The effects of reductions in the numbers of animals used for the potency assay of the diphtheria and tetanus components of adsorbed vaccines by the methods of the European pharmacopoeia.
Forty eight assays of adsorbed diphtheria vaccine and seven assays of adsorbed tetanus vaccine using either a lethal challenge (38 assays) or a serum neutralization test (17 assays) were evaluated for the effects of reductions in the number of animals used at each dilution on the potency values and 95% confidence intervals. The results were assessed in the light of the requirements of the European Pharmacopoeia and the WHO. In the majority of assays, 50% of the number of animals presently required would have sufficed for the determination of a potency within the limits of confidence stipulated by the European Pharmacopoeia and the WHO. Therefore it is concluded that a simplified assay with a reduced number of animals is suitable for the routine potency testing of the diphtheria and tetanus components of the combined vaccines and the monovalent that were examined. Flexibility in the national and international requirements in respect of the numbers of animals used at each dilution is suggested for the routine potency assay of the diphtheria and tetanus components of adsorbed vaccines.


Multiple-injection affinity capillary electrophoresis to estimate binding constants of receptors to ligands.
Multiple-injection affinity capillary electrophoresis (MIACE) is used to determine binding constants (Kb) between receptors and ligands using as model systems vancomycin and teicoplanin from Streptomyces orientalis and Actinoplanes teichomyceticus, respectively, and their binding to D-Ala-D-Ala peptides and carbonic anhydrase B (CAB. EC 126.96.36.199) and the binding of the latter to arylsulfonamides. A sample plug containing a non-interacting standard is first injected followed by multiple plugs of sample containing the receptor and then a final injection of sample containing a second standard. Between each injection of sample, a small plug of buffer is injected which contains an increasing concentration of ligand to effect separation between the multiple injections of sample. Electrophoresis is then carried out in an increasing concentration of ligand in the running buffer. Continued electrophoresis results in a shift in the migration time of the receptor in the sample plugs upon binding to their respective ligand. Analysis of the change in the relative migration time ratio (RMTR) or electrophoretic mobility (mu) of the resultant receptor-ligand complex relative to the non-interacting standards, as a function of the concentration of ligand yields a value for Kb. The MIACE technique is a modification in the ACE method that allows for the estimation of binding affinities between biological interactions on a timescale faster than that found for standard ACE. In addition sample volume requirements for the technique are reduced compared to traditional ACE assays. These findings demonstrate the advantage of using MIACE to estimate binding parameters between receptors and ligands.


Voltage-clamp analysis of the sodium and potassium currents in skeletal muscle fibres treated with 4-aminopyridine.
External application of low concentrations of 4-aminopyridine blocks potassium currents without affecting sodium currents in pieces of single frog skeletal muscle fibres. The blockade of potassium currents was voltage-dependent, being partially relieved on depolarization.


Aplasia cutis congenita of the scalp: how much therapy is necessary in large defects?
To show that local antibiotic management and a regular inspection of aplasia cutis congenita of the skull can give an excellent result. This case reports a girl born with aplasia cutis congenita of the skull presenting with a large aplasia of the epidermis, dermis, subcutaneous tissue and galea, including a bone defect without any additional risk factor, e.g. early eschar formation, cerebrospinal fluid leakage or uncommon dural blood vessels. A primarily conservative treatment with local wet and antibiotic dressings together with a systemic antibiotic treatment for the first 2 wk led to an excellent result and thus prevented untimely operative and peri-operative procedures. Here we have shown that conservative treatment might be an option, even if the wound diameter is greater than 1 cm(2), to prevent infants from any untimely operative procedure with an elevated operative risk if any additional risk factors are excluded.


Phase Angle Is a Stronger Predictor of Hospital Outcome than Subjective Global Assessment-Results from the Prospective Dessau Hospital Malnutrition Study.
This prospective cohort study of 16,943 consecutive patients compared phase angle (PhA, foot-to-hand at 50 kHz) and subjective global assessment (SGA) to predict outcomes length of hospital stay (LOS) and in-hospital mortality in patients at risk of malnutrition (NRS-2002 ≥ 3). In 1505 patients, the independent effects on LOS were determined by competing risk analysis and on mortality by logistic regression. In model I, including influence factors age, sex, BMI, and diagnoses, malnourished (SGA B and C) patients had a lower chance for a regular discharge (HR 0.74; 95%CI 0.69−0.79) and an increased risk of mortality (OR 2.87; 95%CI 1.38−5.94). The association of SGA and outcomes regular discharge and mortality was completely abrogated when PhA was added (model II). Low PhA reduced the chance of a regular discharge by 53% in patients with a PhA ≤ 3° (HR 0.47; 95%CI 0.39−0.56) as compared to PhA > 5°. Mortality was reduced by 56% for each 1° of PhA (OR 0.44; 95%CI 0.32−0.61). Even when CRP was added in model III, PhA ≤ 3° was associated with a 41% lower chance for a regular discharge (HR 0.59; 95%CI 0.48−0.72). In patients at risk of malnutrition, the objective measure PhA was a stronger predictor of LOS and mortality than SGA.


Hypochlorous acid-induced heme damage of hemoglobin and its inhibition by flavonoids.
Hypochlorous acid (HOCl), produced by activated neutrophils, is highly reactive oxidizing and chlorinating agent of biologically relevant molecules. Hemoglobin (Hb), present in large amounts inside red blood cells, is the main target for HOCl in these cells. In this work heme damage of hemoglobin induced by hypochlorous acid in the absence and presence of some popular dietary flavonoids, catechin, epigallocatechin gallate and quercetin has been studied by stopped-flow spectrophotometry. Hypochlorous acid, being in a large molar excess to hemoglobin, initiates modifications of the heme group of oxy- as well as of methemoglobin, which eventually leads to heme damage. Flavonoids present at concentrations comparable with that of hemoglobin inhibit these processes. The kinetics of the reactions of the investigated flavonoids with HOCl has been also studied and the rate constants of the order of 10(5)M(-1)s(-1) have been found. It is concluded that under conditions used in this study the inhibition of Hb heme damage by flavonoids results from the competition of these compounds with hemoglobin towards HOCl and/or from the formation of Hb-flavonoid complex in which heme group is more resistant against HOCl-induced damage.


Varied clinical presentation and outcome of SARS-CoV-2 infection in liver transplant recipients: Initial experience at a single center in Madrid, Spain.
Which are the consequences of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in liver transplant (LT) recipients? We attempted to address this question by reviewing our single-center experience during the first 2 months of the pandemics at a high incidence area. Nineteen adult patients (5 females) were diagnosed by May 5, 2020. Median age was 58 (range 55-72), and median follow-up since transplantation was 83 (range 20-183) months. Cough (84.2%), fever (57.9%), and dyspnea (47.4%) were the most common symptoms. Thirteen patients (68.4%) had pneumonia in x-ray/CT scan. Hydroxychloroquine was administered in 11 patients, associated with lopinavir/ritonavir and interferon β in 2 cases each. Immunomodulatory therapy with tocilizumab was used in 2 patients. Immunosuppression (IS) was halted in one patient and modified in only other two due to potential drug interactions. Five (26.3%) patients were managed as outpatient. Two patients (10.5%) died, 10 (52.6%) were discharged home, and 2 (10.5%) were still hospitalized after a median follow-up of 41 days from the onset of symptoms. Baseline IS regimen remained unchanged in all surviving recipients, with good liver function. Our preliminary experience shows a broad spectrum of disease severity in LT patients with COVID-19, with a favorable outcome in most of them without needing to modify baseline IS.


Development of polymorphic microsatellite markers in Camellia chekiangoleosa (Theaceae) using 454-ESTs.
A set of microsatellite markers for Camellia chekiangoleosa was developed and characterized using 454 sequencing technology to study the population genetic structure and the diversity of germplasm collections. Eighteen polymorphic microsatellite markers were identified and tested in 150 individuals from three natural populations of C. chekiangoleosa. Alleles numbered from two to seven, and the observed and expected heterozygosities ranged from 0.100 to 0.760 and 0.133 to 0.809, respectively. These markers will potentially be conducive to further genetic studies on C. chekiangoleosa.


Electrogenerated chemiluminescence. 68. Detection of sodium ion with a ruthenium(II) complex with crown ether moiety at the 3,3'-positions on the 2,2'-bipyridine ligand.
We show here how a Ru(bpy)2(CE-bpy) complex can be used for the ECL detection of sodium ion in both aqueous and nonaqueous media. Recognition of Na+ by the crown ether moiety in CE-bpy results in a significant increase in the ECL emission intensity of the complex.


A virus between families: nucleotide sequence and evolution of Strawberry latent ringspot virus.
Several clones of golden ginger mint (Mentha x gracilis, 'Variegata') were found infected with Strawberry latent ringspot virus (SLRSV). The virus was purified and cloned and the complete nucleotide sequence of a mint isolate was obtained. RNA 1 consists of 7,496 nucleotides excluding the poly-A tail and encodes a polyprotein with signature enzymatic motifs found in other picorna-like plant viruses. RNA 2 consists of 3,842 nucleotides excluding the poly-A tail, encoding a polyprotein that is processed to a putative movement protein and the two coat proteins of the virus. A satellite RNA of 1,117 nucleotides was associated with this isolate encoding for a putative protein of 31 kDa. Phylogenetic analysis revealed that SLRSV shares characteristics with members of the Cheravirus, Fabavirus, Comovirus and Sadwavirus genera indicative of the uniqueness of SLRSV. The close relationship of SLRSV with these genera led to the examination of aphid and beetle transmission of the virus with, however, negative results.


Central catecholamine depletion inhibits peripheral lymphocyte responsiveness in spleen and blood.
Experimental and clinical evidence has demonstrated extensive communication between the CNS and the immune system. To analyse the role of central catecholamines in modulating peripheral immune functions, we injected the neurotoxin 6-hydroxydopamine (6-OHDA) i.c.v. in rats. This treatment significantly reduced brain catecholamine content 2, 4 and 7 days after injection, and in the periphery splenic catecholamine levels were reduced 4 days after treatment. Central catecholamine depletion induced an inhibition of splenic and blood lymphocyte proliferation and splenic cytokine production and expression (interleukin-2 and interferon-gamma) 7 days after injection. In addition, central treatment with 6-OHDA reduced the percentage of spleen and peripheral blood natural killer (CD161 +) cells, and T-cytotoxic (CD8 +) cells in peripheral blood. The reduction in splenocyte proliferation was not associated with a glucocorticoid alteration but was completely abolished by prior peripheral sympathectomy. These data demonstrate a crucial role of central and peripheral catecholamines in modulating immune function.


