Contraceptive counseling in 2 urban cities in Ghana and the extent of shared decision-making.
Increased use of contraception is associated with reduced maternal mortality worldwide; however, an unmet need remains high in many places, including Ghana. The quality of care provided by family planning practitioners influences contraceptive use; one way to improve the quality of care is to adopt a client-centered approach to counseling, including engaging in shared decision-making. In Ghana, little is currently known about the extent of shared decision-making between clients and providers in contraceptive counseling encounters. The purpose of this study was to explore the extent of shared decision-making during contraceptive counseling in 2 cities in Ghana. This was a cross-sectional study across 6 urban family planning clinics in Accra and Kumasi, Ghana. We recorded, transcribed, and analyzed 20 family planning patient-provider interactions using the "Observing PatienT InvOlvemeNt" (OPTION) scale. This scale has 12 domains, which are scored on a 5-point scale, from 0 ("the behavior is not observed") to 4 ("the behavior is observed and executed at a high standard"); the scores of each domain are summed up for a total score ranging from 0 to 48. In these encounters, the mean total scores for each interaction ranged from a low of 9.25/48 to a high of 21.5/48. Although providers were thorough in sharing medical information with clients, they did not actively involve clients in the decision-making process and did not generally elicit client preferences. Across the 12 domains, the mean total score was 34.7%, which is below the 50% that would correspond with a "baseline skill level," suggesting there are very low levels of shared decision-making currently occurring. In these 20 patient-provider encounters, counseling was mainly a sharing of medical information from the provider with the client, without the provider eliciting information from the client about her preferences for method characteristics, side effects, or method preference. Family planning counseling in these settings would benefit from increased shared decision-making to engage patients in their contraceptive choice.


Development of muscle fiber types in the prenatal rat hindlimb.
Immunohistochemistry was used to examine the expression of embryonic, slow, and neonatal isoforms of myosin heavy chain in muscle fibers of the embryonic rat hindlimb. While the embryonic isoform is present in every fiber throughout prenatal development, by the time of birth the expression of the slow and neonatal isoforms occurs, for the most part, in separate, complementary populations of fibers. The pattern of slow and neonatal expression is highly stereotyped in individual muscles and mirrors the distribution of slow and fast fibers found in the adult. This pattern is not present at the early stages of myogenesis but unfolds gradually as different generations of fibers are added. As has been noted by previous investigators (e.g., Narusawa et al., 1987, J. Cell Biol. 104, 447-459), all of the earliest generation (primary) muscle fibers initially express the slow isoform but some of these primary fibers later lose this expression. In this study we show that loss of slow myosin in these fibers is accompanied by the expression of neonatal myosin. This switch in isoform expression occurs in all primary fibers located in specific regions of particular muscles. However, in other muscles primary fibers which retain their slow expression are extensively intermixed with those that switch to neonatal expression. Later generated (secondary) muscle fibers, which are interspersed among the primary fibers, express neonatal myosin, although a few of them in stereotyped locations later switch from neonatal to slow myosin expression. Many of the observed changes in myosin expression occur coincidentally with the arrival of axons in the limb or the invasion of axons into individual muscles. Thus, although both fiber birth date and intramuscular position are grossly predictive of fiber fate, neither factor is sufficient to account for the final pattern of fiber types seen in the rat hindlimb. The possibility that fiber diversification is dependent upon innervation is tested in the accompanying paper (K. Condon, L. Silberstein, H.M. Blau, and W.J. Thompson, 1990, Dev. Biol. 138, 275-295).


The role of antibody in the inhibition of the growth of Meth.A tumour in syngeneic experiments in vivo and in vitro.
An in vitro technique for detecting anti-tumour responses was studied and shown to involve non- T cells. Further examination of the effector mechanism revealed that tumour inhibition was antibody mediated, probably through complement dependent lysis; ADCC was considered unlikely. The amount of antibody involved was small, as shown by indirect immunofluorescence labelling of tumour cells, but was nevertheless effective in vitro and in causing regression of tumours in vivo. These findings may have important implications for the manipulation of the host responses to tumours.


Efficient Production of Melanin by Aureobasidium Melanogenum Using a Simplified Medium and pH-Controlled Fermentation Strategy with the Cell Morphology Analysis.
Natural melanin is a biopolymer with wide application prospects in medicine, food, cosmetics, environmental protection, agriculture, and so on. Microbial fermentation is an important and effective way to produce melanin. In this study, Aureobasidium melanogenum, known as black yeast with cellular pleomorphism, was used for the production of melanin. Based on the characteristic of A. melanogenum secreting melanin under oligotrophic stress, a simple medium containing only glucose, MgSO4·7H2O, and KCl was constructed for the production of melanin. The melanin titer of 6.64 ± 0.22 g/L was obtained after 20 days of fermentation without pH control. The cell morphological changes of A. melanogenum during the production of melanin were recorded, and the results showed that chlamydospore might be the most favorable cell morphology for melanin synthesis. Then, different fermentation strategies with cell morphology analysis were developed to further improve the production of melanin in a 5-L fermenter. Results showed that the maximum titer of melanin reached 18.50 g/L by using the fermentation strategy integrating pH control, ammonium salt addition, and H2O2 stimulation, which increased by 178.6% than that of the strategy without pH control. Furthermore, the melanin obtained from the fermentation broth was characterized as eumelanin containing an indole structure. This study provided a potentially feasible fermentation strategy for the industrial production of melanin.


Infant epidermal skin physiology: adaptation after birth.
Functional and structural skin adaptation is a dynamic process which starts immediately after birth in humans and in mammalian skin in general. This adjustment to the extrauterine dry environment is accomplished in the first year of postnatal life of humans. To assess the dynamic changes in vivo after birth in the molecular composition and skin physiology parameters compared with older children and adults. The molecular composition of the stratum corneum (SC) and the water profile were investigated noninvasively by in vivo Raman confocal microscopy as a function of depth. Functional parameters including transepidermal water loss (characterizing epidermal permeability barrier), capacitance (as an indirect parameter for SC hydration) and skin surface pH were assessed noninvasively. The measurements were performed in 108 subjects divided into six age groups: full-term newborns (1-15 days), babies aged 5-6 weeks, babies aged 6±1 months, children aged 1-2 years, children aged 4-5 years and adults aged 20-35 years. We showed that skin acidification is still under development during the first weeks of life. While the basal epidermal barrier is competent immediately after birth, the SC is less hydrated in the first 2 weeks of postnatal life. Similar continuous decreasing water content towards the surface for all age groups was observed, whereas this gradient was lower for the newborns. Dynamic changes in the amounts of the natural moisturizing factor constituents were revealed in the period of infancy. We demonstrated the relation of formation of an acidic pH as well as underlying mechanisms in the induction of a fully hydrated SC over the first weeks of human life as a dynamic functional adaptation.


Proteinuria predicts 10-year cancer-related mortality in patients with type 2 diabetes.
We conducted a cohort study to determine if proteinuria predicts cancer-related mortality in type 2 diabetic subjects. Between July 1996 and June 2003, we enrolled 646 type 2 diabetic subjects. Participants were followed-up until December 31, 2008. The vital status was ascertained by linking records with computerized death certificates in Taiwan. During a median follow-up of 10.4 years, 158 subjects had died, including 59 from cancers. Subjects with proteinuria had a hazard ratio (HR) of 2.77 (95% CI 1.82-4.21) for all-cause mortality and 1.99 (95% CI 1.00-3.94) for cancer-related mortality after adjustment for demographic factors and medical conditions. Specifically, proteinuria showed a trend of increased colon cancer death. The presence of proteinuria significantly improved the predictive ability of cancer-related mortality (increase in concordance statistics or area under the ROC curve=0.03). Patients with both proteinuria and estimated glomerular filtration rate <60 ml/min per 1.73 m² showed higher HR for all-cause mortality than patients with proteinuria only (adjusted HRs (95% CI), 4.01 (2.42-6.67) vs. 2.69 (1.51-4.79), both p<0.01). Proteinuria can predict 10-year all-cause and cancer-related mortalities independently in type 2 diabetic subjects, over and above the established risk factors associated with type 2 diabetes.


Increased endogenous cytokinin in the Arabidopsis amp1 mutant corresponds with de-etiolation responses.
The high-cytokinin Arabidopsis thaliana (L.) Heynh. mutant amp1 of has been further characterised. We extend our previous work on the cytokinin level in the amp1 mutant and show that it contains high levels of endogenous cytokinins in both light- and dark-grown plants, and that amp1 can be phenocopied in the dark by growing wild-type plants on exogenous cytokinin. Previously we showed that dark-grown amp1 plants display de-etiolated characteristics such as short unhooked hypocotyls, opened cotyledons and formation of leaves. We further show that amp1 mutants display de-etiolated plastid morphology and increased levels of transcripts of light-regulated genes, indicating that in this mutant light-mediated processes are partially induced in the dark. The amp1 mutant also shows a reduced level of expression of several light-regulated genes compared with the wild type when grown in light, and has an altered dark-adaptation response when compared with the wild type. These results demonstrate an association between high cytokinin levels and de-etiolation, and we infer that cytokinin itself or a cytokinin-mediated process is involved in regulation of etiolation. The map location, phenotypes and de-etiolation responses in the amp1 mutant are different from those of previously described de-etiolated mutants such as det1, det2, cop1 and cop9. We propose a model in which cytokinin acts as a component of the induction of photomorphogenic processes via a signal transduction pathway which is independent of light.


Estimation of post-mortem interval: A comparison between cerebrospinal fluid and vitreous humour chemistry.
Accurate estimation of post-mortem interval is of great importance in medico-legal autopsy cases. The present study is intended to compare the accuracy of estimating post-mortem interval from biochemical parameters of vitreous humour and cerebrospinal fluid (CSF). Both the fluids were collected from 100 medico-legal autopsies with known time of death and analysed for potassium, glucose, sodium, calcium, urea and creatinine. The current study found that vitreous humour is a better fluid in comparison to CSF for estimation of post-mortem interval. It is also observed that among the statistically significant parameters in both the fluids, level of potassium and sodium in vitreous humour are giving more accurate results in comparison to their corresponding parameters in CSF while accuracy of glucose is more or less same in both the fluid. Nevertheless potassium concentration in vitreous humour is a single best time honoured parameter to estimate post-mortem interval.


