles respectively. Risk of infection with STH was 60% significantly lower in the richest wealth quintile compared to the poorest (Prevalence Ratio [PR] = 0.4843, 95% CI = 0.2704-0.8678, p = 0.015). Open defecators were more likely to harbour STH than those who did not (PR = 1.7878, 95% CI = 1.236-2.5846, p = 0.00201). Pit latrine and water closet toilets each approximately reduced STH infection by 50% (p < 0.05). Preventive chemotherapy for all age groups, health education and provision of basic amenities especially toilets are needed in order to achieve the goal toward the 2020 target of STH control.


Control charts demonstrated limited utility for the monitoring of lung function in asthma.
Statistical process control charts have been advocated for use in monitoring of lung function in asthma. We aimed to evaluate their application in asthma using existing data from a randomized trial. Patients on optimal inhaled corticosteroid/bronchodilator therapy (n=81) were randomized to continue the same or change to corticosteroid alone. Baseline statistical control was assessed from 20 days of electronically recorded lung function (peak expiratory flow [PEF], forced expiratory volume in one second [FEV(1)]). The ability to detect lung function changes was assessed during 10 days after randomization. PEF measurements were in statistical control during baseline for only 59-79% of patients for different combinations of five control chart rules (e.g., Rule 1: >3 standard deviations outside mean and 95% expected to be in control), with similar proportions for FEV(1). After randomization, among those previously in statistical control, Rule 1 signaled lower FEV(1) for 35% of patients randomized to treatment change compared with 6% continuing baseline treatment (P=0.004). Control charts performed poorly for PEF, signaling a decrease for 8% of patients randomized to treatment change compared with 11% continuing treatment (P=0.7). A paradoxical increase was signaled for many patients continuing treatment. Control charts may not be suitable for use in many patients with asthma.


Diagnosing tuberculosis in a resource-poor setting: the value of a trial of antibiotics.
Diagnosis of smear-negative but culture-positive pulmonary tuberculosis in resource-poor settings is difficult. To determine the value of assessing response to a trial of antibiotics in the identification of patients with positive cultures but negative Ziehl-Neelsen (ZN) smears, we compared clinicians' diagnoses with culture in 334 consecutive adults with suspected tuberculosis in rural South Africa; 142 patients (43%) had culture-positive pulmonary tuberculosis. Diagnosis by ZN smear alone was insensitive (61%) but highly specific (94%). Only half of the smear-negative but culture-positive cases were correctly identified by failing to respond to a broad spectrum antibiotic. The remainder responded to therapy and were discharged. Diagnostic sensitivity therefore increased to 80%, but specificity fell to 78%. A more rigorous algorithm may improve diagnosis of smear-negative pulmonary tuberculosis in resource-poor settings.


'Demand characteristics' in body-size estimation in anorexia nervosa.
We asked 24 patients with anorexia nervosa and 30 normal controls to estimate their body-size several times, each time using different instructions. The degree of over-estimation was found to vary predictably with the wording of the instructions. Informing the subject that she had made an error without specifying the direction of the error resulted in reduced over-estimation on a subsequent trial, for both anorexics and controls. 'Internally directed' instructions were associated with a greater degree of over-estimation than 'external' instructions in both groups, but particularly in anorexic subjects. Our results indicate the necessity of controlling the 'demand characteristics' of such experiments.


