Infection rate and spontaneous seroreversion of anti-hepatitis C virus during the natural course of hepatitis C virus infection in the general population.
Hepatitis C virus (HCV) infection is common worldwide but there are different prevalence rates in different countries. Data on the incidence of HCV in the general population are scarce. Spontaneous viral clearance occurs in 10-25% of infected individuals after acute infection yet controversy exists regarding the frequency of spontaneous clearance during the natural course of HCV infection in the general population. Anti-HCV prevalence, HCV infection rate, and the kinetics of anti-HCV were studied in a cross section of the general population of central Italy. Anti-HCV prevalence (EIA-3 Ortho, RIBA-3 Ortho Chiron) was estimated in 3884 randomly selected individuals. Infection rate and antibody kinetics were estimated in 2032 participants for whom a second blood sample was taken after a median follow up of seven years. HCV-RNA determination by polymerase chain reaction was performed on follow up sera. The overall confirmed anti-HCV prevalence was 2.4%. Two participants seroconverted for anti-HCV, giving an overall infection rate of 1.4 cases per 10 000 person years (95% confidence interval 0.2-5.2 per 10 000 person years). Of the 36 individuals confirmed as anti-HCV positive at enrollment, seven (19.4%) showed complete seroreversion. Seven (87%) of the eight individuals with indeterminate results at enrollment were serologically non-reactive at the end of follow up. Of the 25 participants confirmed to be anti-HCV positive at both enrollment and follow up, 23 (92.0%) with stable serological profiles tested positive for HCV-RNA at the end of follow up. There is still a permanent risk, although low, of HCV spread in the general population in an area of low level endemicity. In this setting, a wide spectrum of modifications of viral and antibody patterns can be observed in HCV infected patients.


Improvement of metabolic syndrome markers through altitude specific hiking vacations.
To study the influence of a 3-week hiking vacation at moderate (1700 m) and low altitude (LA) (200 m) on key-markers of the metabolic syndrome, 71 male volunteers (age 36-66 yr old) with the metabolic syndrome [according to the National Cholesterol Education Program's Adult Treatment Panel III (NCEP-ATP III) - or World Health Organization (WHO) - definition] participated in the study and were randomly assigned into a moderate altitude (MA) group (1700 m, no. 36) and a low altitude (LA) group (200 m, no. 35). The 3-week vacation program included 12 moderate- intensity guided hiking tours [4 times/week, 55-65% heart rate maximum (HRmax)] with a total exercise time of 29 h plus moderate recreational activities. Both study groups had a comparable and balanced nutrition with no specific dietary restrictions. Anthropometric, metabolic and cardiovascular parameters were measured 10-14 days before vacation, several times during the 3-week vacation, 7-10 days and 6-8 weeks after return. All participants tolerated the vacation without any adverse effects. Body weight, body fat, waist-circumference, fasting glucose, total cholesterol, LDL-cholesterol (LDL-C), plasma fibrinogen, resting systolic and diastolic blood pressure were significantly decreased over time in both study groups. In the LA group, fasting insulin and homeostasis model assessment (HOMA)-index were significantly decreased one week after return. Relative cycle ergometry performance was significantly increased after return compared to baseline. In both study groups, waist-to-hip ratio (WHR), 2-h oral glucose tolerance test (OGTT), HDL-cholesterol (HDL-C), and triglycerides remained unchanged. The 3-week vacation intervention at moderate and LA had a positive influence on all key-markers of the metabolic syndrome. No clinically relevant differences could be detected between the study groups. A hiking vacation at moderate and LA can be recommended for people with stable, controlled metabolic and cardiovascular risk factors.


Hemostatic and Thrombotic Considerations in the Diagnosis and Management of Childhood Arterial Ischemic Stroke: A Narrative Review.
Although rare in children, arterial ischemic stroke (AIS) is associated with increased mortality and neurological morbidity. The incidence of AIS after the neonatal period is approximately 1-2/100,000/year, with an estimated mortality of 3-7%. A significant proportion of children surviving AIS experience life-long neurological deficits including hemiparesis, epilepsy, and cognitive delays. The low incidence of childhood AIS coupled with atypical clinical-presentation and lack of awareness contribute to delay in diagnosis and consequently, the early initiation of treatment. While randomized-clinical trials have demonstrated the efficacy and safety of reperfusion therapies including thrombolysis and endovascular thrombectomy in appropriately-selected adult patients, similar data for children are unavailable. Consequently, clinical decisions surrounding reperfusion therapy in childhood AIS are either extrapolated from adult data or based on local experience. The etiology of childhood AIS is multifactorial, often occurring in the setting of both acquired and congenital risk-factors including thrombophilia. While multiple studies have investigated the association of thrombophilia with incident childhood AIS, its impact on stroke recurrence and therefore duration and intensity of antithrombotic therapy is less clear. Despite these limitations, a significant progress has been made over the last decade in the management of childhood AIS. This progress can be attributed to international consortiums, and in selected cohorts to federally-funded clinical trials. In this narrative review, the authors have systematically appraised the literature and summarize the hemostatic and thrombotic considerations in the diagnosis and management of childhood AIS focusing on the evidence supporting reperfusion therapies, relevance of thrombophilia testing, and duration and drug choices for secondary-prophylaxis.


[Clinical characteristics of 11 patients with chronic lymphocytic leukemia with t (14;19) (q32;q13)].
Objective: To analyze the clinicopathological characteristics of 11 cases of chronic lymphocytic leukemia (CLL) with t (14;19) (q32;q13) . Methods: The case data of 11 patients with CLL with t (14;19) (q32;q13) in the chromosome karyotype analysis results of the Blood Diseases Hospital, Chinese Academy of Medical Sciences from January 1, 2018, to July 30, 2022, were retrospectively analyzed. Results: In all 11 patients, t (14;19) (q32;q13) involved IGH::BCL3 gene rearrangement, and most of them were accompanied by +12 or complex karyotype. An immunophenotypic score of 4-5 was found in 7 patients and 3 in 4 cases. We demonstrated that CLLs with t (14;19) (q32;q13) had a mutational pattern with recurrent mutations in NOTCH1 (3/7), FBXW7 (3/7), and KMT2D (2/7). The very-high-risk, high-risk, intermediate-risk, and low-risk groups consisted of 1, 1, 6, and 3 cases, respectively. Two patients died, 8 survived, and 2 were lost in follow-up. Four patients had disease progression or relapse during treatment. The median time to the first therapy was 1 month. Conclusion: t (14;19) (q32;q13), involving IGH::BCL3 gene rearrangement, is a rare recurrent cytogenetic abnormality in CLL, which is associated with a poor prognosis.


Incidence of suboptimal response to tumor necrosis factor antagonist therapy in inflammatory bowel disease in newly industrialised countries: The EXPLORE study.
Incidence of inflammatory bowel disease (IBD) is increasing in newly industrialised countries (NICs); however, data on suboptimal response to anti-tumor necrosis factor (anti-TNF) agents are limited. To assess incidence and indicators of suboptimal response to first anti-TNF therapy in IBD patients in NICs. A chart review was conducted in ten countries from Asia-Pacific (APAC), Latin America (LatAm), and Russia and the Middle East (RME) regions among patients diagnosed with ulcerative colitis (UC) or Crohn's disease (CD), initiating anti-TNF therapy in 2010-2015. The cumulative incidence of suboptimal response to anti-TNF therapy was assessed using the following indicators: dose escalation or discontinuation, augmentation with non-biologic therapy, IBD-related hospitalization, or surgery. The study included 1,674 patients (570 UC; 1,104 CD). At 24 months, 32.9% of UC (APAC: 45.1%; LatAm: 38.2%; RME: 23.8%) and 41.2% of CD patients (APAC: 54.1%; LatAm: 42.5%; RME: 29.5%) had experienced suboptimal response. The most frequent first indicator was non-biologic therapy augmentation in LatAm (41.7%), IBD-related hospitalization in RME (UC: 50.7%; CD:37.3%) and in APAC for CD (39.1%), and anti-TNF discontinuation in APAC for UC (38.3%). Suboptimal response to anti-TNF agents is common in IBD patients in NICs. Observed regional differences in the incidence and indicators may reflect local practice and anti-TNF restrictions in IBD management. NCT03090139.


Prenatal diagnosis of de novo mosaic deletion 13q associated with multiple abnormalities.
A case of prenatal diagnosis of de novo mosaic deletion of the long arm of chromosome 13 (del(13)(q13.3)) is presented. Routine scanning in a 27-year-old primigravida at 25 weeks' gestation showed fetal bilateral hydronephrosis. Detailed anomaly scanning in our tertiary referral center further demonstrated posterior meningoencephalocele, sloping forehead, microcephaly, syndactyly and hypoplastic thumbs. Both genetic amniocentesis and cordocentesis revealed a mosaic karyotype, 46,XY/46,XY,del(13)(q13.3). Sonographic findings were confirmed by postmortem autopsy and additional abnormalities such as agenesis of corpus callosum, hypoplastic cerebellum and macroglossia were diagnosed. Detailed sonography in this case thus revealed multiple malformations that prompted fetal karyotyping at 25 weeks' gestation.


Alu-LTR real-time nested PCR assay for quantifying integrated HIV-1 DNA.
An improved Alu-long terminal repeat (LTR) polymerase chain reaction (PCR) assay is described for the quantification of integrated HIV-1 DNA in infected cells. The method includes generation of an infected cell line containing numerous randomly distributed HIV-1 integrated DNA for the construction of the DNA standard and a two-step real-time PCR assay in which the first-round PCR amplifies the DNA sequence between the HIV-1 LTR and the nearest chromosomal Alu element, and the nested PCR specifically amplifies PCR products from the first-round PCR. This assay allows us to quantify proviral DNA with both accuracy and high sensitivity (six proviruses within 50,000 cell equivalents) and exhibits a broad range of quantification spanning 5 log10 provirus copies. This Alu-LTR-based real-time nested PCR assay may be particularly useful to quantify integrated HIV-1 DNA in patients. It may also allow for the precise study of integration of HIV-1 DNA or HIV-1 based lentiviral vectors and may be a valuable tool to test future inhibitors of integration.


Characterization and molecular cloning of a bovine lentivirus related to human immunodeficiency virus.
An infectious virus which causes persistent lymphocytosis, lymphadenopathy, lesions in the central nervous system (CNS), progressive weakness and emaciation was previously isolated from the leukocytes of cattle. Our present studies show that this virus encodes a reverse transcriptase (RT) with Mg2+ cation preference, replicates and induces syncytia in a variety of embryonic bovine tissues in vitro, and has a morphology most similar to the human immunodeficiency virus (HIV). Moreover, serologic analyses have demonstrated a conservation of epitopes between the major core protein of this bovine retrovirus and HIV. Shared antigenic determinants were also observed with other pathogenic retroviruses of the lentivirus subfamily. To resolve the phylogenetic relationship of this virus, proviral molecular clones were derived and used to determine the nucleotide sequence of the highly conserved RT domain. The sequence data and serologic analyses together show that this bovine retrovirus is a novel lentivirus related to HIV and other lentiviruses. We propose that this virus be tentatively named bovine immunodeficiency-like virus (BIV) to reflect its genetic relationship and biological similarity to HIV.