An Inducible, Large-Intestine-Specific Transgenic Mouse Model for Colitis and Colitis-Induced Colon Cancer Research.
Animal models are an important tool to understand intestinal biology. Our laboratory previously generated C57BL/6-Tg(Car1-cre)5Flt transgenic mice (CAC) with large-intestine-specific Cre recombinase (Cre) expression as a model to study colon health. To expand the utility of the CAC mouse model by determining the impact of chemically induced colitis on CAC transgene expression. CAC mice were crossed to Rosa reporter mice (Rosa26R (flox/flox) ) with a lox-STOP-lox signal controlling β-galactosidase (βgal) expression and then further crossed with Apc(CKO/CKO) mice in some experiments to delete Apc alleles (Apc (Δ580) ). Initially, 8-week-old CAC(Tg/WT);Rosa26R (flox/WT) ;Apc (Δ580/WT) mice were treated with dextran sulfate sodium (DSS) in drinking water (5 days, 0, 0.65, 1.35, or 2.0 %). Colon tissue damage and βgal labeling were analyzed 10 day after stopping DSS. Next, 8-week-old CAC(Tg/WT);Rosa26R(flox/flox) mice were treated with 0 or 1.35 % DSS, and colonic βgal labeling was assessed at 30 day post-DSS treatment. Finally, 10-week-old CAC(Tg/WT);Apc (Δ580/WT) mice were treated with DSS (0 or 2 %) for 5 days and colonic tumors were analyzed at 20 weeks. CAC(Tg/WT);Rosa26R (flox/WT) ;Apc (Δ580/WT) mice had a DSS dose-dependent increase in colon epithelial damage that correlated with increased epithelial βgal labeling at 10 days (r (2) = 0.9, β = 0.75). The βgal labeling in CAC(Tg/WT);Rosa26R(flox/flox) mice colon remained high at 30 days, especially in the crypts of the healed ulcer. DSS also increased colon tumor incidence and multiplicity in CAC(Tg/WT);Apc (Δ580/WT) mice. DSS-mediated epithelial damage induces a persistent, Cre-mediated recombination of floxed alleles in CAC mice. This enables the examination of gene function in colon epithelium during experimental colitis and colitis-induced colon cancer.


Antibody therapy for Alzheimer's disease.
The economic, social and emotional impact of Alzheimer's dementia is increasing dramatically as greater numbers live to advanced ages. The dearth of effective therapies has led to innovative approaches to treat the disease. This review summarizes the rationale, progress and setbacks regarding the use of antibody-based therapies to treat Alzheimer's disease and discusses future directions for this approach in Alzheimer's and other disorders.


CircRNA-SCAF8 promotes vascular endothelial cell pyroptosis by regulating the miR-93-5p/TXNIP axis.
To investigate the role and mechanism of circRNA-SR-related CTD associated factor 8 (SCAF8) in regulating endothelial cell pyroptosis in high glucose environment. Human umbilical vein endothelial cells (HUVECs) were cultured and divided into six groups. The normal control group and high glucose control group were cultured in cell culture medium with 5 and 33 mmol/L glucose, respectively. The RNA control group, circRNA-SCAF8 inhibition group, miR-93-5p overexpression group and miR-93-5p inhibition group were added with non-functional siRNA, circRNA-SCAF8 inhibitor, miR-93-5p overexpression molecule and miR-93-5p inhibitor in high glucose environment, respectively. Cell viability and pyroptosis were detected by cell counting kit-8 (CCK-8) assay, flow cytometry and Hoechst 33342/propidium iodide fluorescence double staining. Western blotting and enzyme-linked immunosorbent assay were used to detect the expression of pyroptosis-related factors including apoptosis-associated speck-like protein containing a CARD (ASC), cysteine aspartic acid specific protease-1 (caspase-1) and Gasdermin D (GSDMD), NOD like receptor protein 3 (NLRP-3), thioredoxin interacting proteins (TXNIP), IL-18 and IL-1β. The expression of circRNA-SCAF8, miR-93-5p and TXNIP was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Fluorescence in situ hybridization (FISH) was used to locate circRNA-SCAF8 and miR-93-5p. Dual luciferase assay was used to verify the targeted regulatory relationship between miR-93-5p and upstream and downstream molecules. Compared with the RNA control group, the cell survival rate of circRNA-SCAF8 inhibition group and miR-93-5p overexpression group increased (both P<0.01), the pyroptosis decreased (both P<0.01), and the expressions of pyroptosis-related factors such as TXNIP, NLRP-3, caspase-1, GSDMD, ASC, IL-18 and IL-1β were significantly decreased (all P<0.05). The expression of miR-93-5p was significantly increased after inhibition of circRNA-SCAF8 (P<0.01), and the expression of circRNA-SCAF8 tended to decrease after overexpression of miR-93-5p, but with no statistical significance (P>0.05). Dual luciferase assay showed that miR-93-5p downre-gulated circRNA-SCAF8 expression by binding to the 3 ´ UTR region of circRNA-SCAF8, and miR-93-5p downregulated TXNIP expression by binding to the 3 ´ UTR region of TXNIP. FISH showed that circRNA-SCAF8 and miR-93-5p were both located in the cytoplasm and were highly associated in the cells. qRT-PCR showed that the relative expression of TXNIP increased or decreased after overexpression or inhibition of miR-93-5p compared with the RNA control group, respectively (both P<0.05), suggesting that miR-93-5p could regulate TXNIP gene expression. CircRNA-SCAF8/miR-93-5p/TXNIP axis is involved in the regulation of pyroptosis in HUVECs under high glucose.


Fine antigenic specificity and genetic restriction of lysozyme-specific suppressor T cell factor produced by radiation leukemia virus-transformed suppressor T cells.
Culture supernatants obtained from a radiation leukemia virus-transformed, hen egg-white lysozyme (HEL)-specific, suppressor T cell line are able, when injected into mice, to specifically suppress the anti-HEL antibody response. Suppression is observed on both primary and secondary anti-HEL antibody responses evaluated by direct and developed hemolytic plaque assays. Culture supernatants from this HEL-specific suppressor T cell line do not suppress the antibody response induced by a structurally related lysozyme, demonstrating the presence in the culture supernatant of a suppressor factor endowed with fine antigenic specificity. The suppressor factor is able to selectively suppress the anti-HEL antibody response induced by the N-terminal C-terminal peptide of the HEL molecule indicating that the fine specificity of this factor is restricted to an antigenic epitope present in this region of the HEL molecule. The suppressive activity is restricted by genes located within the H-2 complex and analysis of the suppression induced in recombinant mice demonstrates that the interaction between HEL-specific suppressor T cell factor and its cellular target requires identity in the I-J region of the H-2 complex.


Gut microbiome causal impacts on the prognosis of breast cancer: a Mendelian randomization study.
Growing evidence has shown that gut microbiome composition is associated with breast cancer (BC), but the causality remains unknown. We aimed to investigate the link between BC prognosis and the gut microbiome at various oestrogen receptor (ER) statuses. We performed a genome-wide association study (GWAS) to analyse the gut microbiome of BC patients, the dataset for which was collected by the Breast Cancer Association Consortium (BCAC). The analysis was executed mainly via inverse variance weighting (IVW); the Mendelian randomization (MR) results were verified by heterogeneity tests, sensitivity analysis, and pleiotropy analysis. Our findings identified nine causal relationships between the gut microbiome and total BC cases, with ten and nine causal relationships between the gut microbiome and ER-negative (ER-) and ER-positive (ER+) BC, respectively. The family Ruminococcaceae and genus Parabacteroides were most apparent among the three categories. Moreover, the genus Desulfovibrio was expressed in ER- BC and total BC, whereas the genera Sellimonas, Adlercreutzia and Rikenellaceae appeared in the relationship between ER + BC and total BC. Our MR inquiry confirmed that the gut microbiota is causally related to BC. This further explains the link between specific bacteria for prognosis of BC at different ER statuses. Considering that potential weak instrument bias impacts the findings and that the results are limited to European females due to data constraints, further validation is crucial.


Change of causative organisms under antifungal treatment in immunosuppressed patients with HIV-infections.
Oral and esophageal candidosis are very common in HIV-infected patients. Due to the lack of efficacy of topical antimycotics in advanced stages of HIV-infection oral azoles are mainly used for treatment. Azoles most often used are ketoconazole and fluconazole. While Candida albicans clearly is the most frequently encountered yeast before and after treatment other species can be found somewhat less frequently after treatment. This especially applies to Candida glabrata. Candida spp. other than C. albicans obviously may cause manifest oral candidosis. This shift of microbes deserves the more interest as they are less susceptible to azole drugs.


Defect of SLC38A3 promotes epithelial-mesenchymal transition and predicts poor prognosis in esophageal squamous cell carcinoma.
Solute carrier family 38 (SLC38s) transporters play important roles in amino acid transportation and signaling transduction. However, their genetic alterations and biological roles in tumors are still largely unclear. This study aimed to elucidate the genetic signatures of SLC38s transporters and their implications in esophageal squamous cell carcinoma (ESCC). Analyses on somatic mutation and copy number alterations (CNAs) of SLC38A3 were performed as described. Immunohistochemistry (IHC) assay and Western blot assay were used to detect the protein expression level. MTS assay, colony formation assay, transwell assay and wound healing assay were used to explore the malignant phenotypes of ESCC cells. Immunofluorescence assay was used to verify the colocalization of two indicated proteins and immunopreciptation assay was performed to confirm the interaction of proteins. Our findings revealed that SLC38s family was significantly disrupted in ESCC, with high frequent CNAs and few somatic mutations. SLC38A3 was the most frequent loss gene among them and was linked to poor survival and lymph node metastasis. The expression of SLC38A3 was lower in tumor tissues compared to that in normal tissues, which was also significantly associated with worse clinical outcome. Further experiments revealed that depletion of SLC38A3 could promote EMT in ESCC cell lines, and the interaction of SLC38A3 and SETDB1 might lead to the reduced transcription of Snail. Pharmacogenomic analyses demonstrated that fifteen inhibitors were showed significantly correlated with SLC38A3 expression. Our investigations have provided insights that SLC38A3 could act as a suppressor in EMT pathway and serve as a prognostic factor and predictor of differential drug sensitivities in ESCC.


