Production of human antibodies by in vitro immunization using a fusion protein containing the transcriptional transactivator of HIV-1.
Antigen-specific activation of human B cells represents a key step for the production of monoclonal antibodies. Several approaches have been developed over the last thirty years in order to improve the process of lymphocyte activation in vitro. In the present study, we investigated whether the transcriptional transactivator (Tat) of human immunodeficiency virus, which possesses numerous biological activities, is able to trigger antibody secretion when incubated with human peripheral blood mononuclear cells. No such effect was observed when using Tat as a free protein. However, we found a significant IgM antibody production when Tat was previously fused to a double domain, called ZZ, derived from protein A of Staphylococcus aureus. The effect was also observed when the fusion protein, called ZZTat101, was incubated with purified B cells, indicating that the phenomenon does not require T-cell help. Antibody secretion was observed in the absence of cytokines that are usually used during in vitro immunization experiments, indicating that ZZTat101 provides the signals required for the initiation of the immune response. Antibody secretion was observed using a ZZTat mutant, containing only the Tat residues 22 to 57, called ZZTat22-57, indicating that this region is sufficient to initiate the immune response. In contrast, the effect was not found with a ZZTat22-57 mutant devoid of the seven Tat cysteines located between residues 22 and 37, demonstrating that these residues play a crucial role in the phenomenon. Our results pave the way to the development of a new in vitro immunization method based on antigens associated with ZZTat.


Are obese children truly unfit? Minimizing the confounding effect of body size on the exercise response.
To test the hypothesis that obese children are unfit (i.e., have abnormal responses to exercise testing consistent with reduced levels of habitual physical activity), we used new analytic strategies in studies of 18 obese children performing cycle ergometry. The subject's weight (mean +/- SD) was 168 +/- 24% that predicted by height, and the age range was 9 to 17 years. Size-independent measures of exercise (e.g., the ratio of oxygen uptake (VO2) to work rate during progressive exercise and the temporal response of VO2, carbon dioxide output (VCO2), and minute ventilation (VE) at the onset of exercise) were used. The ability to perform external mechanical work was corrected for VO2 at unloaded pedaling (change in maximum oxygen uptake (delta VO2max) and in anaerobic threshold (delta AT). On average, obese children's responses were in the normal range: delta VO2max, 104 +/- 41% (+/- SD) predicted (by age); delta AT, 85 +/- 51%; ratio of change in VE to change in VCO2, 111 +/- 21% and ratio of change in VO2 to change in work rate, 100 +/- 24%, but six of the obese children had values of delta VO2max or delta AT that were more than 2 SD below normal. In addition, obese children did not have increased delta VO2max or delta AT with age as observed in nonobese children. Although the response time of VO2 was normal (99 +/- 32% of predicted), those for both VCO2 and VE were prolonged. We conclude that the finding of obesity in a child is not a reliable indicator of poor fitness but that testing cardiorespiratory responses to exercise can be used to identify subjects with serious impairment and to individualize therapy.


Antitumor effects of recombinant human tumor necrosis factor against human tumor xenografts transplanted into nude mice.
Antitumor activities of recombinant human tumor necrosis factor (rH-TNF) against human tumor xenografts in nude mice were studied. Thirteen human tumor xenografts serially transplanted into nude mice were used for experiments; five gastric, two breast, two gallbladder, one colon and one esophageal carcinoma, one liposarcoma and one squamous carcinoma of the neck. They were inoculated into the subcutaneous tissue of BALB/c nu/nu nude mice and the treatment was started when the estimated tumor weight reached 100-300 mg. rH-TNF was administered intratumorally at schedule of qd X 5 or q3d X 5. rH-TNF showed a marked antitumor activity against various human tumors. The hemorrhagic necrosis was observed in all types of the human tumor xenografts (100 per cent), and the complete regression of the tumor was noted in 4 of 11 tumors (36.4 per cent). On the contrary, intraperitoneal rH-TNF exhibited little antitumor effect. The additive effect in the combination of TNF and Mitomycin C was observed against two Mitomycin C resistant gastric tumors.


Erythropoiesis in vitro. III. The role of potassium ions in erythroid colony formation.
The role of potassium as an essential promotor of erythroid progenitor growth (BFU-e & CFU-e) from normal murine hematopoietic tissues was studied. Dialyzed fetal calf serum, over a wide range of concentrations, was shown to reduce the numbers of BFU-e and CFU-e that could be cultured from normal murine bone marrow. A dose-dependent addition of 1 M KC1 restored erythroid progenitor colony growth to the levels generally seen when normal, non-dialyzed fetal calf serum was used. Furthermore, when [K] was increased in some human urinary and sheep plasma erythropoietin preparations, the number of erythroid progenitor cells cultured also increased. This influence is crucial to the differentiation of committed stem cells into the erythroid pathway and must therefore be considered in the development of serum-free growth media.


Gender differences in cardiothoracic surgery letters of recommendation.
To investigate whether or not gender influences letters of recommendation for cardiothoracic surgery fellowship. From applications to an Accreditation Council Graduate Medical Education cardiothoracic surgery fellowship program between 2016 and 2021, applicant and author characteristics were examined with descriptive statistics, analysis of variance, and Pearson χ2 tests. Linguistic software was used to assess communication differences in letters of recommendation, stratified by author and applicant gender. An additional higher-level analysis was then performed using a generalized estimating equations model to examine linguistic differences among author-applicant gender pairs. Seven hundred thirty-nine recommendation letters extracted from 196 individual applications were analyzed; 90% (n = 665) of authors were men and 55.8% (n = 412) of authors were cardiothoracic surgeons. Compared with women authors, authors who are men wrote more authentic (P = .01) and informal (P = .03) recommendation letters. When writing for women applicants, authors who are men were more likely to display their own leadership and status (P = .03) and discuss women applicants' social affiliations (P = .01), like occupation of applicant's father or husband. Women authors wrote longer letters (P = .03) and discussed applicants' work (P = .01) more often than authors who are men. They also mentioned leisure activities (P = .03) more often when writing for women applicants. Our work identifies gender-specific differences in letters of recommendation. Women applicants may be disadvantaged because their recommendation letters are significantly more likely to focus on their social ties, leisure activities, and the status of the letter writer. Author and reviewer awareness of gender-biased use of language will aid in improvements to the candidate selection process.


Application of an open-chamber multi-channel microfluidic device to test chemotherapy drugs.
The use of precision medicine for chemotherapy requires the individualization of the therapeutic regimen for each patient. This approach improves treatment efficacy and reduces the probability of administering ineffective drugs. To ensure accurate decision-making in a timely manner, anticancer drug efficacy tests must be performed within a short timeframe using a small number of cancer cells. These requirements can be satisfied via microfluidics-based drug screening platforms, which are composed of complex fluidic channels and closed systems. Owing to their complexity, skilled manipulation is required. In this study, we developed a microfluidic platform, to accurately perform multiple drug efficacy tests using a small number of cells, which can be conducted via simple manipulation. As it is a small, open-chamber system, a minimal number of cells could be loaded through simple pipetting. Furthermore, the extracellular matrix gel inside the chamber provides an in vivo-like environment that enables the localized delivery of the drugs to spontaneously diffuse from the channels underneath the chamber without a pump, thereby efficiently and robustly testing the efficacy and resistance of multiple drugs. We demonstrated that this platform enabled the rapid and facile testing of multiple drugs using a small number of cells (~ 10,000) over a short period of time (~ 2 days). These results provide the possibility of using this powerful platform for selecting therapeutic medication, developing new drugs, and delivering personalized medicine to patients.


Validity of Commercial Activity Trackers in Children With Congenital Heart Disease.
Increasing physical activity levels is a high priority to optimize long-term health in children with congenital heart disease (CHD). Commercial activity trackers have been validated in adults and are increasingly used to measure and promote physical activity in pediatric populations, but they have not been validated in children. In 30 children with CHD aged 10-18 years, we assessed the validity of physical activity form the wrist-based Fitbit Charge HR (Fitbit, San Francisco, CA) against hip-based ActiGraph (ActiGraph LLC, Pensacola, FL) accelerometers under free living conditions for 7 days. We assessed the association between devices using intraclass correlation coefficients (ICCs) and Bland-Altman plots. Receiver operating curves were used to identify Fitbit step cut points. There was a strong association between the 2 devices for daily steps across 138 analyzed person-days (ICC = 0.855; P < 0.001), but poorer agreement for time spent in physical activity intensities (ICCs < 0.7). Daily Fitbit steps of ≥ 12,500 identified meeting physical activity guidelines defined as ≥ 60 minutes of moderate-to-vigorous physical activity per day. Fitbit devices recorded more steps than accelerometers (-2242 steps per day, 95% limits of agreement of -7738 to 3253). Between-device differences were greater in boys vs girls. Fitbit devices were worn for longer than accelerometers (-36 minutes per day, 95% limits of agreement, -334 to 261), but overall differences in wear time explained little of the variance in step differences (7%, P = 0.048). Commercial activity trackers provide opportunities to remotely monitor physical activity in children with CHD, but absolute values might differ from accelerometers. These findings are important because of the increasing emphasis on physical activity promotion and monitoring in children with cardiovascular risk factors.


Forced expiratory flows and volumes in a Swedish cohort of healthy term infants.
The use of pulmonary function tests (PFTs) in infants has increased during the last decades, making the need for equipment- and ethnic-specific reference data mandatory for appropriate interpretation of the results. Our aim was to investigate how well the already published reference equations for infant spirometry fit a healthy population of Swedish infants. We performed forced tidal and raised volume expiratory maneuvers in healthy infants using Jaeger BabyBody equipment. PFT data were collected from 91 healthy infants aged between 3 months to 2 years at 143 occasions. Mean (standard deviation) z-scores were 0.68(1.33) for maximal flow at functional residual capacity (V'max FRC), -0.15(0.96) for forced vital capacity (FVC), 0.40(1.33) for the forced expired volume in the initial 0.5 seconds (FEV0.5 ) and 0.52(0.93) for the ratio FEV0.5 /FVC, respectively. Z-scores for all indices but FEV0.5 /FVC were highly dependent on length. We have shown that the use of previously published reference equations may result in an age-related misinterpretation of lung function measure in a Swedish infant population.


Automation of protein assays with a microcomputer interfaced to a spectrophotometer.
We have developed a system for semi-automatic protein quantitation assays based on an inexpensive, general-purpose microcomputer. A commercial curve fitting program was modified for direct data input from the spectrophotometer via an analog to digital converter. The software provides specific prompts for protein assays resulting in a rapid and user friendly, semi-automated system. A listing of the protein assay results are printed, or stored to a magnetic disk, along with a graphic representation of the standard curve.