Ketotifen inhibits the cutaneous but not the airway responses to platelet-activating factor in man.
We studied the effect of ketotifen, an oral antiallergic and antihistaminic drug, on the airway and cutaneous responses to platelet-activating factor (PAF) in a double-blind, randomized, and crossover study in six normal subjects. Ketotifen (three doses of 2 mg taken during a 14-hour period before PAF) did not alter PAF-induced bronchoconstriction and did not prevent the accompanying flushing and coughing. The transient neutropenia (74.5 +/- 4.8% fall; p less than 0.001) and rebound neutrophilia (104 +/- 55% rise) induced by PAF were not affected by ketotifen. On the day placebo was received, airway responsiveness to methacholine increased after PAF exposure with the concentration needed to cause a 40% fall in baseline partial expiratory flow rate (PC40), decreasing from 69.2 mg/ml (geometric standard error of the mean, 2.69) to 23.3 mg/ml (2.34) on day 3 (p less than 0.001). Ketotifen had no effect, because on the day ketotifen was administered, mean PC40 also decreased from 52.7 mg/ml (2.5) to 21.5 mg/ml (2.14) (p less than 0.01). In the skin, ketotifen reduced the flare area (from 8.05 +/- 3.60 to 1.14 +/- 0.29 cm2; p less than 0.05) and the wheal volume (from 0.068 +/- 0.010 to 0.045 +/- 0.008 cc; p = 0.02) induced by intradermal PAF (200 ng). Cutaneous responses to histamine (1 microgram) were significantly inhibited. Thus, the bronchoconstriction and bronchial hyperresponsiveness induced by PAF are not inhibited by ketotifen. Ketotifen inhibits PAF-induced wheal and flare in the skin, which is probably histamine dependent. The airway effects of PAF are unlikely to be mediated by histamine release.


Knowledge-based potentials in protein design.
Knowledge-based potentials are statistical parameters derived from databases of known protein properties that empirically capture aspects of the physical chemistry of protein structure and function. These potentials play a key role in protein design by improving the accuracy of physics-based models of interatomic interactions and enhancing the computational efficiency of the design process by limiting the complexity of searching sequence space. Recently, knowledge-based potentials (in isolation or in combination with physics-based potentials) have been applied to the modification of existing protein function, the redesign of natural protein folds and the complete design of a non-natural protein fold. In addition, knowledge-based potentials appear to be providing important information about the global topology of amino acid interactions in natural proteins. A detailed study of the methods and products of these protein design efforts promises to greatly expand our understanding of proteins and the evolutionary process that created them.


Potential impact of (18)FDG-PET/CT on surgical approach for operable squamous cell cancer of middle-to-lower esophagus.
Fluorodeoxyglucose-positron emission tomography (PET)/computed tomography (CT) is reported to have a significant advantage over CT for staging esophageal cancer (EC). However, whether PET/CT may play a useful role in guiding surgical approach remains undetermined. Patients with potentially resectable squamous cell EC were randomized into either PET/CT group or CT group. The surgical data and survival outcomes were compared. Compared to the CT group, the right-sided approach was more frequently used (42.6% versus 25.5%, P=0.065) in the PET/CT group in order to allow surgical access to radiographically suspicious lymph nodes inaccessible from the left, thus enabling the removal of more involved lymph nodes (2.83 versus 1.76; P=0.039) as well as their stations (1.65 versus 1.08; P=0.042). Although the overall survival between the two groups was similar, the PET/CT group had a longer disease-free survival (DFS) than the CT group (27.1 months versus 18.9 months; P=0.019), especially in the subgroup of node-positive patients (22.5 months versus 13.5 months; P=0.02). Preoperative imaging arm was the only prognostic factor found to independently influence DFS. For patients with middle-to-lower EC, surgical approaches directed by PET/CT may increase the likelihood of complete resection and affect DFS.


Loading Behaviors Do Not Match Loading Abilities Postanterior Cruciate Ligament Reconstruction.
Strategies that underload the surgical limb after anterior cruciate ligament reconstruction (ACLr) are observed in submaximal tasks. It is not known what underlies these strategies in early rehabilitation. The purpose of this study was to determine if underloading can be attributed to the inability to meet task demands with and without attention to limb loading or learned behavior. Twenty individuals (110.6 [18.1] days) post-ACLr and 20 healthy individuals (CTRL) participated in this study. Participants performed standing, sit-to-stand, and squat tasks under natural, instructed, and feedback conditions. Limb-loading symmetry was calculated as the between-limb ratio of vertical ground reaction force impulse during each task. General Linear Model repeated-measures analysis, 2 (group) × 3 (condition), determined the effects of group and condition on limb-loading symmetry for each task. Significant interactions were observed for each task (all P < 0.001). Compared with CTRL, ACLr exhibited greater asymmetry during natural (deficits: standing, 10%, P = 0.001; sit-to-stand, 25%, P < 0.001; squat, 15%, P < 0.001) and instructed (deficits: sit-to-stand, 13%, P = 0.001; squat, 8%, P = 0.04), but not feedback conditions. The CTRL maintained symmetry across conditions and tasks. Anterior cruciate ligament reconstruction exhibited greater asymmetry in natural compared with instructed (deficits: standing, 11%, P < 0.001; sit-to-stand, 14%, P < 0.001; squat, 8%, P = 0.001) and feedback (deficits: standing, 10%, P = 0.001; sit-to-stand, 21%, P < 0.001; squat, 15%, P < 0.001) conditions. The presence of loading asymmetries in natural but not feedback conditions indicates that individuals 3 months post-ACLr shift loading away from surgical limb despite the ability to meet task demands which may be suggestive of nonuse behavior. Even when instructed to load symmetrically, individuals continued to exhibit some degree of asymmetry.


Ubiquitin-mediated mechanisms of translational control.
mRNAs translation to proteins constitutes an important step of cellular gene expression that is highly regulated in response to different extracellular stimuli and stress situations. The fine control of protein synthesis is carried out both qualitatively and quantitatively, depending on the cellular demand at each moment. Post-translational modifications, in turn regulated by intracellular signaling pathways, play a key role in translation regulation. Among them, ubiquitination, whose role is becoming increasingly important in the control of translation, determines a correct balance between protein synthesis and degradation. In this review we focus on the role of ubiquitination (both degradative K48-linkage type and non-degradative K63-linkage type and monoubiquitination) in eukaryotic translation, both at the pre-translational level during the biogenesis/degradation of the components of translational machinery as well as at the co-translational level under stressful conditions. We also discuss other ubiquitin-dependent regulatory mechanisms of mRNA protection and resumption of translation after stress removal, where the ubiquitination of ribosomal proteins and associated regulatory proteins play an important role in the global rhythm of translation.


Evaluation of the clinical educational environment based on the DREEM model from the viewpoint of the OR students.
The most important part of learning happens in the clinic. To determine the differences between the real educational environment and the desirable environment, the DREEM1 model is used. The present study was conducted to evaluate the clinical educational environment based on the DREEM model from the viewpoint of the OR2 students of the Lorestan University of Medical Sciences. The present descriptive-analytical cross-sectional study evaluated the viewpoint of 118 students of operation room (OR) technology using the DREEM (Dundee Ready Educational Environment Measure) questionnaire in the hospitals affiliated with the Lorestan University of Medical Sciences during the second semester of 2020. The DREEM questionnaire has 50 statements and is divided into five sections, which are rated on a five-point Likert scale (0-4). The data were analyzed using frequency distribution tables, mean and standard deviation indices, Mann-Whitney and Kruskal-Wallis tests. Data analysis was conducted using IBM SPSS Statistics 22.0. IBM Corp.; 2013. and the level of significance was set at 0.05. The total perception of the students of the clinical educational environment was good and excellent in 73.8% of the cases and of the subscales was also good (50.8%-63.2%). There was a significant relation between the total score of students' perception of the clinical educational environment and age (Z = 5.618, P < 0.001), semester (χ2= 43.929, df = 3, P < 0.001), internship hospital (χ2= 12.948, df = 6, P = 0.044) and also the mean score of the subscales (P < 0.001). As the GPA3 and interest in the major increased, the mean score of total perception and its subscales also increased except for perception from the educators (P < 0.05). According to the results, the studied students had a positive perception of the clinical educational environment. It is recommended that the scientific foundation of the educators, the physical facilities of the operating rooms, and social communication should be strengthened to improve the care, treatment, and educational services. It will be useful to use the results to improve the accreditation level of medical centers.


Prenatal exposure to common plasticizers: a longitudinal study on phthalates, brain volumetric measures, and IQ in youth.
Exposure to phthalates, used as plasticizers and solvents in consumer products, is ubiquitous. Despite growing concerns regarding their neurotoxicity, brain differences associated with gestational exposure to phthalates are understudied. We included 775 mother-child pairs from Generation R, a population-based pediatric neuroimaging study with prenatal recruitment, who had data on maternal gestational phthalate levels and T1-weighted magnetic resonance imaging in children at age 10 years. Maternal urinary concentrations of phthalate metabolites were measured at early, mid-, and late pregnancy. Child IQ was assessed at age 14 years. We investigated the extent to which prenatal exposure to phthalates is associated with brain volumetric measures and whether brain structural measures mediate the association of prenatal phthalate exposure with IQ. We found that higher maternal concentrations of monoethyl phthalate (mEP, averaged across pregnancy) were associated with smaller total gray matter volumes in offspring at age 10 years (β per log10 increase in creatinine adjusted mEP = -10.7, 95%CI: -18.12, -3.28). Total gray matter volumes partially mediated the association between higher maternal mEP and lower child IQ (β for mediated path =-0.31, 95%CI: -0.62, 0.01, p = 0.05, proportion mediated = 18%). An association of higher monoisobutyl phthalate (mIBP) and smaller cerebral white matter volumes was present only in girls, with cerebral white matter volumes mediating the association between higher maternal mIBP and lower IQ in girls. Our findings suggest the global impact of prenatal phthalate exposure on brain volumetric measures that extends into adolescence and underlies less optimal cognitive development.