Eustachian Tube Function in 6-Year-Old Children with and without a History of Middle Ear Disease.
To test the hypothesis that eustachian tube opening efficiency, measured as the fractional gradient equilibrated (FGE), is lower in 6-year-old children with no middle ear disease but a well-documented history of recurrent acute otitis media, as compared with children with a negative disease history (control). Cross-sectional study. Tertiary care pediatric hospital. Bilateral eustachian tube function was evaluated in 44 healthy 6-year-old children (19 boys, 29 white). None had middle ear disease at the time of testing, but 23 had a history of recurrent acute otitis media. Twenty-one had no significant past otitis media. Eustachian tube function was measured with a pressure chamber protocol that established negative middle ear gauge pressures (referenced to the chamber pressure) and recorded that pressure before and after a swallow. FGE was calculated as the change in middle ear gauge pressure with swallowing divided by the preswallow pressure. Between-group comparisons of the preswallow pressures and FGEs were made with a 2-tailed Student's t test. FGE was independent of the preswallow middle ear gauge pressure. For the 39 and 44 evaluable ears in the control and recurrent acute otitis media groups, the mean preswallow pressures were -194 daPa (95% confidence interval [95% CI] = -211 to -177) versus -203 (95% CI = -216 to -190; P > .40), and FGEs were 0.32 (95% CI = 0.21-0.43) vs 0.16 (95% CI = 0.08-0.24; P = .016), respectively. In children with past recurrent acute otitis media, residual eustachian tube opening inefficiency is maintained after they have "outgrown" their middle ear disease.


Proteoglycan synthesis in normal and Lowe syndrome fibroblasts.
Lowe (oculocerebrorenal) syndrome (LS) is an X-linked disorder characterized by congenital cataracts, generalized hypotonia, mental retardation, and renal Fanconi syndrome. The basic defect remains unknown, but the possibility that fibroblasts express reduced sulfation of glycosaminoglycans has been studied in several laboratories. A mechanism involving overproduction of an enzyme (nucleotide pyrophosphatase) active against adenosine 3'-phosphate, 5'-phosphosulfate (PAPS) has been postulated. Decreased synthesis of normally sulfated glycosaminoglycans was also reported. We measured the synthesis of proteoglycans and glycosaminoglycans by incorporation of [3H]glucosamine and Na2(35)SO4 into cultured fibroblasts from four LS patients and related it directly to the synthesis in six normal fibroblast cultures. We found that the rate of synthesis varied greatly among the normal cultures (cv, 30%), but not significantly between LS and the normal. The LS fibroblasts' ability to sulfate glycosaminoglycans was assayed as the amount of 3H-glycosaminoglycan eluting at low ionic strength on anion exchange chromatography, the amount of non-sulfated disaccharide present in chondroitinase digests of labeled proteoglycans, and the ratio of 35S to 3H incorporation into proteoglycans. Each parameter suggested that the LS cells were synthesizing normally sulfated glycosaminoglycans (e.g. % delta Di-0S, 21 +/- 6 in normal; 27 +/- 6 in LS). The cells' ability to sulfate glycosaminoglycans was tested under conditions of markedly stimulated glycosaminoglycan synthesis, by treating the cultures with a beta-D-xyloside. LS and normal cells responded to the treatment by elevating the rate of synthesis of normally sulfated glycosaminoglycans (3.5-6-fold in normal, 3-7-fold in LS). Nucleotide pyrophosphatase activities were found to be elevated in each of our four LS cell strains as in the previous studies, excluding genetic heterogeneity as an explanation for our findings. We conclude that LS fibroblasts do not express defects in sulfation of glycosaminoglycans or in synthesis of proteoglycans.


Functional properties of DOM in a stream draining blanket peat.
The functional properties of dissolved organic matter (DOM) from Rough Sike, a stream draining blanket peat in the northern Pennines, UK, were investigated using a series of 12 standardised assays. Nine stream samples were collected at different discharges during 2003--2006, and DOM concentrates obtained by low temperature rotary evaporation. Suwannee River Fulvic Acid was used as a quality control standard in the assays. Dissolved organic matter in high-discharge samples was more light-absorbing at 280 and 340 nm and adsorbed more strongly to alumina, than DOM characteristic of low streamflow, but was less fluorescent and hydrophilic, and poorer in proton-dissociating groups. No significant differences were found in light absorption at 254 nm, copper- or benzo(a)pyrene binding, or photochemical fading. Combination of the Rough Sike data with previously-published results for other streams and a lake yields totals of 20-23 values per assay, for a range of DOM types. For the combined data, variability in all the assays is significant (p < 0.001), as judged by comparison with variations in repeat measurements on the quality control standard. Analysis of the combined data shows that DOM hydrophilicity and adsorption are well-predicted by linear relationships with the extinction coefficient at 340 nm (E340), while good quadratic relationships exist between E340 and both buffering capacity and fluorescence.


A versatile chemical tool for the preparation of conductive graphene-based polymer nanocomposites.
A general route for the functionalization of graphene and graphene derivatives with a low molecular weight polymer by thiol-radical reactions is reported. Polymer-modified graphene is employed as a filler for high density polyethylene to generate materials with good electrical conductivity.


Identification of high-confidence human poly(A) RNA isoform scaffolds using nanopore sequencing.
Nanopore sequencing devices read individual RNA strands directly. This facilitates identification of exon linkages and nucleotide modifications; however, using conventional direct RNA nanopore sequencing, the 5' and 3' ends of poly(A) RNA cannot be identified unambiguously. This is due in part to RNA degradation in vivo and in vitro that can obscure transcription start and end sites. In this study, we aimed to identify individual full-length human RNA isoforms among ∼4 million nanopore poly(A)-selected RNA reads. First, to identify RNA strands bearing 5' m7G caps, we exchanged the biological cap for a modified cap attached to a 45-nt oligomer. This oligomer adaptation method improved 5' end sequencing and ensured correct identification of the 5' m7G capped ends. Second, among these 5'-capped nanopore reads, we screened for features consistent with a 3' polyadenylation site. Combining these two steps, we identified 294,107 individual high-confidence full-length RNA scaffolds from human GM12878 cells, most of which (257,721) aligned to protein-coding genes. Of these, 4876 scaffolds indicated unannotated isoforms that were often internal to longer, previously identified RNA isoforms. Orthogonal data for m7G caps and open chromatin, such as CAGE and DNase-HS seq, confirmed the validity of these high-confidence RNA scaffolds.


Effect of tobacco craving cues on memory encoding and retrieval in smokers.
Previous studies have shown that cue-elicited tobacco craving disrupted performance on cognitive tasks; however, no study has examined directly the effect of cue-elicited craving on memory encoding and retrieval. A distinction between encoding and retireval has been reported such that memory is more impaired when attention is divided at encoding than at retrieval. This study tested the hypothesis that active imagery of smoking situations would impair encoding processes, but have little effect on retrieval. Imagery scripts (cigarette craving and neutral content) were presented either before presentation of a word list (encoding trials) or before word recall (retrieval trials). A working memory task at encoding and free recall of words were assessed. Results indicated that active imagery disrupted working memory on encoding trials, but not on retrieval trials. There was a trend toward impaired working memory following craving scripts compared with neutral scripts. These data support the hypothesis that the cognitive underpinnings of encoding and retrieval processes are distinct.


Magnetic dispersive solid-phase extraction using ZSM-5 zeolite/Fe2O3 composite coupled with screen-printed electrodes based electrochemical detector for determination of cadmium in urine samples.
A novel, simple, fast, sensitive and environmentally friendly approach is presented to determine cadmium in urine samples, combining magnetic dispersive solid-phase extraction (MDSPE) for sample preparation and screen-printed carbon electrodes (SPCEs) for square-wave anodic stripping voltammetry. This association involves the miniaturization of sample preparation and measurement process. Firstly, cadmium was extracted directly from urine samples employing a ZSM-5/Fe2O3, then, the composite enriched with cadmium was deposited onto the SPCE and finally covered with a suitable electrolyte for electrochemical detection. Thereby, the elution and detection of cadmium were carried out in a single step. To optimize experimental parameters affecting MDSPE, a two-step multivariate strategy has been employed. The method has been evaluated under optimized extraction/elution conditions (i.e., type of sorbent, ZSM-5/Fe2O3; amount of sorbent, 10 mg; sample pH, 6.8; extraction time, 5.5 min; and HCl concentration, 0.5 M) using standard addition calibration. Standard addition calibration curves gave a good linearity in the range from 0 to 30 μg L-1 with correlation coefficients ranging from 0.997 to 0.998 (N = 7). The limit of detection, evaluated empirically and statistically, ranged from 0.5 to 1.0 μg L-1 and from 0.4 to 0.8 μg L-1, respectively, which are lower than the threshold level established by the Ministry of Labour and Social Affairs (Spain) and World Health Organization for normal cadmium content in urine (i.e., 3.4 and 4.0 μg L-1, respectively). The repeatability of the proposed method was evaluated at 5 and 20 μg L-1 spiking levels obtaining coefficients of variation ranged between 12 and 15% (n = 6). A certified reference material (REC-8848/Level II) was analyzed to assess method accuracy finding 92% and 1.3 μg L-1 as the recovery (trueness) and standard deviation values, respectively. Finally, the method was applied to spiked urine samples, obtaining good agreement between spiked and found concentrations (recovery ranged from 89 to 98% and CV values ranged from 7% to 14%). Therefore, this is a new and successful contribution to the portable total analytical systems.


Global Asthma Network Phase I Surveillance: Geographical Coverage and Response Rates.
Background-The Global Asthma Network (GAN) Phase I is surveying school pupils in high-income and low- or middle-income countries using the International Study of Asthma and Allergies in Childhood (ISAAC) methodology. Methods-Cross-sectional surveys of participants in two age groups in randomly selected schools within each centre (2015-2020). The compulsory age group is 13-14 years (adolescents), optionally including parents or guardians. Six to seven years (children) and their parents are also optional. Adolescents completed questionnaires at school, and took home adult questionnaires for parent/guardian completion. Children took home questionnaires for parent/guardian completion about the child and also adult questionnaires. Questions related to symptoms and risk factors for asthma and allergy, asthma management, school/work absence and hospitalisation. Results-53 centres in 20 countries completed quality checks by 31 May 2020. These included 21 centres that previously participated in ISAAC. There were 132,748 adolescents (average response rate 88.8%), 91,802 children (average response rate 79.1%), and 177,622 adults, with >97% answering risk factor questions and >98% answering questions on asthma management, school/work absence and hospitalisation. Conclusion-The high response rates achieved in ISAAC have generally been maintained in GAN. GAN Phase I surveys, partially overlapping with ISAAC centres, will allow within-centre analyses of time-trends in prevalence.