Characteristics and proteomic analysis of pyrene degradation by Brevibacillus brevis in liquid medium.
Polycyclic aromatic hydrocarbons (PAHs) are widely spread in various ecosystems and are of great concern due to their potential toxicity, mutagenicity and carcinogenicity. Bioremediation has been proposed as an effective approach to remove PAHs. In this study, the physiological responses and proteome of Brevibacillus brevis under exposure to pyrene, a four-ring compound from PAHs family, were investigated. The changes of cell viability of B. brevis were observed during the degradation of pyrene by means of flow cytometry. The results indicated that pyrene stimulated superoxide dismutase (SOD) activity from 93.9 to 100.6 U mg-1 prot, whereas inhibited catalase (CAT) activity from 29.1 to 20.3 U mg-1 prot. The main compositions of B. brevis changed during pyrene degradation, with the proportion of unsaturated fatty acids increased by 13.4%. In addition, we performed a proteomic approach (two-dimensional gel electrophoresis and MALDI-TOF/TOF-MS) in order to explore how B. brevis survived upon treatment with pyrene. It was showed that the expression of 13 proteins increased whereas 10 other decreased after pyrene-treatment. The differentially expressed proteins were identified and the results indicated that they were involved in multiple biological processes including energy metabolism, biosynthesis, transmembrane transport and oxidative stress. Overall, these findings offered a new insights into the cellular response strategy developed by B. brevis to overcome the pyrene stress.


Patterns of influenza B circulation in Brazil and its relevance to seasonal vaccine composition.
Data on the burden of disease and circulation patterns of influenza B lineages for Brazil are limited. This review aims to describe the pattern of influenza B occurrence in Brazil to have a better understanding of its epidemiology and its relevance when considering seasonal influenza vaccine composition. A review of the data including analysis of international and local surveillance data as well as information from online search of databases using Medical Subject Headings terms in conjunction with screening of abstracts from scientific events was performed. Based on international epidemiologic surveillance data, moderate levels of influenza B disease (19%; 2006-2014) were observed. Of these nine years, it was possible to compare data from three years (2007, 2008 and 2013) which have information on the circulating influenza B lineage. Co-circulation of influenza B lineages was observed in all these three influenza seasons, of which, during one season, a high degree of mismatch between the vaccine lineage and the predominant circulating lineage (91.4% [2013]) was observed. Local surveillance data reveal a distinct and dynamic distribution of respiratory viruses over the years. Data from published literature and abstracts show that influenza B is a significant cause of disease with an unpredictable circulation pattern and showing trends indicating reemergence of the B/Victoria lineage. The abstracts report notable levels of co-circulation of both influenza B lineages (2000-2013). Mismatch between the Southern hemisphere vaccine and the most prevalent circulating viruses in Brazil were observed in five influenza seasons. The evidence on co-circulation of two influenza B lineages and mismatched seasons in Brazil indicates the benefit of quadrivalent influenza vaccines in conferring broader seasonal influenza protection. Additionally, improving influenza surveillance platforms in Brazil is important for monitoring disease trends and the impact of introducing seasonal influenza vaccination.


Short-term retentive values of zirconium oxide posts cemented with glass ionomer and resin cement: an in vitro study and a case report.
This paper presents the results of an investigation focusing on the retention of all-ceramic posts. The aim of the study was to evaluate the influence of two luting agents--a glass-ionomer cement (Ketac Cem) and a resin cement (Panavia F)--on the retention of a prefabricated ceramic post (Cerapost) after different pretreatment techniques in vitro. Forty freshly extracted incisors and canines were treated endodontically. The root canals were filled with laterally condensed gutta-percha and sealer. The teeth were assigned to 1 of 4 groups, each containing 10 specimens: sandblasted/nonsandblasted post cemented with glass-ionomer cement and sandblasted/nonsandblasted post cemented with resin cement. Tensile tests were carried out. The specimens were also quantitatively analyzed under a scanning electron microscope. The retentive strengths of posts, sandblasted and nonsandblasted, and cemented with Panavia F resin cement, were significantly increased as compared to those cemented with Ketac Cem. Both the luting agent and the post-surface pretreatment had a significant influence on the retention of the ceramic post.


Treatment of female androgenetic alopecia with cimetidine.
Ten white women with moderate to severe androgenetic alopecia were treated with cimetidine 300 mg by mouth five times a day. Duration of therapy ranged from 1.5-9 months, with a median of 5 months. Seven patients (70%) showed good to excellent regrowth of hair. No major side effects were noted. The patients were followed up for 5 months. Cimetidine presumably has the ability to block androgen action. Cimetidine is not a first-line drug for the treatment of androgenetic alopecia in women and should be used only in selected cases.


Topological transition and Majorana zero modes in 2D non-Hermitian chiral superconductor with anisotropy.
We study a non-Hermitian chiral topological superconductor system on two dimensional square lattice, from which we obtained a rich topological phase diagram and established an exact relationship between topological charge flow of exceptional points in generalized Brillouin zone and change of topological properties. Its rich topological phase diagram is the result of competition between anisotropy and non-Hermitian effect. This system belongs to class D according to AZ classification of non-Hermitian systems. Each topological phase can be characterized by a 2DZnumber, which indicates the number of chiral edge modes, and two 1DZ2numbers, which indicate the existence of zero modes at edge dislocations.


[Morphometrical study on inhibitory effect of vitamin B6 and banana-stem extract on calcium crystallization].
To search for the anticrystallization mechanism by vitamin B(6) and banana stem extract. Thirty-six male China-1 mice were divided into 4 groups: normal, crystallization, vitamin B(6) and banana-stem extract groups. Calcium oxalate crystallization was induced by 1% glycol and 1% ammonium chloride in the last 3 groups. Vitamin B(6) was then given to the 3rd group and banana stem extract to the 4th group for 3 weeks. At the end of 3 weeks, all the mice were sacrificed and the kidneys were taken for morphometrical studies using image analysis system and assayed for calcium oxalate. Morphometrical area density and numerical density of crystallization was significantly less in the vitamin B(6) group and the banana-stem extract group as compared with the crystallization group, being the least in the banana-stem extract group. Oxalate in renal tissue was also significantly reduced. Both vitamin B(6) and banana stem extract had no effect on renal tissue calcium. Vitamin B(6) and banana-stem extract may be useful agents in the treatment of patients with hyperocaluric urolithiasis.


The occurrence of oral-mucosal lesions among employees of a steelmill--a preliminary study.
A study has been conducted on 198 employees of a steel-mill industry. These participants were examined for mucosal lesions in the oral cavity and including the commissures and vermillion borders of the lips. The prevalences of 16 types of lesions are reported.


Natural history of cervical human papillomavirus lesions does not substantiate the biologic relevance of the Bethesda System.
The newly introduced Bethesda System proposes to replace the terms dysplasia, carcinoma in situ (CIS), or cervical intraepithelial neoplasia (CIN) with two terms, low-grade squamous intraepithelial lesion (SIL) or high-grade SIL. We tested the biologic relevance of the Bethesda System (ie, its ability to establish a close correlation between biologic behavior and the different grades of lesions) in a series of 528 women with genital human papillomavirus (HPV)-associated precancerous lesions prospectively followed for 10 years. The cervical biopsies were reclassified as either low-grade or high-grade SIL, and the data obtained by colposcopy, Papanicolaou smear, and HPV typing, as well as the biological behavior of the lesions, were analyzed in these two groups. Altogether, 77.4% (376 of 486) of the lesions were classified as low-grade SIL lesions; the rest (22.6%) belonged to the high-grade SIL category. In the low-grade SIL category, 46.8% of the women were 24 years old or younger, as compared with 37.3% in the high-grade SIL group. The colposcopic appearance was normal significantly more frequently in the low-grade SIL lesions (22.1%) than in the high-grade category (8.5%) (P less than .001). A single Papanicolaou smear was inadequate to distinguish between low-grade and high-grade SIL, as evidenced by almost identical distributions of Papanicolaou smear class I and II cells in both categories. Noteworthy was the discovery of normal Papanicolaou smears in 8.2% of the high-grade SIL lesions.(ABSTRACT TRUNCATED AT 250 WORDS)


The use of thiopurines for the treatment of inflammatory bowel diseases in clinical practice.
Thiopurines are the most commonly used immunomodulatory drugs in inflammatory bowel diseases. To evaluate the use, the therapeutic and safety profiles of thiopurines in a large sample of IBD patients. We reviewed 3641 case histories of IBD patients. Thiopurines were prescribed in 582 patients (16.0%); the analysis was performed on the 553 (267 ulcerative colitis, 286 Crohn's disease) with exhaustive clinical data. The main indications for treatment were steroid-dependence (328/553, 59.3%) and steroid-resistance (113/553, 20.7%). Thiopurines were started when CD were younger than UC patients (p<0.001) but earlier from diagnosis in UC than in CD patients (p=0.003). Efficacy was defined as optimal (258/553, 46.6%), partial (108/553, 19.5%), absent (85/553, 15.4%) and not assessable (102/553, 18.4%). Efficacy was independent of disease type, location/extension or duration and age at starting. Side effects were observed in 151/553 (27.3%) patients, leading to drug discontinuation in 101 (18.3%). 15 out of the 130 (11.5%) patients who took thiopurines for more than 4 years relapsed, more frequently in CD than in UC (OR=3.67 95% C.I. 0.98-13.69; p=0.053). Thiopurines confirm their clinical usefulness and acceptable safety profile in managing complicated IBD patients. The majority of patients treated for longer than 4 years maintain response. No clinical and demographic predictive factors for efficacy and side effects were identified.


Dietary amino acids and blood pressure: a cohort study of patients with cardiovascular disease.
Dietary protein has been variably reported to either lower or raise blood pressure. The purpose of this study was to determine whether intakes of specific amino acids differentially associate with blood pressure. Observational cohort study by secondary analysis of clinical trial data. Study of low-fat versus Mediterranean-style diets in patients with prevalent cardiovascular disease. Dietary amino acids. Systolic and diastolic blood pressure. Dietary nutrients and cardiovascular risk factors were assessed at baseline, 3 and 6 months, and then every 6 months for 2 years. Baseline blood pressure was 119 ± 16 (SD)/72 ± 10 (SD) mm Hg (n = 92) and dietary protein intake was 80 ± 31 g/d. Independent amino acid variables (quartiles of intake) were analyzed by generalized estimating equation models with prespecified covariates for time-varying systolic and diastolic blood pressure. The odds of each 1-SD higher systolic or diastolic blood pressure (ie, 16 and 10 mm Hg, respectively) were increased per quartile of intake for methionine (ORs of 1.29 [95% CI, 1.14-1.46] and 1.21 [95% CI, 1.05-1.39], respectively) and alanine (ORs of 1.17 [95% CI, 1.05-1.30] and 1.22 [95% CI, 1.07-1.38], respectively). Quartiles of intake for threonine (ORs of 0.84 [95% CI, 0.74-0.96] and 0.87 [95% CI, 0.75-1.01], respectively) and histidine (ORs of 0.92 [95% CI, 0.86-1.00] and 0.89 [95% CI, 0.82-0.97], respectively) had inverse associations with the same degree of difference in blood pressure. Modest sample-size biases toward the chance of false-negative results; potential for residual confounding; colinearity between amino acids may obscure relevant relationships to blood pressure; associational findings do not establish causality. Intakes of methionine and alanine were associated positively with higher blood pressure, whereas intakes of threonine and histidine had inverse associations. These amino acids merit further study for advancing dietary approaches to blood pressure reduction.