Evidence for a role of phospholipase C-gamma1 in the pathogenesis of bipolar disorder.
Several studies have indicated that patients with bipolar disorder (BD) who respond well to lithium prophylaxis constitute a biologically distinct subgroup. Lithium is thought to stabilize mood by acting at the phosphoinositide cycle. We have investigated a polymorphism located in the gene (PLCG1) that codes for a gamma-1 isozyme of phospholipase (PLC), an enzyme that plays an important role in the phosphoinositide second messenger system. A population-based association study and a family-based linkage study were carried out on patients who were considered excellent responders to lithium prophylaxis. Response to lithium was evaluated prospectively with an average follow-up of 14.4 +/- 6.8 years. The PLCG1 polymorphism was investigated in 136 excellent lithium responders and 163 controls. In addition, the segregation of this marker was studied in 32 families ascertained through lithium-responsive bipolar probands. The allele distributions between lithium-responsive bipolar patients and controls were different, with a higher frequency of one of the PLCG1 polymorphisms in patients (chi2 = 8.09; empirical P = 0.033). This polymorphism, however, confers only a small risk (OR = 1.88, CI 1.19-3.00). Linkage studies with the same marker yielded modest support for the involvement of this gene in the pathogenesis of BD when unilineal families were considered (Max LOD = 1.45; empirical P = 0.004), but not in the whole sample. Our results provide preliminary evidence that a PLC isozyme may confer susceptibility to bipolar disorder, probably accounting for a fraction of the total genetic variance. Whether this polymorphism is implicated in the pathogenesis of BD or in the mechanism of lithium response remains to be determined.


The araBAD operon of Salmonella typhimurium LT2. II. Nucleotide sequence of araA and primary structure of its product, L-arabinose isomerase.
The nucleotide sequence of gene araA of Salmonella typhimurium LT2 has been determined. The gene encodes an L-arabinose isomerase (EC 126.96.36.199) of 500 amino acid residues with a calculated Mr of 55814. The ATG start codon of araA is 10 bp distal to the TAA termination codon of araB. A presumed ribosome-binding site (RBS) "TAAGGA" 7 bp from the ATG codon overlaps the stop codon of araB. L-Arabinose isomerase was purified and the amino acid composition is in agreement with that predicted from the DNA sequence. The NH2-terminus of the protein is modified as the sequence cannot be analyzed by the automated Edman degradation. Amino acid composition analyses of both NH2-terminal and C-terminal cyanogen bromide (CNBr) cleaved peptides and partial amino acid sequence of the C-terminal peptide are consistent with the deduced amino acid sequence.


Synthesis and Characterization of Silica, Silver-Silica, and Zinc Oxide-Silica Nanoparticles for Evaluation of Blood Biochemistry, Oxidative Stress, and Hepatotoxicity in Albino Rats.
Evaluation of nanoparticles (NPs) for biomedical applications has received a lot of attention for detailed study on pharmacokinetics prior to clinical application. In this study, pure C-SiO2 (crystalline silica) NPs and SiO2 nanocomposites with silver (Ag) and zinc oxide (ZnO) were prepared by utilizing different synthesis routes such as sol-gel and co-precipitation techniques. The prepared NPs showed highly crystalline nature as confirmed by X-ray diffraction analysis where average crystallite sizes of 35, 16, and 57 nm for C-SiO2, Ag-SiO2, and ZnO-SiO2 NPs, respectively, were calculated. Fourier transform infrared analysis confirmed the presence of functional groups related to the chemicals and procedures used for sample preparation. Due to agglomeration of the prepared NPs, the scanning electron microscope images showed large particle sizes when compared to their crystalline sizes. The optical properties of the prepared NPs such as absorption were obtained with UV-Vis spectroscopy. For in vivo biological evaluation, albino rats, both male and female, kept in different groups were exposed to NPs with 500 μg/kg dose. Hematological, serum biochemistry, histo-architecture, oxidative stress biomarkers, and antioxidant parameters in liver tissues along with various biomarkers for the evaluation of erythrocytes were estimated. The results on hemato-biochemistry, histopathological ailments, and oxidative stress parameters exhibited 95% alteration in the liver and erythrocytes of C-SiO2 NPs-treated rats while 75 and 60% alteration in the liver tissues of rats due to exposure to Ag-SiO2 and ZnO-SiO2 NPs, respectively, when compared with the albino rats of the control (untreated) group. Therefore, the current study showed that the prepared NPs had adverse effects on the liver and erythrocytes causing hepatotoxicity in the albino rats in respective order C-SiO2 > Ag SiO2 > ZnO-SiO2. As the C-SiO2 NPs appeared to be the most toxic, it has been concluded that coating SiO2 on Ag and ZnO reduced their toxicological impact on albino rats. Consequently, it is suggested that Ag-SiO2 and ZnO-SiO2 NPs are more biocompatible than C-SiO2 NPs.


Potassium promoted macro-mesoporous Co3O4-La0.88Sr0.12CoO3-δ nanotubes with large surface area: A high-performance catalyst for soot removal.
The construction of porous perovskite nanotubular materials with a good intrinsic activity, as well as a greater dispersion of the active sites is an effective strategy to obtain a high-performance catalyst used in soot removal. Thence, macro-mesoporous Co3O4-La0.88Sr0.12CoO3-δ nanotubes with large specific surface area (154.4 m2·g-1) from the acid etching of the porous La0.6Sr0.4CoO3-δ nanotubes, are supported by 5% K through bubbling method following calcination for soot combustion. The relationship between the specific surface area and K dispersion and their effect on the activity are studied by a series of isothermal kinetic measurements combined with the characterizations and activity evaluation results. It can be found that the greater the amount is of K+ incorporated into perovskite lattice, the better the dispersion of K, as well as the La2O2CO3 formed on the catalyst surface, thus leading to the enhanced performance in the soot catalytic combustion. As a result, the 5% K supported macro-mesoporous Co3O4-La0.88Sr0.12CoO3-δ nanotubes after acid etching show good activity and stability, where the T50 is 338 °C (5% O2 + 500 ppm NO + 6% H2O) with a good CO2 selectivity (above 99%), the activate energy is 78.1 kJ·mol-1, and the turnover frequency is 5.14 × 10-4 s-1.


Comparing Acute Kidney Injury Reports Among Antibiotics: A Pharmacovigilance Study of the FDA Adverse Event Reporting System (FAERS).
A study using the US FDA Adverse Event Reporting System (FAERS) found significant acute kidney injury (AKI) reporting associations with vancomycin, fluoroquinolones, penicillin combinations, and trimethoprim-sulfamethoxazole. Other antibiotics may also lead to AKI, but no study has systemically compared AKI reporting associations for many available antibiotics. The objective of this study was to evaluate the reporting associations between AKI and many available antibiotics using FAERS. FAERS reports from 1 January 2015 to 31 December 2017 were included in the study. The Medical Dictionary for Regulatory Activities (MedDRA) was used to identify AKI cases. Reporting odds ratios (RORs) and corresponding 95% confidence intervals (CIs) for the reporting associations between antibiotics and AKI were calculated. A reporting association was considered statistically significant when the lower limit of the 95% CI was > 1.0. A total of 2,042,801 reports (including 20,138 AKI reports) were considered. Colistin had the greatest proportion of AKI reports, representing 25% of all colistin reports. AKI RORs (95% CI) for antibiotics were, in descending order: colistin 33.10 (21.24-51.56), aminoglycosides 17.41 (14.49-20.90), vancomycin 15.28 (13.82-16.90), trimethoprim-sulfamethoxazole 13.72 (11.94-15.76), penicillin combinations 7.95 (7.09-8.91), clindamycin 6.46 (5.18-8.04), cephalosporins 6.07 (5.23-7.05), daptomycin 6.07 (4.61-7.99), macrolides 3.60 (3.04-4.26), linezolid 3.48 (2.54-4.77), carbapenems 3.31 (2.58-4.25), metronidazole 2.55 (1.94-3.36), tetracyclines 1.73 (1.26-2.36), and fluoroquinolones 1.71 (1.49-1.97). This study found 14 classes of antibiotics having significant reporting associations with AKI. Among the antibiotics evaluated in this study, colistin had the highest AKI ROR and moxifloxacin had the lowest.


Formation and thermodynamic stability of intermolecular (R*R*Y) DNA triplex in GAA/TTC repeats associated with Freidreich's ataxia.
It is well established that GAA/TTC base triplet expansion is the cause of degenerative disorder in Freidreich's Ataxia. It is also known that these repeat sequences fold back to form the unusual intramolecular triple helix structures in DNA of the type Pyrimidine *Purine *Pyrimidine or Purine *Purine*Pyrimidine. In this paper we report on the stability of Purine *Purine*Pyrimidine intermolecular triple helix DNA containing GAA/TTC repeats under physiological conditions. Using the oligonucleotides 5' TCGC GAA GAA GAA GAA GAA CGCT 3', 5'-AGCG TTC TTC TTC TTC TTC GCGA-3' for duplex and 5'-AAG AAG AAG AAG AAG -3' as triplex forming strand (TFO), we have established the formation of triplex by UV-melting temperature (Tm), stoichiometry of mixing and circular dichroic spectra. This was further confirmed by gel-retardation assay. The thermodynamic parameters Delta G, Delta H and Delta S for both duplex and triplex formation were determined at different salt concentrations. The results suggest the formation of a stable intermolecular, anti - parallel triplex in GAA/TTC repeat sequences where each repeat unit contains two A*A*T and one G*G*C triplet. The therapeutic agents and TFOs, which competitively inhibit the in-vivo intra-molecular triplex by formation of a more stable inter-molecular triplex, could be used to reverse the transcription deficit in GAA/TTC expansions in Frataxin gene.


Natural killer cell reactivity: activation and cytolysis mechanism models, involving heat shock protein, haemopoietic histocompatibility, major histocompatibility complex and complement molecules.
The close association of heat shock protein (HSP), haemopoietic histocompatibility (Hh), major histocompatibility complex (MHC), and complement genes on the same chromosomal region, and the fact that all these genes are inherited on the whole in each haplotype of an individual, might indicate some evolutionary and functional correlations among them. Several data suggest for HSP70 molecules a possible role as a molecular target recognizable by natural killer (NK) cells. HSP70 sequences from both prokaryotic and eukaryotic organisms reveal that about half of the amino acid residues are identical and many of the remaining residues are similar. I here assume that NK reactivity might start, early in the immunogenesis process, as a effect of the interaction between HSP70 molecules and a hypothetical HSP receptor of yet immature non-cytolytic NK cells. To this receptor, an HSP molecule might act as an activator or an inhibitor depending on whether its amino acid residues are reactive or not with it, respectively. Later in the immunogenesis process, murine Hh or human equivalent molecules, dominantly expressed in bone marrow target cells, might select the non-reactive NK clones of an individual, inducing them to mature and express a lytic machinery. As a consequence of the NK maturation, proliferating hemopoietic target cells expressing only or mainly activator HSPs on their surface might undergo NK cytolysis. This might explain the NK lysis of apparently normal cells found in human foetal marrow; moreover, this might explain in some way the F1 hybrid resistance phenomenon. The NK reactivity of an individual would be further modulated by the expression on the NK surface of particular receptors (CD94, p58) specific for defined MHC molecules (Cw1, Cw3, Bw6, B7) on the target cells. Such a specific interaction would induce an 'NK effector inhibition'. The NK reactivity mechanism might have been further evolutionarily modified and adapted by the involvement of other NK receptors, such as CD11b (specific for the C3b factor of the complement) and CD16 (specific for the IgG Fc piece). Cooperation among HSP, MHC, CD11b, CD16, C3b and Fc allows us to propose original models of the activation and cytolysis mechanisms in the NK cytotoxicity and antibody-dependent cell cytotoxicity phenomena.