Protein-protein interactions in fatty acid elongase complexes are important for very-long-chain fatty acid synthesis.
Fatty acid elongase (FAE), which catalyzes the synthesis of very-long-chain fatty acids (VLCFAs), is a multiprotein complex; however, little is known about its quaternary structure. In this study, bimolecular fluorescence complementation and/or yeast two-hybrid assays showed that homo-interactions were observed in β-ketoacyl-CoA synthases (KCS2, KCS9, and KCS6), Eceriferum2-like proteins [CER2 and CER2-Like2 (C2L2)], and FAE complex proteins (KCR1, PAS2, ECR, and PAS1), except for CER2-Like1 (C2L1). Hetero-interactions were observed between KCSs (KCS2, KCS9, and KCS6), between CER2-LIKEs (CER2, C2L2, and C2L1), and between FAE complex proteins (KCR1, PAS2, ECR, and PAS1). PAS1 interacts with FAE complex proteins (KCR1, PAS2, and ECR), but not with KCSs (KCS2, KCS9, and KCS6) and CER2-LIKEs (CER2, C2L2, and C2L1). Asp308 and Arg309-Arg311 of KCS9 were essential for the homo-interactions of KCS9 and hetero-interactions between KCS9 and PAS2 or ECR. Asp339 of KCS9 is involved in its homo- and hetero-interactions with ECR. Complementation analysis of the Arabidopsis kcs9 mutant by the expression of amino acid-substituted KCS9 mutant genes showed that Asp308 and Asp339 of KCS9 are involved in the synthesis of C24 VLCFAs from C22. This study suggests that protein-protein interaction in FAE complexes is important for VLCFA synthesis and provides insight into the quaternary structure of FAE complexes for efficient synthesis of VLCFAs.


Charge movement and SR calcium release in frog skeletal muscle can be related by a Hodgkin-Huxley model with four gating particles.
Charge movement currents (IQ) and calcium transients (delta[Ca2+]) were measured simultaneously in frog skeletal muscle fibers, voltage clamped in a double vaseline gap chamber, using Antipyrylazo III as the calcium indicator. The rate of release of calcium from the SR (Rrel) was calculated from the calcium transients using the removal model of Melzer, W., E. Rios, and M. F. Schneider (1987. Biophys. J. 51:849-863.). IQ and delta [Ca2+] were calculated for 100 ms depolarizing test pulses to membrane potentials from -30 to +20 mV. To eliminate an inactivating component of Rrel, each test pulse was preceded by a large, fixed prepulse to +20 mV. The resulting Rrel records, which represent the noninactivating component of Rrel, were compared with integral of IQdt.(Q), the total charge that moves. The voltage dependence of the steady state Rrel was steeper then that of Q and shifted to the right. During depolarization, the Rrel waveform was similar to that of Q but was delayed by several ms, while, during repolarization, Rrel preceded Q. All of these results could be explained with a Hodgkin-Huxley type model for E-C coupling in which four voltage sensors in the t-tubule membrane which give rise to IQ must all be in their activating positions for the calcium release channel in the SR membrane to open. his model is consistent with the structural architecture of the triadic junction in which four dihydropyridine receptors (the voltage sensors for E-C coupling) in the t-tubule membrane are closely associated with each ryanodine receptor(the calcium release channel) in the SR membrane [Block, B. A., T. Imagawa, K. P. Campbell, and C. Franzini-Armstrong. 1988. J.Cell. Biol. 107:2587-2600.]). Some aspects of this work have appeared in abstract form (Simon, B. J., and D. Hill. 1991. Biophys. J.59:64a. ([Abstr.]).


Mob/oriT, a mobilizable site-specific recombination system for unmarked genetic manipulation in Bacillus thuringiensis and Bacillus cereus.
Bacillus thuringiensis and Bacillus cereus are two important species in B. cereus group. The intensive study of these strains at the molecular level and construction of genetically modified bacteria requires the development of efficient genetic tools. To insert genes into or delete genes from bacterial chromosomes, marker-less manipulation methods were employed. We present a novel genetic manipulation method for B. thuringiensis and B. cereus strains that does not leave selection markers. Our approach takes advantage of the relaxase Mob02281 encoded by plasmid pBMB0228 from Bacillus thuringiensis. In addition to its mobilization function, this Mob protein can mediate recombination between oriT sites. The Mob02281 mobilization module was associated with a spectinomycin-resistance gene to form a Mob-Spc cassette, which was flanked by the core 24-bp oriT sequences from pBMB0228. A strain in which the wild-type chromosome was replaced with the modified copy containing the Mob-Spc cassette at the target locus was obtained via homologous recombination. Thus, the spectinomycin-resistance gene can be used to screen for Mob-Spc cassette integration mutants. Recombination between the two oriT sequences mediated by Mob02281, encoded by the Mob-Spc cassette, resulted in the excision of the Mob-Spc cassette, producing the desired chromosomal alteration without introducing unwanted selection markers. We used this system to generate an in-frame deletion of a target gene in B. thuringiensis as well as a gene located in an operon of B. cereus. Moreover, we demonstrated that this system can be used to introduce a single gene or an expression cassette of interest in B. thuringiensis. The Mob/oriT recombination system provides an efficient method for unmarked genetic manipulation and for constructing genetically modified bacteria of B. thuringiensis and B. cereus. Our method extends the available genetic tools for B. thuringiensis and B. cereus strains.


Quality of life of adults with workplace exacerbation of asthma.
A cross-sectional study collecting demographic, work history, disease, and quality-of-life (QOL) data from adults with asthma was explored for a relationship between workplace exacerbation of asthma (WEA) and QOL. The study population of adults with asthma was drawn from adults affiliated with Fallon Community Health Plan, a health maintenance organization serving Massachusetts. The sample consisted of 598 adults with asthma. Based on univariate analyses, study participants with WEA had a statistically significant higher Total QOL score, indicating a worse quality of life, than participants whose asthma was not work-related (2.43 vs. 1.74, P < or = 0.001), and also higher scores on the instrument's four subscales for Breathlessness, Mood Disturbance, Social Disruptions, and Health Concerns. After controlling for covariates using multiple linear regression, the relationship between WEA and the Total QOL score was statistically significant (P = 0.0004) with a coefficient of 0.54. The coefficient for WEA was also statistically significant based on regression models for all the subscales with the exception of the Breathlessness score (P = 0.08). In summary, WEA was associated with a worse QOL. Ideally, employees and employers would work together to minimize the conditions at work that contribute to WEA, which should decrease the frequency of WEA and related degradation of QOL.


Shoulder abduction and external rotation restoration with nerve transfer.
In upper brachial plexus palsy patients, loss of shoulder function and elbow flexion is obvious as the result of paralysed muscles innervated by the suprascapular, axillary and musculocutaneus nerve. Shoulder stabilisation, restoration of abduction and external rotation are important as more distal functions will be affected by the shoulder situation. Between 2005 and 2011, eleven patients with upper type brachial plexus palsy were operated on with triceps nerve branch transfer to anterior axillary nerve branch and spinal accessory nerve transfer to the suprascapular nerve for shoulder abduction and external rotation restoration. Nine patients met the inclusion criteria for the study. All patients were men with ages ranged from 21 to 35 years (average, 27.4 years). The interval between injury and surgery ranged from 4 to 11 months (average, 7.2 months). Atrophy of the supraspinatus, infraspinatus and deltoid muscle and subluxation at the glenohumeral joint was obvious in all patients preoperatively. During the pre-op examination all patients had at least muscle grading 4 on the triceps muscle. The mean post-operative value of shoulder abduction was 112.2° (range: 60-170°) while preoperatively none of the patients was able for abduction (p<0.001). The mean post-operative value of shoulder external rotation was 66° (range: 35-110°) while preoperatively none of them was able for external rotation (p<0.001). Postoperative values of shoulder abduction were significantly better that those of external rotation (p=0.0004). The postoperative average muscle grading for shoulder abduction according the MRC scale was 3.6±0.5 and for the shoulder external rotation was 3.2±0.4. Combined nerve transfer by using the spinal accessory nerve for suprascapular nerve neurotisation and one of the triceps nerve branches for axillary nerve and teres minor branch neurotisation is an excellent choice for shoulder abduction and external rotation restoration.


Characteristics of benign lymphoadenosis of oral mucosa.
To investigate the pathological characteristics and carcinogenesis mechanism of benign lymphoadenosis of oral mucosa (BLOM). The expressions of Ki-67, CD34 and apoptosis were evaluated by immunohistochemical SP staining in 64 paraffin-embedded tissue samples. Of them, 9 were from BLOM with dysplasia, 15 from BLOM without dysplasia, 15 from oral squamous cell carcinoma (OSCC), 15 from oral precancerosis, and 10 from normal tissues. Cell proliferation, apoptosis and angiogenesis of tissue samples were also analyzed. The expression of Ki-67 in BLOM with dysplasia, oral precancerosis and OSCC was significantly higher than in BLOM without dysplasia and normal mucosa. The microvascular density (MVD) in BLOM with and without dysplasia, oral precancerosis, and OSCC was significantly higher than in normal mucosa. Apoptosis in BLOM and oral precancerosis was significantly higher than in OSCC and normal mucosa. Benign lymphoadenosis of oral mucosa has potentialities of cancerization.


Interactions of a series of coumarins with reactive oxygen species. Scavenging of superoxide, hypochlorous acid and hydroxyl radicals.
Sixteen plant-derived or synthetic coumarins with various hydroxyl and other substitutions were tested for their ability to inhibit lipid peroxidation and to scavenge hydroxyl radicals, superoxide radicals and hypochlorous acid. Seven unsubstituted or monosubstituted coumarins were essentially inactive in all tests except for ability to scavenge OH with rate constants approximately greater than 1 x 10(9) M-1. sec-1. Of the remaining nine, six containing dihydroxy substitutions were effective inhibitors of Fe3+-ascorbate-dependent microsomal lipid peroxidation (IC50 less than 20 microM), with ortho-dihydroxy + one additional substitution optimal (IC50 less than 10 microM). ortho-Dihydroxylated coumarins were pro-oxidant (enhanced OH generation) in the Fe3+-EDTA-H2O2 deoxyribose system but decreased OH' generation in the Fe3+-ascorbate-H2O2 deoxyribose system, indicating that these compounds can both chelate iron ions and also readily donate electrons for redox cycling of Fe3+. The meta-dihydroxycoumarin did not show this behaviour, but was an effective scavenger of hypochlorous acid, a property shared by only one other compound. Several other coumarins with one or more hydroxyl substituents were also capable of effectively removing superoxide anions (IC50 3.7-72 microM), although some could not be quantified due to direct rapid reduction of cytochrome c. We conclude that several compounds, notably 5,7-dihydroxy-4-methylcoumarin, possess beneficial biochemical profiles of interest in relation to pathophysiological processes dependent upon reactive oxygen species.


COVID-19 and Cardiovascular Health.
In late February 2020, a time when severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, or COVID-19) still felt like an abstraction in the United States, New York City's first infected patient was admitted to Mount Sinai Hospital's emergency room. Working a few doors down was Sean Pinney, the Director of Advanced Heart Failure and Transplantation. Little did he know, but "that night was the beginning of hell," he said.