Oral cancer in Puerto Rico.
All cancer cases diagnosed in Puerto Rico are analyzed and registered in the Central Cancer Registry. The incidence of oral cancer in Puerto Rico is fourth-highest in the world. (In 1972 oral cancer accounted 72% of all cancers in males in Puerto Rico.) A review of the data revealed important contrasts with reported statistics from the continental United States (e.g., 75.6% of the cases in Puerto Rico were initially treated by radiation, compared with 15.3% of the cases in Western New York States.


In vitro (organ culture) studies of the toxicity of specific A-gliadin peptides in celiac disease.
Specific peptides of known amino acid sequence were prepared from alpha-gliadin (A-gliadin) by cleavage of the protein with cyanogen bromide and chymotrypsin and purification of the resulting peptides. The three peptides derived from the cyanogen bromide cleavage spanned the complete sequence of A-gliadin (266 residues). Four peptides derived from chymotryptic digestion covered the N-terminal sequence through residue 68. These peptides were tested for toxicity in celiac disease by organ culture of biopsied small intestinal tissues taken from patients with active celiac disease. Enterocyte height was used as a measure of peptide effect on cultured tissues. Five of seven peptides tested significantly inhibited increase of enterocyte height in the cultures and were considered toxic on this basis. The largest common sequences among the toxic peptides were -pro-ser-gln-gln- and -gln-gln-gln-pro-; these sequences were absent from the nontoxic peptides. The relationship of these sequences to the damaging effect of gliadins on the small intestinal mucosa in celiac disease remains to be investigated.


Engaging the pyridine-DMAD zwitterion in a novel strategy for the selective synthesis of highly substituted benzene and cyclopentenedione derivatives.
[reaction: see text] The pyridine-mediated reaction of dimethyl acetylenedicarboxylate and cyclobutene-1,2-diones affords selective access to either hexasubstituted benzene derivatives or cyclopentenedione derivatives depending on the concentration of pyridine.


The role of circulating viral and tumour DNA in the diagnosis and management of HPV associated anogenital cancers, a systematic review and meta-analysis.
Human papillomavirus associated anogenital cancers are a significant global burden. The detection of biomarkers (circulating tumour DNA; ctDNA or circulating HPV DNA; cHPV DNA) in blood referred to as "liquid biopsy" may support the early diagnosis and monitoring of affected individuals. A systematic review, including meta-analysis of studies available in the literature on the utilization of ctDNA and cHPV DNA as diagnostic, predictive, and monitoring biomarker tests of HPV associated anogenital cancers was performed following the criteria of PRISMA. A total of 31 studies were eligible for systematic review; 20 used cHPV DNA in cervical cancers; 7 used ctDNA in cervical cancer; 5 used cHPV DNA in anal cancer; no eligible studies on vulva, vaginal or penile cancer were available. The meta-analysis identified low sensitivity (0.36) and high specificity (0.96) of cHPV DNA as diagnostic for cervical cancer. Comparatively, there was high sensitivity (0.95) and specificity (1.0) of cHPV DNA for the diagnosis of anal cancer. cHPV DNA and/or ctDNA in cervical cancer were prognostic markers associated with poor clinical outcomes. Additionally, in anal cancer the post treatment detection of cHPV DNA was informative in the prediction of treatment response or progression-free survival. ctDNA and cHPV DNA are promising diagnostic and prognostic biomarkers for the detection of anogenital disease. Evolution and refinement of molecular tools is likely to improve performance further. Additionally the comparative absence of studies in the vulval, vaginal and penile context warrants further exploration and research.


Complex and spatially segregated auditory inputs of the mouse superior colliculus.
Although the visual circuits in the superior colliculus (SC) have been thoroughly examined, the auditory circuits lack equivalent scrutiny. SC neurons receiving auditory inputs in mice were characterized and three distinguishable types of neurons were found. The auditory pathways from external nuclei of the inferior colliculus (IC) were characterized, and a novel direct inhibitory connection and an excitation that drives feed-forward inhibitory circuits within the SC were found. The direct excitatory and inhibitory inputs exhibited distinct arbourization patterns in the SC. These findings suggest functional differences between excitatory and inhibitory sensory information that targets the auditory SC. The superior colliculus (SC) is a midbrain structure that integrates auditory, somatosensory and visual inputs to drive orientation movements. While much is known about how visual information is processed in the superficial layers of the SC, little is known about the SC circuits in the deep layers that process auditory inputs. We therefore characterized intrinsic neuronal properties in the auditory-recipient layer of the SC (stratum griseum profundum; SGP) and confirmed three electrophysiologically defined clusters of neurons, consistent with literature from other SC layers. To determine the types of inputs to the SGP, we expressed Channelrhodopsin-2 in the nucleus of the brachium of the inferior colliculus (nBIC) and external cortex of the inferior colliculus (ECIC) and optically stimulated these pathways while recording from SGP neurons. Probing the connections in this manner, we described a monosynaptic excitation that additionally drives feed-forward inhibition via circuits intrinsic to the SC. Moreover, we found a profound long-range monosynaptic inhibition in 100% of recorded SGP neurons, a surprising finding considering that only about 15% of SGP-projecting neurons in the nBIC/ECIC are inhibitory. Furthermore, we found spatial differences in the cell body locations as well as axon trajectories between the monosynaptic excitatory and inhibitory inputs, suggesting that these inputs may be functionally distinct. Taking this together with recent anatomical evidence suggesting an auditory excitation from the nBIC and a GABAergic multimodal inhibition from the ECIC, we propose that sensory integration in the SGP is more multifaceted than previously thought.


Sonographic findings of healthy volunteers infected with dengue virus.
Sonography has historically been used in developing countries to help diagnose dengue infection during epidemics of dengue hemorrhagic fever in endemic areas and to predict the clinical course. In this article, we describe the sonographic findings in subjects infected with attenuated, monovalent strains of dengue virus. As part of a major research protocol to validate challenge strains of dengue virus for use in vaccines, 12 subjects were infected with 1 of 4 strains of dengue virus, and 3 subjects received placebo. The challenge was followed by an observation period. During this time, they were imaged regardless of the development of symptoms. Seven of 12 subjects infected with dengue virus showed sonographic evidence of subclinical plasma leakage, including perihepatic and perisplenic ascites, pericardial effusion, and gallbladder wall thickening. None of the 3 placebo recipients developed effusions. Sonographic evidence of fluid collection was seen in over half of subjects infected with dengue virus who did not show any evidence of dengue hemorrhagic fever. These findings shed light on possible mechanisms of plasma leakage and its role in the pathogenesis of dengue fever and dengue hemorrhagic fever.


Ultrasonographic evaluation of pelvic organ support during pregnancy.
To investigate whether pregnancy affects levator hiatus dimensions and the position and mobility of the bladder neck and the levator ani muscle in nulliparous pregnant women. In the present study, 274 nulliparous pregnant women were examined at 21 weeks and 37 weeks of gestation using three-dimensional and four-dimensional transperineal ultrasonography at rest, during contraction, and during Valsalva maneuver. Levator hiatus dimensions were the anteroposterior diameter, the transverse diameter, and the area measured in rendered images. Positions of the bladder neck and levator plate were analyzed in the midsagittal plane, and mobility was calculated as displacement of the bladder neck or levator plate from rest to contraction or from rest to Valsalva. A significant increase for all levator hiatus dimension measurements was found from 21 weeks to 37 weeks of gestation. The most marked change was found for levator hiatus area at rest and during Valsalva maneuver, in which the mean area was increased by 17.1% (11.7-13.7 cm(2)) and 21.4% (15.4-18.7 cm(2)), respectively. Bladder neck mobility changed significantly during pregnancy. The most marked change was seen from rest to contraction (mean -14 mm, standard deviation 0.4). An increase of all hiatal dimensions as well as bladder neck mobility was found from 21 weeks to 37 weeks of gestation in nulliparous pregnant women. The findings indicate that the changes in pelvic organ support are not solely caused by delivery, but also by physiologic changes during pregnancy. II.


SerpinB3 and Yap Interplay Increases Myc Oncogenic Activity.
SerpinB3 has been recently described as an early marker of liver carcinogenesis, but the potential mechanistic role of this serpin in tumor development is still poorly understood. Overexpression of Myc often correlates with more aggressive tumour forms, supporting its involvement in carcinogenesis. Yes-associated protein (Yap), the main effector of the Hippo pathway, is a central regulator of proliferation and it has been found up-regulated in hepatocellular carcinomas. The study has been designed to investigate and characterize the interplay and functional modulation of Myc by SerpinB3 in liver cancer. Results from this study indicate that Myc was up-regulated by SerpinB3 through calpain and Hippo-dependent molecular mechanisms in transgenic mice and hepatoma cells overexpressing human SerpinB3, and also in human hepatocellular carcinomas. Human recombinant SerpinB3 was capable to inhibit the activity of Calpain in vitro, likely reducing its ability to cleave Myc in its non oncogenic Myc-nick cytoplasmic form. SerpinB3 indirectly increased the transcription of Myc through the induction of Yap pathway. These findings provide for the first time evidence that SerpinB3 can improve the production of Myc through direct and indirect mechanisms that include the inhibition of generation of its cytoplasmic form and the activation of Yap pathway.


Prognostic factors affecting the risk of thoracic progression in extensive-stage small cell lung cancer.
The efficacy of combined modality therapy is evaluated for patients with extensive-stage (ES) small cell lung cancer (SCLC). This study evaluated prognostic factors affecting the risk of thoracic progression in ES-SCLC patients likely to undergo thoracic radiotherapy combined chemotherapy. A retrospective review of ES-SCLC patients who had received systemic chemotherapy at our hospital was performed. Tumor size, metastatic sites, and laboratory data at diagnosis were evaluated as potential prognostic factors. In ES-SCLC patients without pleural dissemination, the rate of thoracic progression after initial chemotherapy was assessed. Eighty-three of 96 consecutive ES-SCLC patients were analyzed. The overall response rate was 55 %, median progression free survival was 5.0 months (mo), and overall survival (OS) was 9.2 mo. Tumor size (19.4 mo for ≤3 cm vs. 8.5 mo for >3 cm, p = 0.017) and the number of metastatic sites (12.9 mo for single sites vs. 7.1 mo for multiple sites, p = 0.015) were prognostic factors, in addition to known prognostic factors such as performance status and the levels of LDH and sodium. Cox proportional hazard model showed that the OS was significantly worse in patients with large (>3 cm) primary tumor size {HR 2.44 [95 % confidential interval (CI) 1.05-5.68], p = 0.038} and multiple metastatic sites [HR 1.81 (95 % CI 1.08-3.04), p = 0.026]. In 51 cases without pleural dissemination, the number of metastatic sites was associated with thoracic progression after initial chemotherapy (65 % for single sites vs. 36 % for multiple sites, p = 0.036). Large tumor size and multiple metastatic sites at diagnosis significantly predicted poor survival in ES-SCLC patients. Based on the high rate of thoracic progression in ES-SCLC patients with single site of distant metastasis, we should consider thoracic radiotherapy combined with chemotherapy for this population.


