Tumores rari et inusitati. Primary squamous-cell carcinoma of the caecum: report of a case and current theories of aetiology.
A case of squamous cell carcinoma of the caecum is reported, confirmed by light and electron microscopy. The tumour was associated with pelvic irradiation ten years previously. Current theories of aetiology of squamous-cell carcinoma of the large bowel are reviewed.


Decision Trees for Predicting the Physiological Responses of Rabbits.
The thermal environment inside a rabbit house affects the physiological responses and consequently the production of the animals. Thus, models are needed to assist rabbit producers in decision-making to maintain the production environment within the zone of thermoneutrality for the animals. The aim of this paper is to develop decision trees to predict the physiological responses of rabbits based on environmental variables. The experiment was performed in a rabbit house with 26 rabbits at eight weeks of age. The experimental database is composed of 546 observed data points. Sixty decision tree models for the prediction of respiratory rate (RR, mov.min-1) and ear temperature (ET, °C) of rabbits exposed to different combinations of dry bulb temperature (tdb, °C) and relative humidity (RH, %) were developed. The ET model exhibited better statistical indices than the RR model. The developed decision trees can be used in practical situations to provide a rapid evaluation of rabbit welfare conditions based on environmental variables and physiological responses. This information can be obtained in real time and may help rabbit breeders in decision-making to provide satisfactory environmental conditions for rabbits.


Pseudo-obstruction of the bowel in pregnancy: case reports.
Two patients with intestinal obstruction in pregnancy are presented and discussed. In neither case was there any obvious cause for the intestianl obstruction. Pseudo-obstruction of the bowel has not previously been described as a complication of pregnancy.


The Continuing Effect of COVID-19 Pandemic on Physical Well-Being and Mental Health of ICU Healthcare Workers in Turkey: A Single-Centre Cross-Sectional Later-Phase Study.
This study aimed to evaluate the effect of COVID-19 pandemic on physical well-being and mental health of ICU healthcare workers (HCWs). A total of 51 ICU HCWs working at a tertiary care hospital were included in this cross-sectional study conducted before (January 2019-January 2020) and during (January 2021-April 2021) COVID-19 pandemic. Data on sociodemographic and work-related characteristics, COVID 19 history and current mental health issues via Hospital Anxiety-Depression Scale (HADS), Pittsburgh Sleep Quality Index (PSQI), Eating Attitudes Test (EAT-40), Suicidal Ideation Scale (SIS) and Maslach Burnout Inventory (MBI) were recorded. Overall, 62.7% of participants were nurses, heavy workload (working ≥200 h/month) was reported by 76.5% of participants and previous history of COVID-19 was confirmed by 62.7%. Current mental health issues involved poor sleep quality in majority (96.1%) of participants, anxiety (51.0%), depression (51.0%) in at least half of them and a moderate degree of emotional exhaustion Heavy workload was associated with more remarkable decrease in sleep duration (median change: -0.5 vs. -1.0 h/day, P = .020), Vit B12 (median change: 60[-48-293] vs. -65[-371-262] pg/mL, P < .001) and Vit D (median change: -1.6[-13.1-20] vs. -9.7[-39.7-21.8] ng/mL, P = .004) during pandemic, while working hours per month were also significantly higher in those with versus without anxiety (264[150-390] vs. 240[150-264] h, P = .003) and with versus without depression (264[150-390] vs. 240[150-264] h, P = .037). Our findings indicate high prevalence of mental health issues including anxiety and depression as well as poor sleep quality and emotional burnout among ICU HCWs, particularly those with heavy workload.


Crosstalk between huntingtin and syntaxin 1A regulates N-type calcium channels.
We have identified a novel interaction between huntingtin (htt) and N-type calcium channels, a channel key in coupling calcium influx with synaptic vesicle exocytosis. Htt is a widely expressed 350-kDa cytosolic protein bearing an N-terminal polyglutamine tract. Htt is proteolytically cleaved by calpains and caspases and the resultant htt N-terminal fragments have been proposed to be biologically active; however, the cellular function of htt and/or the htt fragments remains enigmatic. We show that N-terminal fragments of htt (consisting of exon1) and full-length htt associate with the synaptic protein interaction (synprint) region of the N-type calcium channel. Given that synprint has previously been shown to bind syntaxin 1A and that this association elicits inhibition of N-type calcium channels, we tested whether htt(exon1) affects the modulation of these channels. Our data indicate that htt(exon1) enhances calcium influx by blocking syntaxin 1A inhibition of N-type calcium channels and attributes a key role for htt N-terminal fragments in the fine tuning of neurotransmission.