National Institutes of Health funding among vascular surgeons is rare.
The National Institutes of Health (NIH) is an essential source of funding for vascular surgeons conducting research. NIH funding is frequently used to benchmark institutional and individual research productivity, help determine eligibility for academic promotion, and as a measure of scientific quality. We sought to appraise the current scope of NIH funding to vascular surgeons by appraising the characteristics of NIH-funded investigators and projects. In addition, we also sought to determine whether funded grants addressed recent Society for Vascular Surgery (SVS) research priorities. In April 2022, we queried the NIH Research Portfolio Online Reporting Tools Expenditures and Results (RePORTER) database for active projects. We only included projects that had a vascular surgeon as a principal investigator. Grant characteristics were extracted from the NIH Research Portfolio Online Reporting Tools Expenditures and Results database. Principal investigator demographics and academic background information were identified by searching institution profiles. There were 55 active NIH awards given to 41 vascular surgeons. Only 1% (41/4037) of all vascular surgeons in the United States receive NIH funding. Funded vascular surgeons are an average of 16.3 years out of training; 37% (n = 15) are women. The majority of awards (58%; n = 32) were R01 grants. Among the active NIH-funded projects, 75% (n = 41) are basic or translational research projects, and 25% (n = 14) are clinical or health services research projects. Abdominal aortic aneurysm and peripheral arterial disease are the most commonly funded disease areas and together accounted for 54% (n = 30) of projects. Three SVS research priorities are not addressed by any of the current NIH-funded projects. NIH funding of vascular surgeons is rare and predominantly consists of basic or translational science projects focused on abdominal aortic aneurysm and peripheral arterial disease research. Women are well-represented among funded vascular surgeons. Although the majority of SVS research priorities receive NIH funding, three SVS research priorities are yet to be addressed by NIH-funded projects. Future efforts should focus on increasing the number of vascular surgeons receiving NIH grants and ensuring all SVS research priorities receive NIH funding.


Systemic anti-inflammatory therapy aided by double-headed nanoparticles in a canine model of acute intraocular inflammation.
Novel approaches circumventing blood-ocular barriers in systemic drug delivery are lacking. We hypothesize receptor-mediated delivery of curcumin (CUR) across intestinal and ocular barriers leads to decreased inflammation in a model of lens-induced uveitis. CUR was encapsulated in double-headed polyester nanoparticles using gambogic acid (GA)-coupled polylactide-co-glycolide (PLGA). Orally administered PLGA-GA2-CUR led to notable aqueous humor CUR levels and was dosed (10 mg/kg twice daily) to adult male beagles (n = 8 eyes) with induced ocular inflammation. Eyes were evaluated using a semiquantitative preclinical ocular toxicology scoring (SPOTS) and compared to commercial anti-inflammatory treatment (oral carprofen 2.2 mg/kg twice daily) (n = 8) and untreated controls (n = 8). PLGA-GA2-CUR offered improved protection compared with untreated controls and similar protection compared with carprofen, with reduced aqueous flare, miosis, and chemosis in the acute phase (<4 hours). This study highlights the potential of PLGA-GA2 nanoparticles for systemic drug delivery across ocular barriers.


Helicase structure and mechanism.
Structural information on helicase proteins has expanded recently beyond the DNA helicases Rep and PcrA, and the hepatitis C virus RNA helicase to include UvrB, members of the DEA(D/H)-box RNA helicase family, examples of DnaB-related helicases and RuvB. The expanding database of structures has clarified the structural 'theme and variations' that relate the different helicase families. Furthermore, information is emerging on the functions of the conserved helicase motifs and their participation in the mechanisms by which these proteins catalyze the remodeling of DNA and RNA in ATP-dependent activities.


Immigrants' experiences of maternity care in Japan.
Language and cultural differences can negatively impact immigrant women's birth experience. However, little is known about their experiences in Japan's highly homogenous culture. This cross-sectional study used survey data from a purposive sampling of immigrant women from 16 hospitals in several Japanese prefectures. Meeting the criteria and recruited to this study were 804 participants consisting of 236 immigrant women: Chinese (n = 83), Brazilian (n = 62), Filipino (n = 43), South Korean (n = 29) and from variety of English speaking nations (n = 19) and 568 Japanese women. The questionnaire was prepared in six languages: Japanese (kana syllables), Chinese, English, Korean, Portuguese, and Tagalog (Filipino). Associations among quality of maternity care, Japanese literacy level, loneliness and care satisfaction were explored using analysis of variance and multiple linear regression. The valid and reliable instruments used were Quality of Care for Pregnancy, Delivery and Postpartum Questionnaire, Rapid Estimate of Adult Literacy in Medicine Japanese version, the revised UCLA Loneliness Scale-Japanese version and Care satisfaction. Care was evaluated across prenatal, labor and delivery and post-partum periods. Immigrant women scored higher than Japanese women for both positive and negative aspects. When loneliness was strongly felt, care satisfaction was lower. Some competence of Japanese literacy was more likely to obstruct positive communication with healthcare providers, and was associated with loneliness. Immigrant women rated overall care as satisfactory. Japanese literacy decreased communication with healthcare providers, and was associated with loneliness presumably because some literacy unreasonably increased health care providers' expectations of a higher level of communication.


Intraobserver reliability of posturography in patients with vestibular neuritis.
The aim of the study was to establish the intraobserver reliability of a posturographic method in patients (n = 34) with vestibular neuritis. Intraclass correlation coefficients (relative reliability) for all parameters and test positions (ALL(mean)) ranged from 0.71 (95% CI: 0.41-0.85) to 0.92 (95% CI: 0.84-0.96). Absolute reliability (coefficient of variation) ranged between 3.1% (95% CI: 2.60-8.67) and 42.3% (95% CI: 40.7-74.5). Reliability of single test positions is much lower. The posturographic system showed good relative and satisfactory absolute intraobserver reliability for ALL(mean).


Transient increase in blood thrombogenicity may be a critical mechanism for the occurrence of acute myocardial infarction.
Although the mechanism for the occurrence of acute myocardial infarction (MI) has been investigated by many pathological and clinical studies, it has not been adequately clarified yet. Although the disruption of vulnerable plaque is a well-known cause of acute MI, there are many silent plaque disruptions detected in the coronary artery by intravascular imaging studies. Therefore, many vulnerable plaques may disrupt and heal without causing acute MI. Some additional mechanisms other than the disruption of vulnerable plaque would be essential for the onset of acute MI. On the other hand, blood thrombogenicity would change dynamically due to circadian rhythms and many other factors. The combination of plaque and blood thrombogenicity would play an important and determinant role for the onset of acute MI.


Studies of Metabolism Using (13)C MRS of Hyperpolarized Probes.
First described in 2003, the dissolution dynamic nuclear polarization (DNP) technique, combined with (13)C magnetic resonance spectroscopy (MRS), has since been used in numerous metabolic studies and has become a valuable metabolic imaging method. DNP dramatically increases the level of polarization of (13)C-labeled compounds resulting in an increase in the signal-to-noise ratio (SNR) of over 50,000 fold for the MRS spectrum of hyperpolarized compounds. The high SNR enables rapid real-time detection of metabolism in cells, tissues, and in vivo. This chapter will present a comprehensive review of the DNP approaches that have been used to monitor metabolism in living systems. First, the list of (13)C DNP probes developed to date will be presented, with a particular focus on the most commonly used probe, namely [1-(13)C] pyruvate. In the next four sections, we will then describe the different factors that need to be considered when designing (13)C DNP probes for metabolic studies, conducting in vitro or in vivo hyperpolarized experiments, as well as acquiring, analyzing, and modeling hyperpolarized (13)C data.


[Results of the DOSAK tumor register].
The German-Swiss-Austrian Group on Maxillofacial Tumors (DOSAK) has been performing observational studies in oral cancer. Since 1989, approximately 1600 cases of tumor of the head and neck per years has been collected in a central tumor registry. The database consists of more than 16,000 patients from 71 clinics; almost two-thirds are primary cases of squamous cell carcinoma. The data show great differences in patho-anatomical findings, therapy concepts, and five-year survival rates among the hospitals.


Late acquisition of the rTCA carbon fixation pathway by Chlorobi.
The reverse tricarboxylic acid (rTCA) cycle is touted as a primordial mode of carbon fixation due to its autocatalytic propensity and oxygen intolerance. Despite this inferred antiquity, however, the earliest rock record affords scant supporting evidence. In fact, based on the chimeric inheritance of rTCA cycle steps within the Chlorobiaceae, even the use of the chemical fossil record of this group is now subject to question. While the 1.64-billion-year-old Barney Creek Formation contains chemical fossils of the earliest known putative Chlorobiaceae-derived carotenoids, interferences from the accompanying hydrocarbon matrix have hitherto precluded the carbon isotope measurements necessary to establish the physiology of the organisms that produced them. Overcoming this obstacle, here we report a suite of compound-specific carbon isotope measurements identifying a cyanobacterially dominated ecosystem featuring heterotrophic bacteria. We demonstrate chlorobactane is 13C-depleted when compared to contemporary equivalents, showing only slight 13C-enrichment over co-existing cyanobacterial carotenoids. The absence of this diagnostic isotopic fingerprint, in turn, confirms phylogenomic hypotheses that call for the late assembly of the rTCA cycle and, thus, the delayed acquisition of autotrophy within the Chlorobiaceae. We suggest that progressive oxygenation of the Earth System caused an increase in the marine sulfate inventory thereby providing the selective pressure to fuel the Neoproterozoic shift towards energy-efficient photoautotrophy within the Chlorobiaceae.


Mammalian eIF4E2-GSK3β maintains basal phosphorylation of p53 to resist senescence under hypoxia.
Hypoxia modulates senescence, but their physiological link remains unclear. Here, we found that eIF4E2, a hypoxia-activated translation initiation factor, interacted with GSK3β to maintain phosphorylation of p53, thus resisting senescence under hypoxia. RNA-binding protein RBM38 interacted with eIF4E to inhibit the translation of p53, but GSK3β-mediated Ser195 phosphorylation disrupted the RBM38-eIF4E interaction. Through investigation of RBM38 phosphorylation, we found that the eIF4E2-GSK3β pathway specifically regulated proline-directed serine/threonine phosphorylation (S/T-P). Importantly, peptides e2-I or G3-I that blocking eIF4E2-GSK3β interaction can inhibit the basal S/T-P phosphorylation of p53 at multiple sites, therby inducing senescence through transcriptional inhibition. Additionally, a nanobody was screened via the domain where eIF4E2 bound to GSK3β, and this nanobody inhibited S/T-P phosphorylation to promote senescence. Furthermore, hypoxia inhibited eIF4E2-GSK3β pathway by mediating S-Nitrosylation of GSK3β. Blocking eIF4E2-GSK3β interaction promoted liver senescence under hypoxia, thus leading to liver fibrosis, eventually accelerating N, N-diethylnitrosamine (DEN)-induced tumorigenesis. Interestingly, eIF4E2 isoforms with GSK3β-binding motif exclusively exist in mammals, which protect zebrafish heart against hypoxia. Together, this study reveals a mammalian eIF4E2-GSK3β pathway that prevents senescence by maintaining basal S/T-P phosphorylation of p53, which underlies hypoxia adaptation of tissues.