Loss of TRIM24 promotes IL-10 expression via CBP/p300-dependent IFNβ1 transcription during macrophage activation.
As an anti-inflammatory cytokine, interleukin 10 (IL-10) plays a vital role in preventing inflammatory and autoimmune pathologies while also maintaining immune homeostasis. IL-10 production in macrophages is tightly regulated by multiple pathways. TRIM24, a member of the Transcriptional Intermediary Factor 1 (TIF1) family, contributes to antiviral immunity and macrophage M2 polarization. However, the role of TRIM24 in regulating IL-10 expression and its involvement in endotoxic shock remains unclear. In vitro, bone marrow derived macrophages cultured with GM-CSF or M-CSF were stimulated with LPS (100ng/ml). Murine models of endotoxic shock were established by challenging the mice with different dose of LPS (i.p). RTPCR, RNA sequencing, ELISA and hematoxylin and eosin staining were performed to elucidate the role and mechanisms of TRIM24 in endotoxic shock. The expression of TRIM24 is downregulated in LPS-stimulated bone marrow-derived macrophages (BMDMs). Loss of TRIM24 boosted IL-10 expression during the late stage of LPS-stimulation in macrophages. RNA-seq analysis revealed the upregulation of IFNβ1, an upstream regulator of IL-10, in TRIM24 knockout macrophages. Treatment with C646, a CBP/p300 inhibitor, diminished the difference in both IFNβ1 and IL-10 expression between TRIM24 knockout and control macrophages. Loss of TRIM24 provided protection against LPS-induced endotoxic shock in mice. Our results demonstrated that inhibiting TRIM24 promoted the expression of IFNβ1 and IL-10 during macrophage activation, therefore protecting mice from endotoxic shock. This study offers novel insights into the regulatory role of TRIM24 in IL-10 expression, making it a potentially attractive therapeutic target for inflammatory diseases.


Development of a molecular method to identify hepatitis E virus in water.
Hepatitis E virus (HEV) causes an infectious form of hepatitis associated with contaminated water. By analyzing the sequence of several HEV isolates, a reverse transcription-polymerase chain reaction method was developed and optimized that should be able to identify all of the known HEV strains. When tested under laboratory conditions, this method was able to detect low levels of five diverse HEV variants. In addition, internal controls were constructed so that any PCR inhibition could be detected. Finally, virus-spiked environmental water samples were analyzed successfully with these assays.


The molecular link between oxidative stress, insulin resistance, and type 2 diabetes: A target for new therapies against cardiovascular diseases.
Type 2 Diabetes Mellitus (T2D) is a chronic disease with a pandemic incidence whose pathogenesis has not yet been clarified. Raising evidence highlighted the role of oxidative stress in inducing insulin resistance, pancreatic beta-cell dysfunction, and leading to cardiovascular disease (CVD). Therefore, understanding the link between oxidative stress, T2D and CVD may help to further understand the pathological processes beyond this association, to personalize the algorithm of the cure, and to find new therapeutic targets. Here, we discussed the role of oxidative stress and the decrease of antioxidant defenses in the pathogenesis of T2D. Furthermore, some aspects of hypoglycemic therapies and their potential role as antioxidant agents were examined, which might be pivotal in preventing CVD in T2D patients.


Covalent immobilization of cellulases onto a water-soluble-insoluble reversible polymer.
The covalent immobilization of a commercial preparation of cellulase on a reversibly soluble-insoluble enteric polymer Eudragit S-100 by carbodiimide coupling was carried out. The characteristics of covalent Eudragit cellulase were evaluated using Fourier transform infrared (FTIR) spectra, circular dichroism (CD) spectra, and fluorescence spectra. FTIR, CD, and fluorescence measurements also revealed that the cellulases were covalently bonded to the supports. Covalent Eudragit cellulase had binding efficiency of 81.08% which was higher than the noncovalent Eudragit cellulase 56.83%. The relative activity of the native cellulase and covalent Eudragit cellulase increased and reached the maximum (at pH 5.0, 50°C) and then decreased with further increases in pH and temperature. The covalent Eudragit cellulase shows higher stability especially at higher pH and temperature. The K (m) value of covalent Eudragit cellulase (4.78 g·L(-1)) was decreased compared to that of the native cellulase (2.89 g·L(-1)). The affinity of the cellulase to its substrate was increased when it was immobilized on Eudragit S-100.


Integrated LC-MS metabolomics with dual derivatization for quantification of FFAs in fecal samples of hepatocellular carcinoma patients.
FFAs display pleiotropic functions in human diseases. Short-chain FAs (SCFAs), medium-chain FAs, and long-chain FAs are derived from different origins, and precise quantification of these FFAs is critical for revealing their roles in biological processes. However, accessing stable isotope-labeled internal standards is difficult, and different chain lengths of FFAs challenge the chromatographic coverage. Here, we developed a metabolomics strategy to analyze FFAs based on isotope-free LC-MS-multiple reaction monitoring integrated with dual derivatization. Samples and dual derivatization internal standards were synthesized using 2-dimethylaminoethylamine or dansyl hydrazine as a "light" label and N,N-diethyl ethylene diamine or N,N-diethyldansulfonyl hydrazide as a "heavy" label under mild and efficient reaction conditions. General multiple reaction monitoring parameters were designed to analyze these FFAs. The limit of detection of SCFAs varied from 0.5 to 3 nM. Furthermore, we show that this approach exhibits good linearity (R2 = 0.99374-0.99929), there is no serious substrate interference, and no quench steps are required, confirming the feasibility and reliability of the method. Using this method, we successfully quantified 15 types of SCFAs in fecal samples from hepatocellular carcinoma patients and healthy individuals; among these, propionate, butyrate, isobutyrate, and 2-methylbutyrate were significantly decreased in the hepatocellular carcinoma group compared with the healthy control group. These results indicate that the integrated LC-MS metabolomics with isotope-free and dual derivatization is an efficient approach for quantifying FFAs, which may be useful for identifying lipid biomarkers of cancer.


Associations between composite dietary antioxidant index and estimated 10-year atherosclerotic cardiovascular disease risk among U.S. adults.
Atherosclerotic cardiovascular disease (ASCVD) remains the leading cause of death and disability both in U.S. and worldwide. Antioxidants have been proved critical in mitigating the development of atherosclerosis. This study aimed to investigate the associations between composite dietary antioxidant index (CDAI) and estimated 10-year ASCVD risk among U.S. adults. Data extracted from the National Health and Nutrition Examination Survey were analyzed. A total of 10,984 adults aged 18 years and above were included in this study. CDAI was calculated based on the dietary intake reported in their 24-h recall interviews. The estimated 10-year ASCVD risk was calculated via Pooled Cohort Equations (PCE). After adjusting potential confounders, it was indicated that CDAI score was negatively correlated with 10-year ASCVD risk (OR 0.97, 95% CI 0.95-0.99). Stratify CDAI score by quartile, results showed that participants in the second, third, and fourth quartiles had lower ASCVD odds ratio (Q2: OR 0.87, 95% CI 0.69-1.09; Q3: OR 0.78, 95% CI 0.62-0.98; Q4: OR 0.74, 95% CI 0.59-0.94) than those in the first quartile (Q1, lowest CDAI score group), which was confirmed by the trend test as well (p < 0.05). Subgroup analyses stratified by sex, age, race/ethnicity, and smoking status did not show significant effect modification. Higher dietary antioxidants intake is associated with lower ASCVD risk among U.S. adults, for which policymakers and healthcare professionals may consider increasing the consumption of antioxidant-rich foods as a preventive strategy for ASCVD.


Coexpression of the receptor-associated protein gephyrin changes the ligand binding affinities of alpha 2 glycine receptors.
The inhibitory glycine receptor (GlyR) is a ligand-gated chloride channel protein, whose ligand binding alpha subunit occurs in several isoforms in the mammalian central nervous system. Here we show that coexpression of the GlyR-associated protein gephyrin changes the agonist and antagonist binding affinities of GlyRs generated by alpha 2 subunit expression in 293 kidney cells. Thus, a receptor-associated protein modifies the functional properties of a neurotransmitter receptor. This may contribute to an optimization of the postsynaptic neurotransmitter response.


Direct electron transfer of Cytochrome c at mono-dispersed and negatively charged perylene-graphene matrix.
Mono-dispersed 3,4,9,10-perylene tetracarboxylic acid (PTCA) functionalized graphene sheets (PTCA-graphene) were fabricated by a chemical route and dispersed well in aqueous solution. PTCA-graphene with plenty of -COOH groups as electrostatic absorbing sites were beneficial to the loading of Cytochrome c (Cyt c). Cyt c, which was tightly immobilized on the PTCA-graphene modified glassy carbon electrode, maintained its natural conformation. Direct electron transfer of Cyt c and the electro-catalytic activity towards the reduction of H2O2 were also achieved. It has been substantiated that PTCA-graphene is a preferable biocompatible matrix for Cyt c.


HIV-1 genotype and phenotype correlate with virological response to abacavir, amprenavir and efavirenz in treatment-experienced patients.
To assess the safety and efficacy of three new drugs in patients with antiretroviral failure and to correlate retrospectively baseline factors with virological response. Open-label, 48-week, single-arm, multi-center phase II trial conducted at nine US university or government clinics and private practices. Patients with HIV-1 RNA > or =500 copies/ml despite > or =20 weeks of treatment with at least one protease inhibitor received abacavir 300 mg twice a day, amprenavir 1200 mg twice a day and efavirenz 600 mg once a day. Other antiretrovirals were prohibited until week 16 except for substitutions for possible abacavir hypersensitivity. HIV RNA at weeks 16 and 48. A total of 101 highly treatment-experienced patients enrolled; 60 were naive to non-nucleoside analog reverse transcriptase inhibitors (NNRTI). HIV RNA < 400 copies/ml was attained in 25 out of 101 (25%) patients at 16 weeks (35% of NNRTI-naive and 10% of -experienced patients) and 23 (23%) patients at 48 weeks (33% of naive and 7% of experienced patients). CD4 cells increased by a median of 15 x 10(6) and 43 x 10(6) cells/l at weeks 16 and 48, respectively. Drug-related rash occurred in 50 out of 99 (51%) of patients, and 17 out of 99 (17%) permanently discontinued one or more drugs as a result. Lower baseline viral load, fewer NNRTI-related mutations, absence of decreased abacavir (> or =4-fold) and efavirenz (> or =10-fold) susceptibility, and greater number of drugs to which virus was susceptible were associated with virological response at week 16. Abacavir, amprenavir and efavirenz durably reduced HIV RNA and increased CD4 cell counts in a subset of treatment-experienced adults. Baseline viral load and some genotypic and phenotypic markers of resistance correlated with HIV RNA response.