Adolescent leisure reading and its longitudinal association with prosocial behavior and social adjustment.
Reading is a popular leisure activity for children, teenagers, and adults. Several theories agree that reading might improve social cognition, but the empirical evidence remains tentative, with research on adolescents especially lacking. We employed a very large, and nationally representative, longitudinal dataset from the National Educational Panel Study (NEPS) in Germany to examine this hypothesis. Specifically, we tested whether reading prospectively predicted future self-reported prosocial behavior and social adjustment in adolescents, controlling for a number of covariates. Two-way cross-lagged panel analyses probed the longitudinal relationship between leisure reading and these social outcomes from Grade 6 to Grade 9. In addition, we examined the effect of cumulative reading experience across Grades 5-8 on future social outcomes, using structural equation modeling. We also explored the unique contributions of cumulative reading experience in different literary genres (classic literature, popular literature, nonfiction, comic books). Cumulative reading in general did not predict future prosocial behavior and social adjustment. However, cumulative reading of modern classic literature was positively associated with later prosocial behavior and social adjustment. PROTOCOL REGISTRATION: The stage 1 protocol for this Registered Report was accepted in principle on 08 November 2021. The protocol, as accepted by the journal, can be found at: https://doi.org/10.17605/OSF.IO/KSWY7 .


Does Mycobacterium tuberculosis have plasmids?
Evidence of the presence of plasmids in Mycobacterium tuberculosis is lacking, whereas they are widespread in some other mycobacterial species. We examined, by agarose gel electrophoresis, a total of 197 clinical isolates of M. tuberculosis, mostly resistant to one or more antibiotics, and were able to detect bands of apparently extrachromosomal DNA at a low level in some isolates. These presumptive plasmids could not be isolated by CsCl/ethidium bromide gradient ultracentrifugation, and may consist of unusual forms of DNA. The possible existence of single stranded plasmid DNA is discussed.


PNMT-containing catecholaminergic neurons are not necessarily adrenergic.
The intermediolateral cell column (IML) in the spinal cord is densely innervated by the phenylethanolamine-N-methyltransferase (PNMT)-containing neurons of the ventrolateral medulla (C1 cell group). The present study used a sensitive HPLC-EC assay for catecholamines to quantitate epinephrine levels in the region of the spinal cord containing the IML. In dissections of thoracic cord enriched in IML, as well as total thoracic cord, epinephrine levels were below the limit of sensitivity of the assay, indicating that epinephrine levels were less than 0.2 pg/mg tissue. In the region of the IML, epinephrine levels were less than 0.05% of the norepinephrine content. Thus, no epinephrine could be detected in the IML even though the IML contains a dense network of nerve terminals that stain immunohistochemically for PNMT and the other enzymes required for epinephrine biosynthesis. These results suggest that nerve terminals that contain all the enzymes required for epinephrine biosynthesis are not necessarily adrenergic.


IgA vasculitis in adults: the performance of the EULAR/PRINTO/PRES classification criteria in adults.
In 2010, EULAR/PRINTO/PRES proposed new classification criteria for paediatric IgA vasculitis (IgAV) that have a higher diagnostic sensitivity than the 1990 ACR criteria. These criteria have so far not been evaluated in adults, in whom IgAV is considered as a rare disease. Our main objective was to compare the diagnostic performance of EULAR/PRINTO/PRES and ACR classification criteria in adult IgAV. Adult IgAV cases fulfilling the 2012 revised International Chapel Hill Consensus Conference Nomenclature of Vasculitides (ICHCCNV) definition of IgAV at a secondary/tertiary rheumatology referral centre were critically reviewed in a partially retrospective and partially prospective manner. First, we compared the diagnostic sensitivity of ACR and EULAR/PRINTO/PRES criteria in this group of patients. Second, the diagnostic specificity of ACR and EULAR/PRINTO/PRES was determined by applying these criteria to a control group of patients with other systemic vasculitides. Between 1 January 2010 and 31 December 2014 350 new cases of systemic vasculitis were identified. IgAV was diagnosed in 129, and other systemic vasculitides in 221 (123 had large, six medium and 92 small vessel vasculitis) cases according to ICHCCNV. The diagnostic sensitivity and specificity of the IgAV EULAR/PRINTO/PRES criteria were 99.2 % (95 % CI 95.4-99.9 %) and 86.0 % (95 % CI 80.7-90.3 %), and of the ACR criteria 86.8 % (95 % CI 79.7-92.1 %) and 81.0 % (95 % CI 75.2-85.9 %), respectively with an inter-criteria agreement of 77.5 % (95 % CI: 70.8-84.1 %). In the adult population the EULAR/PRINTO/PRES IgAV classification criteria had a higher sensitivity and specificity than the ACR criteria.


Genome-wide association scan identifies a prostaglandin-endoperoxide synthase 2 variant involved in risk of knee osteoarthritis.
Osteoarthritis (OA), the most prevalent form of arthritis in the elderly, is characterized by the degradation of articular cartilage and has a strong genetic component. Our aim was to identify genetic variants involved in risk of knee OA in women. A pooled genome-wide association scan with the Illumina550 Duo array was performed in 255 controls and 387 cases. Twenty-eight variants with p < 1 x 10(-5) were estimated to have probabilities of being false positives <or=0.5 and were genotyped individually in the original samples and in replication cohorts from the UK and the U.S. (599 and 272 cases, 1530 and 258 controls, respectively). The top seven associations were subsequently tested in samples from the Netherlands (306 cases and 584 controls). rs4140564 on chromosome 1 mapping 5' to both the PTGS2 and PLA2G4A genes was associated with risk of knee OA in all the cohorts studied (overall odds ratio OR(mh) = 1.55 95% C.I. 1.30-1.85, p < 6.9 x 10(-7)). Differential allelic expression analysis of PTGS2 with mRNA extracted from the cartilage of joint-replacement surgery OA patients revealed a significant difference in allelic expression (p < 1.0 x 10(-6)). These results suggest the existence of cis-acting regulatory polymorphisms that are in, or near to, PTGS2 and in modest linkage disequilibrium with rs4140564. Our results and previous studies on the role of the cyclooxygenase 2 enzyme encoded by PTGS2 underscore the importance of this signaling pathway in the pathogenesis of knee OA.


Effects of model orientation on the visuomotor imitation of arm movements: the role of mental rotation.
An experiment was conducted to examine whether visual perspective has an effect on visuomotor imitation. The experiment presented varied visual perspectives in terms of spatial disparity between a model and an observer (model-observer-disparity) on six levels. Female participants were asked to imitate different arm movements presented in videos by animated models. These arm movements were elbow-extension-flexion movements with one (lower complexity) or three (higher complexity) reversals of movement direction. The results showed that model-observer-disparity affects spatial trajectories and velocities in the performance of motor imitations. The movements' complexity did not affect orientation dependence. Due to the non-linearity of the data, it is questionable if orientation dependence can be at least partially explained by mental rotation processes or differential ideomotor effects. According to these results, high model-observer-disparity should be avoided when using visual instructions in visuomotor imitation.


Synthesis of ecdysone-14C and ecdysterone-14C from cholesterol-14C in cockroaches (Periplaneta americana) without molting glands.
The synthesis of molting hormone of cockroaches of which the prothoracic glands had previously been extirpated was investigated after injection of cholesterol-14C. It could be proved by means of radio thin-layer chromatography of extracts that ecdysterone and in small amounts ecdysone are synthesized in Periplaneta americana larvae without prothoracic glands. The results demonstrate that other tissues are also able to perform the molting hormone besides the molting gland itself.


Vascularized ulnar nerve graft for reconstruction of a large defect of the median or radial nerves after severe trauma of the upper extremity.
We report 3 cases of successful vascularized ulnar nerve graft for reconstructing a large median or radial nerve defect after severe trauma of the upper extremity.


Protective effects of SUN N8075, a novel agent with antioxidant properties, in in vitro and in vivo models of Parkinson's disease.
SUN N8075 is a novel antioxidant with neuroprotective properties. This study was designed to elucidate its neuroprotective effects against 6-hydroxy dopamine (6-OHDA)-induced cell death and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced neurotoxicity (known as in vitro and in vivo models of Parkinson's disease, respectively). In the in vitro study, on human neuroblastoma SH-SY5Y cells, SUN N8075 decreased the hydrogen peroxide (H2O2)-induced production of reactive oxygen species and protected against 6-OHDA-induced cell death. In the in vivo study, SUN N8075, when injected intraperitoneally (i.p.) twice with a 5-h interval, inhibited lipid peroxidation (viz. the production of thiobarbituric acid reactive substance) in the mouse forebrain at 1 h after the second injection. Mice were injected i.p. with MPTP (10 mg/kg) four times at 1-h intervals, and brains were analyzed 7 days later. SUN N8075 at 30 mg/kg (i.p., twice) exhibited a protective effect against the MPTP-induced decrease in tyrosine hydroxylase (TH)-positive fibers in the striatum. Moreover, SUN N8075 at 10 and 30 mg/kg (i.p., twice) had a similar protective effect against the MPTP-induced decrease in TH-positive cells in the substantia nigra. Further, SUN N8075 30 mg/kg (i.p. twice) markedly suppressed the MPTP-induced accumulation of 8-hydroxy-deoxyguanosine (8-OHdG) in the striatum. These findings indicate that SUN N8075 exerts protective effects, at least in part via an anti-oxidation mechanism, in these in vitro and in vivo models of Parkinson's disease.


Calcium-dependent lipolytic acyl-hydrolase activity in purified plant mitochondria.
Ageing of isolated potato mitochondria induced by CaCl2 resulted in rapid enzymatic hydrolysis of the membrane phospholipids with the liberation of free fatty acids. The enzyme responsible for this effect was identified as a membrane bound lipolytic acyl-hydrolase which was unmasked by CaCl2. The presence of this lipolytic acyl-hydrolase induced severe functional impairments in the mitochondrial oxidative and phosphorylative properties.


A re-examination of mitotic activity in the early chick embryo.
As a result of extensive mitotic index analysis in colchicine-arrested chick embryos during gastrulation, it was ascertained that the primitive streak is a region of elevated mitotic index as compared to the surrounding tissue. Along the cephalo-caudal axis, the embryo displays two large peaks of mitotic index, one at the posterior end of the primitive streak and the other just anterior to Hensen's node. The length of the various phases of the mitotic period was determined in vitro by time-lapse filming, and the colchicine-arrested mitotic indices in vivo and in vitro were determined and compared for various regions. Some observations regarding the orientation of mitotic spindles and abnormal mitosis in vitro are also included, and the relevance of the above observations to early embryonic development is discussed.