Role of M2 and M3 muscarinic acetylcholine receptor subtypes in activation of bladder afferent pathways in spinal cord injured rats.
To evaluate the role of M2 and M3 muscarinic acetylcholine receptor (mAChR) subtypes in the activation of bladder afferent pathways in rats with chronic spinal cord injury (SCI). Adult female Sprague-Dawley rats were spinalized at the T9 level. Continuous cystometry was performed under awake conditions 2 or 4 weeks after SCI. The effects of intravesical administration of an mAChR agonist (oxotremorine-methiodide), a nonselective antagonist (atropine), an M2-selective antagonist (methoctramine), and an M3-selective antagonist (darifenacin) were examined. After cystometry, the bladder was removed and separated into the mucosa and detrusor, and the M2 and M3 mAChR mRNA expression in the mucosa was determined using real-time quantitative polymerase chain reaction. At 2 and 4 weeks after SCI, intravesical administration of a nonselective mAChR agonist (25 μM oxotremorine-methiodide) increased the area under the curve of nonvoiding contractions, although the intercontraction interval of voiding contractions and maximal voiding pressure did not change. This effect was blocked by atropine and methoctramine (10 μM) but not by darifenacin (50 μM). However, mAChR antagonists alone (10-50 μM) had no effect on cystometric parameters. M2 mAChR mRNA expression was increased in the mucosa of SCI rats compared with that in normal rats. Our results suggest that the M2 mAChR subtype plays an important role in bladder afferent activation that enhances detrusor overactivity in SCI rats. However, because mAChR antagonists alone did not affect any cystometric parameters, the muscarinic mechanism controlling bladder afferent activity might not be involved in the emergence of detrusor overactivity in SCI.


Physicochemical interpretation of the pharmacokinetics of percutaneous absorption.
Theoretical expressions are derived to predict the amount of a drug reaching the dermal capillaries as a function of simple physicochemical parameters. The mathematical approach involves the solution of the diffusion equation for the drug in the skin using the technique of Laplace transformation. First-order kinetics are assumed for the absorption of drug from the vehicle into the skin and for uptake into the capillaries. The relative importance of these different processes (absorption from vehicle, diffusion through the skin, and capillary removal) involved in the percutaneous penetration of a drug is discussed, and their influence upon the time course of the absorption phenomenon is illustrated.


Extrahepatic portal hypertension due to congenital obstruction of the portal vein and associated gross hepatic lobulation.
A 10-233k old girl presented with splenomegaly and recurrent hematemesis from esophageal varices. Splenoportography revealed a dilated extrahepatic portion of the portal vein with nonvisualization of its intrahepatic tributaries. The child died following an episode of hematemesis and was found to have a dilated portal vein which ended blindly. In addition, there was abnormal lobulation of the inferior surface of the liver which was not cirrhotic. The portal vascular anomaly, which presumably was responsible for the portal hypertension, was probably due to failure of communication between the embryonic vitelline veins or to atresia of the portal vein secondary to pressure from the abnormal hepatic lobulation in utero. It would appear that congenital factors may be significant in the etiology and pathogenesis of some cases of extrahepatic portal hypertension in early life and recognition of such developmental anomalies is of importantance in management.


Balance, risk of falls, risk factors and fall-related costs in individuals with diabetes.
Sensory loss and impaired balance are considered risk factors of incident falls. The aim of this study was to assess the relationship between degree of foot sensation and balance, risk of falls, incidence of fall-related injuries and costs in a cohort of patients with diabetes. (Non)-neuropathic subjects participating in the Rotterdam Diabetic Foot Study were followed prospectively. Subjects underwent sensory testing of the feet (39 item Rotterdam Diabetic Foot Study Test Battery (RDF-39)); balance was assessed at the second follow-up (Brief-BESTest) as were data on incident falls. Medical records and financial data were abstracted to estimate fall-related morbidity and in-hospital costs. A higher RDF-39 score, cerebral artery disease, type 2 diabetes, height and age were predictors of the Brief-BESTest total score. 41/296 patients (13.9%) reported two or more falls during follow-up. Predictors for recurrent falls were a higher RDF-39 score (aOR: 1.124, p < 0.0005), male gender (aOR: 0.319, p = 0.016), age (aOR: 0.938, p = 0.003) and type 2 diabetes (aOR: 3.157, p = 0.100). Thirty-one patients used medical resources (median US$ 440.45 (IQR: 179-1162). Degree of sensory loss correlates significantly with an increased imbalance and risk of falls. The RDF-39 may be used as stratification tool in medical decision-making and patient information.


[A case report: hirudotherapy as a treatment modality in the microsurgery].
Medicinal leeches are known to inject more than 100 bioactive substances in the tissue whilst sucking blood. They are being successfully used for the treatment of diseases and symptoms, especially in plastic surgery. We report the case of a 7-year old boy, who had lost the first phalanx of the middle finger of the right hand after an accident. The detached digit was replanted in the plastic surgery clinic, and treated within a period of 15 days with medicinal leeches (Hirudo medicinalis). Soon after the initiation of the hirudotherapy, the revascularisation of the re-attached phalanx, as well as the resolving of tissue oedema and pain was observed. Turkish physicians should be aware of the advantages of hirudotherapy in plastic and reconstructive surgery, especially in cases of detached digits and flaps.


Molecular characterization of the gut microbiome in egyptian patients with remitting relapsing multiple sclerosis.
Multiple Sclerosis is the most common chronic demyelinating disease of the central nervous system, representing the main cause of non-traumatic disabling disorders in young adults. The etiology of Multiple Sclerosis is not fully appreciated, although strong evidence points to genetic and environmental factors. The role of the gut microbiome in Multiple Sclerosis pathogenesis is a rapidly emerging area of study in that field. The aim of the present study is to identify the gut microbiome that are likely related to Multiple Sclerosis as well as their possible role in the susceptibility and the course of the disease. Thirty Egyptian patients with relapsing remitting multiple sclerosis, who presented to the Multiple Sclerosis Clinic of Alexandria University Hospital were enrolled in the study. These were diagnosed according to the McDonald 2017 criteria. A cross matching control group of 20 healthy subjects of similar age and sex was included. Stool specimens were taken from each for detection of gut microbiome profile by quantitative SYBR Green Real Time PCR assay. The present study demonstrate that RRMS patients have a distinct gut microbiome compared to healthy controls, with certain gut microbes showing decreased or increased abundance in Multiple Sclerosis patients compared to controls. Multiple Sclerosis patients have significantly higher B. fragilis than the normal control. Although the level of Prevotella, and Lactobacilli appear much less in MS patients than the control, the difference was not statistically significant. The same for C. perfringes which was higher in Multiple Sclerosis patients. The overall results of the study agree with previous studies reporting that patients with Multiple Sclerosis in Egypt exhibit microbial gut dysbiosis. However, the results of individual bacteria may differ, according to the age, treatment, level of physcial exercise and type of therapy at the time of enrollment.


Possible Abscopal Effect Observed in Frontal Meningioma After Localized IMRT on Posterior Meningioma Resection Cavity Without Adjuvant Immunotherapy.
Background: Localized radiation therapy (RT) is known to infrequently cause off-target or "abscopal" effects at distant metastatic lesions. The mechanism through which abscopal effects occur remains unknown, but is thought to be caused by a humoral immune response to tumor-specific antigens generated by RT. Combination treatment regimens involving RT and immunotherapy to boost the humoral immune response have demonstrated synergistic effects in promoting and accelerating abscopal effects in metastatic cancer. Nevertheless, abscopal effects, particularly after RT alone, remain exceedingly rare. Case Presentation: We report the case of an 84-year-old man with an atypical meningioma, who demonstrated a radiographically significant response to an untreated second intracranial lesion, likely also a meningioma, after intensity-modulated radiation therapy (IMRT) to a separate, detatched resection cavity. Serial annual MRI imaging starting at 2- to 3.5-year (most recent) post-IMRT follow-up demonstrated a persistent decrease in both tumor size and surrounding edema in the untreated second lesion, suggestive of a possible abscopal effect. Conclusions: We describe here the first report of a potential abscopal effect in meningioma, summarize the limited literature on the topic of abscopal effects in cancer, and detail the existing hypothesis on how this phenomenon may occur and possibly relate to the development of future treatments for patients with metastatic disease.


Relationship between mean platelet volume and retinopathy in patients with type 2 diabetes mellitus.
To evaluate the effect of mean platelet volume (MPV) on diabetic retinopathy in patients with type 2 diabetes mellitus. In this study, ocular findings and MPV values were retrospectively reviewed in 192 patients with type 2 diabetes mellitus. The patients were classified into four groups according to ocular findings, as follows: group 1, diabetic patients without diabetic retinopathy (n = 70); group 2, diabetic patients with non-proliferative diabetic retinopathy (n = 64); group 3, diabetic patients with proliferative diabetic retinopathy (n = 58); and group 4, healthy controls (n = 100). A significant difference was found in MPV values between groups 2 and 4 (P = 0.001), between groups 3 and 4 (P = 0.001), and between groups 1 and 4 (P = 0.004). No significant difference was found in MPV values between groups 1 and 2 (P = 0.241) and between groups 2 and 3 (P = 0.460); whereas there was a statistically significant difference between groups 1 and 3 (P = 0.015). The three diabetic groups (groups 1, 2, and 3) were compared with each other. While there was a statistically significant difference between groups 1 and 3 (P = 0.015), there was no significance between groups 2 and 3 (P = 0.46), and between group 1 and 2 (P = 0.241). Logistic regression analysis found a 1.40-fold increase in the risk of retinopathy development (OR: 1.404; P = 0.002) and a 1.46-fold increase in the risk of proliferative diabetic retinopathy (OR: 1.466; P = 0.002) as the MPV value increased. In diabetic patients, the risk of retinopathy development increases with higher MPV values.


Trends in insurance coverage for bariatric surgery and the impact of evidence-based reviews.
The recent increase in demand for bariatric surgery has placed mounting economic pressure on insurance companies and other third-party payers (TPPs). As a result, some of the TPPs have responded by excluding or limiting their coverage of all or certain types of bariatric surgical procedures, and cite as their reason, a lack of evidence that supports the safety and efficacy of such procedures. Over the years, so-called "evidence-based reviews" have been used to back these claims. Some of these reviews have significant flaws and limitations that are discussed.