Expanding the spectrum of chronic necrotising (semi-invasive) aspergillosis: a series of eight cases presenting as radiologically solid lung nodules mimicking malignancy.
To clinicopathologically characterise a subset of aspergillosis cases characterised by radiologically solid lung nodules. We reviewed our archives for lung biopsies/resections with fungal hyphae on histology. Cases classifiable as a well-established form of aspergillosis were excluded. Unclassifiable cases with radiologically solid lung nodules and histological evidence of fungal hyphae consistent with Aspergillus were analysed in detail. Of 134 lung biopsies/resections with fungal hyphae, eight presented as a solid lung nodule on imaging and could not be classified as a well-recognised form of aspergillosis. All patients were non-immunocompromised/minimally immunocompromised adult smokers. Imaging showed solid, predominantly solitary, spiculated, subpleural, hypermetabolic upper lobe nodules in emphysematous lungs. Malignancy was suspected clinically in all cases. Histologically, the nodules were necrotising granulomas containing fungal hyphae consistent with Aspergillus. On follow-up, none of the nodules recurred or progressed to invasive disease. Aspergillosis can present as a radiologically solid, spiculated lung nodule suspicious for malignancy.


Aneuploidy of glandular epithelial cells in histologically normal prostate glands.
The percentage of aneuploidy in normal prostate glands from subjects 13-38 years old and 45-66 years old ranged from 0-78% and 0-63%, respectively. In contrast to adults, aneuploidy was absent in fetal and postnatal prostates. It is concluded that aneuploidy is a fundamental attribute of histologically normal adult prostate glands.


Patient-derived tumor models: a suitable tool for preclinical studies on esophageal cancer.
Esophageal cancer (EC) is the tenth most common cancer worldwide and has high morbidity and mortality. Its main subtypes include esophageal squamous cell carcinoma and esophageal adenocarcinoma, which are usually diagnosed during their advanced stages. The biological defects and inability of preclinical models to summarize completely the etiology of multiple factors, the complexity of the tumor microenvironment, and the genetic heterogeneity of tumors severely limit the clinical treatment of EC. Patient-derived models of EC not only retain the tissue structure, cell morphology, and differentiation characteristics of the original tumor, they also retain tumor heterogeneity. Therefore, compared with other preclinical models, they can better predict the efficacy of candidate drugs, explore novel biomarkers, combine with clinical trials, and effectively improve patient prognosis. This review discusses the methods and animals used to establish patient-derived models and genetically engineered mouse models, especially patient-derived xenograft models. It also discusses their advantages, applications, and limitations as preclinical experimental research tools to provide an important reference for the precise personalized treatment of EC and improve the prognosis of patients.


Relation of late potentials to ejection fraction and wall motion abnormalities in acute myocardial infarction.
A prospective study was performed to determine the relation between quantitative signal-averaged parameters and ejection fraction (EF) and wall motion abnormalities determined by radionuclide ventriculography in patients with acute myocardial infarction (AMI). In 50 patients with AMI, signal-averaging of the surface QRS complex (200 beats; filter frequencies of 40 to 250 Hz and 80 to 250 Hz) was performed and radionuclide ventriculograms were recorded 8 +/- 5 days after AMI. Twenty-five of these patients (50%) had anterior wall AMI, 20 (40%) had inferior wall AMI and 5 (10%) had non-Q-wave AMI. The duration of the low-amplitude signals of less than 40 microV, the signal-averaged QRS complex and the root-mean-square voltage of the terminal 40 ms were determined. In addition to EF determinations, wall motion abnormalities were assessed for the presence or absence of dyskinetic, akinetic and hypokinetic segments. A wall motion score was constructed by separating the left and right ventricles into 21 segments in the anterior, left anterior oblique and lateral views. On the basis of the presence or absence of late potentials, the patients were separated into 2 groups: group I comprised 15 patients (30%) with late potentials and group II 35 patients (70%) without late potentials. The low-amplitude signals (49 +/- 12 vs 24 +/- 8 ms) and the signal-averaged QRS complex (122 +/- 20 vs 96 +/- 15 ms) were significantly longer and the root-mean-square voltage (13.8 +/- 4.9 vs 54.3 +/- 27.4 microV) significantly lower in group I than in group II.(ABSTRACT TRUNCATED AT 250 WORDS)


Coping with subjective and objective stress during a pandemic pregnancy: implications for maternal mental health.
The COVID-19 pandemic heightened prenatal maternal stress, a risk factor for poorer maternal and infant health. There was substantial variability, however, in the extent to which the stress of pandemic pregnancy influenced maternal mental health. Some of this variability may have been due to the different coping strategies used to manage pandemic stress. In this cross-sectional study of 7,383 pregnant women in the U.S. (M = 25.69 ± 8.71 weeks gestational age) recruited during the first and second U.S. waves of the COVID-19 pandemic, we examined associations of objective stressors, the subjective experience of stress, and the use of four coping strategies with anxiety and depressive symptoms. Spiritual coping, planning/preparation, and avoidant coping were associated with increased subjective and objective stress and with greater mood and anxiety symptoms, whereas coping by positive appraisal was associated with modestly lower subjective stress and with lower mood and anxiety symptoms. We also found small interactions of stress and coping in predicting mood and anxiety symptoms, suggesting that fit between coping strategy and type of stress influences coping outcomes. Specific coping strategy used as well as the fit between coping strategy and stress type may determine whether coping buffers or exacerbates mood and anxiety symptoms. The small magnitude of these associations suggests that individual-level coping may be insufficient in the face of the overwhelming nature of the stress accompanying a global pandemic. This work adds to our understanding of coping with pregnancy stress in the context of population-level stressors (i.e., a pandemic or large-scale disaster).


The effects of low pH and high water temperature on oxidative stress and cell damage in juvenile olive flounder Paralichthys olivaceus: comparison of single and combined environmental conditions.
The use of fossil fuels by anthropogenic activities causes ocean acidification and warming, and these changes in the marine environment can negatively affect the metabolism, growth, and survival of fish. In the present study, we evaluated the ability of olive flounder Paralichthys olivaceus to cope with future marine environmental changes by investigating the oxidative stress (cortisol, HSP70), antioxidant enzyme (superoxide dismutase; SOD, catalase; CAT) activity, and apoptosis (caspase-3) after exposure to control conditions (20 °C and pH 8.1), warming (30 °C) and acidification (pH 7.5) conditions, and a combined environment (30 °C and pH 7.5) for 28 days. Under warming conditions, increased oxidative stress, activity of antioxidant enzymes, and apoptosis were observed. Acidifying conditions showed negative effects at the beginning of exposure, but these effects were offset over time. Even in a combined environment of acidification and warming, negative effects were seen only at the beginning of exposure and were not sustained. In conclusion, the effects of acidification on oxidative stress, antioxidant response, and apoptosis in P. olivaceus did not exceed the effects of warming. These results suggest that P. olivaceus can cope with the predicted future acidifying environment.


Nanoparticle-Coupled Topical Methotrexate Can Normalize Immune Responses and Induce Tissue Remodeling in Psoriasis.
Methotrexate (MTX) is an antiproliferative drug used for treating inflammatory diseases, including psoriasis. Nevertheless, its use in localized therapy is hindered because of poor transdermal penetration. We show that MTX coupled with gold nanoparticles (GNPs) demonstrates superior antiinflammatory efficacy than MTX alone in an imiquimod-induced mouse model, significantly reducing γδ T cells, CD4+ T cells, and neutrophils. Furthermore, it was well tolerated upon systemic and topical administration. In an AGR129 human xenograft mouse model, two-week topical treatment with MTX-GNPs inhibited skin hyperplasia significantly better than topical calcipotriol-betamethasone and led to profound tissue remodeling, involving the upregulation of extracellular matrix reorganization and the downregulation of cornification and keratinization processes. The number of resident T cells in the grafts, as well as interleukin-17 production, drastically decreased upon MTX-GNP treatment. While both MTX and MTX-GNPs directly prevented the proliferation and induced apoptosis of T cells, the suppression of cytokine production was a shared mechanism of GNP and MTX-GNPs. In conclusion, MTX-GNPs influence immune and stromal components of the skin, leading to the potent inhibition of pathogenesis in preclinical psoriasis. MTX-GNPs surpass the efficacy of conventional MTX and standard of care, emerging as a non-steroidal, topical alternative for psoriasis treatment.


Antibacterial conductive self-healing hydrogel wound dressing with dual dynamic bonds promotes infected wound healing.
In clinical applications, there is a lack of wound dressings that combine efficient resistance to drug-resistant bacteria with good self-healing properties. In this study, a series of adhesive self-healing conductive antibacterial hydrogel dressings based on oxidized sodium alginate-grafted dopamine/carboxymethyl chitosan/Fe3+ (OSD/CMC/Fe hydrogel)/polydopamine-encapsulated poly(thiophene-3-acetic acid) (OSD/CMC/Fe/PA hydrogel) were prepared for the repair of infected wound. The Schiff base and Fe3+ coordination bonds of the hydrogel structure are dynamic bonds that can be repaired automatically after the hydrogel network is disrupted. Macroscopically, the hydrogel exhibits self-healing properties, allowing the hydrogel dressing to adapt to complex wound surfaces. The OSD/CMC/Fe/PA hydrogel showed good conductivity and photothermal antibacterial properties under near-infrared (NIR) light irradiation. In addition, the hydrogels exhibit tunable rheological properties, suitable mechanical properties, antioxidant properties, tissue adhesion properties and hemostatic properties. Furthermore, all hydrogel dressings improved wound healing in the infected full-thickness defect skin wound repair test in mice. The wound size repaired by OSD/CMC/Fe/PA3 hydrogel + NIR was much smaller (12%) than the control group treated with Tegaderm™ film after 14 days. In conclusion, the hydrogels have high antibacterial efficiency, suitable conductivity, great self-healing properties, good biocompatibility, hemostasis and antioxidant properties, making them promising candidates for wound healing dressings for the treatment of infected skin wounds.


A qualitative study of active travel amongst commuters and older adults living in market towns.
Being physically active is associated with better health, but rates of physical inactivity are high amongst adults in England. Active travel, defined as making routine journeys in physically active ways, has been identified as a potential solution. There is a lack of research into how modal shift towards active travel can be encouraged in market towns. The aims of the current study are to understand how new cycling and walking infrastructure and community activation projects might support modal shift to active travel amongst commuters and older adults in market towns. This was a qualitative study using focus groups, 'go-along' interviews, and in-depth interviews as the main methods of data collection. Thirty-three participants (12 commuters and 21 older adults) took part across six focus groups. Eight of these also completed a go-along interview (4 walking, 4 cycling). Data were analysed using the Framework method of thematic analysis. Market towns have existing advantages for active travel, being relatively compact with most routine destinations within easy reach. The barriers to active travel faced by older adults and commuters in market towns are similar to those in cities; poor infrastructure remains the key barrier. Poorly maintained paths are hazardous for older pedestrians, and low-or-no lighting and lack of well-connected, delineated cycle routes deter both commuters and older adults. One factor which does appear qualitatively different to cities is participants' perception that the social norms of cycling differ in market towns. Policies to promote active travel in market towns are most likely to be effective when they include measures targeted at both individual behaviour change and population level measures like large-scale infrastructure improvements. Initiatives to change the social norms around cycling may be required to increase active travel rates.