Liver fibrosis score predicts mortality in heart failure patients with preserved ejection fraction.
Heart failure with preserved ejection fraction (HFpEF) has several pathophysiological aspects, including stiffness and/or congestion of multiple organs. Poor prognosis is expected in heart failure patients with liver stiffness, which has recently been assessed by non-alcoholic fatty liver disease fibrosis score (NFS; based on aspartate aminotransferase to alanine aminotransferase ratio, platelet counts, and albumin). We aimed to investigate the impact of NFS on prognosis of HFpEF patients, with consideration for the peripheral collagen markers such as procollagen type III peptide (PIIIP), type IV collagen 7S, and hyaluronic acid. We performed a prospective observational study. Consecutive 492 hospitalized HFpEF patients were divided into four groups based on their NFS: first-fourth quartiles (n = 123). The fourth quartile group had the highest levels of PIIIP, type IV collagen 7S, hyaluronic acid, and B-type natriuretic peptide (P<0.001 each). In addition, there were significant positive correlations between PIIIP, type IV collagen 7S, hyaluronic acid, B-type natriuretic peptide, and NFS (P < 0.001 each). In the follow-up period (mean 1107 days), 93 deaths occurred. All-cause mortality increased in all four quartiles (8.1%, 12.2%, 23.6%, and 31.7%, P < 0.001). In the multivariable Cox proportional hazard analysis, NFS was an independent predictor of all-cause mortality in the HFpEF patients. NFS, a novel indicator of liver fibrosis, correlates with circulating systemic markers of fibrosis and congestion and is associated with higher all-cause mortality in HFpEF patients. NFS can be calculated simply and may be a useful tool to assess liver stiffness and prognosis in HFpEF patients.


Fibrinogen-fibrin system regulators from bloodsuckers.
Thrombin inhibitors from bloodsucking leeches and insects that block conversion of fibrinogen to fibrin are considered. Regulatory mechanisms influencing the fibrinogen-fibrin system in leeches include fibrinogen degradation, inhibition of factor XIIIa, and lysis of fibrin clots. The review also summarizes recent data on plasminogen activator from the vampire bat Desmodus rotundus and a role of fibrin as its cofactor.


Competition of tamoxifen with thyroxine for TBG binding: ligand binding assay and computational data.
Tamoxifen, a nonesteroidal antiesterogen, is widely used in the treatment of breast cancer. Recently, the effect of tamoxifen on thyroid function has caused considerable concern, yet the results of different studies are controversial and the precise mechanism of such influence is obscure. In view of the fact that some drugs such as furosemide, diclofenac and mefenamic acid, based on the structural similarities to thyroxine could compete for binding to thyroxine binding globulin (TBG) and appears that there are some structural similarities between tamoxifen and thyroxine, one can hypothesize that tamoxifen is also able to compete for TBG binding and thereby affecting thyroid function tests. In this study, we designed an in vitro binding assay as well as computational methods using MOPAC 7 package for evaluation of competitive potency of tamoxifen for TBG binding in comparison with well-known TBG competitors (including furosemide, mefenamic acid and diclofenac). The result of competition assay and Scatchard analysis revealed that tamoxifen does not bind to TBG at the T4 binding site, thus it is not a thyroxine competitor. Computational results also indicated that structural characteristics of tamoxifen are significantly different from those of T4 and its well-known competitors. In conclusion, the probability of competition between tamoxifen and T4 is ruled out by these results.


T lymphocyte lines induce autoimmune encephalomyelitis, delayed hypersensitivity and bystander encephalitis or arthritis.
Lines of rat T lymphocytes responsive to the basic protein of myelin (BP) or to the purified protein derivative of Mycobacterium tuberculosis (PPD) were inoculated i.v. into recipient rats. As reported previously, the anti-BP line cells, but not the anti-PPD line cells spontaneously accumulated in the central nervous system and caused encephalomyelitis. However, the anti-PPD line cells could be induced to enter the brain and cause bystander encephalitis by intracerebral inoculation of PPD. Anti-PPD or anti-BP line cells could mediate delayed-type hypersensitivity skin reactions or bystander arthritis elicited by specific antigen. The lines did not cause specific cytolysis in vitro. Susceptibility to delayed-type hypersensitivity or bystander disease was long lasting in rats inoculated with anti-PPD line cells, while rats inoculated with anti-BP line cells were susceptible for only a few days. Thus, lines of T lymphocytes can mediate a variety of pathological reactions directed by the presence of specific antigen, self or foreign.


The Sphygmomanometer Pain Test: A Simple Method for Identifying Patients at Risk of Excessive Pain after Total knee Arthroplasty.
Despite the overall success of total knee arthroplasty (TKA), a relatively large proportion of patients remain dissatisfied with the outcome. We hypothesized that patients with a lower threshold for pain were more likely to have worse outcomes after TKA. Forty-eight consecutive patients with primary knee osteoarthritis graded a standardized painful stimulus caused by inflating a sphygmomanometer placed on the forearm, on a visual analog scale (VAS) scale before their TKA. We compared the VAS scores to patients' Knee Society scores and Knee Society function scores 2 years after TKA. Patients with a severe VAS score (>74 mm) had significantly worse Knee Society scores compared to patients with mild (0-44 mm) and moderate (45-74 mm) VAS scores (55 ± 20.5 vs 81.5 ± 11.1 and 84.8 ± 13, respectively, P = .04) and worse Knee Society Function scores (34 ± 20.7 vs 75.2 ± 17.3 and 77 ± 17.4, respectively, P = .027) at 2 years after TKA. Patients with a lower threshold for pain, as determined by a standardized painful stimulus, are more likely to have lower Knee Society pain and function scores after TKA.


Exploring the potential antidepressant mechanisms of puerarin: Anti-inflammatory response via the gut-brain axis.
Puerarin has been shown to have a good antidepressant effect, and our previous study found that it can remedy stress-induced dysbiosis. However, its gut microbiota-related mechanism has not been fully elucidated. Therefore, this study aimed to investigate the potential link between puerarin on gut microbiota and inflammatory responses in depressed rats. A chronic unpredictable mild stress (CUMS) rat model of depression was established, open field test (OFT), sucrose preference test (SPT) and forced swimming test (FST) were used to evaluate its antidepressant effect. 16S rRNA sequencing was performed to identify the rat fecal microflora. At the same time, inflammatory cytokines, colonic histopathological changes, and brain-derived neurotrophic factor (BDNF), nuclear factor kappa-B (NF-κB), inhibitor a of NF-κB (IκB-α) protein expression were detected. Puerarin attenuated CUMS-induced depressive-like behavior and gut microbiota dysregulation in rats, significantly reducing the abundance of harmful bacteria such as Desulfovibrio, Verrucomicrobiae, and Verrucomicrobia. In addition, puerarin can also reduce the pro-inflammatory factors and increase the level of anti-inflammatory factors in depressed rats, improve the damaged colon tissue, enhance the expression of BDNF and IκB-α in the hippocampus and inhibit the expression of NF-κB. Direct evidence that puerarin improves depressive-like behaviors via gut microbiota is lacking. The underlying mechanism of puerarin's antidepressant-like effect is closely related to the bidirectional communication of the microbiota-gut-brain axis by regulating the inflammatory response.


Experiences with tick exposure, Lyme disease, and use of personal prevention methods for tick bites among members of the U.S. population, 2013-2015.
Consistent and effective use of personal prevention methods for tickborne diseases, including Lyme disease (LD), is dependent on risk awareness. To improve our understanding of the general U.S. population's experiences with tick exposure and use of personal prevention methods, we used data from ConsumerStyles, a web-based, nationally representative questionnaire on health-related topics. Questions addressed tick bites and LD diagnosis in the last year, use of personal prevention methods to prevent tick bites, and willingness to receive a theoretical LD vaccine. Of 10,551 participants surveyed over three years, 12.3 % reported a tick bite for themselves or a household member in the last year, including 15.4 % of participants in high LD incidence (LDI) states, 16.3 % in states neighboring high LDI states, and 9.4 % in low LDI states. Participants in high LDI states and neighboring states were most likely to use personal prevention methods, though 46.6 % of participants in high LDI states and 53.9 % in neighboring states reported not using any method. Participants in low LDI states, adults ≥ 75 years of age, those with higher incomes, and those living in urban housing tended to be less likely to practice personal prevention methods. Likeliness to receive a theoretical LD vaccine was high in high LDI (64.5 %), neighboring (52.5 %), and low LDI (49.7 %) states. Targeted educational efforts are needed to ensure those in high LDI and neighboring states, particularly older adults, are aware of their risk of LD and recommended personal prevention methods.


Early and long-term wear of 'fast-set' conventional glass-ionomer cements.
Fast setting high viscous glass-ionomer cements have been introduced to reach the set state sooner and to improve early mechanical properties. The aim of this study was to investigate whether 'fast-set' cements have improved the early three-body wear. The three-body wear of four glass-ionomer cements was measured after 1, 4, 7, 30, 90, and 365 days. The chemical composition of the cements was determined with EDAX. The particle size of the powders was determined with a Mastersizer 2000. The three-body wear of Fuji IX Fast was only slightly less than that of 'regular-set' Fuji IX, but Ketac Molar Quick showed a higher wear compared to the regular-set Ketac Molar. The wear rate of all four glass-ionomer cements decreased over a period of 1 year and could be described by second order reaction kinetics and depended on polyalkenoic acid content, chemical composition, and the size of the glass particles. Based on the results of this laboratory study on long-term wear, the glass-ionomer cements investigated may compete with composites, but the early wear characteristics should be further improved. Furthermore, a better understanding of the wear and reaction behavior of glass-ionomer cements is obtained.


Nedocromil sodium and sodium cromoglycate in patients aged over 50 years with asthma.
We conducted a double-blind parallel-group study to compare the efficacy of inhaled nedocromil sodium and inhaled sodium cromogylcate in patients aged 50 years or over with asthma. Seventy-seven patients were randomized, 38 to receive nedocromil sodium (4 mg q.d.s.) and 39 to receive sodium cromoglycate (10 mg q.d.s.) over a 16-week period. Eight patients withdrew from the study. Both patient groups showed a reduction in inhaled bronchodilator usage during the treatment period. No statistically significant differences were seen in diary card PEF recordings or symptom scores. Both clinic PEF and FEV1 were significantly greater (P < 0.05) in the sodium cromoglycate group at week 12. Patients considered the treatment to be very or moderately effective in 59% of the nedocromil sodium group and 50% of the sodium cromoglycate group. Thus, the study suggests that there is no difference in response to nedocromil sodium or sodium cromoglycate in this older group of patients with asthma who are already on moderate doses of inhaled steroids.