Dietary intake of advanced glycation endproducts and risk of hepatobiliary cancers: A multinational cohort study.
Advanced glycation endproducts (AGEs) may contribute to liver carcinogenesis because of their proinflammatory and prooxidative properties. Diet is a major source of AGEs, but there is sparse human evidence on the role of AGEs intake in liver cancer etiology. We examined the association between dietary AGEs and the risk of hepatobiliary cancers in the European Prospective Investigation into Cancer and Nutrition prospective cohort (n = 450 111). Dietary intake of three AGEs, Nε -[carboxymethyl]lysine (CML), Nε -[1-carboxyethyl]lysine (CEL) and Nδ -[5-hydro-5-methyl-4-imidazolon-2-yl]-ornithine (MG-H1), was estimated using country-specific dietary questionnaires linked to an AGEs database. Cause-specific hazard ratios (HR) and their 95% confidence intervals (CI) for associations between dietary AGEs and risk of hepatocellular carcinoma (HCC), gallbladder and biliary tract cancers were estimated using multivariable Cox proportional hazard regression. After a median follow-up time of 14.9 years, 255 cases of HCC, 100 cases of gallbladder cancer and 173 biliary tract cancers were ascertained. Higher intakes of dietary AGEs were inversely associated with the risk of HCC (per 1 SD increment, HR-CML = 0.87, 95% CI: 0.76-0.99, HR-CEL = 0.84, 95% CI: 0.74-0.96 and HR-MH-G1 = 0.84, 95% CI: 0.74-0.97). In contrast, positive associations were observed with risk of gallbladder cancer (per 1 SD, HR-CML = 1.28, 95% CI: 1.05-1.56, HR-CEL = 1.17; 95% CI: 0.96-1.40, HR-MH-G1 = 1.27, 95% CI: 1.06-1.54). No associations were observed for cancers of the intra and extrahepatic bile ducts. Our findings suggest that higher intakes of dietary AGEs are inversely associated with the risk of HCC and positively associated with the risk of gallbladder cancer.


An analysis of applicant competitiveness to general surgery, surgical subspecialties, and integrated programs.
General surgery was once the gateway into a career in surgery. Over time, surgical subspecialties developed separate residency programs, and recently, integrated programs have emerged. It is unknown what impact the presence of surgical subspecialties and integrated programs have had on general surgery. Our objective was to evaluate match trends and quantify competitiveness of the general surgery, integrated programs, and surgical subspecialties matches. National Residency Matching Program match data and applicant characteristics from 2010 through 2020 were analyzed for US senior allopathic applicants. Integrated programs were defined as plastic and vascular surgery, and surgical subspecialties were defined as otolaryngology, orthopedic surgery, and neurosurgery. Trends were evaluated using linear regression, programs were compared on 10 metrics by Wilcoxon rank-sum tests, and a logistic regression was used to rank each specialty match. The number of US senior applicants per position to integrated programs decreased and approached that of general surgery and surgical subspecialties, but the median number of applicants per position to general surgery was lower than to surgical subspecialties or integrated programs (1.21 interquartile range). Our logistic regression showed United States Medical Licensing Examination scores, research experience, Alpha Omega Alpha Honor Society membership, and graduation from a top medical school to be the most important factors in the match, and our weighted rank score found general surgery (2.85) to be less competitive than surgical subspecialties (1.92) or integrated programs (1.17). Throughout the last decade, integrated programs and surgical subspecialties have matched more competitive applicants based on the most significant predictors of the match. Moving forward, it is important that general surgery strives to attract the best and brightest out of medical school.


Nailing femoral shaft fracture with postless distraction technique: a new technique enabled by shape-conforming pad.
Femoral shaft fractures are usually treated with nailing using a traction table and a perineal post, but this may occasionally result in various groin-related complications, including pudendal nerve neurapraxia. Although most of them are transient, complication rates of up to 26% are reported. Recently, postless distraction technique has been described for elective hip arthroscopy. In this study we compared post and postless distraction technique in femoral shaft fracture nailing in terms of (1) quality of reduction, (2) outcome, and (3) complications. We reviewed 50 patients treated with postless distraction nailing technique for femoral shaft fractures and compared them with our historical case series (95 patients). The following data were collected for all patients: age, gender, weight, height, diagnoses (fractures were classified according to the 2018 revision of AO classification), type and size of nail surgical timing, Trendelenburg angles during surgery, quality of reduction according to Baumgaertner and Thoresen classifications, Modified Harris Hip Scores at 6 months, and perineal complications. Median age was 53 years, and median weight was 70 kg (range 50-103 kg). We found no significant difference in terms of quality of reduction (72 versus 74% "excellent" reduction for subtrochanteric fractures, while 81 versus 79% "excellent" reduction for femoral shaft fractures) and functional outcomes (Modified Harris Hip Score 74 versus 79). One patient in the control group had a failure of the fixation, and one patient in the postless group had a deep infection. Two patients in the control group reported pudendal nerve neurapraxia for 4 months, while none reported complication linked to the postless technique. Our results using the postless distraction technique show a sufficient distraction to allow reduction and internal fixation of the femoral fracture with a standard femoral nail. IV.