[Patient-centeredness].
Patient-centeredness has developed into the guiding principle of healthcare policy over the last decade. However, its practical implementation is hindered by numerous problems and opposing interests. To define and elucidate the term patient-centeredness, describe evidence-based measurement tools and outline recommendations for practical implementation. A narrative literature review was carried out in Medline, Cochrane Library, PsyINfo and CINHAL. Based on the results the concept of Patient-Reported Experience Measures (PREM) is explained and a comprehensive PREM system is developed. Patient-centeredness is a not yet clearly defined theoretical construct. Patient-centeredness covers up to 16 different aspects ranging from patient involvement to the transition of care. Patient centeredness is most frequently measured via PREMs. Contrary to other countries, there are only a limited number of validated PREMs available in German and measurement qualities are frequently unclear or insufficient. No comprehensive PREM system has been developed for the German language so far. Important aspects of surgical care are not captured by currently available German language PREMs. Contrary to other countries no comprehensive PREM system is available in the German language. Currently available German PREMs do not enable the adequate assessment of surgical aspects of care. Important organizational, regulatory, methodological, and financial aspects must be addressed before patient-centeredness can be implemented into every clinical practice in German-speaking countries.


Maternal Psychiatric Disorder and the Risk of Autism Spectrum Disorder or Intellectual Disability in Subsequent Offspring.
Psychiatric disorders are more common in the mothers of children with autism spectrum disorder (ASD) or intellectual disability (ID) after the birth of their child. We aimed to assess the relationship between women's psychiatric contacts and subsequent offspring with ASD/ID. We linked three Western Australian registers to investigate pre-existing maternal outpatient psychiatric contacts and the odds of ASD/ID in a subsequent child. Women with a previous outpatient psychiatric contact were more than twice as likely to have a child with ASD [OR 2.07 (95 % CI 1.7, 2.6)] or ID [OR 2.31 (2.1, 2.6)]. Further research exploring the effect on pregnancy outcomes of medications prescribed to women with psychiatric disorders is implicated.


The Role of Attachment in Gambling Behaviors and Gambling Disorder: A Systematic Review.
In recent years, a growing number of attachment-based studies have contributed to the understanding of both substance and behavioral addictions. Although gambling is a form of addictive behavior widespread all over the world, both among young people and adults, the evidence on the association between attachment-related phenomena and gambling has not yet been systematized in literature. The aim of the present study, therefore, is to provide a systematic literature review aimed at summarizing the empirical evidence on this topic. Following the updated 2020 PRISMA guidelines, a systematic search in four electronic scientific databases (Scopus, PubMed, PsycInfo and Web of Science) was conducted. After removing duplicates, 146 records were double-screened, with 12 articles meeting the inclusion criteria. Additionally, by means of a backward search a further article was selected. Altogether, 13 articles were selected for the present systematic review. With few exceptions, the results underline the significant role played by attachment-related phenomena in gambling behaviors, highlighting that specific attachment contexts have a different influence on gambling, also depending on whether gamblers are youths or adults. In particular, while secure attachment has proven to be a protective factor for the onset of gambling behavior, insecure attachment has emerged to be a vulnerability factor in two ways. On the one hand, it directly favors gambling behaviors; on the other, it affects coping strategies and the individual's ability to identify and regulate emotions, which in turn predict gambling. Limitations, strengths, and implications of the present systematic review are discussed.


Unraveling the Pivotal Role of Bradykinin in ACE Inhibitor Activity.
Historically, the first described effect of an angiotensin converting enzyme (ACE) inhibitor was an increased activity of bradykinin, one of the substrates of ACE. However, in the subsequent years, molecular models describing the mechanism of action of ACE inhibitors in decreasing blood pressure and cardiovascular risk have focused mostly on the renin-angiotensin system. Nonetheless, over the last 20 years, the importance of bradykinin in regulating vasodilation, natriuresis, oxidative stress, fibrinolysis, inflammation, and apoptosis has become clearer. The affinity of ACE appears to be higher for bradykinin than for angiotensin I, thereby suggesting that ACE inhibitors may be more effective inhibitors of bradykinin degradation than of angiotensin II production. Data describing the effect of ACE inhibition on bradykinin signaling support the hypothesis that the most cardioprotective benefits attributed to ACE inhibition may be due to increased bradykinin signaling rather than to decreased angiotensin II signaling, especially when high dosages of ACE inhibitors are considered. In particular, modulation of bradykinin in the endothelium appears to be a major target of ACE inhibition. These new mechanistic concepts may lead to further development of strategies enhancing the bradykinin signaling.


Anisotropic magnetic property, magnetostriction, and giant magnetocaloric effect with plateau behavior in TbMn2Ge2 single crystal.
The ternary RMn2Ge2 (R = rare earth) intermetallic compounds have attracted great attention due to their interesting magnetic behaviors and magnetotransport responses. Here, we reported our observation of anisotropic magnetic property, magnetostriction, and magnetocaloric effect (MCE) in TbMn2Ge2 single crystal. Below the transition temperature of Tb magnetic sublattices ([Formula: see text] ~ 95 K), strong Ising-like magnetocrystalline anisotropy is observed with an out-of-plane ferromagnetic moments 5.98 μB/f.u. along the easy c axis, which is two orders of magnitude larger than that of field along a axis. Above [Formula: see text], a field-induced metamagnetic transition is observed from the spin-flip of Mn sublattices. During this transition, remarkable magnetostriction effect is observed, indicating of strong spin-lattice coupling. The responses of Tb and Mn sublattices to the magnetic field generate a giant magnetic entropy change ([Formula: see text]) and large values of relative cooling power (RCP) and temperature-averaged entropy change (TEC). The calculated maximum magnetic entropy change ([Formula: see text]), RCP, and TEC(10) with magnetic field change of 7 T along c axis reach 24.02 J kg-1 K-1, 378.4 J kg-1, and 21.39 J kg-1 K-1 near [Formula: see text], which is the largest among RMn2Ge2 families. More importantly, this giant MCE shows plateau behavior with wide window temperatures from 93 to 108 K, making it be an attractive candidate for magnetic refrigeration applications.


Oximetry-derived perfusion index for intraoperative identification of successful thoracic sympathectomy.
Endoscopic thoracic sympathectomy treats patients with hyperhidrosis. Laser Doppler flow and changes in palmar temperature have been advocated for intraoperative monitoring. The pulse oximetry-derived perfusion index (PI) is used to quantify pulsatile blood flow at the pulse oximeter. Upper limb sympathectomy is associated with increased flow to the ipsilateral extremity; thus we postulate that it will increase PI. We evaluated changes in intraoperative PI as a monitor of successful thoracic sympathectomy. After institutional review board approval and informed consent, 10 adult patients undergoing bilateral endoscopic thoracic sympathectomy under general anesthesia were studied. Finger pulse-oximetry probes were placed on each hand, and reference probes on each earlobe. Hemodynamic variables and PI were continuously monitored. Data were collected immediately before and every minute after sympathectomy for 5 minutes. Anesthetic management remained constant throughout. A successful sympathectomy was defined by a twofold increase in PI on the ipsilateral arm. Data were analyzed with analysis of variance and Student's t tests; a p < 0.05 was considered significant. Baseline oximetric waveforms were adequate in all subjects. Right sympathectomy was associated with a 372% increase in PI (p < 0.0001), and left sympathectomy with a 316% increase in PI (p < 0.029). This occurred as early as 1 minute after transection of the sympathetic chain. The PI in the reference probes as well as the hemodynamics remained constant. All patients had postoperative resolution of their hyperhidrosis symptoms. In patients with hyperhidrosis of the upper extremity, the intraoperative PI derived from pulse oximetry is an additional indicator of successful thoracic sympathectomy.


Effect of ion exchange resins on the drug release from matrix tablets.
Ion exchange resins were incorporated into hydroxypropylmethylcellulose (HPMC) matrix tablets to modify the release of oppositely charged drugs. The drug release from HPMC tablets containing drug-resin complexes was significantly slower than from HPMC tablets containing drug without resin. A physical mixture of drug and ion exchange resin (cationic drug, propranolol HCl, with the cation exchange resin, Amberlite IRP 69, or the anionic drug, sodium diclofenac, with the anion exchange resin, cholestyramine (Duolite ATP-143)) resulted in almost the same drug release as tablets containing preformed drug-resin complexes. Upon contact with the dissolution medium, a gel layer formed rapidly around the solid tablet core and the complex between the drug and the resin formed in situ within the gelled regions. No effect of pH of the dissolution medium (0.1 N HCl or pH 7.4 phosphate buffer) or resin counterion was observed with the strong cation exchanger, Amberlite IRP 69. The resin was dissociated at both pH-values, allowing drug binding. With the weak cation exchange resin, Amberlite IRP 88, in situ complex formation and retardation was only observed in pH 7.4 buffer but not in 0.1 N HCl because of the non-ionization of the carboxyl groups. The drug release depended also on the amount and particle size of the resin particles and the type of carrier. The use of smaller resin particles eliminated the burst release seen with larger resin particles. Upon comparing different carrier materials, a rapid formation of the gel layer was important for the in situ complex formation. The drug release was in the order of Gelucire 54/02 (glyceryl palmitostearate) > polyethylene oxide 400K > HPMC K15M.


Liver function in Huntington's disease assessed by blood biochemical analyses in a clinical setting.
Huntington's disease (HD) is a dominantly inherited, progressive neurological disorder caused by a CAG repeat elongation in the huntingtin gene. In addition to motor-, psychiatric- and cognitive dysfunction, peripheral disease manifestations in the form of metabolic changes and cellular dysfunction are seen. Blood levels of a wide range of hormones, metabolites and proteins have been analyzed in HD patients, identifying several changes associated with the disease. However, a comprehensive panel of liver function tests (LFT) has not been performed. We investigated a cohort of manifest and premanifest HD gene-expansion carriers and controls, using a clinically applied panel of LFTs. Here, we demonstrate that the level of alkaline phosphatase is increased in manifest HD gene-expansion carriers compared to premanifest HD gene-expansion carriers and correlate with increased disease severity indicated by the Unified Huntington's disease rating scale-Total Functional Capacity Score (UHDRS-TFC). For gamma-glutamyl transferase, elevated levels were more frequent in the manifest groups than in both the HD gene-expansion negative controls and premanifest HD gene-expansion carriers. Finally, the manifest HD gene-expansion carriers displayed moderate increases in total cholesterol and blood glucose relative to the premanifest HD gene-expansion carriers, as well as increased C-reactive protein relative to HD gene-expansion negative controls. Our results show that LFT values are elevated more frequently in manifest compared to premanifest HD gene-expansion carriers and controls. The majority of the manifest HD gene-expansion carriers receive medication, and it is possible that this can influence the liver function tests performed in this study.