Plasmodium berghei oocysts possess fatty acid synthesis and scavenging routes.
Malaria parasites carry out fatty acid synthesis (FAS) in their apicoplast organelle via a bacterially related (type II) enzymatic pathway. In the vertebrate host, exoerythrocytic Plasmodium stages rely on FAS, whereas intraerythrocytic stages depend on scavenging FA from their environment. In the mosquito, P. falciparum oocysts express and rely on FAS enzymes for sporozoite formation, but P. yoelii oocysts do not express, nor depend on, FAS enzymes and thus rely on FA scavenging to support sporogony. In P. berghei, FAS enzymes are similarly expendable for sporogony, indicating it conforms to the P. yoelii scenario. We show here that P. berghei, unexpectedly, expresses FAS enzymes throughout oocyst development. These findings indicate that P. berghei can employ FAS alongside FA scavenging to maximise sporogony and transmission, and is more similar to P. falciparum than previously assumed with respect to FA acquisition by the oocyst. The ability of oocysts to switch between FAS and scavenging could be an important factor in the non-competitive relationship of resource exploitation between Plasmodium parasites and their mosquito vectors, which shapes parasite virulence both in the insect and vertebrate.


Pre-treatment of peripheral blood progenitor cells with macrophage inflammatory protein-1alpha induces prolonged survival of early progenitor cells over 6 weeks of long-term culture.
It has previously been shown that a combination of macrophage inflammatory protein-1alpha (MIP-1alpha) and interleukin (IL)-3 maintained human bone marrow (BM)-derived long-term culture-initiating cells (LTC-IC) for at least 8 weeks in vitro. We investigated colony- and cobblestone area-formation potential of peripheral blood progenitor cells (PBPC) at week 6 of long-term culture (LTC) in the absence of exogenous MIP-1alpha. but using cells which had been pre-incubated in the presence of MIP-1alpha for 40 h in liquid culture. The recovery of colony-forming cells (CFC) and cobblestone-area-forming cells (CAFC) after pre-incubation of PBPC with MIP-1alpha was up to threefold higher (P<0.05, n=5) than that of untreated controls. These results demonstrate that short-term pre-treatment of PBPC with MIP-1alpha induces long-lasting survival effects on early PBPC-derived progenitors in vitro.


Relationships between acrylamide and glycidamide hemoglobin adduct levels and allergy-related outcomes in general US population, NHANES 2005-2006.
Acrylamide-induced immunotoxicity and allergic dermatitis have been reported in animal experiments and clinical reports, respectively. However, epidemiological evidence from the general population is limited. The purpose of the present study was to estimate the associations between acrylamide exposure and allergy-related outcomes in the general US population. A total of 6982 subjects were selected from the National Health and Nutrition Examination Survey 2005-2006 (NHANES). Internal exposure was measured by the hemoglobin adducts of acrylamide (HbAA) and its metabolite glycidamide (HbGA). Allergy-related outcomes including asthma, hay fever, allergy, itchy rash, sneeze, wheeze and eczema were obtained by self-administered questionnaires. Allergic sensitization was assessed by the total immunoglobulin E (IgE) levels. The associations of HbAA and HbGA quartiles with allergy-related outcomes were calculated using logistic regression models with multivariable adjustments. Analyses were additionally stratified according to age, gender and serum cotinine levels. When setting quartile 1 of HbAA as reference, the odds ratios (ORs) [95% confidence intervals (CIs)] of quartile 2 to 4 for eczema were 1.18 (0.79-1.76), 1.14 (0.73-1.78) and 1.58 (1.14-2.18), respectively (ptrend = 0.002). Individuals at the highest quartile of HbGA had significantly elevated likelihoods of itchy rash (OR = 1.37, 95% CI = 1.02-1.83, ptrend = 0.032) and eczema (OR = 1.45, 95% CI = 1.06-1.97, ptrend = 0.044). The stratification analyses indicated various results in different subgroups. This study indicated significant associations between HbAA and HbGA levels and the likelihoods of allergy-related outcomes in the general US population, depending on age, gender and smoke exposure status. These findings suggested potential public health concerns for the widespread exposure to acrylamide.


Reminiscence therapy-based care program relieves post-stroke cognitive impairment, anxiety, and depression in acute ischemic stroke patients: a randomized, controlled study.
Cognitive and mental impairments are common health problems in acute ischemic stroke (AIS) patients. In this study, we aimed to assess the benefit of a reminiscence therapy-based care (RTBC) program on cognitive impairment restoration, anxiety, and depression reduction in AIS patients. Totally 130 AIS patients were recruited in this randomized, controlled study and randomly assigned to the RTBC group or control group in 1:1 ratio for 12-month intervention. Mini-Mental State Examination (MMSE), Montreal cognitive assessment (MoCA), Hospital Anxiety and Depression Scale for anxiety/depression (HADS-A/HADS-D), and Zung self-rating anxiety/depression scale (SAS/SDS) were assessed at month 0 (M0), M3, M6, M9, and M12. Meanwhile, patients' satisfaction was also evaluated at M3, M6, M9, and M12. RTBC increased MMSE score and MoCA score and reduced cognitive impairment patients' percentage assessed by MoCA score at M12 compared with control. RTBC reduced HADS-A score at M12, but not anxiety patients' percentage or severity by HADS-A at M12; besides, RTBC significantly lowered the SAS score at M9 and M12, and anxiety patients' percentage and severity by SAS at M12 compared with control. RTBC reduced HADS-D score at M9 and M12 (while statistically non-significant), but not depression patients' percentage or severity by HADS-D at M12; it decreased SDS score at M9 and M12, but not depression patients' percentage or severity by SDS at M12 compared with control. Additionally, RTBC obsessed higher patients' satisfaction at M3, M6, and M12 compared with control. RTBC could help reduce cognitive impairment, anxiety, and depression in post-stroke management for AIS patients.


A retrospective study on antibacterial treatments for koalas infected with Chlamydia pecorum.
Chlamydiosis remains the leading infectious disease and is one of the key factors responsible for the dramatic reduction of koala populations in South-East Queensland (SEQ) and New South Wales (NSW) regions of Australia. Possible infection outcomes include blindness, infertility, painful cystitis, and death if left untreated. Studies have reported the treatment efficacy of chloramphenicol and doxycycline, which are the two most commonly administered treatments in diseased koalas, in clinical settings. However, none have directly compared the treatment efficacy of these antibacterials on koala survival. A retrospective study was essential to identify any relationships between the demographical information, and the animals' responses to the current treatment regimens. Associations were explored between six explanatory (sex; maturity; location; clinical signs, treatment; treatment duration) and two outcome variables (survival; post-treatment PCR). Results showed that female koalas had a statistical trend of lower odds of surviving when compared to males (OR = 0.36, p = 0.05). Koalas treated with chloramphenicol for ≥ 28 days had greater odds of surviving than when treated for < 28 days (OR = 8.8, p = 0.02), and those koalas administered doxycycline had greater odds of testing PCR negative when compared to chloramphenicol treatments (OR = 5.45, p = 0.008). There was no difference between the antibacterial treatments (chloramphenicol, doxycycline, and mixed/other) and the survival of koalas. Female koalas had greater odds of exhibiting UGT signs only (OR = 4.86, p < 0.001), and also greater odds of having both ocular and UGT clinical signs (OR = 5.29, p < 0.001) when compared to males. Of the koalas, 28.5% initially had no clinical signs but were PCR positive for C. pecorum. This study enables further understanding of the complex nature between chlamydial infection and response to antibacterial treatment.


Iron influences the abundance of the iron regulatory protein Cir1 in the fungal pathogen Cryptococcus neoformans.
The GATA-type, zinc-finger protein Cir1 regulates iron uptake, iron homeostasis and virulence factor expression in the fungal pathogen Cryptococcus neoformans. The mechanisms by which Cir1 senses iron availability, although as yet undefined, are important for understanding the proliferation of the fungus in mammalian hosts. We investigated the influence of iron availability on Cir1 and found that the abundance of the protein decreases upon iron deprivation. This destabilization was influenced by reducing conditions and by inhibition of proteasome function. The combined data suggest a post-translational mechanism for the control of Cir1 abundance in response to iron and redox status.


Distribution of pre-pro-glucagon and glucagon-like peptide-1 receptor messenger RNAs in the rat central nervous system.
Glucagon-like peptide-1 (GLP-1) is derived from the peptide precursor pre-pro-glucagon (PPG) by enzymatic cleavage and acts via its receptor, glucagon-like peptide-1 receptor (GLP-1R). By using riboprobes complementary to PPG and GLP-1R, we described the distribution of PPG and GLP-1R messenger RNAs (mRNAs) in the central nervous system of the rat. PPG mRNA-expressing perikarya were restricted to the nucleus of the solitary tact or to the dorsal and ventral medulla and olfactory bulb. GLP-1R mRNA was detected in numerous brain regions, including the mitral cell layer of the olfactory bulb; temporal cortex; caudal hippocampus; lateral septum; amygdala; nucleus accumbens; ventral pallium; nucleus basalis Meynert; bed nucleus of the stria terminalis; preoptic area; paraventricular, supraoptic, arcuate, and dorsomedial nuclei of the hypothalamus; lateral habenula; zona incerta; substantia innominata; posterior thalamic nuclei; ventral tegmental area; dorsal tegmental, posterodorsal tegmental, and interpeduncular nuclei; substantia nigra, central gray; raphe nuclei; parabrachial nuclei; locus ceruleus, nucleus of the solitary tract; area postrema; dorsal nucleus of the vagus; lateral reticular nucleus; and spinal cord. These studies, in addition to describing the sites of GLP-1 and GLP-1R synthesis, suggest that the efferent connections from the nucleus of the solitary tract are more widespread than previously reported. Although the current role of GLP-1 in regulating neuronal physiology is not known, these studies provide detailed information about the sites of GLP-1 synthesis and potential sites of action, an important first step in evaluating the function of GLP-1 in the brain. The widespread distribution of GLP-1R mRNA-containing cells strongly suggests that GLP-1 not only functions as a satiety factor but also acts as a neurotransmitter or neuromodulator in anatomically and functionally distinct areas of the central nervous system.