Maternal height and the risk of cesarean delivery in nulliparous women.
To evaluate the relationship between maternal height of < 155 cm and the risk of caesarean delivery due to cephalo-pelvic disproportion (CPD) among nulliparous women. Retrospective cohort study A total of 660 term (GA > or = 37 weeks), uncomplicated singleton nulliparous pregnant women were enrolled on admission to labour room. The patients were divided into two groups based on maternal height, < or = 155 cm (440 cases) as control and < 155 cm (220 cases) as study group. The medical records of these women were reviewed. Various baseline clinical characteristics were collected. Intrapartum characteristics and maternal and neonatal outcomes were recorded. The rate of caesarean section of all indications was only slightly higher among study group than control group (16.4% and 13.7% respectively, p = 0.514). No significant difference was observed in the rate of caesarean delivery due to CPD (7.3% and 10.5% in control and study group respectively, p = 0.376). The rate of caesarean delivery due to CPD was highest among those with height < 150 cm (p < 0.001). Mean birth weight was significantly lower among study group than control group (2,927.7 +/- 368.1 g and 3,068.4 +/- 358.5 g respectively, p < 0.001). Low birth weight (< 2,500 g) was significantly higher among study group than control group (10.9% and 3.2% respectively, p < 0.001). Term singleton nulliparous pregnant women with maternal height < 155 cm were not associated with a greater likelihood of caesarean section for CPD. However, mean birth weight was significantly lower and low birth weight was significantly increased among mothers with short stature.


Differential anthocyanin accumulation in radish taproot: importance of RsMYB1 gene structure.
RsMYB1a was the crucial MYB, and RsbHLH4 is the essential partner in regulating the anthocyanin biosynthesis in radish. There are four color types of radish according to whether or not the anthocyanin accumulates in the skin and flesh of taproot. Red radishes accumulate a substantial amount of anthocyanins in both the skin and flesh. It is well known that the MYB-bHLH-WD40 transcription factor(s) complex regulates the biosynthesis of anthocyanin in plants. Here in, four candidate MYB and bHLH genes, RsMYB1a, RsMYB1b, RsbHLH2 and RsbHLH4, were isolated from red radish 'Hongxin 1'. The expression of RsbHLH4 and the two structural genes RsANS and RsUFGT was significantly positively correlated with anthocyanin contents. The expression of RsMYB1a was also highly correlated with anthocyanin accumulation, particularly when the white flesh sample of 'Hongxin 1-1' was excluded. The transient expression of RsMYB1a in the radish cotyledon and leaf induced anthocyanin accumulation with even stronger promoting role when expression in combination with RsbHLH4. These results suggested that RsMYB1a was the crucial MYB, and that RsbHLH4 is an essential partner in regulating the biosynthesis of anthocyanins in radish. The low or undetectable RsbHLH4 expression paralleled the lack of anthocyanin accumulation in the white flesh of 'Hongxin 1-1' and 'Shaguan 1'. Assays demonstrated that RsMYB1a interacted with RsbHLH4 and activated the expression of RsbHLH4. Notably, all the dark red radish cultivars have a longer RsMYB1a genomic DNA sequence, while the short and nonfunctional RsMYB1a is present in non-red cultivars. The length of the first intron and the presence of an early stop codon of RsMYB1 might underlie the differential anthocyanin accumulation in the radish taproot.


The effect of incubation time distribution on the extinction characteristics of a rabies epizootic.
The continuous model of Anderson et al. (1981), Nature 289, 765-771, is successful in describing certain characteristics of rabies epizootics, in particular, the secondary recurrences which follow the initial outbreak; however, it also predicts the occurrence of exponentially small minima in the infected population, which would realistically imply extinction of the virus. Here we show that inclusion of a more realistic distribution of incubation times in the model can explain why extinction will not occur, and we give explicit parametric estimates for the minimum infected fox density which will occur in the model, in terms of the incubation time distribution.


Ultrastructural study of the long-term development of two experimental cutaneous calcinoses (topical calciphylaxis and topical calcergy) in the rat.
Skin calcification induced by topical calciphylaxis was provoked by a subcutaneous injection of iron chloride in rats previously sensitized by dihydrotachysterol. A cutaneous topical calcergy was induced by an injection of potassium permanganate. An electron-microscopical study of the long-term evolution of both these models of calcification was made. After the initial stages, mineralization of the connective tissue continued by a secondary nucleation process without matrix vesicles. The mineral composed of needle-like structures, apatite in nature, was mainly deposited between and around collagen fibrils, and showed various arrangements in calcified plaques. Intrafibrillar calcification was rarely observed and appeared only in the later stages. The extension of calcified deposits then stopped. Finally, there was a fragmentation of the mineralized area which was progressively surrounded by uncalcified collagen fibrils. A demineralization process, caused by cells such as macrophages and multinucleated giant cells, rather than a resorption of the calcified deposits, was noted. It is important to emphasize that, in both models of ectopic calcification, an evolution toward ectopic ossification was never observed, which is perhaps due to the absence of extensive resorption mechanisms.


5-Nitro-2,3-bis-(thio-phen-2-yl)quinoxaline.
The title compound, C16H9N3O2S2, was synthesized via a condensation reaction in refluxing acetic acid. The dihedral angles between the mean plane of the quinoxaline unit and the thienyl rings are 35.16 (5)° and 24.94 (3)°.


Isotope ratio by HRGC-MS of citrus Junos tanaka (yuzu) essential oils: m/z 137/136 of terpene hydrocarbons.
The isotope ratios of monoterpene hydrocarbons in Citrus junos Tanaka (yuzu) essential oils from different origins were determined by ordinary high-resolution gas chromatography-mass spectrometry (HRGC-MS). Both intensities of the molecular mass peaks (m/z 136) and of the isotope peaks (m/z 137) of monoterpene hydrocarbons were measured by single-ion monitoring with an MS analysis. The isotope ratios (m/z 137/136) of the ten monoterpene hydrocarbons commonly contained in citrus essential oils, alpha-pinene, beta-pinene, sabinene, myrcene, alpha-phellandrene, alpha-terpinene, limonene, gamma-terpinene, beta-phellandrene and terpinolene, were determined in yuzu samples of the highest commercial quality from 42 different production districts. Statistical treatment of these data by the t-test and sign test revealed significant differences of the isotope effects in each yuzu sample. It is suggested that this technique will be applicable for evaluating the quality, genuineness and origin of citrus fruits and their products. The isotope fingerprints were also demonstrated in several citrus fruits other than the yuzu samples.


Modified technique of bilateral gluteus maximus transposition for reconstruction of sphincter for pediatric traumatic fecal incontinence.
The aim of the study was to report our 13-year experience of 14 children with gluteus maximus muscular transposition to treat post-trauma fecal incontinence and discuss our technical modifications to this surgical procedure. Fourteen children (median age: 9.9years) with complete fecal incontinence after traumas received this procedure from December 1998 to February 2011. The major modification of the surgery was that we transposed two thick muscular bundles about 2cm in diameter bilaterally. They surrounded the middle portion of rectum to act as sphincters. We used dynamic defecography and anorectal endosonography to observe the functions of the transposed muscles. Wexner scores, fecal incontinence quality of life questionnaire and self-rated health measurement scale scores had been used to evaluate their life quality. The median follow-up time was 6.3years. Twelve children reported prominently improved fecal controls with reduced stool frequency. Postoperative dynamic defecography and anorectal endosongraphy vividly demonstrated the satisfactory voluntary contractile and relaxed states of the reconstructed muscle. Wexner scores were significantly improved both 1year and 2years after the procedure (P<.05). Two year after surgery, rectal maximum systolic pressure, contraction duration and maximum systolic volume significantly increased compared with the results before and 1year after surgery (P<.05) CONCLUSIONS: This modified technique of bilateral gluteus maximus transposition for reconstruction of sphincter efficiently improved fecal control and life quality for pediatric patients with traumatic fecal incontinence.


Associations of Unintended Pregnancy With Maternal and Infant Health Outcomes: A Systematic Review and Meta-analysis.
Unintended pregnancy is common in the US and is associated with adverse maternal and infant health outcomes; however, estimates of these associations specific to current US populations are lacking. To evaluate associations of unintended pregnancy with maternal and infant health outcomes during pregnancy and post partum with studies relevant to current clinical practice and public health in the US. Cochrane Central Register of Controlled Trials and Database of Systematic Reviews, PsycINFO, SocINDEX, and MEDLINE databases (January 1, 2000, to June 15, 2022) and manual review of reference lists. Epidemiologic studies relevant to US populations that compared key maternal and infant health outcomes for unintended vs intended pregnancies and met prespecified eligibility criteria were included after investigators' independent dual review of abstracts and full-text articles. Investigators abstracted data from publications on study methods, participant characteristics, settings, pregnancy intention, comparators, confounders, and outcomes; data were validated by a second investigator. Risk of bias was independently dual rated by investigators using criteria developed by the US Preventive Services Task Force. Results of studies controlling for confounders were combined by using a profile likelihood random-effects model. Prenatal depression, postpartum depression, maternal experience of interpersonal violence, preterm birth, and infant low birth weight. Thirty-six studies (N = 524 522 participants) were included (14 cohort studies rated good or fair quality; 22 cross-sectional studies); 12 studies used large population-based data sources. Compared with intended pregnancy, unintended pregnancy was significantly associated with higher odds of depression during pregnancy (23.3% vs 13.9%; adjusted odds ratio [aOR], 1.59 [95% CI, 1.35-1.92]; I2 = 85.0%; 15 studies [n = 41 054]) and post partum (15.7% vs 9.6%; aOR, 1.51 [95% CI, 1.40-1.70]; I2 = 7.1%; 10 studies [n = 82 673]), interpersonal violence (14.6% vs 5.5%; aOR, 2.22 [95% CI, 1.41-2.91]; I2 = 64.1%; 5 studies [n = 42 306]), preterm birth (9.4% vs 7.7%; aOR, 1.21 [95% CI, 1.12-1.31]; I2 = 1.7%; 10 studies [n = 94 351]), and infant low birth weight (7.3% vs 5.2%; aOR, 1.09 [95% CI, 1.02-1.21]; I2 = 0.0%; 8 studies [n = 87 547]). Results were similar in sensitivity analyses based on controlling for history of depression for prenatal and postpartum depression and on study design and definition of unintended pregnancy for relevant outcomes. Studies provided limited sociodemographic data and measurement of confounders and outcomes varied. In this systematic review and meta-analysis of epidemiologic observational studies relevant to US populations, unintended pregnancy, compared with intended pregnancy, was significantly associated with adverse maternal and infant outcomes. PROSPERO Identifier: CRD42020192981.