Hormonal Treatment Effect on Sexual Distress in Transgender Persons: 2-Year Follow-Up Data.
As far as we know, no studies to date have investigated the psychobiological correlates of sexual distress (SD) nor the impact of hormonal treatment (HT) on SD in transgender persons. To evaluate the psychobiological correlates of SD and assess the effects of HT on SD in transgender persons without gender-affirming surgery. A consecutive series of 301 transgender persons (160 transwomen and 141 transmen) was considered for the cross-sectional study, and a subset of 72 subjects was studied in a 2-year follow-up. A physical examination was performed. Blood samples were drawn for determination of cortisol levels. Subjects completed psychometric measures. During 2 years of HT, the evaluation of SD was prospectively repeated. Psychobiological correlates of SD in transgender population. Changes in SD during gender affirming hormonal treatment. Knowing how hormonal treatment influence SD will help care providers when counseling transgender people. To the authors' knowledge, this is the first study prospectively evaluating the impact of gender affirming hormonal treatment on sexual distress in transgender individuals. The main limitations are represented by the small size of the sample and the use of questionnaires validated only in the cisgender population. SD showed a positive correlation with body uneasiness (P < .0001) and with dissatisfaction toward gender-related body parts or shapes (all P < .05). In addition, SD correlated positively with general psychopathology (P < .0001), alexithymia, social anxiety, and humiliation scales (all P < .05). In transmen, SD was positively associated with autism levels (P < .005), as well as with cortisol levels (P < .02). A significant correlation between SD and perceived discrimination was observed in transwomen (P < .05). In transwomen, SD was positively associated with hair density and negatively with breast growth (both P < .05). Finally, in transmen, a negative correlation was found between SD and hair density (P < .05). When the impact of HT on SD was evaluated, a significant reduction of SD was observed across time in both transwomen and transmen (P = .001 and P = .01, respectively). The present results support the efficacy of HT in reducing SD in transgender persons. Ristori J, Cocchetti C, Castellini G, et al. Hormonal Treatment Effect on Sexual Distress in Transgender Persons: 2-Year Follow-Up Data. J Sex Med 2020;17:142-151.


Disinfection of respirator tubing: a comparison of chemical versus hot water machine-assisted processing.
A comparison of machine-assisted chemical disinfection, using a glutaraldehyde solution, and machine-assisted hot water disinfection was made by placing a large inoculum (approximately 10(7) colonies of Pseudomonas aeruginosa or Acinetobacter calcoaceticus var. anitratus) in sets of tubing before processing. Machine-assisted chemical processing proved to be the most efficient method with a disinfection failure rate of 6 per cent (two positive tubes out of 35). Machine-assisted hot water processing had a disinfection failure rate of 83 per cent (44 of 53 tubes were not successfully disinfected). Some of the tubes that were disinfection failures were dried in a hot air cabinet after the initial sampling, which further reduced the colony counts in the tubing but did not totally eliminate growth except in one tube. The colony counts after drying, showed that A. anitratus was 15 per cent more sensitive to hot air drying than Ps. aeruginosa, but this was not a statistically significant reduction.


Who are the real bird brains? Qualitative differences in behavioral flexibility between dogs (Canis familiaris) and pigeons (Columba livia).
Pigeons given a simultaneous spatial discrimination reversal, in which a single reversal occurs at the midpoint of each session, consistently show anticipation prior to the reversal as well as perseveration after the reversal, suggesting that they use a less effective cue (time or trial number into the session) than what would be optimal to maximize reinforcement (local feedback from the most recent trials). In contrast, rats (Rattus norvegicus) and humans show near-optimal reversal learning on this task. To determine whether this is a general characteristic of mammals, in the present research, pigeons (Columba livia) and dogs (Canis familiaris) were tested with a simultaneous spatial discrimination mid-session reversal. Overall, dogs performed the task more poorly than pigeons. Interestingly, both pigeons and dogs employed what resembled a timing strategy. However, dogs showed greater perseverative errors, suggesting that they may have relatively poorer working memory and inhibitory control with this task. The greater efficiency shown by pigeons with this task suggests they are better able to time and use the feedback from their preceding choice as the basis of their future choice, highlighting what may be a qualitative difference between the species.


Adaptation maintains population homeostasis in primary visual cortex.
Sensory systems exhibit mechanisms of neural adaptation, which adjust neuronal activity on the basis of recent stimulus history. In primary visual cortex (V1) in particular, adaptation controls the responsiveness of individual neurons and shifts their visual selectivity. What benefits does adaptation confer on a neuronal population? We measured adaptation in the responses of populations of cat V1 neurons to stimulus ensembles with markedly different statistics of stimulus orientation. We found that adaptation served two homeostatic goals. First, it maintained equality in the time-averaged responses across the population. Second, it maintained independence in selectivity across the population. Adaptation scaled and distorted population activity according to a simple multiplicative rule that depended on neuronal orientation preference and on stimulus orientation. We conclude that adaptation in V1 acts as a mechanism of homeostasis, enforcing a tendency toward equality and independence in neural activity across the population.


Incidence and Predictors of Relapse After Weaning Immune Suppressive Therapy in Cardiac Sarcoidosis.
Cardiac sarcoidosis (CS) is a relapsing-remitting disease, and immune suppression (IS) is the mainstay of therapy. Predictors of relapse for patients with CS in remission are not well characterized. We assessed incidence of relapse in consecutive patients with CS treated with high-dose steroids and/or steroid-sparing agents (SSA) in our center from 2000 to 2020. Remission was defined as reaching maintenance therapy (no IS, SSA, and/or prednisone ≤5 mg/d) for ≥1 month. Relapse was defined as recurrence of CS syndrome requiring IS intensification: heart failure, ventricular arrhythmia, decrease in left ventricular ejection fraction, or increased disease burden on imaging. Among 68 patients, the mean age was 50.7±9.0 years; 25 (37%) were women, and 32 (47%) were Black. In total, 59 patients (87%) reached remission. Over a median follow-up of 39.5 months (interquartile range 17.6, 92.5), 28 (48%) relapsed. Greater percentage of late gadolinium enhancement (LGE) on pretreatment magnetic resonance imaging corresponded with increased likelihood of relapse (odds ratio 1.396 per 5% increase [95% confidence interval (CI) 1.04 to 1.88]; p = 0.028). LGE ≥11% predicted elevated risk of relapse (adjusted odds ratio 4.998 [1.34 to 18.64]; p = 0.017). Shorter time to relapse was observed with isolated CS (adjusted hazard ratio 4.084 [1.44,11.56]; p = 0.008) and LGE ≥11% (adjusted hazard ratio 3.007 [1.01, 8.98]; p = 0.049). Approximately 1 in 2 patients with CS in remission experienced relapse. Greater burden of LGE on cardiac magnetic resonance imaging and isolated CS are associated with greater risk of relapse. Future work is needed to refine risk stratification for relapse and to optimize surveillance strategies on the basis of the burden of disease.


Geometric orientation by humans: angles weigh in.
Human participants were trained to navigate to two geometrically equivalent corners of a parallelogram-shaped virtual environment. The unique shape of the environment combined three distinct types of geometric information that could be used in combination or in isolation to orient and locate the goals: the angular amplitudes of the corners, the relative wall length relationships, and the principal axis of symmetry. In testing, participants were placed in manipulated versions of the training environment that tested which types of geometry they had encoded and how angular information weighed in against the other two geometric properties. The test environments were (a) a rectangular environment that removed the angular information, (b) a rhombic environment that removed wall length information and drastically reduced the principal axis, and (c) a reverse-parallelogram-shaped environment that placed angular information against both wall length and principal axis information. Participants chose accurately in the rectangular and rhombus environments, despite the removal of one of the cues. In the conflict test, participants preferred corners with the correct angular amplitudes over corners that were correct according to both wall length relationships and the principal axis. These results are comparable to recent findings with pigeons and suggest that angles are a salient orientation cue for humans.


Evaluation of the conjoint efficacy in Chinese medicine with the longitudinal latent variable linear mixed model.
Chinese medicine (CM) clinical efficacy evaluation research involves the longitudinal multivariate measurement which means that patients are measured repeatedly and each patient is measured by several indicators on each fixed cross-section. Although each indicator can be evaluated separately with a longitudinal linear mixed model, it is important to consider all the endpoints together especially when researchers pay special attention to the change of the conjoint efficacy for several indicators in one patient. In this article, we introduce a latent variable linear mixed model to the CM conjoint efficacy evaluation and discuss why and how to analyze the longitudinal multivariate endpoint data in the clinical CM efficacy evaluation research. It may lead to the new insight of using such methodology in the field of conjoint efficacy evaluating of CM study. And with the definition of syndrome and symptom in the CM theory, the applied discussion brings the insight of CM syndrome evaluating in future. We illustrate this methodology using an example of CM efficacy evaluation from an ischemic stroke research.


Efficacy Profile and Safety of Very Low-Dose Rituximab in Patients with Graves' Orbitopathy.
Background: Rituximab (RTX), a chimeric human-murine anti-CD20 monoclonal antibody, has been used for treatment of active moderate-severe Graves' orbitopathy (GO) since 2004 as second-line therapy in patients unresponsive to intravenous steroids. We conducted an open-label prospective study (EUDRACT 2012-001980-53) in which patients were treated with a single infusion of only 100 mg RTX to analyze the efficacy and safety of this low dose. Methods: Seventeen patients, of whom nine had disease that was unresponsive to intravenous methylprednisolone and eight with newly diagnosed GO, were enrolled. Disease activity was assessed with the clinical activity score (CAS) and severity with a composite ophthalmic score. Long-term surgical treatment and quality of life were also assessed, as well as treatment-related adverse events. Results: Mean baseline CAS was 4.56 ± 0.96 and decreased to 1.25 ± 1.14 at 24 weeks (p = 0.001). Disease inactivation occurred within 24 weeks in >90% of patients and was unrelated to disease duration. Severity improved in about 60% of patients, with no relapses. All patients showed peripheral depletion of CD20+ and CD19+ cells at the end of RTX infusion (60 minutes). Two patients required surgical orbital decompression because of optic neuropathy (ON). Among adverse events observed, there was one patient who developed a cytokine release syndrome. Conclusions: A dose of 100 mg RTX is effective in patients with active moderate-severe GO. Low doses are better tolerated, expose patients to immune suppression for a shorter period of time, and are extremely cost effective, compared with higher doses. This dose, consistently with all other immunosuppressants, does not prevent the progression of GO to dysthyroid ON.