Homoharringtonine, a clinically approved anti-leukemia drug, sensitizes tumor cells for TRAIL-induced necroptosis.
One hallmark of cancer cells is their ability to evade physiologic signals causing regulated cell death (RCD). Correspondingly, TRAIL-based therapies to eliminate human cancer cells via enforced induction of apoptosis have been established and represent a promising approach in anti-cancer research. However, due to frequently appearing intrinsic or acquired resistances of tumor cells against apoptosis, TRAIL-based apoptotic strategies for the treatment of cancer patients have shown limited efficacy. As a potential alternative, regulated necrosis (and necroptosis triggered e.g. by TRAIL receptors 1/2) has recently gained considerable attention. Regulated necrosis represents a mode of RCD molecularly distinct from apoptosis whose potential in anti-cancer therapy is almost uncharacterized. Since in most cancer cells survival pathways counteract the effects of TRAIL-induced RCD, sensitizers such as cycloheximide (CHX) are frequently added in cell culture to overcome this problem. Unfortunately, those sensitizers are cytotoxic and therefore not suitable for the treatment of cancer patients. Here, we have alternatively employed homoharringtonine (HHT), a plant alkaloid which was recently approved by the U. S. Food and Drug Administration to treat patients with chronic myeloid lymphoma. We show that HHT is an efficient sensitizer for TRAIL-induced necroptosis in multiple human cancer cell lines. In addition, HHT-enhanced TRAIL-mediated necroptosis occurs via the same signaling pathways (involving RIPK1/RIPK3/MLKL) as CHX-enhanced necroptosis. Importantly, consecutive treatment schedules of necroptosis and apoptosis in either combination revealed remarkable additive effects not reached by repetitive apoptotic treatments alone. Taken together, our data demonstrate that HHT can replace harmful substances such as CHX to sensitize human cancer cells to TRAIL-induced necroptosis. Thus, HHT represents a promising enhancer in TRAIL-based necroptotic anti-cancer therapies also in patients.


Combination therapy with itraconazole and terbinafine for phaeohyphomycosis caused by Exophiala spinifera: A case report and literature review.
Exophiala spinifera is a rare dematiaceous fungus causing cutaneous, subcutaneous and disseminated phaeohyphomycosis (PHM). Standard antifungal therapy for PHM is still uncertain. Here, we report a case of a Chinese male with PHM caused by E. spinifera, who received significant clinical improvement after the treatment with oral itraconazole and terbinafine. With the aim of evaluating the antifungal therapy for PHM caused by E. spinifera, a detailed review was performed.


The Role of Postmastectomy Radiation Therapy in Patients With Breast Cancer Responding to Neoadjuvant Chemotherapy.
When surgery is the first line of breast cancer treatment, numerous randomized clinical trials and meta-analyses have demonstrated that postmastectomy radiation therapy (PMRT) improves locoregional control and survival for many women with axillary lymph node-positive disease. In contrast, there are no randomized data regarding the use of PMRT in women who receive neoadjuvant chemotherapy (NAC) first followed by mastectomy. This has led to controversy regarding which patient with breast cancer will benefit from PMRT after NAC, particularly in women with clinically node-positive axillary disease that responds well and is down staged to pathologically negative disease at surgery (ypN0). We review the current evidence on this topic, which forms the underlying basis for the ongoing phase III clinical trial-National Surgical Adjuvant Breast and Bowel Project (NSABP) B51/RTOG 1304-that is examining the role of regional nodal irradiation in patients with clinical N1 disease that responds to NAC and becomes ypN0 at surgery.