Barbara E. Waud, M.D. (1931-), an early woman clinician scientist and professor of anesthesiology at University of Massachusetts Medical School.
After the first successful public demonstration of modern anesthesia in 1846, most female anesthetists were nurses by trade since none were yet allowed to attend medical school to become physicians. The turn of the twentieth century, however, brought about greater opportunity for female physician-anesthetists. We explore the life and career of Barbara E. Waud (1931-), a pioneering woman physician and researcher in the field of anesthesiology. Waud chose to pursue a career in medicine at a time when most women did not even attend college, and for most of her training and practice, she was the only woman in her department. Personal interviews with Waud, her daughter, and her colleagues highlight her rebellious and resilient nature that helped her overcome the obstacles put forth by male colleagues, and the judgment she received from female acquaintances for being a working mother. Waud's impressive career of dedicated clinical practice and ground-breaking research spanned four decades and inspired generations of physicians.


The enhancement role of Matrigel on HBV infection in HepG2-NTCP cells.
The hepatoma cell lines stably expressing sodium taurocholate cotransporting polypeptide (NTCP), the receptor of hepatitis B virus (HBV) infection, serve as important infection models for studying viral biology and drug discovery. However, the efficiency of infection greatly varies. In this study, we studied the effects and potential mechanisms of Matrigel® hESC-qualified (M-hq), a biological basement membrane matrix commonly used in cell culture, on promotion HBV in vitro infection in HepG2-NTCP cells. For the first time, our findings demonstrate that M-hq could enhance the infection efficiency of cell culture-derived HBV with no impact on the cell viability, the HBV transcription and response to antiviral treatments. The infection enhancement is reproducible and is suggested to occur at HBV attachment step. Our study suggests that this novel system is applicable for studying HBV biology and new drugs.


[Mathematical foundations for biophysical methods in fetal monitoring. II. Cardiotocography].
The principles of mathematical analysis of cardiotocographic records are described, together with the methods of calculation of the baseline, oscillations, short-term and long-term variability, and cardiotocographic analysis in frequency range.


Impacts of long-standing illness and chronic illness on working hours and household income in a longitudinal UK study.
Individuals within households encounter a variety of events including development of a disability or chronic illness. We used data from the Understanding Society, 2009-2016, to determine whether there are changes to working hours or household income as a result of an individual developing an illness. After adjusting for a variety of sociodemographic characteristics, there were few associations observed between one's own individual illness status and household income. There was a clear trend of reduction of weekly working hours with increasing severity and chronicity of the individuals' illness or disease. Individuals who were not ill, but lived in an household with an ill person worked about 30-min less per week, b = -0.69, 95% confidence interval (CI)=(-1.09, -0.30), while those with a limiting long-standing illness and a chronic disease worked 3.5 h less per week, b = -3.64, 95% CI=(-4.21, -3.08), compared to individuals with no illness in their household. Individuals with a limiting illness only had lower incomes, b = -0.04, 95% CI=(-0.07, -0.004) compared to individuals with no household illness. These associations were not greatly changed with the inclusion of reception of benefits or being cared for. Interactions were observed by gender, age being cared for and reception of benefits. Additionally, there were differences were observed by working age groups and between those who lived alone and those who did not. The findings suggest that while there is a reduction of working hours among individuals with an illness or who have an ill person in their home, household income is resilient to the experience of an illness, in the United Kingdom. However, this appeared to differ by household composition, i.e. whether individuals were of working age or whether they lived alone. Identification of households at highest risk of income reduction may serve to inform policy and appropriate distribution of services and support.


Ultrafast Roaming Mechanisms in Ethanol Probed by Intense Extreme Ultraviolet Free-Electron Laser Radiation: Electron Transfer versus Proton Transfer.
Ultrafast H2+ and H3+ formation from ethanol is studied using pump-probe spectroscopy with an extreme ultraviolet (XUV) free-electron laser. The first pulse creates a dication, triggering H2 roaming that leads to H2+ and H3+ formation, which is disruptively probed by a second pulse. At photon energies of 28 and 32 eV, the ratio of H2+ to H3+ increases with time delay, while it is flat at a photon energy of 70 eV. The delay-dependent effect is ascribed to a competition between electron and proton transfer. High-level quantum chemistry calculations show a flat potential energy surface for H2 formation, indicating that the intermediate state may have a long lifetime. The ab initio molecular dynamics simulation confirms that, in addition to the direct emission, a small portion of H2 undergoes a roaming mechanism that leads to two competing pathways: electron transfer from H2 to C2H4O2+ and proton transfer from C2H4O2+ to H2.


Micronized cellulose from citrus processing waste using water and electricity only.
Along with a water-soluble fraction rich in pectin, the hydrodynamic cavitation of citrus processing waste carried out in water demonstrated directly on semi-industrial scale affords an insoluble fraction consisting of micronized cellulose of low crystallinity ("CytroCell"). Lemon and grapefruit CytroCell respectively consist of 100-500 nm wide cellulose nanorods, and of 500-1000 nm wide ramified microfibrils extending for several μm. These findings establish a technically viable route to low crystallinity micronized cellulose laying in between nano- and microcellulose, using water and electricity only.


Protect effect of bicyclol on cisplatin-induced nephrotoxicity in mice.
This study investigated the protective effects of bicyclol against cisplatin-induced nephrotoxicity and the possible mechanisms in mice. Bicyclol (250 mg/kg, p.o., 5 days) showed significant protection as evidenced by the decrease of elevated serum creatine and blood urea nitrogen, and improvement of histopathological injury induced by cisplatin. The formation of kidney malondialdehyde with a concomitant reduction of reduced glutathione were also inhibited by bicyclol, while the activities of kidney superoxide dismutase, catalase and glutathione peroxidase were all increased, respectively. Bicyclol also inhibited the increase of kidney and serum nitric oxide induced by cisplatin. In addition, induction of induced nitric oxide synthase and nitrotyrosine were suppressed by bicyclol. Bicyclol suppressed cisplatin-induced extracelluar signal regulated kinases 1/2 and p38 mitogen-activated protein kinase activation in the kidney of mice. Results obtained demonstrate that bicyclol pre-administration can prevent the nephrotoxicity induced by cisplatin.


Quantitative autoradiography of hemicholinium-3 binding sites in human amygdala.
We report the binding properties and subnuclear localization of [3H]hemicholinium-3 binding sites in human amygdala using quantitative autoradiography. Specific binding was saturable and high affinity (apparent Kd 2-11 nM). Binding was highest in the basolateral nucleus which receives dense cholinergic innervation from the basal forebrain. Binding closely approximated acetylcholinesterase reactivity. These data support [3H]hemicholinium-3 as a quantitative marker for cholinergic terminals in human brain.


Drug-eluting bead transarterial chemoembolization with medium-sized versus small-sized CalliSpheres microspheres in unresectable primary liver cancer.
The optimal microparticle size for drug-eluting beads transarterial chemoembolization (DEB-TACE) remains unknown. This retrospective cohort study analyzed the efficacy and safety of CalliSpheres microsphere embolization in the treatment of unresectable hepatocellular carcinoma (HCC) to determine the influence of particle size on the results. Forty-two patients with unresectable HCC were enrolled in this retrospective study from January 2018 to January 2020. Patients received DEB-TACE with CalliSpheres of 100-300 μm (small-size, n = 15) or 300-500 μm (medium-size, n = 27). The tumor response was evaluated via enhanced CT or MRI at 1 month, 3 months, and 6 months after treatment, based on the Modified Response Evaluation Criteria in Solid Tumors. Adverse events after DEB-TACE were recorded. Complete response, partial response, stable disease, and progressive disease were recorded in 20%, 20%, 33.3%, 26.7%, respectively, of patients in the small-size group and 3.7%, 25.9%, 44.4%, 25.9% of patients in the medium-size group, respectively. No significant difference was found between the two groups (p = 0.516). Major adverse events, including grade three liver toxicity (n = 4) and liver abscess (n = 3), occurred significantly more in the small-size group, while none were reported in the medium size group (p < 0.05). DEB-TACE with medium-size (300-500 μm) CalliSpheres microspheres had similar efficacy and a better safety profile than DEB-TACE with small-size (100-300 μm) CalliSpheres, indicating that medium-size microspheres may be a better choice for unresectable primary liver cancer.


Hyperspectral Imaging for Determining Pigment Contents in Cucumber Leaves in Response to Angular Leaf Spot Disease.
Hyperspectral imaging technique was employed to determine spatial distributions of chlorophyll (Chl), and carotenoid (Car) contents in cucumber leaves in response to angular leaf spot (ALS). Altogether, 196 hyperspectral images of cucumber leaves with five infection severities of ALS were captured by a hyperspectral imaging system in the range of 380-1,030 nm covering 512 wavebands. Mean spectrum were extracted from regions of interest (ROIs) in the hyperspectral images. Partial least square regression (PLSR) models were used to develop quantitative analysis between the spectra and the pigment contents measured by biochemical analyses. In addition, regression coefficients (RCs) in PLSR models were employed to select important wavelengths (IWs) for modelling. It was found that the PLSR models developed by the IWs provided the optimal measurement results with correlation coefficient (R) of prediction of 0.871 and 0.876 for Chl and Car contents, respectively. Finally, Chl and Car distributions in cucumber leaves with the ALS infection were mapped by applying the optimal models pixel-wise to the hyperspectral images. The results proved the feasibility of hyperspectral imaging for visualizing the pigment distributions in cucumber leaves in response to ALS.


Thyroid Cancer Incidence and Mortality by Socioeconomic Level in the State of São Paulo, Brazil 2001-2017.
Thyroid cancer is rising largely due to greater detection of indolent or slow-growing tumors; we sought to compare the incidence and mortality profiles of thyroid cancer in the State of São Paulo by socioeconomic status (SES). Data on thyroid cancer cases diagnosed from 2003 to 2017 in the Barretos Region and from 2001 to 2015 in the municipality of São Paulo were obtained from the respective cancer registries. Corresponding death data were obtained from a Brazilian public government database. Age-standardized rates were calculated and presented as thematic maps. The rates were also calculated by SES and spatial autocorrelation was assessed by global and local indices. There were 419 cases of thyroid cancer and 21 deaths in Barretos, contrasting with the highly populated São Paulo, with 30 489 cases and 673 deaths. The overall incidence rates in São Paulo (15.9) were three times higher than in Barretos (5.7), while incidence rates in women were close to five times higher in Barretos and four times higher in São Paulo than in men. Mortality rates were, in relative terms, very low in both regions. A clear stepwise gradient of increasing thyroid cancer incidence with increasing SES was observed in São Paulo, with rates in very high SES districts four times those of low SES (31.6 vs 8.1). In contrast, the incidence rates in Barretos presented little variation across SES levels. Thyroid cancer incidence varied markedly by SES in São Paulo, with incidence rates rising with increasing socioeconomic index. Overdiagnosis is likely to account for a large proportion of the thyroid cancer burden in the capital.