Spontaneous mutations in hlyD and tuf genes result in resistance of Dickeya solani IPO 2222 to phage ϕD5 but cause decreased bacterial fitness and virulence in planta.
Lytic bacteriophages able to infect and kill Dickeya spp. can be readily isolated from virtually all Dickeya spp. containing environments, yet little is known about the selective pressure those viruses exert on their hosts. Two spontaneous D. solani IPO 2222 mutants (0.8% of all obtained mutants), DsR34 and DsR207, resistant to infection caused by lytic phage vB_Dsol_D5 (ΦD5) were identified in this study that expressed a reduced ability to macerate potato tuber tissues compared to the wild-type, phage-susceptible D. solani IPO 2222 strain. Genome sequencing revealed that genes encoding: secretion protein HlyD (in mutant DsR34) and elongation factor Tu (EF-Tu) (in mutant DsR207) were altered in these strains. These mutations impacted the DsR34 and DsR207 proteomes. Features essential for the ecological success of these mutants in a plant environment, including their ability to use various carbon and nitrogen sources, production of plant cell wall degrading enzymes, ability to form biofilms, siderophore production, swimming and swarming motility and virulence in planta were assessed. Compared to the wild-type strain, D. solani IPO 2222, mutants DsR34 and DsR207 had a reduced ability to macerate chicory leaves and to colonize and cause symptoms in growing potato plants.


A serine proteinase in the penetration glands of the hexacanths of Hymenolepis diminuta (Cestoda, Cyclophyllidea).
Histochemically demonstrable activity of a serine proteinase was detected in the penetration glands of Hymenolepis diminuta hexacanths. At the optimal pH of 8.4 the enzyme hydrolyzed N-blocked L-aminoacyl- and N-blocked L-peptidyl-naphthylamides bearing L-arginine at the P1 subsite. The proteinase did not require either Ca2+ or Mg2+ for its activity and was insensitive to 1 mM EGTA and 1 mM EDTA. Organic fluorophosphates inhibited it, whereas thiol-blocking compounds did not. At operative pH values of 4.8 and 3.8 generated during electrophoresis in a stacking and a resolving gel, respectively, the proteinase migrated toward the cathode. When examined for proteolytic activity at the optimal pH of 8.4, the separated enzyme produced a single band of gelatinolysis in a gelatin-containing polyacrylamide gel. During in vitro maintenance of the hexacanths, the secretion from their penetration glands formed a mucous cyst surrounding the individual larvae. The cyst was resistant to and protected the hexacanths from the proteolytic activity of trypsin, papain, and proteinases extracted from the gut of the beetle Tenebrio molitor (the host). Hexacanths extracted from the hemocoel of T. molitor at 24 and 48 h after infection were surrounded by similar mucous cysts. Consequently, roles in penetration and protection for the secretion from the penetration glands are postulated.


Aβ5-x Peptides: N-Terminal Truncation Yields Tunable Cu(II) Complexes.
The Aβ5-x peptides (x = 38, 40, 42) are minor Aβ species in normal brains but elevated upon the application of inhibitors of Aβ processing enzymes. They are interesting from the point of view of coordination chemistry for the presence of an Arg-His metal binding sequence at their N-terminus capable of forming a 3-nitrogen (3N) three-coordinate chelate system. Similar sequences in other bioactive peptides were shown to bind Cu(II) ions in biological systems. Therefore, we investigated Cu(II) complex formation and reactivity of a series of truncated Aβ5-x peptide models comprising the metal binding site: Aβ5-9, Aβ5-12, Aβ5-12Y10F, and Aβ5-16. Using CD and UV-vis spectroscopies and potentiometry, we found that all peptides coordinated the Cu(II) ion with substantial affinities higher than 3 × 1012 M-1 at pH 7.4 for Aβ5-9 and Aβ5-12. This affinity was elevated 3-fold in Aβ5-16 by the formation of the internal macrochelate with the fourth coordination site occupied by the imidazole nitrogen of the His13 or His14 residue. A much higher boost of affinity could be achieved in Aβ5-9 and Aβ5-12 by adding appropriate amounts of the external imidazole ligand. The 3N Cu-Aβ5-x complexes could be irreversibly reduced to Cu(I) at about -0.6 V vs Ag/AgCl and oxidized to Cu(III) at about 1.2 V vs Ag/AgCl. The internal or external imidazole coordination to the 3N core resulted in a slight destabilization of the Cu(I) state and stabilization of the Cu(III) state. Taken together these results indicate that Aβ5-x peptides, which bind Cu(II) ions much more strongly than Aβ1-x peptides and only slightly weaker than Aβ4-x peptides could interfere with Cu(II) handling by these peptides, adding to copper dyshomeostasis in Alzheimer brains.


High Expression of Tumor HLA-DR Predicts Better Prognosis and Response to Anti-PD-1 Therapy in Laryngeal Squamous Cell Carcinoma.
HLA-DR is expressed in epithelial and several types of tumor cells. However, the correlation between tumor-expressed HLA-DR (teHLA-DR) and patient outcome as well as its regulation on the tumor microenvironment (TME) of laryngeal squamous cell carcinoma (LSCC) are yet to be elucidated. Hematoxylin and eosin (HE) staining were performed to define the tumor nest and stroma of LSCC tissue microarrays. teHLA-DR tumor cell, CD4+ and CD8+ tumor-infiltrating T lymphocytes (TITLs) were obtained and analyzed through double-labeling immunofluorescence and immunohistochemical staining. The recurrence-free (RFS) and overall survival (OS) curves were plotted using the Kaplan-Meier method and tested by the log-rank test method. Expression of teHLA-DR+ tumor cells and infiltration of T lymphocytes and their corresponding subgroups were analyzed by flow cytometry using fresh LSCC tissue samples. Our research discovered elevated expressions of multiple MHC-II-related genes in tumor compared to the adjacent normal tissue samples of LSCC patients. We also found that patients in the teHLA-DR high-expression group (teHLA-DRhigh) tend to have less tumor recurrence and better survival outcomes compared to those in the teHLA-DRlow group. Intriguingly, teHLA-DR+ tumor cells had significantly higher PD-L1 and PD-L2 expression and their TME showed increased infiltrated T lymphocytes (TITLs). Flow cytometry analysis and IHC staining indicated that CD4+ TITLs but not CD3+ total TITLs or CD8+ TITLs were significantly enriched in teHLA-DR+ tumors. teHLA-DR may be a predictive marker for favorable prognosis and response to anti-PD-1/PD-L1 therapy of LSCC, possibly due to the increased CD4+ TITLs in the TME.


Intramolecular labelling of sucrose made by leaves from [14C)carbon dioxide or [3-14C]serine.
Pea leaves were illuminated in air containing 150 or 1000p.p.m. of 14CO2 for various times. Alternatively, segments of wheat leaves were supplied with [3-14C]serine for 40 min in the light in air with 145, 326 or 944p.p.m. of 12CO2. Sucrose was extracted from the leaf material, hydrolysed with invertase, and 14C in the pairs of carbon atoms C-3+C-4, C-2+C-5 and C-1+C-6 in the glucose moiety was measured. The results obtained after metabolism of 14CO2 were consistent with the operation of the photosynthetic carbon-reduction cycle; the effects of CO2 concentration on distribution of 14C in the carbon chain of glucose after metabolism of [3-14C]serine is more easily explained by metabolism through the glycollate pathway than by the carbon-reduction cycle.


Retinal nerve fiber layer thinning: a window into rapid eye movement sleep behavior disorders in Parkinson's disease.
Retinal nerve fiber layer (RNFL) thinning occurs in Parkinson's disease (PD) and other neurodegenerative diseases. Idiopathic RBD (iRBD) is a well-established prodromal hallmark of synucleinopathies and occurs secondary to many neurodegenerative diseases, including PD. The aim of this study is to determine whether or not retinal structures are altered with the onset of rapid eye movement (REM) sleep behavior disorders (RBD). In all, a total of 63 patients with PD, 14 patients with idiopathic RBD, and 26 sex- and age-matched healthy controls were enrolled and underwent optical coherence tomography measurements (HD-OCT (Zeiss) ) for the average and every quadrant of RNFL thickness. The REM Sleep Behavior Disorder Screening Questionnaire (RBDSQ) was used to classify PD patients with clinically probable RBD (PD + pRBD) or without probable RBD (PD - pRBD). Patients with iRBD were identified by polysomnography. For patients with RBD (idiopathic or secondary to PD), we found a significant decrease in RNFL thickness compared with groups without RBD (PD - pRBD and healthy controls) (all p < 0.05). Average RNFL thickness in patients with iRBD is significantly thinner than in healthy controls (p < 0.05). In PD, the average RNFL thickness was dramatically thinner in the PD + pRBD group than the PD - pRBD group (p < 0.005). Compared with healthy controls, RNFL thickness was slightly thinner in the drug-naive PD group but not the PD group with drug treatment. Multiple linear regression analysis showed that RBDSQ score was negatively associated with average and inferior RNFL variation in PD (all p < 0.005). The findings show that RNFL was slightly but significantly thinner in idiopathic RBD. In PD, RNFL thickness may vary depending on the presence of RBD.


Notch and disease: a growing field.
Signals through the Notch receptors are used throughout development to control cellular fate choices. Our intention here is to provide an overview of the involvement of Notch signaling in human disease, which, keeping pace with the known biology of the pathway, manifests itself in a pleiotropic fashion. A pathway with such broad action in normal development, a profound involvement in the biology of adult stem cells and intricate and complex controls governing its activity, poses numerous challenges. We provide an overview of Notch related pathologies identified thus far and emphasize aspects that have been modeled in experimental systems in order to understand the underlying pathobiology and, hopefully, help the definition of rational therapeutic avenues.


Guidelines for the practice of utilization review: essential but lacking.
Utilization review (UR) has developed to the point where it soon will be, if it is not already, a separate profession. Yet it lacks practice guidelines and principles to guide practitioners. In the absence of such, practitioners may inappropriately use those of their original professions or other guidelines not in conformity with the objectives of UR. Practice guidelines for UR should arise out of professional consensus following extensive discussions. As a step in that direction, a number of proposed guidelines are presented.


Conformational comparison in the growth hormone family.
1. The method of Kubota et al. [Biochim. biophys. Acta 701, 242-252 (1982)] was applied to several members of the growth hormone family in order to examine their conformational homology. 2. The method neither detects differences between rat, cow, sheep, horse and alpaca hormones, nor between monkey and human hormones. 3. Lack of homology between primate and non-primate growth hormones was found in segments 42-49 and 184-191. The first fragment could be linked to species-specificity.


Excitotoxic lesions of the lateral parabrachial nucleus do not prevent cholecystokinin-induced suppression of milk intake in rats.
We examined the effect of lateral (visceral) and medial (gustatory) parabrachial nucleus (PBN) lesions on the suppression of milk intake induced with cholecystokinin (CCK) in 24 h food-deprived rats. Irrespective of CCK dose, each type of PBN lesion significantly elevated the consumption of milk. Neither medial nor lateral PBN lesions had any discernible influence on the anorectic action of CCK, however. In combination with the results from a previous study using the same rats (Brain Res. 894 (2001) 288), the present results support the view that the role of the lateral PBN in CCK-induced anorexia depends on the nature of the food used at test.