Comparison of tissue plasminogen activator-antibiotic locks with heparin-antibiotic locks in children with catheter-related bacteraemia.
An accepted pathogenesis of catheter-related bacteraemia (CRB) is the seeding of microorganisms from the intraluminal biofilm of central venous catheters. Antibiotic locks (ABL) are solutions containing high concentrations of antimicrobials with or without anticoagulants that aim to destroy the biofilm. In this study, two different ABL solutions, tissue plasminogen activator (TPA)-based and heparin-based ABL, used in conjunction with systemic antibiotics, were prospectively compared in the treatment of CRB. A total of 42 children on chronic haemodialysis with 11,016 catheter-days were observed for signs and symptoms of CRB over a period of 10 months. Twenty-four CRBs were diagnosed in 18 children (2.2 CRB/1000 catheter-days) and were treated with the protocol. Symptoms of CRB resolved in 83% within 48 h of treatment. None of the infected catheters required early emergent exchange or removal for poorly controlled CRB. Six children had recurrence of CRB within 6 weeks, of which four required catheter exchange. There was no specific microorganism or type of CRB that predisposed to higher recurrence rates. The mean infection-free survival of the catheters following TPA-ABL treatment was shorter than that following heparin-ABL treatment, but was not statistically significant by the log-rank test (126.8 +/- 81.6 days versus 154.5 +/- 70.4 days). Both TPA-ABL and heparin-ABL used in conjunction with systemic antibiotics can effectively clear CRB without significant late recurrence at 6 weeks. Early use of ABL for management of CRB can potentially decrease the need for catheter removal, thus salvaging vascular access sites.


Longitudinal white matter and cognitive development in pediatric carriers of the apolipoprotein ε4 allele.
We have previously demonstrated cross-sectional differences in magnetic resonance imaging (MRI) measurements of white matter myelin and gray matter in infants with or without the apolipoprotein ε4 allele, a major genetic risk factor for late-onset Alzheimer's disease (AD). In this study, we sought to compare longitudinal MRI white matter myelin and cognitive-behavioral changes in infants and young children with and without this allele. Serial MRI and cognitive tests were obtained on 223 infants and young children, including 74 ε4 carriers and 149 non-carriers, 2-68 months of age, matched for age, gestational duration, birth weight, sex ratio, maternal age, education, and socioeconomic status. Automated brain mapping algorithms and non-linear mixed models were used to characterize and compare trajectories of white matter myelin and cognitive-behavioral test scores. The APOE ε4 carriers had statistically significant differences in white matter myelin development, in the uncinate fasciculus, temporal lobe, internal capsule and occipital lobe. Additionally, ε4 carriers had a slightly greater rate of development in early learning composite a surrogate measure of IQ representative of expressive language, receptive language, fine motor, and visual skills, but displayed slightly lower non verbal development quotient scores a composite measure of fine motor and visual skills across the entire age range. This study supports the possibility that ε4 carriers have slightly altered rates of white matter and cognitive development in childhood. It continues to raise questions about the role of APOE in human brain development and the relevance of these developmental differences to the predisposition to AD.


Laterobasal membranes from intestinal epithelial cells: isolation free of intracellular membrane contaminants.
A simplified method for isolating highly purified laterobasal membranes (LBM) from enterocytes is based on treatment of membranes with 8 mM CaCl2 concentration in order to aggregate intracellular membrane contaminants. The resultant LBM showed an average 15-fold enrichment and constituted 8% of the original K-stimulated phosphatase in the initial crude homogenate. It showed typical LBM migration on counter-current distribution (CCD) and was essentially free of contamination with endoplasmic reticulum and Golgi membranes. This method is highly efficient and yields sufficient purified LBM to allow comprehensive analysis of enterocyte membrane events.


Alcohol consumption upon direct-acting antiviral therapy for hepatitis C among persons with human immunodeficiency virus in the United States.
Direct-acting antivirals (DAA) are highly effective against hepatitis C virus (HCV) infection among persons with human immunodeficiency virus (PWH). However, alcohol use post-DAA treatment poses a continued threat to the liver. Whether the focus on liver health alone during HCV treatment can impact alcohol consumption is unclear. Therefore, we examined the change in alcohol use among HCV-coinfected PWH who received DAA therapy by non-addiction medical providers. In our longitudinal clinical cohort study, we identified HCV-coinfected PWH who received interferon-free DAA therapy between January 2014 and June 2019 in the Centers for AIDS Research Network of Integrated Clinical Systems. The Alcohol Use Disorders Identification Test-Consumption (AUDIT-C) was the alcohol screening instrument. We used mixed-effects logistic regression models to estimate the longitudinal change in alcohol use upon DAA therapy. Among 738 HCV-coinfected PWH, 339 (46 %) reported any alcohol use at the end of HCV treatment, including 113 (15 %) with high-risk use (i.e., AUDIT-C ≥3 for women, ≥4 for men). Concurrently, 280 (38 %) PWH noted active drug use, and 357 (48 %) were currently smoking. We observed no changes in the odds of any alcohol or high-risk alcohol use over time with DAA therapy. Findings were similar in the PWH subgroup with a history of alcohol use before DAA treatment. For PWH with HCV, alcohol use did not change following interferon-free DAA treatment by non-addiction medical providers. Thus, clinicians should consider integrating targeted alcohol use interventions into HCV care to motivate reduced alcohol consumption and safeguard future liver health.