Radioimmunoassay and the hormones of thyroid function.
Radioimmunoassay (RIA) has provided the tools for wide-reaching investigations that have changed and continue to change many important concepts of thyroid physiology and pathosphysiology. The RIA vor human thyrotropin (TSH) was developed in 1965; development of the RIA for triiodothyronine (T3), thyroxine (T4), thyroxine-binding globulin (TBG), and, recently, thyrothropin-releasing hormone (TRH) and thyroglobulin (Tg) followed. The capacity to measure nanogram and picogram concentrations with relative ease and speed has permitted the demonstration of dynamic relationships of the intrathyroidal and circulating thyroid hormones to each other and to the pituitary and hypothalamic regulating hormones. Evidence for the presence of cross-influences between TRH and other hypothalamic regulating hormones on the secretion of pituitary hormones has accumulated. The impact of the new information on clinical practice is now becoming evident. There is new appreciation of the value of assaying serum T3 and TSH concentrations in the clinical management of patients with disturbed function of the thyroid, pituitary, or hypothalamus. The necessary components for RIA performance can be purchansed separately or in kit form from commercial sources. With appropriate quality-control procedures, precise, sensitive, and reliable data can be generated. Awareness of the specific technical problems relating to the RIA of these hormones is absolutely necessary to assure reliable results. The availability of kits or their components permits the performance of these studies in the community hospital and in reliable commercial-service laboratories.


Dynamic tracking of variant frequencies depicts the evolution of mutation sites amongst SARS-CoV-2 genomes from India.
With the exponential spread of the coronavirus disease 2019 (COVID-19) pandemic across the world within the 12 months, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains are continuously trying to adapt themselves to the host environment by random mutations. When doing so, some variants with evolutionary advantages such as better human to human transmissibility potential might get naturally selected. This short communication demonstrates how the mutation frequency patterns are evolving in 2457 SAR-CoV-2 strains isolated from COVID-19 patients across diverse Indian states. We have identified 19 such variants showing contrasting mutational probabilities in the span of 7 months. Out of these, 14 variants are showing increasing mutational probabilities suggesting their propagation with time due to their unexplored evolutionary advantages. However, the mutational probabilities of five variants have significantly decreased in June onward as compared to March/April, suggesting their termination with time. Further in-depth investigation of these identified variants will provide valuable knowledge about the evolution, infection strategies, transmission rates, and epidemiology of SARS-CoV-2.


Development of sensitive magnetic nanoparticle assisted rapid sandwich assay(s-MARSA) to monitor Parkinson's disease and Schizophrenia pharmacotherapy.
Parkinson's disease and Schizophrenia fall under low dopamine neurodegenerative and high dopamine psychiatric disorders respectively. Pharmacological interventions to correct mid-brain dopamine concentrations sometimes overshoots the physiological dopamine levels leading to psychosis in Parkinson's disease patients and, extra-pyramidal symptoms in schizophrenia patients. Currently no validated method is available to monitor side effects in such patients, Apolipoprotein E is one of the CSF biomarkers identified in the recent past that shows an inverse relation to mid-brain dopamine concentration. In this study, we have developed s-MARSA for the detection of Apolipoprotein E from ultra-small volume (2 μL) of CSF. s-MARSA exhibits a broad detection range (5 fg mL-1 to 4 μg mL-1) with a better detection limit and could be performed within an hour utilizing only a small volume of CSF sample. The values measured by s-MARSA strongly correlates with the values measured by ELISA. Our method has advantages over ELISA in having a lower detection limit, a broader linear detection range, shorter analysis time, and requiring a low volume of CSF samples. The developed s-MARSA method holds promise for the detection of Apolipoprotein E with clinical utility for monitoring pharmacotherapy of Parkinson's and Schizophrenia patients.