Atrial receptors, vasopressin and blood volume in the dog.
Recent work has clarified the relationship between stimulation of left atrial receptors and plasma vasopressin concentration (pAVP) and has allowed a rational explanation of a number of previously anomalous findings. There is now good evidence that mitral obstruction causes a decrease in pAVP and that the decreases in pAVP can occur within a normal range of pAVP in anaesthetized and unanaesthetized animals. A stimulus which is localised to the left atrial receptors also causes a decrease in pAVP and it is likely that this is due to stimulation of the complex unencapsulated endings in the atrium, with myelinated afferent fibres. Evidence is lacking that changes in the stimulus to ventricular receptors or to cardio-pulmonary receptors with C-fibre afferents influences pAVP. The diuretic response to left atrial distension is two-fold, an increase in free water clearance and a natriuresis. The increase in free water clearance is due to the decrease in pAVP; the cause of the natriuresis is unknown. The changes in pAVP occur rapidly in response to atrial distension (within 5 min). The stimulus provided to atrial receptors by atrial distension and the decrease in pAVP is maintained for at least 90 min. pAVP is also modulated in response to small changes in blood volume (+/- 10%). The changes in pAVP that occur over this range of blood volume are likely to be in the range of 1-10 pg/ml and to have their effects on renal water excretion rather than on vascular resistance. The much larger changes in pAVP which occur with greater degrees of blood loss, and which can affect vascular resistance are likely to be produced by changes in the stimulus to other receptors, but a low input from atrial receptors may be permissive for these stimuli to be effective. More work is needed to clarify the relationship between inputs from different receptor types.


Endoproteolytic cleavage of HIV-1 gp160 envelope precursor occurs after exit from the trans-Golgi network (TGN).
Endoproteolytic processing of human immunodeficiency virus type 1 (HIV-1) gp160 membrane glycoprotein precursor into gp 120 and gp41 is necessary for formation of infectious HIV particles [1]. We have studied the intracellular site of this processing using inhibition of transport at reduced temperature (20 degrees C). That reduced temperature (20 degrees C) inhibits the intracellular transport also in Jurkat-tat cells was demonstrated using the Semliki Forest virus p62 precursor processing as model. In HIV-1 infected Jurkat-tat cells the proteolytic processing of gp 160 precursor did not occur when the protein was accumulated in the TGN at 20 degrees C temperature. When the temperature was shifted to 37 degrees C the HIV-1 gp 160, which had accumulated in the TGN at the reduced temperature, was proteolytically processed. The processing of gp 160 was inhibited when the temperature reversion was carried out in the presence of brefeldin A (BFA) or aluminium fluoride (ALFn) indicating that the exit from the TGN is required for the proteolytic cleavage of HIV-1 gp160 precursor. The results suggest that the processing of gp 160 takes place at a yet unidentified transport step which is distal to the TGN/20 degrees C block site.


The Activitrax rate responsive pacemaker system.
Bipolar Medtronic Activitrax rate responsive pacemakers were implanted in 31 patients for ventricular (28) or atrial (3) pacing. Mean follow-up was 16 months (range 10 to 26). Twenty pacemakers were implanted after catheter ablation of the His bundle, 7 for sick sinus syndrome. 1 for atrioventricular block and 3 for sick sinus syndrome with atrioventricular block. A rate response value was selected that gave a pacing rate of about 100 pulses/min during walking. Of the 31 patients, all had 24-hour ambulatory electrocardiographic monitoring with diary, 11 walked a 20-minute circuit, including a flight of stairs, and 20 had a treadmill exercise test. In 9 patients the pacing rate could be compared with the underlying sinus rate during exercise and was seen to match it very closely. In 12 patients the pacing rate during car driving was found to be similar to the sinus rate of 5 volunteers under similar conditions (mean minimum and maximum rate was 80 and 99 pulses/min, respectively). No pacing-induced arrhythmias were seen during ambulatory electrocardiographic monitoring. At high pacing rates slightly irregular pacing intervals were sometimes observed, which was due to polarization sensing. Sporadically, 1 pacing interval shortened to the upper rate value, because of a known and now resolved timing anomaly. Neither anomaly was of clinical consequence and the first could be resolved by reprogramming.(ABSTRACT TRUNCATED AT 250 WORDS)


Contribution of the C-terminal dipeptide of transforming growth factor-alpha to its activity: biochemical and pharmacologic profiles.
We have used a matrix of biological (two distinct guinea-pig stomach contractile smooth muscle preparations) and biochemical (human placental membrane receptor binding and phosphorylation) assays to evaluate the activity profiles of epidermal growth factor-urogastrone (EGF-URO, mouse and human), transforming growth factor-alpha (TGF-alpha, human) and the TGF-alpha derivative lacking the C-terminal dipeptide, Leu49-Ala50, TGF-alpha-(1-48). In the longitudinal muscle (LM) bioassay, the relative potencies of the peptides were: EGF-URO greater than TGF-alpha greater than TGF-alpha-(1-48), with relative activity ratios (EC50S) of approximately 1:3:16. In the LM assay system, TGF-alpha-(1-48) was a partial agonist. In the circular muscle (CM) bioassay, the relative order of potencies was: TGF-alpha- greater than EGF-URO greater than TGF-alpha-(1-48), with EC50S of about 1:2:7. In the CM assay, all three peptides were full agonists, even though EGF-URO caused an intense desensitization of the tissue whereas TGF-alpha and TGF-alpha-(1-48) did not. The relative affinities of the peptides in the placenta membrane binding assay, EGF-URO greater than TGF-alpha greater than TGF-alpha-(1-48), were in good qualitative and quantitative agreement with the LM (but not the CM) bioassay, with relative KDS in the proportions of about 1:3:17. In the phosphorylation assay, using either the phosphorylated EGF-URO receptor or calpactin-II as an index of receptor kinase activation, the relative potencies of the peptides, EGF-URO greater than TGF-alpha greater than TGF-alpha-(1-48), were also qualitatively in accord with the relative potencies measured in the LM and ligand binding assays (but not in the CM preparation); however, quantitatively, the relative potency ratios (EC50S) observed in the phosphorylation assay (1:2:3) were somewhat out of keeping with the relative values observed in the LM and ligand binding assays. All three peptides were full agonists in the phosphorylation assay. Our data point to the importance of the C-terminal dipeptide, Leu49-Ala50 of TGF-alpha in terms of the binding affinity and intrinsic activity of this polypeptide; and our work provides further evidence for the distinct nature of the EGF-URO/TGF-alpha receptor system present in the CM bioassay preparation. The biological/biochemical activity profiles documented for the three polypeptides can serve as a basis for the further evaluation of other synthetic and naturally occurring members of the EGF-URO/TGF-alpha family of polypeptides.


On 'Fluorescence and the structure of proteins. I. Effects of substituents on the fluorescence of indole and phenol compounds' by Robert W. Cowgill.
This note on the work of Robert W. Cowgill in 1963 (Archives Biochemistry and Biophysics 100, 36-44), addressing the effect of substituents in indole and phenol compounds as models for tryptophan and tyrosine, intends to frame it within the foundations of protein fluorescence and its applications, considering its relevance and impact.


Comparative analysis of buffalo, local and continental cattle carcasses with the European Union classification system in Greece.
The carcasses produced in Greece from Greek Buffalo, indigenous and local crossbred cattle were evaluated using the European Union (EU) bovine classification system and compared with carcasses of continental breeds reared in the country. The main carcass production (67.8%) came from animals born in Greece with a relatively low percentage from local breeds (17%). The lightest carcass weight was found in Greek Brachyceros [145.2 ± 2.3 kg (males); 146.0 ± 3.1 kg (females)], while among locals the Greek Blonde rendered the heaviest carcass weights [303.2 ± 2.9 kg (males); 265.3 ± 3.5 kg (females)]. Muscle conformation scoring was the lowest in Buffalo [4.1 ± 0.1 (males); 4.9 ± 0.0 (females)] and the highest for males of Charolais and Blonde d'Aquitaine (11.2 ± 0.0 and 10.7 ± 0.0, respectively). The meat-purposed breeds produced carcasses of higher quality than dual-purposed, local breeds and dairy Holstein-Friesian. With the discriminant analysis, the correct classification of carcasses varied from 53.4% (males) to 47.8% (females) with high assignment percentage of Greek Buffalo males (81.6%), Simmental males (80.7%) and Holstein-Friesian females (98.5%).


Use telehealth as needed: telehealth substitutes in-person primary care and associates with the changes in unplanned events and follow-up visits.
Telehealth rapidly expanded since the outbreak of the COVID-19 pandemic. This study aims to understand how telehealth can substitute in-person services by 1) estimating the changes in non-COVID emergency department (ED) visits, hospitalizations, and care costs among US Medicare beneficiaries by visit modality (telehealth vs. in-person) during the COVID-19 pandemic relative to the previous year; 2) comparing the follow-up time and patterns between telehealth and in-person care. A retrospective and longitudinal study design using US Medicare patients 65 years or older from an Accountable Care Organization (ACO). The study period was April-December 2020, and the baseline period was March 2019 - February 2020. The sample included 16,222 patients, 338,872 patient-month records and 134,375 outpatient encounters. Patients were categorized as non-users, telehealth only, in-person care only and users of both types. Outcomes included the number of unplanned events and costs per month at the patient level; number of days until the next visit and whether the next visit happened within 3-, 7-, 14- and 30-days at the encounter level. All analyses were adjusted for patient characteristics and seasonal trends. Beneficiaries who used only telehealth or in-person care had comparable baseline health conditions but were healthier than those who used both types of services. During the study period, the telehealth only group had significantly fewer ED visits/hospitalizations and lower Medicare payments than the baseline (ED 13.2, 95% CI [11.6, 14.7] vs. 24.6 per 1,000 patients per month and hospitalization 8.1 [6.7, 9.4] vs. 12.7); the in-person only group had significantly fewer ED visits (21.9 [20.3, 23.5] vs. 26.1) and lower Medicare payments, but not hospitalizations; the both-types group had significantly more hospitalizations (23.0 [21.4, 24.6] vs. 17.8). Telehealth was not significantly different from in-person encounters in number of days until the next visit (33.4 vs. 31.2 days) or the probabilities of 3- and 7-day follow-up visits (9.2 vs. 9.3% and 21.8 vs.23.5%). Patients and providers treated telehealth and in-person visits as substitutes and used either depending on medical needs and availability. Telehealth did not lead to sooner or more follow-up visits than in-person services.