Community-based HIV testing through a general health check event in a high HIV-prevalent multicultural area in Rotterdam, The Netherlands: a pilot study on feasibility and acceptance.
HIV testing is crucial for finding the remaining cases in a declining HIV epidemic in The Netherlands; providing HIV testing in non-traditional settings may be warranted. We conducted a pilot study to determine the feasibility and acceptability of a community-based HIV testing (CBHT) approach with general health checks to improve HIV test uptake. CBHT's main conditions were low-threshold, free-of-charge, general health check, and HIV education. We interviewed 6 community leaders, 25 residents, and 12 professionals/volunteers from local organizations to outline these main conditions. Walk-in test events were piloted at community organizations, providing HIV testing along with body mass index (BMI), blood pressure, blood glucose screening, and HIV education (October 2019 to February 2020). Demographics, HIV testing history, risk perception, and sexual contact were collected via questionnaires. To evaluate the pilots' feasibility and acceptance, we utilized the RE-AIM framework and predefined goals, incorporating quantitative data from the test events and qualitative input from participants, organizations, and staff. A total of 140 individuals participated (74% women, 85% non-Western, median age 49 years old). The number of participants during the seven 4-h test events ranged from 10 to 31. We tested 134 participants for HIV, and one was found positive (positivity 0.75%). Almost 90% of the participants were never tested or > 1 year ago, and 90% perceived no HIV risk. One-third of the participants had one or more abnormal test results on BMI, blood pressure, or blood glucose. The pilot was well-rated and accepted by all parties. The staff had concerns about waiting time, language problems, and privacy. Participants hardly indicated these concerns. This CBHT approach is feasible, acceptable, and well-suited for testing not (recently) tested individuals and detecting new cases. Besides reducing HIV-associated stigma and increasing HIV test acceptance, offering multiple health tests may be appropriate as we frequently observed multiple health problems. Whether this laborious approach is sustainable in the micro-elimination of HIV and should be deployed on a large scale is questionable. CBHT like ours may be suitable as a supplement to more sustainable and cost-effective methods, e.g., proactive HIV testing by general practitioners and partner notification.


Effect of herd management on the contamination of night holding areas (correos) and infections with gastrointestinal nematodes of N'Dama cattle in The Gambia.
The densities of infective nematode larvae (L3/m2) in the night holding places (locally called "correos") of 2 traditionally kept N'Dama herds were estimated at weekly intervals throughout an entire rainy season. Herd 1 moved correos every 3 weeks whereas herd 2 remained in the same area for most of the rainy season. Removal to a new correo was invariably accompanied by a drastic drop of L3/m2. Conversely, L3/m2 increased rapidly up to values of more than 1,000 when the herds used the same night holding place for more than 3 weeks. Calves kept in herds with frequent changes of the correo showed significantly lower nematode egg counts and higher growth rates during the rainy season, combined with a reduced weight loss during the following dry season. The results of this study indicate that a regular frequent change of the correo is an effective method of reducing nematode infection risk and increasing calf growth and that this might be a sustainable part of an integrated strategic programme to control gastrointestinal nematode infections wherever correos are used.


Maternal physical activity and sedentary behaviour before and during in vitro fertilization treatment: a longitudinal study exploring the associations with controlled ovarian stimulation and pregnancy outcomes.
To evaluate the association of objectively measured physical activity (PA) and sedentary behaviour before and during in vitro fertilization (IVF) with controlled ovarian stimulation (COS) and pregnancy outcomes. This longitudinal study involved 107 infertile women undergoing IVF treatment. PA and sedentary behaviour were measured for 14 consecutive days using accelerometry as follows: (1) before IVF treatment, (2) during IVF at the implantation time, immediately after embryo transfer, and (3) after positive pregnancy test. Total screen time was assessed by questionnaires. COS results were measured as the number of oocytes and embryos obtained, and the study outcomes included positive hCG, clinical pregnancy, and live birth. Compared with baseline activity levels, women significantly reduced their PA and increased sedentary behaviour during IVF (p ≤ 0.001). Higher average PA, light PA, and ratio between breaks in every ≥ 30-min blocks of sedentary time showed positive associations, while sedentary time, number, and time accumulated in blocks of ≥ 30 min of sedentary time associated negatively with oocyte and embryo counts (all p < 0.05). Women with high total screen time during non-work days (≥ 7 h) obtained 4.7 oocytes (p = 0.005) and 2.8 embryos (p = 0.008) less in COS. PA and sedentary behaviour before and during IVF did not affect the positive hCG, clinical pregnancy, and live birth outcomes. Our study results suggest that higher time spent in PA and lower time spent in sedentary behaviour before entering assisted reproduction is associated with better COS outcomes, while activity levels before and during IVF do not affect the implantation, pregnancy, and live birth outcomes.


Development of Emulgel Delivery of Mupirocin for Treatment of Skin Infection.
To design controlled release topical delivery of mupirocin for the treatment of skin infection. Mupirocin is an antibacterial drug. Mupirocin works to kill the bacteria, which include strains of Staphylococcus aureus and Streptococcus pyogenes. It is also used for the treatment of inflammation of a hair follicle. The half-life of mupirocin is only 20-40 min. It has very slight solubility in water. Patent literature had shown work on ointment, antibiotic composition, nasal and topical composition. Emulgel is a duel control release system for the topical delivery of hydrophobic drugs. The objective was to formulate emulgel with controlled delivery of mupirocin using Sepineo P 600. Soya oil, tween 80 and polyethylene glycol 400 (Oil:Surfactant:Cosurfactant) were used for emulsion formulation. Emulgel was optimized by 32 factorial design. Sepineo P 600 and hydroxy propyl methyl cellulose K4M were used as independent variables. Drug excipient compatibility analysis was carried out by FTIR, UV and DSC spectra. Emulgel was evaluated for its physical characterization, in vitro release, ex vivo release, antimicrobial and anti-inflammatory study. DSC, UV and FTIR analysis confirmed drug excipient compatibility. FE SEM showed a size range between 228-255 nm. Zeta potential was found to be -25.1 mV, which showed good stability of the emulsion. Design expert software showed F2 as an optimized batch. Release studies indicated that the controlled release of drugs forms Sepineo P 600 gel due to its higher gelling capacity. Batch F2 showed comparable results with marketed formulation against Staphylococcus aureus. For batch F2, 40 μg/ml was the minimal inhibitory concentration. Antimicrobial and anti-inflammatory study proved successful development of stably controlled release mupirocin emulgel.


Not all inflammation in the right lower quadrant is appendicitis: a case report of Escherichia coli O157:H7 with a review of the literature.
Although significant work has been presented on this subject in pediatric, infectious disease, and epidemiologic literature, there is a noteworthy lack of information on Escherichia coli O157:H7 in any surgical journals. As this disease can present with signs and symptoms often ascribed to the acute abdomen, it is imperative that the general surgeon, pediatric surgeon, and colorectal surgeon are all familiar with this infection and its clinical ramifications. A case report followed by a review of the literature is presented.


Survival-related DNA repair phenomena in cultured rat-kangaroo cells.
Cultured cells (line PtK-2) from the marsupial mammal rat-kangaroo, or potoroo (Potorous tridactylis), which photoreactivate (PR) both UV-induced dimers and lethality, excise few dimers, and are only slightly sensitized by post-UV exposure to caffeine, were subjected to caffeine and hydroxyurea (HU) treatments during the 30-min PR period. It was found that neither caffeine nor HU inhibited PR of lethality as measured by colony-forming ability. Further, the cells exhibited no photoprotective properties and 3 mM caffeine potentiated the same slight survival decrease in both photoreactivated and unphotoreactivated cells. It is evident that caffeine does not inhibit PR or the survival-related dark repair systems to any great extent, and hence the caffeine-sensitive post-UV dark repair found in this (and possibly other) mammalian cell lines may not be related directly to survival-dependent pyrimidine dimer removal, but instead to lesions or repair processes other than, but not excluding, pyrimidine dimers.


Characterization of water intake and water efficiency in beef cattle1,2.
In the future, water may not be as readily available due to increases in competition from a growing human population, wildlife, and other agricultural sectors, making selection for water efficiency of beef cattle increasingly important. Substantial selection emphasis has recently been placed on feed efficiency in an effort to reduce production costs, but no emphasis has been placed on making cattle more water efficient due to lack of data. Thus, the objective of this study was to calculate water efficiency metrics for cattle and evaluate their relationship to growth, feed intake (FI), and feed efficiency. Individual daily FI and water intake (WI) records were collected on 578 crossbred steers over a 70-d test period. Animals with low water intake ate less feed, had lower gains, and were more water efficient (as defined by water to gain ratio, W/G, and residual water intake, RWI). However, the amount of water consumed by animals had minimal phenotypic relationship with feed efficiency (residual feed intake [RFI], R2 = 0.1050 and feed to gain ratio (F/G) ratio R2 = 0.0726). Cattle that had low DMI consumed less water, had lower gains, had lower RFI, and had higher F/G. The level of feed consumed had minimal relationship with water efficiency. WI, W/G, RWI, and ADG had moderate heritability estimates of 0.39, 0.39, 0.37, and 0.37, respectively. High heritability estimates were observed for DMI and RFI (0.67 and 0.65, respectively). Feed to gain had a low heritability estimate of 0.16. WI had a strong positive genetic correlation with W/G (0.99) and RWI (0.88), thus selecting for decreased WI should also make cattle more water efficient. The genetic correlation between WI and ADG was 0.05; thus, selecting for low WI cattle should have little effect on growth. There is a low to moderate genetic correlation between WI and DMI (0.34). RWI has a positive genetic correlation with W/G ratio (0.89) and F/G ratio (0.42) and is negatively genetically correlated with RFI (-0.57). Water to gain and F/G had a strong positive genetic correlation (0.68). RFI has a positive genetic correlation with W/G ratio (0.37) and F/G (0.88). Minimal antagonisms seem to be present between WI and ADG, although it should be noted that standard errors were large and often not significantly different from zero due to the small sample size. However, care should be taken to ensure that unintended changes do not occur in DMI or other production traits and incorporation of WI into a selection index would likely prove to be the most effective method for selection.


Cardiac-targeted transgenic mutant mitochondrial enzymes: mtDNA defects, antiretroviral toxicity and cardiomyopathy.
Mitochondrial (mt) DNA biogenesis is critical to cardiac contractility. DNA polymerase gamma (Pol gamma) replicates mtDNA, whereas thymidine kinase 2 (TK2) monophosphorylates pyrimidines intramitochondrially. Point mutations in POLG and TK2 result in clinical diseases associated with mtDNA depletion and organ dysfunction. Pyrimidine analogs (NRTIs) inhibit Pol gamma and mtDNA replication. Cardiac "dominant negative" murine transgenes (TGs; Pol gamma Y955C, and TK2 H121N or I212N) defined the role of each in the heart. mtDNA abundance, histopathological features, histochemistry, mitochondrial protein abundance, morphometry, and echocardiography were determined for TGs in "2 x 2" studies with or without pyrimidine analogs. Cardiac mtDNA abundance decreased in Y955C TGs ( approximately 50%) but increased in H121N and I212N TGs (20-70%). Succinate dehydrogenase (SDH) increased in hearts of all mutants. Ultrastructural changes occurred in Y955C and H121N TGs. Histopathology demonstrated hypertrophy in H121N, LV dilation in I212N, and both hypertrophy and dilation in Y955C TGs. Antiretrovirals increased LV mass ( approximately 50%) for all three TGs which combined with dilation indicates cardiomyopathy. Taken together, these studies demonstrate three manifestations of cardiac dysfunction that depend on the nature of the specific mutation and antiretroviral treatment. Mutations in genes for mtDNA biogenesis increase risk for defective mtDNA replication, leading to LV hypertrophy.