Severe congenital neutropenia-associated JAGN1 mutations unleash a calpain-dependent cell death programme in myeloid cells.
Severe congenital neutropenia (SCN) of autosomal recessive inheritance, also known as Kostmann disease, is characterised by a lack of neutrophils and a propensity for life-threatening infections. Using whole-exome sequencing, we identified homozygous JAGN1 mutations (p.Gly14Ser and p.Glu21Asp) in three patients with Kostmann-like SCN, thus confirming the recent attribution of JAGN1 mutations to SCN. Using the human promyelocytic cell line HL-60 as a model, we found that overexpression of patient-derived JAGN1 mutants, but not silencing of JAGN1, augmented cell death in response to the pro-apoptotic stimuli, etoposide, staurosporine, and thapsigargin. Furthermore, cells expressing mutant JAGN1 were remarkably susceptible to agonists that normally trigger degranulation and succumbed to a calcium-dependent cell death programme. This mode of cell death was completely prevented by pharmacological inhibition of calpain but unaffected by caspase inhibition. In conclusion, our results confirmed the association between JAGN1 mutations and SCN and showed that SCN-associated JAGN1 mutations unleash a calcium- and calpain-dependent cell death in myeloid cells.


Functional genomic studies of tick cells in response to infection with the cattle pathogen, Anaplasma marginale.
The coevolution of ticks and the pathogens that they transmit has ensured their mutual survival. In these studies, we used a functional genomics approach to characterize tick genes regulated in response to Anaplasma marginale infection. Differentially regulated genes/proteins were identified by suppression-subtractive hybridization and differential in-gel electrophoresis analyses of cultured IDE8 tick cells infected with A. marginale. Nine of 17 of these genes were confirmed by real-time RT-PCR to be differentially regulated in ticks and/or IDE8 tick cells in response to A. marginale infection. RNA interference was used for functional studies. Six genes, which encode putative selenoprotein W2a, hematopoietic stem/progenitor cells protein-like, proteasome 26S subunit, ferritin, GST, and subolesin control, were found to affect A. marginale infection in IDE8 tick cells. Four genes, which encode putative GST, salivary selenoprotein M, vATPase, and ubiquitin, affected A. marginale infection in different sites of development in ticks. The results of these studies demonstrated that a molecular mechanism occurs by which tick cell gene expression mediates the A. marginale developmental cycle and trafficking through ticks.


Diastereo- and enantioselective direct vinylogous Michael addition of γ-substituted butenolides to 2-enoylpyridines catalyzed by chiral bifunctional amine-squaramides.
The diastereo- and enantioselective direct vinylogous Michael addition reaction of γ-substituted butenolides to 2-enoylpyridines has been achieved. A range of γ,γ-disubstituted butenolide derivatives, bearing two consecutive tri- and tetrasubstituted stereogenic centers, were readily obtained in good yields with excellent stereoselectivities (up to >99 : 1 dr and >99% ee).


Neuropathology of vascular cognitive impairment and vascular dementia.
Understanding of vascular substrates of cognitive decline in the elderly is evolving to include a major emphasis on the impact of small vessel disease (SVD). While existing concepts of multi-infarct dementia and strategic infarct dementia remain valid, they present difficulty in generalizing clinicopathological correlations from patient to patient. The range and significance of lesions that should be included as manifestations of SVD are unresolved, as is their impact on, and association with, neurodegenerative changes. This mini-review summarizes the authors' views on SVD substrates leading to cognitive decline and proposes priorities for pathological investigations of human cerebrovascular mechanisms leading to cognitive decline.


Assessing the intestinal carriage rates of vancomycin-resistant enterococci (VRE) at a tertiary care hospital in Hungary.
Excessive use of antibiotics contributes to the selection of resistant bacteria and intestinal colonization with multiresistant pathogens poses a risk factor for subsequent infections. The present study assessed vancomycin-resistant enterococci (VRE) carriage rates in patients admitted to our tertiary care hospital. Stool samples sent for routine culturing were screened with vancomycin containing solid or broth enrichment media. VRE isolates were identified with matrix-assisted laser desorption/ionization-time of flight mass spectrometry and antibiotic susceptibilities were tested by E-test. Vancomycin resistance genes were detected by polymerase chain reaction. Medical records of carriers were examined for suspected risk factors for colonization. Altogether 3025 stool specimens were analyzed. Solid media identified a VRE carriage rate of 2.2% while broth enrichment detected 5.8%. Seventy percent of the isolates were Enterococcus faecium. VanB genotype was detected in 38.2%, VanA in 37.3%, VanC1 in 22.6%, and VanC2 in 1.9%. All VRE were sensitive to linezolid, daptomycin, and tigecycline. Collective risk factors for carriage were diabetes, normal flora absence, Clostridioides difficile positivity, longer hospital stay, and advanced age. 78.5% of the carriers received antibiotic therapy which was metronidazole in most cases (47.3%). We recommend regular screening of risk groups such as patients with diabetes, history of recent hospitalization, or former C. difficile infection as an imperative step for preventing VRE dissemination.


Ifosfamide induced stress response in human lymphocytes.
Ifosfamide, an isomer of cyclophosphamide, has been shown to be one of the most effective antineoplastic agents for the treatment of human malignancies. There is considerable evidence that the intracellular status of glutathione (GSH) plays a major role in modifying the cytotoxicity of ifosfamide in cells and tissues. We have studied the effects of 4-hydroperoxy-ifosfamide (4-OOH-IF) upon the proliferation of human peripheral blood lymphocytes (PBL) and the intracellular GSH content. The major finding was that occurrence of significant inhibition of [3H]-thymidine incorporation in interleukin-2 (IL-2) expanded PBL after exposure with 4-OOH-IF was accompanied by substantial depletion of intracellular GSH content in these cells. PBL seemed to be more sensitive to this drug induced effect comparing our results obtained in other cells (e.g. Ewing sarcoma, Chinese hamster ovary). In PBL 4-OOH-IF also induced rapid phosphorylation of the small heat shock protein (HSP27) signaling a similar type of stress response as reported for several other agents (e.g. arsenite, phorbol ester, tumor necrosis factor). Reconstitution of the depleted GSH content in PBL after treatment with 4-OOH-IF could be achieved by GSH-monoethylester and mesna within 24 hours of postincubation time. From these results we conclude that human lymphocytes are sensitive targets for ifosfamide induced metabolic stress during treatment. This might have further importance in regard to the immunological function of these cells.


The tecto-olivo-cerebellar pathway in the rat.
The olivo-cerebellar projection to the vermis of cerebellar lobule VII, and the projection of the midbrain tectum onto the inferior olive (IO), were examined in the rat employing, respectively, retrograde and anterograde transport of horseradish peroxidase (HRP). Olivary neurons afferent to lobule VII vermis are located in a restricted region of the contralateral, caudomedial, medial accessory olive (MAO). The termination of the olivary projection demonstrated after HRP placement in the superior colliculus (SC), corresponds closely with this region.


Airway foreign body manifested as a coin lesion.
Foreign body aspiration is a potentially life-threatening event. The nature of the inhaled objects is highly variable, ranging from organic to inorganic material. Although most pills are radiolucent, lanthanum carbonate is radiopaque and may be identified on chest X-rays.


Medicare and Medicaid programs; fire safety requirements for long term care facilities, automatic sprinkler systems. Final rule.
This final rule requires all long term care facilities to be equipped with sprinkler systems by August 13, 2013. Additionally, this final rule requires affected facilities to maintain their automatic sprinkler systems once they are installed.


LMNA cardiomyopathy detected in Japanese arrhythmogenic right ventricular cardiomyopathy cohort.
Arrhythmogenic right ventricular cardiomyopathy (ARVC) is an inherited cardiac disease. While desmosomal gene mutations are considered major causes of ARVC, LMNA mutations have been reported to be possible causes of ARVC. In this study, we performed extensive genetic screening for LMNA mutations in our Japanese ARVC cohort to assess the prevalence and characteristics of LMNA mutation-positive ARVC cases. Our study cohort consisted of 57 ARVC probands. Genetic analyses were performed by using direct sequencing and targeted sequencing of LMNA and four desmosomal genes. We compared clinical features of probands with desmosomal gene mutations to those of probands with LMNA mutations. Among 57 clinically diagnosed ARVC probands, we identified desmosomal gene mutations in 26 probands (45.6%) and two LMNA mutations in two probands. The first LMNA mutation p.M1K was detected in a 62-year-old male proband, while the second mutation p.W514X was found in a 70-year-old male proband. Compared to the 26 probands with desmosomal gene mutations, in the two probands with LMNA mutations, the mean age at diagnosis was significantly higher, and their heart rate at the diagnosis was significantly slower. While both probands with LMNA mutations underwent pacemaker implantation, only one proband with desmosomal mutations received this treatment (2/2 vs. 1/26). Genetic screening for LMNA gene is important for ARVC patients, particularly in patients with bradycardia.


Activation of cytosolic glucocorticoid-receptor complexes in intact WEHI-7 cells does not dephosphorylate the steroid-binding protein.
In order to determine the ratio of phosphates to hormone-binding sites on nonactivated (non-DNA-binding) glucocorticoid receptors in WEHI-7 mouse thymoma cells, we have extracted these receptors from cells grown to a steady state with 32P, labeled them with a saturating concentration of [3H]dexamethasone 21-mesylate, purified them using a monoclonal antibody, and analyzed them by polyacrylamide gel electrophoresis under denaturing and reducing conditions. The complexes contained approximately 5 mol of phosphate/mol of bound steroid. Only half of the phosphates were associated with the approximately 100-kDa protein which is labeled with [3H]dexamethasone 21-mesylate. The remaining phosphates were associated with the approximately 90-kDa non-steroid-binding component of the nonactivated complex. Dual label studies, using [35S]methionine to measure receptor protein and 32P to measure receptor phosphates, have enabled us to determine the phosphate content, relative to receptor protein, of both nonactivated and activated cytosolic complexes generated in intact WEHI-7 cells exposed to triamcinolone acetonide at 37 degrees C. The total amount of phosphate associated with the activated complex is roughly half of that associated with the nonactivated complex, the decrease being accounted for by dissociation of the approximately 90-kDa phosphoprotein which accompanies activation. However, the ratio of 32P to 35S counts associated with the approximately 100-kDa steroid-binding protein is the same for the activated and nonactivated complexes. These results indicate that there is no net change in the phosphorylation of the approximately 100-kDa steroid-binding component of the cytosolic glucocorticoid-receptor complex upon activation in the intact cell.