Protein kinase C activation stabilizes LDL receptor mRNA via the JNK pathway in HepG2 cells.
LDL is the most abundant cholesterol transport vehicle in plasma and a major prognostic indicator of atherosclerosis. Hepatic LDL receptors limit circulating LDL levels, since cholesterol internalized by the liver can be excreted. As such, mechanisms regulating LDL receptor expression in liver cells are appealing targets for cholesterol-lowering therapeutic strategies. Activation of HepG2 cells with phorbol esters enhances LDL receptor mRNA levels through transcriptional and posttranscriptional mechanisms. Here, we show that 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced stabilization of receptor mRNA requires the activity of protein kinase C and is accompanied by activation of the major mitogen activated protein kinase pathways. Inhibitor studies demonstrated that receptor mRNA stabilization is independent of the extracellular signal-regulated kinase or p38(MAPK), but requires activation of the c-Jun N-terminal kinase (JNK). An essential role for JNK in stabilizing receptor mRNA was further confirmed through small interfering RNA (siRNA) experiments and by activating JNK through two protein kinase C-independent mechanisms. Finally, prolonged JNK activation increased steady-state levels of receptor mRNA and protein, and significantly enhanced cellular LDL-binding activity. These data suggest that JNK may play an important role in posttranscriptional control of LDL receptor expression, thus constituting a novel mechanism to enhance plasma LDL clearance by liver cells.


Effect of training on the rating of perceived exertion at the ventilatory threshold.
The purpose of this study was to determine the effect of training on the rating of perceived exertion (RPE) at the ventilatory threshold. College students were assigned to either training (n = 17) or control (n = 10) groups. Trainers completed 18 interval training sessions (five X 5 min cycling at 90-100% VO2max) and 8 continuous training sessions (40 min running or cycling) in 6 weeks. Pre- and post-training, cardiorespiratory, metabolic, and perceptual variables were measured at the ventilatory threshold during graded exercise tests on a cycle ergometer. Ventilatory threshold was that point above which VE X VO2-1 increased abruptly relative to work rate. Post-training means of trained and control subjects were compared using analysis of covariance, with pre-training values as covariates. Following training, the adjusted means for the trained subjects were significantly greater (p less than 0.05) than for controls for VO2max (6%), and for work rate (20%), VO2 (23%), and %VO2max (13%) at the ventilatory threshold. However, adjusted means for RPE at the ventilatory threshold were not significantly different (2%). Both before and after training, exercise at the ventilatory threshold was perceived as 'somewhat hard' to 'hard' (RPE = 13-15) by both groups. The relationship between RPE and %VO2max was altered by training, with trained subjects having a lower RPE at a given %VO2max. It is concluded that RPE at the ventilatory threshold is not affected by training, despite that after training the ventilatory threshold occurs at a higher work rate and is associated with higher absolute and relative metabolic and cardiorespiratory demands.


Identifying DNA methylation biomarkers of cancer drug response.
In the last few years, DNA methylation has become one of the most studied gene regulation mechanisms in carcinogenesis as a result of the cumulative evidence produced by the scientific community. Moreover, advances in the technologies that allow detection of DNA methylation in a variety of analytes have opened the possibility of developing methylation-based tests. A number of studies have provided evidence that specific methylation changes can alter the response to different therapeutic agents in cancer and, therefore, be useful biomarkers. For example, the association of the methylation status of DNA repair genes such as MGMT and MLH1 illustrate the two main mechanisms of response to DNA damaging agents. Loss of methylation of MGMT, and the subsequent increase in gene expression, leads to a reduction in response to alkylating agents as a result of enhanced repair of drug-induced DNA damage. Conversely, the increase in methylation of MLH1 and its resulting loss of expression has been consistently observed in drug-resistant tumor cells. MLH1 encodes a mismatch repair enzyme activated in response to DNA damage; activation of MLH1 also induces apoptosis of tumor cells, and thus loss of its expression leads to resistance to DNA-damaging agents. Other methylation-regulated genes that could serve as biomarkers in cancer therapy include drug transporters, genes involved in microtubule formation and stability, and genes related to hormonal therapy response. These methylation markers have potential applications for disease prognosis, treatment response prediction, and the development of novel treatment strategies.


Application of maize tassel for the removal of Pb, Se, Sr, U and V from borehole water contaminated with mine wastewater in the presence of alkaline metals.
In this study, the removal of Pb(II) from aqueous solutions by tassel powder was studied and optimised. Batch experiments were conducted on simulated solutions using tassel powder adsorbent and the effects of contact time, pH and concentration on the extent of Pb (II) removal was studied. Equilibrium and kinetic models for Pb(II) sorption were developed by considering the effect of contact time and concentration at optimum pH 4 and fixed temperature(25 degrees C). The Freundlich model was found to describe the sorption energetics of Pb(II) on tassel more fully than the Langmuir. A maximum Pb(II) loading capacity of 333.3mg/g on tassel was obtained. The adsorption process could be well described by both the Langmuir and Freundlich isotherms with R(2) values of 0.957 and 0.972, respectively. The kinetic parameters were obtained by fitting data from the effect of contact time on adsorption capacity into the pseudo-first, pseudo-second-order and intra-particle diffusion equations. The kinetics of Pb(II) on tassel surface was well defined using linearity coefficients (R(2)) by pseudo-second-order (0.999), followed by pseudo-first-order (0.795) and lastly intra-particle diffusion (0.6056), respectively. The developed method was then applied to environmental samples taken from borehole waters contaminated with mine wastewater. The removal of Pb (ND-100%), Se (100%), Sr (5.41-59.0%), U (100%) and V (46.1-100%) was attained using tassel. The uptake of the metals from environmental samples was dependent on pH, ionic strength and levels of other competing species.


Cardiac MRI identifies valvular and myocardial disease in a subset of ANO5-related muscular dystrophy patients.
Patients with bi-allelic loss-of-function mutations in the gene ANO5 most commonly present with muscular dystrophy. In some studies, patients with ANO5-related dystrophy (ANO5-RD) had evidence of mild cardiac abnormalities; however, cardiac magnetic resonance imaging (MRI) has not been used for myocardial characterization. Ten patients with genetically confirmed ANO5-RD were enrolled in a phenotyping study to better characterize cardiac involvement. Evaluations included medical history, neurological examination and cardiac evaluations (electrocardiogram, echocardiogram and cardiac MRI). All patients were clinically asymptomatic from a cardiac perspective. Muscle MRI was consistent with previous studies of ANO5-RD with increased T1 signal in the posterior and medial compartments of the upper leg and the posterior compartment of the lower leg. Cardiac studies using echocardiography and cardiac MRI revealed dilation of the aortic root and thickening of the aortic valve without significant stenosis in 3/10 patients. There was evidence of abnormal late gadolinium enhancement (LGE) on cardiac MRI in 2/10 patients. In ANO5-RD, the development of cardiac fibrosis, edema or inflammation as demonstrated by LGE has not yet been reported. Cardiac MRI can characterize cardiac tissue and may detect subtle changes before they appear on echocardiography, with potential prognostic implications.


Inhalable functional mixed-polymer microspheres to enhance doxorubicin release behavior for lung cancer treatment.
Porous poly(cyclohexane-1,4-diyl acetone dimethylene ketal) (PCADK)/poly(d,l-lactide-co-glycolide) (PLGA) mixed-matrix porous microspheres loaded with doxorubicin (Dox) were successfully prepared, and PCADK/PLGA 2/8 was selected as the optimal mixed-matrix proportion. The optimal porous microspheres were characterized by a uniform spherical morphology, an obvious porous surface, an adaptive aerodynamic diameter (2.48 μm), good lung deposition and excellent encapsulation efficiency (77.22 %). The total release of Dox from PCADK/PLGA microspheres was 64.66 %, which was greater than the 46.31 % from the PLGA microsphere group, resulting in the porous PCADK/PLGA microspheres showing a stronger antiproliferative effect than the porous PLGA microspheres. The antiproliferative mechanism was examined through flow cytometry analysis and protein expression level detection, exhibiting enhanced tumor-related protein regulation, improved cell apoptosis induction and increased cycle arrest. Finally, a BALB/c mouse lung cancer model was established to evaluate the in vivo anticancer efficacy, and the PCADK/PLGA microspheres showed significantly stronger anticancer effects than the PLGA microspheres. We envision that employment of this mixed polymer material as a microsphere matrix could be a promising strategy for lung cancer therapy via pulmonary administration.


Urine TGF- beta1 assay validation: spinning down urine samples could diminish TGF-beta1 signal.
TGF-beta1 is a mediator of immune and anti-inflammatory responses. The present research was dedicated to the validation of the urine Quantikine Human TGF-beta1 ELISA assay. A reference interval was developed utilizing 100 urine samples from healthy donors. Samples containing precipitate were centrifuged and supernatants or un-spun specimens were run with TGF-beta1 assay. Supernatant TGF-beta1 concentrations are only a fraction of the un-spun sample values (77% to 29%, even less for the low concentration samples). Validation involving un-centrifuged urine samples demonstrated the following TGF-beta1 assay characteristics: linearity from 23 to 1286 pg/mL, recovery from 95 to 116%; precision: < 17%; analytical specificity 3 pg/mL; reference interval from 0 to 39 pg/mg creatinine; freeze-thaw stability: after first thawing, urine samples could loose from 5 to 40% of TGF-beta1 activity on each following freeze-thaw circle. For TGF-beta1 assay, urine sample preparation should not include a centrifugation step; repeating freeze-thawing should be avoided.


Genetically epilepsy-prone rodents show some changes of ion levels in the brain.
In the present study the water and ion (Na+, K+, Ca2+, Fe3+, Se4+, Mg2+, Mn2+, Mn2, Se4+, Cu2+) content in the brain of genetically epilepsy-prone rats (GEPRs) and of 21-, 45-, and 60-day-old DBA/2 mice were determined, and compared with those measured in normal controls (Sprague-Dawley rats and Swiss mice), to verify whether the predisposition to audiogenic seizures (AGS) may be partially related to changes in the cerebral osmotic and ionic state. Our findings clearly evidenziate two points: a) a more complex shift in brain ionic balance (rather than a peculiar modification in the concentration of a single ion) seems very likely involved in AGS susceptibility; (b) brain Ca2+ and Se4+ amounts, together with the water content, appear to be really important factors to which a role in abnormal seizure predisposition may be attributed.


Accuracy of four digital scanners according to scanning strategy in complete-arch impressions.
Although there are specific and general digital scanning guidelines depending on the system used, it is important to have the necessary flexibility in the acquisition of three-dimensional (3D) images to adapt to any clinical situation without affecting accuracy. The purpose of this in vitro study was to identify and compare the scanning strategy with the greatest accuracy, in terms of trueness and precision, of four intraoral scanners in the impression of a complete dental arch. Four digital scanners were evaluated with a 3D measuring software, using a highly accurate reference model obtained from an industrial scanner as a comparator. Four scanning strategies were applied 10 times on a complete maxillary arch cast inside a black methacrylate box. The data were statistically analyzed using one-way analysis of variance (ANOVA) and post hoc comparisons with Tamhane T2 test. The trueness of the Trios and iTero system showed better results with strategy "D," Omnicam with strategy "B," and True Definition with strategy "C". In terms of precision, both iTero and True Definition showed better results with strategy "D", while Trios showed best results with strategy "A" and Omnicam with strategy "B". There were significant differences between the scanning strategies (p<0.05) with the iTero scanner, but not with the other scanners (p>0.05). The digital impression systems used in the experiment provided sufficient flexibility for the acquisition of 3D images without this affecting the accuracy of the scanner.