A GALNT14 rs9679162 genotype-guided therapeutic strategy for advanced hepatocellular carcinoma: systemic or hepatic arterial infusion chemotherapy.
Although targeted agents are recommended as the first-line treatments for advanced hepatocellular carcinoma (aHCC), systemic chemotherapy or hepatic arterial infusion chemotherapy (HAIC) are still being used in Asian countries. Beside economic considerations, it was found that targeted drugs could not significantly prolong overall survival in aHCC patients with distant metastasis. In addition, chemotherapy could achieve complete response in a small proportion of patients. Here, we aimed to investigate whether combination of three previously identified single nucleotide polymorphism (SNP) predictors (GALNT14-rs9679162, WWOX-rs13338697, and rs6025211) could guide our choice between systemic chemotherapy, HAIC, and targeted agents in aHCC patients. A cohort of 237 real-world aHCC patients (171 receiving systemic chemotherapy followed by various anticancer treatments including sorafenib; 66 receiving HAIC) were included for outcome analysis. By combining the three SNP markers with or without addition of two clinical criteria (tumor diameter <8 cm, neutrophils <80%), small groups of patients were found to harbor high complete response rates to systemic chemotherapy (35.3% if the 3-SNP signature alone matched; 60.0% if clinical criteria also matched). Subsequent sorafenib treatment for chemotherapy non-responders was associated with longer overall survival (P < 0.001). In HAIC-treated patients, GALNT14-rs9679162 genotype "GG" was associated with longer overall survival (P = 0.019, median survival > 10.5 months). In conclusion, pre-test for the 3-SNP signature in aHCC patients could identify potential systemic chemotherapy or HAIC responders. Chemotherapy non-responders still benefited from subsequent sorafenib treatment. Accordingly, we propose a roadmap for aHCC patients when chemotherapy or HAIC is to be used.


Metabolism of exogenous fatty acids, fatty acid-mediated cholesterol efflux, PKA and PKC pathways in boar sperm acrosome reaction.
For understanding the roles of fatty acids on the induction of acrosome reaction which occurs under association of cholesterol efflux and PKA or PKC pathways in boar spermatozoa, metabolic fate of alone and combined radiolabeled 14C-oleic acid and 3H-linoleic acid incorporated in the sperm was compared, and behavior of cholesterol and effects of PKA and PKC inhibitors upon fatty acid-induced acrosome reaction were examined. Semen was collected from a Duroc boar, and the metabolic activities of fatty acids in the spermatozoa were measured using radioactive compounds and thin layer chromatography. Cholesterol efflux was measured with a cholesterol determination assay kit. Participation of fatty acids on the AR through PKA and PKC pathways was evaluated using a specific inhibitor of these enzymes. Incorporation rate of 14C-oleic acid into the sperm lipids was significantly higher than that of 3H-linoleic acid (P < 0.05). The oxidation of 14C-oleic acid was higher in combined radiolabeling rather than in one. The highest amounts of 3H-linoleic acid and 14C-oleic acid were recovered mainly in the triglycerides and phospholipids fraction, and 14C-oleic acid distribution was higher than the 3H-linoleic acid in both labeled (P < 0.05) sperm lipids. In the 3H-linoleic and 14C-oleic acid combined radiolabeling, the incorporation rate of the radioactive fatty acids in all the lipid fractions increased 15 times more than the alone radiolabeling. Boar sperm utilize oleic acid to generate energy for hyperactivation (P < 0.05). Supplementation of arachidonic acid significantly increased (P < 0.05) cholesterol efflux in sperm. When spermatozoa were incubated with PKA or PKC inhibitors, there was a significant reduction of arachidonic acid-induced acrosome reaction (AR) (P < 0.05), and inhibition by PKA inhibitor is stronger than that by PKC inhibitor. Incorporation of unsaturated fatty acids, especially oleic acid, into triglycerides and phospholipids provides prerequisite energy for AR. Cholesterol efflux by arachidonic acid triggers AR. Arachidonic acid activated PKA and PKC pathway participate in induction of the AR.