Free radical scavengers have a preventive effect on heterotopic bone formation following manipulation of immobilized rabbit legs.
The pathogenesis of heterotopic ossification (HO) is still unclear and the preventive measures and therapies are usually insufficient. The authors compared free radical scavengers with placebo in order to assess the magnitude of their inhibitory effect on the development of HO. A standard immobilization-manipulation model was used to induce HO in the hind legs of twenty female New Zealand albino rabbits. The animals were divided into two groups and received daily either placebo or a free radical scavenger (A/A) cocktail in a randomized double-blind fashion. Every four days an X-ray was taken and the thickness and length of new bone formation were measured at the thigh by two investigators independently. Fisher's exact test revealed a significant difference in the development of heterotopic ossification between the placebo group and the A/A group (70% versus none, respectively ; P=0.0031). The ischemia/reperfusion syndrome could be an important precipitating factor in the pathogenesis of heterotopic ossification and free radical scavengers were found to have a significant inhibitory effect on its development in a rabbit model. The results of this experimental model can be an impetus for further research into the prevention of heterotopic bone formation in humans.


Hepatitis B virus infection and the immune response: The big questions.
Clinical events and the host immune response during hepatitis B virus (HBV) infection are intricately linked. Despite decades of research, important questions concerning the immunopathogenesis of chronic HBV infection remain unanswered. For example, it is unclear which immune parameters facilitate persistence, and if HBV can be completely cleared from the human liver. Recent technological breakthroughs now allow researchers to address these seemingly basic, but essential questions surrounding HBV immunity. It will be important to better define the molecular underpinnings of immune cell function and dysfunction during chronic disease and in controlled infection, with particular focus on the liver, as little information is available on the intrahepatic compartment. In the near future, it may be possible to solve some of the controversy surrounding the immune responses to HBV, and establish the features of both the innate and adaptive arms of the immune system required to achieve sustained control of HBV infection.


Acquired B antigen: further studies using synthetic oligosaccharides.
The reactivity of acquired-B red cells with various antisera has been investigated by agglutination inhibition assays using four trisaccharides obtained by chemical synthesis. Two of these had the structure GalNAc alpha 1-3 (LFuc alpha 1-2). Gal and Gal alpha 1-3 (LFuc alpha 1-2) Gal which are characteristic of the A and B determinants respectively and were indeed strong inhibitors of human anti-A and -B antibodies. The other two sugars denoted B-OAc and B-NH2 are derivatives of the B-trisaccharide by substitution of the hydroxyl group on carbon-2 of the alpha-galactose residue with the O-acetyl group (B-OAc) or the amino group (B-NH2) respectively. We have shown that both B and B-NH2 trisaccharides inhibited strongly the agglutination of acquired B red cells by the anti-B reagents (crude or affinity purified) whereas sera containing "anti-acquired B" agglutinins were specifically inhibited by B-NH2 but not by the A, B or B-OAc structures. We have also shown that the agglutination of Tk-activated erythrocytes by the BS-II lectin is specifically inhibited by N-acetylglucosamine but not by B, B-OAc or B-NH2 structures. These results and the observation that anti-acquired B agglutinins cannot be adsorbed on Tk red cells suggest that (i) the "B-like" and the "acquired-B" determinants share a common structure best represented by B-NH2 and (ii) Tk and "acquired-B" antigens are not identical.


(Z)-17 beta-hydroxy-17 alpha-(2-[125I]iodovinyl)-4-estren-3-one: a new specific gamma-emitting ligand for determination of progesterone receptor.
An iodine-125 labeled ligand for progesterone receptor determination was synthesized: (Z)-17 beta-hydroxy-17 alpha-(2-[125I]iodovinyl)-4-estren-3-one ([125I]SH-D 510). The ligand is stable chemically as well as under the conditions of a receptor assay. The relative binding affinity of the nonradioactive compound towards human uterine progesterone receptor was 7.0 for the Z-isomer (promegestone (R5020) 1.0) and 0.95 for the E-isomer. 4 S and 8 S receptor forms were obtained on sucrose density gradient analysis. Progesterone receptors were assayed in 103 human mammary tumour cytosols, using either [3H]promegestone or [125I]SH-D 510. The coefficient of correlation was r = 0.951.


Comparison of the chemical constituents of raw Fructus Aurantii and Fructus Aurantii stir-baked with bran, and the biological effects of auraptene.
Fructus Aurantii (FA) is a Chinese herbal medicine commonly used in clinical practice to improve gastrointestinal motility, treat dyspepsia, and relieve constipation. More than 20 processing methods of FA have been recorded, among which FA stir-baked with bran is the earliest, most time consuming, and the most popular one. Raw FA has a strong ability to promote qi-moving and has middle-energizer-soothing effects; therefore, it is often used to relieve hypochondrium distension and pain, and to relax the stagnation of the liver Qi. FA stir-baked with bran is more effective in nourishing the stomach and curing indigestion. In this study, the chemical composition and differences between raw FA and FA stir-baked with bran were systematically compared. The chemical components that increased after stir-baking FA and bran were separated and their pharmacodynamic characteristics were determined. Lastly, the processing mechanism of FA was further explained. Twelve main chemicals in raw FA and FA stir-baked with bran were compared using high-performance liquid chromatography (HPLC). The main differential components were identified, separated, purified, and then analyzed using pharmacodynamic tests. The intestine-pushing test, in vitro smooth muscle test, and in vitro acetylcholinesterase (AchE) activity test in mice were performed to explain the mechanism of auraptene in improving gastrointestinal motility. Using HPLC, the primary chemical that differed between raw FA and FA stir-baked with bran was identified as auraptene. The processed FA was extracted, separated, and purified to obtain pure auraptene. The intestine-pushing test in mice showed that low (0.6 mg·kg-1) and medium doses (1.2 mg·kg-1) of auraptene could promote peristalsis of the small intestine, whereas a high dose (2.4 mg·kg-1) inhibited peristalsis. In vitro studies on the smooth muscle of mice showed that a low dose of auraptene (0.2 mmol·L-1, 10-800 μL) could promote contraction, whereas a high dose (0.2 mmol·L-1, >1000 μL) had the opposite effect. Auraptene has a mechanism of action similar to that of the acetylcholinesterase inhibitor, neostigmine. Additionally, auraptene could inhibit AchE activity in vitro. Auraptene is the main chemical constituent that differs between raw FA and FA stir-baked with bran. Pharmacodynamic tests showed that auraptene has a cholinergic effect, by virtue of its role as an acetylcholinesterase inhibitor. Moreover, auraptene could dually regulate the gastrointestinal smooth muscle. Auraptene was present in low levels and its content varied in FA stir-baked with bran, depending on the origin and source of FA, and the treatment procedures it was subjected to. In the Chinese Pharmacopoeia, the recommended dose of FA stir-baked with bran is a low dose of 3-10 g, which effectively promotes small-intestinal peristalsis. The mechanism of action is attributed to an increase in the relative content of acetylcholine by the inhibition of AchE activity to promote gastrointestinal motility. The increased levels of auraptene in FA stir-baked with bran are the main reason and the primary purpose for the change in its medicinal properties. This technique, therefore, has potential to be used as one of the main processing mechanisms of raw FA.


Healthcare Utilization After Respiratory Tuberculosis: A Controlled Interrupted Time Series Analysis.
Despite data suggesting elevated morbidity and mortality among people who have survived tuberculosis disease, the impact of respiratory tuberculosis on healthcare utilization in the years following diagnosis and treatment remains unclear. Using linked health administrative data from British Columbia, Canada, we identified foreign-born individuals treated for respiratory tuberculosis between 1990 and 2019. We matched each person with up to four people without a tuberculosis diagnosis from the same source cohort using propensity score matching. Then, using a controlled interrupted time series analysis, we measured outpatient physician encounters and inpatient hospital admissions in the 5 years following respiratory tuberculosis diagnosis and treatment. We matched 1216 individuals treated for respiratory tuberculosis to 4864 non-tuberculosis controls. Immediately following the tuberculosis diagnostic and treatment period, the monthly rate of outpatient encounters in the tuberculosis group was 34.0% (95% confidence interval [CI]: 30.7%, 37.2%) higher than expected, and this trend was sustained for the duration of the post-tuberculosis period. The excess utilization represented an additional 12.2 (95% CI: 10.6, 14.9) outpatient encounters per person over the post-tuberculosis period, with respiratory morbidity a large contributor to the excess healthcare utilization. Results were similar for hospital admissions, with an additional 0.4 (95% CI: .3, .5) hospital admissions per person over the post-tuberculosis period. Respiratory tuberculosis appears to have long-term impacts on healthcare utilization beyond treatment. These findings underscore the need for screening, assessment, and treatment of post-tuberculosis sequelae, as it may provide an opportunity to improve health and reduce resource use.


Cholesterol dysregulation in peripheral blood mononuclear cells of Alzheimer's disease.
Cholesterol and the immune system are involved in Alzheimer's Disease (AD). To investigate the relations among them, we compared the cholesterol content in peripheral blood mononuclear cells (PBMC) of cognitively healthy controls and patients with mild cognitive impairment (MCI) and AD in two independent samples. Free cholesterol content of PBMC was lower in MCI and AD patients, and was modulated by APOE genotype. A decrease of CD8+ and an increase of CD16+ was also found in AD patients. These results suggest that cholesterol levels in PBMCs may represent an early signature of the disease and support the involvement of immune system in AD.


Skin potential and EMG changes induced by cutaneous electrical stimulation. I. Normal man in arousing and non-arousing environment.
Skin potential and EMG responses induced in normal man by electrical stimuli applied to the skin were recorded in the four limbs in order to study somato-sympathetic and somato-motor reflexes. Different patterns of responses were observed in different conditions: alarm, habituation, sensitization and arousal. During alarm, sensitization and arousal, the responses were present in the four limbs; during habituation, the responses were only present in the stimulated and in the contralateral limb. Three sensory thresholds to cutaneous electrical stimulation were identified in habituated subjects: tactile, tingling and pain. Cutaneous and EMG responses appeared at tingling threshold. A relationship between skin potential level and skin potential response was observed.