Developing an implicit measure of disgust propensity and disgust sensitivity: examining the role of implicit disgust propensity and sensitivity in obsessive-compulsive tendencies.
There is increasing evidence that disgust responding occurs at both a primary and secondary level in the form of disgust propensity and disgust sensitivity. The unique contributions of anxiety and disgust need to be established if disgust is to be implicated in the etiology of anxiety disorders such as obsessive-compulsive disorder (OCD). The primary objective of the current study was to develop two separate implicit measures of disgust propensity and sensitivity and to explicate the role of implicit disgust propensity and sensitivity in avoidance behavior and OC tendencies. The current study (N = 33 undergraduate students) utilized a measure of implicit cognition, the Implicit Relational Assessment Procedure (IRAP), to independently analyze disgust propensity and disgust sensitivity. In addition, a series of behavioral approach tasks (BAT) and questionnaires measuring general disgust, obsessive-compulsive (OC) tendencies and general psychopathology were implemented to validate the implicit measures. Disgust sensitivity predicted avoidance behavior on the BATs independent of disgust propensity and anxiety, while disgust propensity did not. Both disgust propensity and sensitivity predicted self-reported OC tendencies and individually predicted obsessing and washing concerns, respectively. Our findings are based on a non-clinical student sample and further research is required for generalization to OCD. The implicit measures appeared to be measuring two separate constructs and had differential relationships with behavior and OC tendencies. Overall, the results support current theories relating to pathological disgust and OCD.


[Relapse of pulmonary tuberculosis in the context of tuberculosis-HIV co-infection in Abidjan (Côte d'Ivoire). Short note].
A cohort study was carried out from October 2001 to March 2002, in order to analyze the main characteristics of 53 adults patients (38 HIV-positive and 15 HIV-negative), who had a relapse of pulmonary tuberculosis. The observance in the first period of the treatment of tuberculosis was regular in 79% of tuberculous patients infected by the HIV (30/38) and in 87% of tuberculous patients who where HIV-negative (13/15) (p = 0.7). The relapse occurred before one year after recovery in 74% among the HIV-positive (28/38) and in 33% among the HIV-negative patients (5/15) (p = 0.00). Most of the HIV-positive patients (74%) had CD4 counts under 200/mm3 and 80% of the HIV-negative patients had more than 500 CD4 counts/mm3 (p = 0.00). After two months of retreatment following the relapse diagnosis, the rate of improvement in patients was about 16% in HIV infected (6/38) and about 67% in HIV-negative patients (10/15) (p = 0.00).


Laparoscopic Versus Open Pre-Treatment Loop Colostomy for Fecal Diversion in Rectal Cancer Patients: Is Laparoscopic Colostomy Better?
Diversion colostomy plays a crucial role in the management of carcinoma rectum in low- and middle-income countries as significant number of patients present with partial intestinal obstruction. The aim of this study was to compare laparoscopic and open approaches for fecal diversion done in patients with adenocarcinoma of the rectum as a pretreatment procedure. The primary end point of our study was time to initiation of neoadjuvant chemo radiation. It was a retrospective study that included all patients diagnosed to have carcinoma rectum and underwent a pretreatment fecal diversion between 2012 and 2014. A total of 55 patients underwent pretreatment diversion colostomy of which 33 were performed via the laparoscopic approach while 22 had open diversion. The time for initiation of neoadjuvant therapy was shorter in the laparoscopic group compared to the open approach (16 days vs. 20.5 days, P = 0.31). The study concluded that pretreatment diversion colostomy using the laparoscopic approach was a safe option in low- and middle-income countries as it was associated with faster recovery and early initiation of neoadjuvant therapy in patients with partially obstructed locally advanced carcinoma rectum.


Accessory Scrotum With Perineal Lipoma: Pathologic Evaluation Including Androgen Receptor Expression.
Accessory scrotum is an unusual developmental anomaly defined as additional scrotal tissue in addition to a normally developed scrotum. The accessory scrotum arises posterior to the normally located scrotum and does not contain a testis. We report a case of an 18-month-old boy with an accessory scrotum attached to a perineal lipoma. We resected both and determined histologically that they were of the same tissue as the scrotum, including the presence of androgen receptor expression. To the best of our knowledge, this is the first case to assess androgen receptor expression in an accessory scrotum using immunostaining.


Iron deficiency in the rat: biochemical studies of brain metabolism.
Studies were performed to determine the effects of iron deficiency on brain metabolism in rats. Concentrations of cytochrome pigments, oxidative phosphorylation, and catalase and monoamine oxidase activities in brain tissue were unaffected by iron deficiency. However, activities of aldehyde oxidase, a key enzyme in the pathway of serotonin degradation, were significantly reduced, and concentrations of serotonin and total 5-hydroxyindole compounds were elevated in brain tissue of iron-deficient animals. Aldehyde oxidase activities and concentrations of 5-hydroxyindole compounds in brain tissues returned to approximately normal values one week after treatment of iron deficient animals with iron dextran.


Micro-computed Evaluation of Canal Transportation and Centering Ability of 5 Rotary and Reciprocating Systems with Different Metallurgical Properties and Surface Treatments in Curved Root Canals.
The aim of this study was to evaluate the transportation and centering ability of 5 different rotary and reciprocating file systems with different metallurgical properties and surface treatments in curved root canals. Fifty mesiobuccal round canals of upper molars with a curvature of 25°-40° were assigned to 5 experimental groups (n = 12) according to the instrumentation system used: ProTaper Next (Dentsply Sirona, York, PA), Reciproc Blue (RCB [VDW, Munich, Germany]), Reciproc (VDW), TruNatomy (TRN [Dentsply Sirona]), and XP-endo Shaper (FKG, La Chaux-de-Fonds, Switzerland). During instrumentation, 5 mL 2.5% sodium hypochlorite was used in each root canal. The final irrigation protocol included 15% EDTA followed by sodium hypochlorite irrigation. The micro-computed tomographic scanning of the samples was performed before and after instrumentation to analyze the transportation and centering ability at 3 canal levels. The results were analyzed with the 1-way analysis of variance test with the corresponding post hoc test. Overall, RCB caused significantly more canal transportation compared with the other techniques (P < .05). There were no significant differences between the other techniques (P > .05). ProTaper Next had a significantly better ability to stay within the central axis of the root canal compared with the Reciproc and RCB techniques (P = .046 and P = .017, respectively). In the apical third, all techniques caused similar apical transportation and centering ability (P > .05). In the middle and cervical parts of the canal, the RCB caused significantly greater canal transportation than the other techniques (P < .05). Under the limitations of this study, all tested techniques had similar transportation and centering abilities in the apical part of the canal. However, the overall results and those in the middle and coronal parts of the canal indicated that reciprocating instruments resulted in more canal transportation and less centered preparations.


Perceived Value of Electronic Medical Records in Community Health Services: A National Cross-Sectional Survey of Primary Care Workers in Mainland China.
To evaluate the degree to which electronic medical records (EMRs) were used in primary care and the value of EMRs as perceived by primary care workers in China. A cross-sectional survey was conducted on 2719 physicians (n = 2213) and nurses (n = 506) selected from 462 community health centres across all regions of mainland China except for Tibet. Regional differences in the responses regarding the functionality of existing EMR systems and the perceived value of EMRs were examined using Chi-square tests and ordinal regression analyses. Less than 59% of the community health centres had adopted EMRs. More than 89% of the respondents believed that it was necessary to adopt EMRs in primary care. Of the existing EMR systems, 50% had access to telehealth support for laboratory, imaging or patient consultation services. Only 38.4% captured data that met all task needs and 35.4% supported referral arrangements. "Management of chronic conditions" was voted (66%) as the top preferred feature of EMRs. Higher levels of recognition of the value of EMRs were found in the relatively more developed eastern region compared with their counterparts in other regions. Rapid EMR adoption in primary care is evident in mainland China. The low level of functionality in data acquisition and referral arrangements runs counter to the requirements for "management of chronic conditions", the most preferred feature of EMRs in primary care. Regional disparities in the realised value of EMRs in primary care deserve policy attention.


Synthesis and characterization of a 5-membered ring cyclic hydroxylamine coupled to triphenylphosphonium to detect mitochondrial superoxide by EPR spectrometry.
As mitochondrial superoxide is becoming an attractive metabolic and pharmacological target, there is an important need for developing analytical tools able to detect superoxide with high sensitivity and specificity. Among EPR-based methods, it has been recently reported that cyclic hydroxylamines offer a high sensitivity to measure superoxide production. Here, we report the synthesis and evaluation of mitoCPH, in which a 5-membered ring hydroxylamine was coupled to a triphenylphosphonium moiety to allow mitochondrial accumulation. MitoCPH efficiently reacted with superoxide with a bimolecular rate constant of 1.5 × 104 M-1 s-1. We assessed the ability of this compound to detect superoxide in PBS buffer, lysates, and in paraquat-stimulated cells. We compared its performance with CMH, a nontargeted 5-membered ring hydroxylamine, and mitoTEMPO-H, a classically used 6-membered ring hydroxylamine targeted to mitochondria. MitoCPH presented a higher sensitivity for superoxide anion detection than commonly used mitoTEMPO-H, both in buffer and in cell lysates. While we have described the ability of mitoCPH to detect superoxide in different cellular media, we cannot exclude other potential contributors to the nitroxide production from this probe. Therefore, mitoCPH should be considered as a mitochondria-targeted probe and its use as selective superoxide probe should be used cautiously.


miR-338-5p Levels and Cigarette Smoking are Associated With Neuropathic Pain Severity in Individuals With Spinal Cord Injury: Preliminary Findings From a Genome-Wide microRNA Expression Profiling Screen.
To identify microRNA biomarkers and clinical factors associated with neuropathic pain after spinal cord injury. Cross-sectional, secondary analysis of baseline data collected from ongoing clinical studies. Using a genome-wide microRNA screening approach, we studied differential microRNA expression in serum from 43 adults with spinal cord injury enrolled in ongoing clinical studies. Least squares regression was used to identify associations between microRNA expression, clinical factors, and neuropathic pain severity. Community-dwelling individuals with spinal cord injury. Participants (N=43) were at least 18 years old with spinal cord injury, with 28 reporting neuropathic pain and 15 reporting no neuropathic pain. Not applicable. Pain presence, type, and intensity were assessed with the International Spinal Cord Injury Pain Basic Data Set. Serum microRNA normalized deep sequencing counts were quantified from blood samples. Participant demographic factors, injury characteristics, medication use, and health habits were collected via questionnaire. miR-338-5p expression and history of cigarette smoking were associated with and explained 37% of the variance in neuropathic pain severity (R2=0.37, F2,18=5.31, P=.02) independent of other clinical factors. No association was identified between miR-338-5p levels and nociceptive pain severity. Our findings suggest that miR-338-5p and cigarette smoking may both play a role in the development or maintenance of neuropathic pain after spinal cord injury. While additional work is needed to confirm these findings, validated target analysis suggests a neuroprotective role of miR-338-5p in modulating neuroinflammation and neuronal apoptosis and that its downregulation may result in maladaptive neuroplastic mechanisms contributing to neuropathic pain after spinal cord injury.