Vitamin D deficiency enhances expression of autophagy-regulating miR-142-3p in mouse and "involved" IBD patient intestinal tissues.
Vitamin D deficiency is an environmental factor involved in the pathogenesis of inflammatory bowel disease (IBD); however, the mechanisms surrounding its role remain unclear. Previous studies conducted in an intestinal epithelial-specific vitamin D receptor (VDR) knockout model suggest that a lack of vitamin D signaling causes a reduction in intestinal autophagy. A potential link between vitamin D deficiency and dysregulated autophagy is microRNA (miR)-142-3p, which suppresses autophagy. In this study, we found that wild-type C57BL/6 mice fed a vitamin D-deficient diet for 5 wk had increased miR-142-3p expression in ileal tissues compared with mice that were fed a matched control diet. Interestingly, there was no difference in expression of key autophagy markers ATG16L1 and LC3II in the ileum whole tissue. However, Paneth cells of vitamin D-deficient mice were morphologically abnormal and had an accumulation of the autophagy adaptor protein p62, which was not present in the total crypt epithelium. These findings suggest that Paneth cells exhibit early markers of autophagy dysregulation within the intestinal epithelium in response to vitamin D deficiency and enhanced miR-142-3p expression. Finally, we demonstrated that treatment-naïve IBD patients with low levels of vitamin D have an increase in miR-142-3p expression in colonic tissues procured from "involved" areas of the disease. Taken together, our findings demonstrate that insufficient vitamin D levels alter expression of autophagy-regulating miR-142-3p in intestinal tissues of mice and patients with IBD, providing insight into the mechanisms by which vitamin D deficiency modulates IBD pathogenesis.NEW & NOTEWORTHY Vitamin D deficiency has a role in IBD pathogenesis, and although the mechanisms surrounding its role remain unclear, it has been suggested that autophagy dysregulation is involved. Here, we show increased ileal expression of autophagy-suppressing miR-142-3p in mice that were fed a vitamin D-deficient diet and in "involved" colonic biopsies from pediatric IBD patients with low vitamin D. miR-142-3p serves as a potential mechanism mediating vitamin D deficiency and reduced autophagy.


High- and low-density lipoprotein cholesterol: correlates in an older population.
A predominantly upper-middle-class white population of 279 men and 345 women with an average age of 66 years was evaluated to determine correlates of high- and low-density lipoprotein cholesterol (HDL and LDL), since these may be the only cholesterol measurements predictive of cardiovascular disease after age 50. Associations of HDL and LDL with age, alcohol use, obesity, smoking, diet, exercise, and medication use were examined using multiple linear regression. Alcohol, average and above-average exercise, postmenopausal estrogen use (women), B vitamin intake, and corticosteroid use were all positively related to HDL levels, and obesity (men only), cigarette smoking, and thyroid hormone use were inversely related. The only statistically significant correlates of LDL were an increase with age (women only) and lower levels in women using thyroid hormone. However, the associations of several behavioral variables with LDL, although not statistically significant, were in the expected direction. These results may suggest the continuing potential for preventive intervention even in an elderly population.


Hepatocytes prepare the soil for liver metastasis.
The liver is the most common organ site of cancer metastasis. Molecular determinants of this organotropism, though, are poorly understood. We recently showed that hepatocytes regulate the formation of a "pro-metastatic" niche in the liver via their release of serum amyloid proteins.