Reversible neurological and brain MRI changes following COVID-19 vaccination: A case report.
Various neurological sequalae have been described following COVID-19 vaccination. Here we describe the first case of untreated post COVID-19 vaccine encephalitis with spontaneous resolution of contrast enhancing hyperintensities on MRI concomitant with clinical improvement. A 59-year-old woman presented with a two-day history of unsteady gait, incoordination, visual symptoms, and lethargy. She had received AZD1222 (AstraZeneca) and mRNA-1273 (Moderna) COVID-19 vaccines at 3 months and 12 days, respectively, before presentation. Brain MRI showed no abnormality on the non-enhanced sequences, but numerous enhancing lesions in the cerebral cortex, deep grey matter, brainstem, and cerebellum. Treatment was expectant, the patient improved clinically over 10 days, and repeat MRI showed near complete resolution of the imaging abnormality. We describe neurological deterioration 12 days after a second dose of COVID-19 vaccine. There was no evidence of edema or demyelinating lesions in the brain on MRI, but there was extensive contrast-enhancement indicating loss of blood-brain barrier (BBB) integrity. This provides a potential in vivo, clinical-imaging correlate of the post-mortem evidence that SARS-CoV-2 spike protein may induce loss of BBB permeability. While this adds to the list of rare adverse neurological reactions to COVID-19 vaccination, the benefits of receiving the vaccine far outweigh these risks.


Dynamic structure of the translocon SecYEG in membrane: direct single molecule observations.
Purified SecYEG was reconstituted into liposomes and studied in near-native conditions using atomic force microscopy. These SecYEG proteoliposomes were active in translocation assays. Changes in the structure of SecYEG as a function of time were directly visualized. The dynamics observed were significant in magnitude (∼1-10 Å) and were attributed to the two large loops of SecY linking transmembrane helices 6-7 and 8-9. In addition, we identified a distribution between monomers and dimers of SecYEG as well as a smaller population of higher order oligomers. This work provides a new vista of the flexible and dynamic structure of SecYEG, an intricate and vital membrane protein.


Characterization of Glomerular Sox9+ Cells in Anti-Glomerular Basement Membrane Nephritis in the Rat.
Mechanisms of glomerular crescent formation and podocyte repair processes are still unclear. Therefore, we investigated the expression of the transcription factor Sox9 as a potential marker of a subpopulation of parietal epithelial cells (PECs) with potential regenerative properties. Glomerular Sox9 expression was characterized in detail in a rat anti-glomerular basement membrane (GBM) nephritis model using immunofluorescence and confocal laser scanning microscopy. In healthy kidneys Sox9 is expressed in a subpopulation of PECs restricted to approximately 20% to 50% of PEC nuclei and was highly conserved in all investigated species. During rat anti-GBM nephritis the number of glomerular Sox9+ cells increased and was associated with proliferation activity. In nephritic glomeruli Sox9 expression was not restricted to Bowman's capsule lining but was also found on cells of the glomerular tuft. Nearly all Sox9+ cells also expressed the PEC marker Pax8, whereas endothelial cells, mesangial cells, macrophages, and T lymphocytes lacked Sox9 expression. At the margins of crescents Sox9+/Pax8+ cells additionally expressed podocyte markers. In contrast, in sclerotic lesions a minority of Sox9+/Pax8+ cells expressed the myofibroblast marker α-smooth muscle actin. In glomerular Sox9+ cells Jagged 1 was up-regulated. During anti-GBM nephritis Sox9+ PECs proliferate and migrate onto the glomerular tuft. Future studies are needed to confirm the origin of Sox9+ cells from PECs and differentiation in both podocytes and/or myofibroblasts.


Pancreatitis and the biliary tree: the continuing problem.
Gallstone-associated pancreatitis continues to have a mortality rate that approaches 10 percent. In a review of 132 fatal cases of acute pancreatitis, no less than a third of the gallstone-associated cases were diagnosed for the first time at autopsy. Early diagnosis of gallstones in these patients remains problematic, but clinical and biochemical factors may aid ultrasonography in defining patients who require endoscopic retrograde cholangiopancreatography. Early operation is advisable in patients with mild disease, but endoscopic papillotomy should be considered in those with severe disease who fail to stabilize after admission. Chronic pancreatitis is frequently associated with cholangiographic evidence of biliary obstruction, and serum alkaline phosphatase concentrations offer a valuable means of monitoring cholestasis. If operation is needed to deal with biliary obstruction, the options are to carry out Roux-Y hepaticojejunostomy or resection of the pancreatic head, the choice being dictated by the indications for direct pancreatic operation.


Recombinant expression and antigenic properties of a 31.5-kDa keratinolytic subtilisin-like serine protease from Microsporum canis.
A secreted 31.5-kDa keratinolytic subtilase (SUB3; AJ431180) is thought to be a Microsporum canis virulence factor and represents a candidate for vaccination trials. In this study, the recombinant keratinase (r-SUB3) was produced by the Pichia pastoris expression system and purified to homogeneity. Recombinant SUB3 displayed identical biochemical properties with the native protease. Experimentally cutaneously infected guinea pigs showed specific lymphoproliferative response towards r-SUB3, while no specific humoral immune response was induced except for one animal. The heterologous expression of SUB3 provides a valuable tool for addressing further investigations on the role of this keratinase in the specific cellular immune response and on its use in vaccination trials in the cat.


Two keratins MW 50,000 and 56,000 are synthesized by psoriatic epidermis.
The keratin polypeptide composition of uninvolved psoriatic epidermis is the same as normal epidermis. In contrast, involved epidermis from psoriasis contains significant amounts of two keratin polypeptides, molecular weight (MW) 56 and 50 kilodaltons (KD). Involved epidermis contains decreased amounts of keratins MW 69, 66.5 and 65.2 KD. Slices of involved psoriatic epidermis incorporate [35S]methionine into all the keratins including those of MW 56 and 50 KD. Unexpectedly, we found that [35S]methionine is incorporated into 56 and 50 KD keratins in uninvolved epidermis even though these keratins are not abundant enough to be seen in Coomassie stained gels. The two-dimensional patterns of iodinated tryptic peptide digests of keratin MW 50 KD from uninvolved, involved and normal epidermis are identical. We conclude that keratins of MW 56 KD and 50 KD are abundant in involved psoriatic epidermis. They are not present in normal or uninvolved psoriatic epidermis, but are rapidly synthesized when normal epidermis (neonatal foreskin) or uninvolved psoriatic epidermis are removed and placed in organ culture. The two keratins may be markers of increased cell proliferation or injury.


Objective measurement of the distal resection margin by MRI of the fresh and fixed specimen after partial mesorectal excision for rectal cancer: 5 cm is not just 5 cm and depends on when measured.
Most studies have directly established the optimal perioperative in situ clearance margin in surgery for rectal cancer from the histologically observed extent of distal spread, neglecting the tissue variability that occurs after resection and fixation of the rectal specimen. To measure the length of the distal resection margin in the fresh and fixed specimen following partial mesorectal excision for rectal cancer using magnetic resonance imaging (MRI) to document tissue shrinkage after surgical removal and fixation. The length of the distal resection margin was measured by MRI of the fresh and fixed specimen and at histopathological examination of the fixed specimen in 10 patients who underwent surgery for upper rectal cancer. In addition, tissue shrinkage was estimated by measuring the total length of the fresh and fixed specimen and distance from the peritoneal reflection anteriorly to the distal cut edge of the specimen. Measured by MRI, the distal resection margin was in the range of 0.6-10.2 cm (mean, 4.6 cm) in the fresh specimen, and 0.5-6.2 cm (mean, 3.2 cm) in the fixed specimen. The tissue shrinkage ratio was a mean of 69% (interquartile range, 61-77%). Taking all ratios from MRI and histopathological examination of tissue shrinkage into account, the collective tissue shrinkage ratio was 70% (95% confidence interval, 67-73%) The length of the distal resection margin was reduced by 30% after surgical removal and fixation of the specimen.


Dangerous reef aquaristics: Palytoxin of a brown encrusting anemone causes toxic corneal reactions.
Although frequently observed in domestic saltwater aquariums, literature on exposure to palytoxin (PTX) of encrusting anemones (Zoanthidea) kept in aquariums is rare. Handling these animals for propagation purposes or during cleaning work can lead to dermal, ocular or respiratory contact with the PTX generated by some Zoanthids. The present study describes a case of ocular exposure to liquid from a Zoanthid, which led to corneal ulcers. The patient also suffered from systemic symptoms of dyspnea and shivering and a suspected rhabdomyolysis, which required monitoring in the Intensive Care Unit. After symptomatic treatment provided insufficient results, the corneal ulcers improved with an amniotic membrane transplantation. A review of the literature regarding ocular exposures to this diverse order of Hexacorallia reveals that severe and systemic symptoms can develop with minimal contact.


Mapping pelvic lymph nodes: guidelines for delineation in intensity-modulated radiotherapy.
To establish guidelines for delineating the clinical target volume for pelvic nodal irradiation by mapping the location of lymph nodes in relation to the pelvic anatomy. Twenty patients with gynecologic malignancies underwent magnetic resonance imaging with administration of iron oxide particles. All visible lymph nodes were outlined. Five clinical target volumes were generated for each patient using modified margins of 3, 5, 7, 10, and 15 mm around the iliac vessels. The nodal contours were then overlaid and individual nodes analyzed for coverage. The volume of normal tissue within each clinical target volume and planning target volume was also measured to aid selection of the margin that could provide maximal nodal, but minimal normal tissue, coverage. In total, 1216 nodal contours were evaluated. The nodal coverage was 56%, 76%, 88%, 94%, and 99% using vessel margins of 3, 5, 7, 10, and 15 mm, respectively. The mean volume of bowel within the planning target volume was 146.9 cm3 with a 7-mm margin, 190 cm3 with a 10-mm margin, and 266 cm3 with a 15-mm margin. Minor modification to the 7-mm margin ensured 99% coverage of the pelvic nodes. Blood vessels with a modified 7-mm margin offer a good surrogate target for pelvic lymph nodes. By making appropriate adjustments, coverage of specific nodal groups may be increased and the volume of normal tissue irradiated decreased. On the basis of these findings, recommended guidelines for outlining pelvic nodes have been produced.


Input estimation for drug discovery using optimal control and Markov chain Monte Carlo approaches.
Input estimation is employed in cases where it is desirable to recover the form of an input function which cannot be directly observed and for which there is no model for the generating process. In pharmacokinetic and pharmacodynamic modelling, input estimation in linear systems (deconvolution) is well established, while the nonlinear case is largely unexplored. In this paper, a rigorous definition of the input-estimation problem is given, and the choices involved in terms of modelling assumptions and estimation algorithms are discussed. In particular, the paper covers Maximum a Posteriori estimates using techniques from optimal control theory, and full Bayesian estimation using Markov Chain Monte Carlo (MCMC) approaches. These techniques are implemented using the optimisation software CasADi, and applied to two example problems: one where the oral absorption rate and bioavailability of the drug eflornithine are estimated using pharmacokinetic data from rats, and one where energy intake is estimated from body-mass measurements of mice exposed to monoclonal antibodies targeting the fibroblast growth factor receptor (FGFR) 1c. The results from the analysis are used to highlight the strengths and weaknesses of the methods used when applied to sparsely sampled data. The presented methods for optimal control are fast and robust, and can be recommended for use in drug discovery. The MCMC-based methods can have long running times and require more expertise from the user. The rigorous definition together with the illustrative examples and suggestions for software serve as a highly promising starting point for application of input-estimation methods to problems in drug discovery.