Identifying the beta-site amyloid precursor protein cleaving enzyme 1 interactome through the proximity-dependent biotin identification assay.
Beta-site amyloid precursor protein (APP) cleaving enzyme 1 (BACE1) is a key drug target against Alzheimer's Disease however, due to its promiscuous proteolytic activity, little is known about its physiological functions. Previous studies have analysed BACE1 cleavage products to examine BACE1 interactions and determine substrates, but these studies cannot establish non-enzymatic (and potentially functional) associations. This study used the biotin identification proximity assay to establish the BACE1 interactome in healthy neuronal cells and identified interactions involved in BACE1 trafficking, post-translational modification and substrates. Furthermore, this method has identified a putative novel role for BACE1 in sex hormone signalling and haem regulation through interaction with the progesterone receptor membrane component 2 (PGRC2). Data are available via ProteomeXchange with identifier PXD021464.


Time-course mass spectrometry data of adipose mesenchymal stem cells acquiring chondrogenic phenotype.
Rabbit adipose mesenchymal stem cells were used for the purpose of studying acquisition of the chondrogenic phenotype over time at 1, 14 and 28 days after in vitro incubation with differentiation media, using nano-liquid chromatography electrospray ionization tandem mass spectrometry analysis. This was part of a preliminary study of the behavior of differentiated adipose stem cells for use in a rabbit model of laryngoplasty. The data comprise .MGF, .RAW, .MZID, and .XLSX, lists of peaks, peptides and proteins identified by nano-flow liquid chromatography electrospray ionization tandem mass spectrometry analysis upon incubation with non-differentiating (ND) or chondrogenic differentiating (CHD) media (ProteomeXchange ID PXD010236). XLSX files contain the following information: day 1 CT (control, N = 3499 proteins), day 14 ND (N = 3106 proteins), day 28 ND (N = 3116 proteins), day 14 CHD (N = 2901 proteins), and day 28 CHD (N = 2876 proteins). Proteins are characterized with respect to their - 10lgP value, percent coverage, number of total as well as unique peptides after trypsin digestion, derivatization method (carbamidomethylation, oxidation, or combined carbamidomethylation + oxidation), average mass, and include a full description.


Characterization of 137 Genomic DNA Reference Materials for 28 Pharmacogenetic Genes: A GeT-RM Collaborative Project.
Pharmacogenetic testing is increasingly available from clinical laboratories. However, only a limited number of quality control and other reference materials are currently available to support clinical testing. To address this need, the Centers for Disease Control and Prevention-based Genetic Testing Reference Material Coordination Program, in collaboration with members of the pharmacogenetic testing community and the Coriell Cell Repositories, has characterized 137 genomic DNA samples for 28 genes commonly genotyped by pharmacogenetic testing assays (CYP1A1, CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4, CYP3A5, CYP4F2, DPYD, GSTM1, GSTP1, GSTT1, NAT1, NAT2, SLC15A2, SLC22A2, SLCO1B1, SLCO2B1, TPMT, UGT1A1, UGT2B7, UGT2B15, UGT2B17, and VKORC1). One hundred thirty-seven Coriell cell lines were selected based on ethnic diversity and partial genotype characterization from earlier testing. DNA samples were coded and distributed to volunteer testing laboratories for targeted genotyping using a number of commercially available and laboratory developed tests. Through consensus verification, we confirmed the presence of at least 108 variant pharmacogenetic alleles. These samples are also being characterized by other pharmacogenetic assays, including next-generation sequencing, which will be reported separately. Genotyping results were consistent among laboratories, with most differences in allele assignments attributed to assay design and variability in reported allele nomenclature, particularly for CYP2D6, UGT1A1, and VKORC1. These publicly available samples will help ensure the accuracy of pharmacogenetic testing.


Kartogenin-enhanced dynamic hydrogel ameliorates intervertebral disc degeneration via restoration of local redox homeostasis.
Over-activation of oxidative stress due to impaired antioxidant functions in nucleus pulpous (NP) has been identified as a key factor contributing to intervertebral disc degeneration (IVDD). While Kartogenin (KGN) has previously demonstrated antioxidant properties on articular cartilage against osteoarthritis, its effects on NP degeneration have yet to be fully understood. This study aimed to investigate the protective effects of KGN on nucleus pulpous cells (NPCs) against an inflammatory environment induced by interleukin (IL)-1β, as well as to explore the therapeutic potential of KGN-enhanced dynamic hydrogel in preventing IVDD. NPCs were isolated from rat caudal IVDs and subjected to treatment with KGN at varying concentrations (ranging from 0.01 to 1 μM) in the presence of IL-1β. The expression of extracellular matrix (ECM) anabolism markers was quantitatively assessed at both the mRNA and protein levels. Additionally, intracellular reactive oxygen species and antioxidant enzyme expression were evaluated, along with the role of nuclear factor erythroid 2-related factor 2 (NRF2). Based on these findings, a dynamic self-healing hydrogel loaded with KGN was developed through interconnecting networks. Subsequently, KGN-enhanced dynamic hydrogel was administered into rat caudal IVDs that had undergone puncture injury, followed by radiographic analysis and immunohistochemical staining to evaluate the therapeutic efficacy. In vitro treatments utilizing KGN were observed to maintain ECM synthesis and inhibit catabolic activities in IL-1β-stimulated NPCs. The mechanism behind this protective effect of KGN on NPCs was found to involve the asctivation of NRF2 and downstream antioxidant enzymes, including glutathione peroxidase 1 and heme oxygenase 1. This was further supported by the loss of both antioxidant and anabolic effects upon pharmacological inhibition of NRF2. Furthermore, a self-healing hydrogel was developed and loaded with KGN to achieve localized and sustained release of the compound. The injection of KGN-enhanced hydrogel effectively ameliorated the degradation of NP ECM and mitigated inflammation in a rat model of puncture-induced IVDD. Our results indicate that KGN exhibits potential as a therapeutic agent for NP degeneration, and that KGN-enhanced dynamic hydrogel represents a novel approach for treating IVDD by restoring redox homeostasis in NP.The translational potential of this article: The dysregulation of oxidant and antioxidant balance has been shown to impede the repair and regeneration of NP, thereby hastening the progression of IVDD following injury. The present investigation has demonstrated that the sustained release of KGN promotes the synthesis of ECM in vitro and mitigates the progression of IVDD in vivo by restoring redox equilibrium, thereby presenting a novel therapeutic candidate based on the antioxidant properties of KGN for the treatment of IVDD.


Relapses or reinfections: analysis of a case of Clostridium difficile-associated colitis by two typing systems.
Immunoblotting and pulsed-field gel electrophoresis of Clostridium difficile isolates were employed to differentiate reinfection by a newly acquired strain from relapse by an original strain in a 10-year-old patient with four episodes of C. difficile-associated colitis. Immunoblot typing demonstrated subserogroup K-1 of serogroup K for the first and second organisms, subserogroup A-1 of serogroup A for the third organism, and subserogroup G-4 of serogroup G for the fourth organism. PFGE analysis revealed consistent results with immunoblot analysis except that the strains from the fourth episode, whose DNA constantly degraded, were nontypable by this method. Five separate isolates of C. difficile from a specimen of each episode showed identical PFGE patterns, indicating that infections of multiple strains probably did not occur in this patient. These typing results suggested that the second episode after a 17-day course of vancomycin therapy represented a relapse by the strain causing the first episode, and that the third and fourth episodes after tapering vancomycin therapy were reinfections by other strains. Both immunoblot and PFGE typing systems are promising tools for analyzing recurrence of C. difficile infection.


Monitoring micropollutants in marine waters, can quality standards be met?
The environmental risks of 33 micropollutants occurring in Belgian coastal zone were assessed as single-substances and as mixtures. Water and sediment samples were taken in harbors, coastal waters and the Scheldt estuary during 2007-2009. Measured environmental concentrations were compared to quality standards such as Predicted No Effect Concentrations (PNECs), Environmental Quality Standards (EQSs), and Ecotoxicological Assessment Criteria (EAC). Out of a total of 2547 samples analyzed, 232 and 126 samples exceeded the EQS and EAC, respectively. Highest risks were observed for TBT, PBDEs, PCBs and the PAHs anthracene, indeno(1,2,3-cd)pyrene, benzo(g,h,i)perylene, benzo(k)fluoranthene, and benzo(b)fluoranthene in the water compartment and for TBT and PCBs in the sediment compartment. Samples taken at all stations during the April 2008 campaign indicate a potential risk of the contaminant mixtures to the aquatic environment (except W06 station). This study argues the need to revise quality standards when appropriate and hence the overall regulatory implication of these standards.


Content of Zinc and Copper in Selected Plants Growing Along a Motorway.
In 2011 a study was carried out analyzing the effects of road traffic on bioaccumulation of zinc and copper in selected species of dicotyledonous plants growing on adjacent grasslands. To do the research the plants were sampled from the 9-km-long Siedlce bypass, a part of the international route E-30. They were collected during the flowering stage, at following distances from the road: 1, 5, 10, 15 m. The content of zinc and copper was determined with the AAS method, with dry mineralisation done before. The highest concentration of the elements, regardless of the distance from the road, was found in Taraxacum spec. Among the tested plants, the lowest zinc content was in Vicia cracca, and the lowest copper content in Rumex acetosa. The limit for copper content was exceeded in Taraxacum spec. and, slightly, in Achillea millefolium growing at the roadside, closest to the roadway.


Nonbenzamidine compounds as selective factor Xa inhibitors.
Nonbenzamidine compounds (imidazole, pyridine, pyrimidine, and thiazole derivatives) as selective serine protease factor Xa inhibitors are discussed.


Bronchoalveolar lavage and clearance of 99m-Tc-DTPA in asbestos workers without evidence of asbestosis.
We performed BAL and measured the clearance of 99m-Tc-DTPA in 20 non-smoking subjects (mean age 50, range 36-68 years) occupationally exposed to asbestos (mean duration 14, range 3-30 years). All had normal lung function and none had clinical or radiological evidence of asbestosis. The mean BAL results were: total cells per ml 737 X 10(3) (360-1210), percentage macrophages 79 (49-96), percentage lymphocytes 13 (1-42), percentage neutrophils 8 (1-40), percentage eosinophils 0 (0-3), asbestos bodies per ml 83 (0-550). Eight subjects showed increased percentage of lymphocytes and four others showed increased percentages of neutrophils when compared with normal ranges in our laboratory. Higher percentages of neutrophils correlated with longer duration of exposure to asbestos (r = 0.54, P less than 0.025), and shorter time since last exposure to asbestos (r = -0.54, P less than 0.025). Four subjects showed faster clearance of 99m-Tc-DTPA than was observed in 31 normal non-smoking control subjects. There was a tendency for faster solute clearance to be associated with greater numbers of BAL macrophages (r = -0.39, P less than 0.10) but there were no significant relationships between solute clearance and other BAL variables. BAL profiles in asbestos workers may be abnormal in the absence of clinical or radiological evidence of asbestosis.