Securing continuation of treatment for children with cancer in times of social unrest and pandemic.
Childhood cancer in Chile reports 500 new cases each year of which 85% are treated in the public health system. Governmental programs ensure access to diagnosis, treatment, follow up and palliative care, whereas Fundación Nuestros Hijos (FNH) provides supportive care for non-covered medical and psychosocial needs. Common financial difficulties in families of children and adolescents with cancer increased considerably when a wave of social unrest arose in October 2019 and the Covid-19 pandemic in March 2020 hit the country, leaving families of children with cancer facing greater challenges. We report here the support activities and interventions carried out by FNH to help the families during the crisis of these months. A socioeconomic survey was conducted among FNH's beneficiary families to know their needs. During these months of acute crisis for many families, support activities and interventions were developed and varied types of aid were allocated to help the families. The main results of the survey in which 525 (70%) of FNH's beneficiary families participated showed that 75% of them had only one breadwinner, and 52% had one unemployed family member. Almost 90% of job loss happened during the months of social unrest and pandemic. Four main interventions: (a) safe transportation, (b) food, (c) heating, (d) internet connectivity, were organized to support important needs of the families and prevent children to miss treatment appointments. Additionally, some families who did not access governmental emergency aid were guided in the process. The aid provided helped the families to relieve some of their needs, facilitated the continuation of treatment during the pandemic, and made the caregivers feel supported and listened.


Calsequestrin distribution, structure and function, its role in normal and pathological situations and the effect of thyroid hormones.
Calsequestrin is the main calcium binding protein of the sarcoplasmic reticulum, serving as an important regulator of Ca(2+). In mammalian muscles, it exists as a skeletal isoform found in fast- and slow-twitch skeletal muscles and a cardiac isoform expressed in the heart and slow-twitch muscles. Recently, many excellent reviews that summarised in great detail various aspects of the calsequestrin structure, localisation or function both in skeletal and cardiac muscle have appeared. The present review focuses on skeletal muscle: information on cardiac tissue is given, where differences between both tissues are functionally important. The article reviews the known multiple roles of calsequestrin including pathology in order to introduce this topic to the broader scientific community and to stimulate an interest in this protein. Newly we describe our results on the effect of thyroid hormones on skeletal and cardiac calsequestrin expression and discuss them in the context of available literary data on this topic.


The turn from just-in-time to just-in-case globalization in and after times of COVID-19: An essay on the risk re-appraisal of borders and buffers.
In this essay, we apply insights from International Economics and Economic Geography to examine how the current COVID-19 crisis may structurally change the international economy. Our key argument is that the current crisis will fundamentally change key economic actors' risk appetite, triggering a renewed risk assessment that will lead to the comeback of buffers and borders across industries. This partial return to regionalization will involve a form of de-globalization that transforms modern just-in-time management into its just-in-case counterpart, because resilience will be priced and discounted for by enterprises and governments alike. We discuss what such a structural change will imply for the International Business of international value chains.


Marketing strategy: a key leverage point for dietitians.
The time is right for planned change in the profession of dietetics. The concept of the nutrition support team offers the clinical dietitian a means of increasing the visibility and effectiveness of dietetic services. Dietitians must take advantage of the renewed interest in nutrition and intervene assertively to improve nutritional care through role expansion. By focusing on a systematic marketing strategy to expand their current role, dietitians can become proactive participants in role change, rather than reactive victims to changes dictated by others. When goals, objectives, and plans are made explicit, definite progress toward desired role changes can be recognized. This case study demonstrates that this change in role is possible within the present state of health case systems and that physicians are willing and even eager to allow dietitians to assume a high degree of responsibility for nutritional care.


Comparative toxicity of aminoglycoside antibiotics towards the lysosomes in a cell culture model.
Cultured rat fibroblasts were used to compare the cellular toxicity of 4 aminoglycosides: accumulation of the antibiotic in the lysosomes, its adverse effect on phospholipases and consequently the intralysosomal accumulation of phospholipids. Gentamicin and Tobramycin are more noxious than Netilmicin and Amikacin. These data, when corrected for the lesser capture of Tobramycin by the kidney proximal tubules, closely match the toxicological evaluation of these aminoglycosides on the basis of animal and clinical studies.