Brackish Groundwaters Contain Plastic- and Cellulose-Degrading Bacteria.
The selected brackish groundwater occurrences in the geotectonic regions of Inner Dinarides of western Serbia (Obrenovačka Banja) and Serbian crystalline core (Lomnički Kiseljak and Velika Vrbnica) were sampled for isolation and identification of plastic- and lignocellulose-degrading bacteria, as well as for the assessment of their enzymatic potential. The examined occurrences belong to the cold and warm (subthermal), weakly alkaline, neutral, and weakly acidic groundwater, and their genetic types are HCO3-Na + K and HCO3-Ca, Mg. The most abundant genera identified by next-generation 16S sequencing of cultivated groundwater samples belong to Aeromonas and Exiguobacterium. Of isolates screened on plastic and lignocellulosic substrates, 85.3% demonstrated growth and/or degrading activity on at least one tested substrate, with 27.8% isolates degrading plastic substrate Impranil® DLN-SD (SD), 1.9% plastic substrate bis(2-hydroxyethyl)terephthalate, and 5.6% carboxymethyl cellulose (CMC). Isolates degrading SD that were identified by 16S rDNA sequencing belonged to genera Stenotrophomonas, Flavobacterium, Pantoea, Enterobacter, Pseudomonas, Serratia, Acinetobacter, and Proteus, while isolates degrading CMC belonged to genera Rhizobium and Shewanella. All investigated brackish groundwaters harbor bacteria with potential in degradation of plastics or cellulose. Taking into account that microplastics contamination of groundwater resources is becoming a significant problem, the finding of plastic-degrading bacteria may have potential in bioremediation treatments of polluted groundwater. Subterranean ecosystems, which are largely untapped resources of biotechnologically relevant enzymes, are not traditionally considered the environment of choice for screening for plastic- and cellulose-degrading bacteria and therefore deserve a special attention from this aspect.


A randomized clinical trial on proximal contacts of posterior composites.
The objective of this study was to investigate clinical changes in proximal contact strength inserting Class II composite resin restorations according to one of three randomly assigned protocols. Seventy-one Class II restorations (MO/DO) were placed by two calibrated operators. Restorations were randomly assigned to one of three groups: one using a circumferential and two a sectional matrix system with separation rings. Proximal contacts were measured by one independent observer with a Tooth Pressure Meter immediately before treatment, and directly after finishing the restoration. Compared to the situation before treatment groups with a sectional matrix system resulted in a statistical significant stronger mean proximal contact strengths (p<0.05), whereas the use of a circumferential matrix system with hand-instrument resulted in a lower proximal contact strength (p<0.05). Class II posterior composite resin restorations placed with a combination of sectional matrices and separation rings resulted in a stronger proximal contact than when a circumferential matrix system was used.


Output space tracking control for above-knee prosthesis.
The control of a knee joint in an active above-knee prosthesis has been designed using the Lyapunov tracking method. A simulation of locomotion was done to prove that the tracking control in output space is a valuable real time control method for artificial legs. The data used for simulation was collected in able-bodied subjects while they walked on a powered treadmill. Human volunteers were braced with an ankle splint (limiting dorsi- and plantar flexion) and with a knee cage (limiting knee movements to the lateral plane). We studied the achieved tracking of the prescribed knee motion, deviations of the thigh movement from the prescribed trajectory, maximal angular deviations from the desired trajectory and the power consumption as functions of a limited maximal knee torque and a damping constant in the knee actuator. We found that the use of output tracking method is suitable for the design of appropriate hardware of an above-knee prosthesis and for real-time control.


Intersection of reproductive coercion and intimate partner violence: cross-sectional influences on women's contraceptive use in Burkina Faso, Côte d'Ivoire and Kenya.
Among nationally representative cross-sections of women in need of contraception from Burkina Faso, Côte d'Ivoire and Kenya, we aimed to: (1) examine the intersection of past-year physical/sexual intimate partner violence (IPV), emotional IPV and reproductive coercion (RC) and (2) assess the impact of physical/sexual IPV, emotional IPV and RC on women's contraceptive use outcomes, including current contraceptive use, method type and covert use. The present analysis uses cross-sectional female data collected in Burkina Faso (December 2020-March 2021), Côte d'Ivoire (October-November 2021) and Kenya (November-December 2020). Burkina Faso, Côte d'Ivoire and Kenya PARTICIPANTS: Analytical samples were restricted to partnered women with contraceptive need who completed a violence module (Burkina Faso n=1863; Côte d'Ivoire n=1105; Kenya n=3390). The exposures of interest-past-year emotional IPV, physical/sexual IPV and RC-were assessed using abridged versions of the Revised Conflict and Tactics Scale-2 and Reproductive Coercion Scale, respectively. Outcomes of interest included current contraceptive use, contraceptive method type (female controlled vs male compliant), and covert contraceptive use, and used standard assessments. Across sites, 6.4% (Côte d'Ivoire) to 7.8% (Kenya) of women in need of contraception experienced RC; approximately one-third to one-half of women experiencing RC reported no other violence forms (31.7% in Burkina Faso to 45.8% in Côte d'Ivoire), whereas physical/sexual IPV largely occurred with emotional IPV. In multivariable models, RC was consistently associated with covert use (Burkina Faso: aOR 2.84 (95% CI 1.21 to 6.64); Côte d'Ivoire: aOR 4.45 (95% CI 1.76 to 11.25); Kenya: aOR 5.77 (95% CI 3.51 to 9.46)). Some IPV in some settings was also associated with covert use (emotional IPV, Burkina Faso: aOR 2.99 (95% CI 1.56 to 5.74); physical/sexual, Kenya: aOR 2.35 (95% CI 1.33 to 4.17)). Across settings, covert use is a critical strategy for women experiencing RC. Country policies must recognise RC as a unique form of violence with profound implications for women's reproductive health.


Novel technique for the treatment of large subcapsular renal hematoma: combined use of percutaneous drainage and urokinase injection.
To evaluate the efficacy and safety of combined use of percutaneous drainage and urokinase injection to treat patients with large subcapsular renal hematoma. We retrospectively analyzed the data of 20 patients with large subcapsular renal hematoma who were treated with percutaneous drainage and urokinase at our institutes between 2002 and 2010. Percutaneous drainage of the hematoma was performed after bleeding stopped. Then, urokinase was injected into the hematoma cavity per our protocol every 3 days. During the treatment and follow-up period, the diameter of hematoma was assessed by ultrasonography and enhanced computed tomography (CT) in all cases. Complete blood count, serum creatinine and blood pressure were evaluated as well. Follow-up periods ranged from 12 to 60 months (mean 24.1 ± 11.5 months). Percutaneous renal hematoma drainage combined with urokinase injection was successfully performed in all the patients. After sufficient drainage, all of the patients were free from their symptoms. The average time to pain relief was 10.8 ± 2.1 days after percutaneous drainage, and the total duration of drainage was 28 ± 5.1 days. Relative to baseline, there was reduction in the diameter of hematoma evaluated by the enhanced CT scan after treatment (from 11.7 ± 3.1 to 2.8 ± 1.0 cm). During long-term follow-up, active bleeding, hypertension, renal function impairment or infectious complications were not found. This study revealed that combined use of percutaneous drainage and urokinase injection was a safe and effective treatment for patient with large subcapsular renal hematoma.


Umbilical cord blood-derived mesenchymal stromal cells promote myeloid-derived suppressor cell proliferation by secreting HLA-G to reduce acute graft-versus-host disease after hematopoietic stem cell transplantation.
Mesenchymal stem cells (MSCs) use multiple mechanisms to constrain both innate and adaptive immune responses to prevent graft-versus-host disease (GVHD). Myeloid-derived suppressor cells (MDSCs), as a heterogeneous population of early myeloid progenitor cells originating from bone marrow, are a naturally occurring immune regulatory population associated with inhibition of ongoing inflammatory responses, indicating their potential for GVHD therapy. There is accumulating evidence that MSCs and MDSCs do not act independently, but rather establish crosstalk. However, the role of MSCs in MDSC expansion and activation in GVHD remains unexplored. In vitro experiments included 2 groups: peripheral blood mononuclear cells (PBMCs) after mobilization and human umbilical cord blood-derived MSCs (UCB-MSCs) co-cultured with PBMCs. The number and functional difference of MDSCs in PBMCs were determined by flow cytometry. The culture supernatants of co-cultured cells were analyzed to identify cytokines involved in MDSC proliferation. The relationship between MSCs and MDSCs was clarified in GVHD and graft-versus-leukemia (GVL) animal models. In vitro experiments confirmed that UCB-MSCs secreted HLA-G protein to promote and maintain the proliferation of MDSCs in peripheral blood after granulocyte colony-stimulating factor mobilization, and UCB-MSCs mediated the function of MDSCs to inhibit the proliferation of T cells and promote the proliferation of regulatory T cells. UCB-MSCs overexpressing HLA-G induced MDSC production in recipient mice, improved the ability of MDSCs to suppress T cells and further reduced acute GVHD (aGVHD) symptoms and survival time without influencing GVL effects. UCB-MSCs expanded MDSCs via HLA-G/Ig-like transcript 4, reducing the severity of aGVHD without affecting GVL. The immunosuppressive potential of MSCs for the treatment of aGVHD significantly affects the development of MDSCs, thereby consolidating the position of MSCs in the prevention and treatment of aGVHD.


Physicochemical and catalytic properties of grafted vanadium species on different mesoporous silicas.
Vanadium modified mesoporous silicas type MCM-41, MCM-48, and SBA-15 are obtained by a solid state method. The samples are characterized and compared at the different steps of their preparation by X-ray diffraction, N(2)-physisorption, FTIR, UV-vis, XPS, and TG-TPR. Samples catalytic activity is tested in ethylacetate oxidation. Formation of various vanadium species, mainly isolated and small oligomeric ones, grafted to the support surface silanol groups, is observed. It is found that the state and the properties of the vanadium species depend on the porous characteristics of the silica host matrix. The nature of the catalytic active center in the ethylacetate oxidation is discussed.