Two new mutations in the porphobilinogen deaminase gene and a screening method using PCR amplification of specific alleles.
Acute intermittent porphyria (AIP) is attributable to defects in the porphobilinogen deaminase (PBGD) gene. Two new mutations have been found in the PBGD gene in Swedish families. The first is a G to A splice mutation in the last position of intron 9. A screening method using allele-specific amplification has been designed for the rapid detection of this mutation. The second mutation is a C to T substitution in exon 10, changing Arg201 to Trp. This mutation can be detected by restriction enzyme cleavage.


Control by the extracellular environment of differentiation pathways in 1003 embryonal carcinoma cells: study at the level of specific intermediate filaments.
1003 is a multipotential embryonal carcinoma (EC) clonal cell line which can be induced to follow different developmental pathways by altering the composition of the culture medium. When grown in serum-containing medium the great majority of 1003 cells remain undifferentiated; they express the ECMA 7 cell-surface embryonic antigen and very low amounts of vimentin. In serum-free medium, most 1003 cells differentiate into neuroepithelial cells. The majority of these cells are still labelled with ECMA 7 antibodies. They contain higher amounts of vimentin than EC cells, but no neurofilament proteins. Neuroepithelial cells then differentiate into neurons through a stage of preneurons containing both vimentin and the 70-K neurofilament protein. Fully differentiated neurons contain 70-K neurofilament protein but no vimentin. The 200-K neurofilament protein is detected later in the neurons. Mesenchymal cells (induced by re-adding serum) express high amounts of vimentin organized in networks. Preneurons , neurons, and mesenchymal cells do not express ECMA 7 antigen.


Identification and characterization of a marine-derived chitinolytic fungus, Acremonium sp. YS2-2.
Chitin is the most abundant biopolymer in marine environments. To facilitate its utilization, our laboratory screened marine-derived fungal strains for chitinolytic activity. One chitinolytic strain isolated from seawater, designated YS2-2, was identified as Acremonium species based on morphological and phylogenetic analyses. Acremonium species are cosmopolitan fungi commonly isolated from both terrestrial and marine environments, but their chitinolytic activity is largely unknown. The extracellular crude enzyme of YS2-2 exhibited optimum chitinolytic activity at pH 6.0-7.6, 23-45°C, and 1.5% (w/v) NaCl. Degenerate PCR revealed the partial cDNA sequence of a putative chitinase gene, chiA, in YS2-2. The expression of chiA was dramatically induced in response to 1% (w/v) colloidal chitin compared to levels under starvation, chitin powder, and glucose conditions. Moreover, the chiA transcript levels were positively correlated with chitinolytic activities under various colloidal chitin concentrations, suggesting that ChiA mediates chitinolytic activity in this strain. Our results provide a basis for additional studies of marinederived chitinolytic fungi aimed at improving industrial applications.


The combination therapy of isomucronulatol 7-O-beta-glucoside (IMG) and CEP-9722 targeting ferroptosis-related biomarkers in non-small cell lung cancer (NSCLC).
NSCLC is a malignant tumor with a high incidence. Ferroptosis presents an essential function in regulating carcinogenesis and tumor progression. However, the ferroptosis-associated prognostic model based on single-cell sequencing of NSCLC remains unexplored. Our study aims to establish a potential predictive model for NSCLC patients and provide available targeted drugs for clinical treatment. The data on NSCLC patients were collected from TCGA and GEO databases to analyze their gene expression profiles. ConsensusCluster was adopted to divide the patients into different groups based on ferroptosis-related genes. Then, the univariable Cox and LASSO analyses were applied to data analysis and model establishment. Single-cell analysis was used to explore the risk score genes in different cell populations and states. The protein levels of these genes were also investigated through the HPA database. Drug sensitivity was evaluated in CellMiner database. CCK8 and colony formation assays were performed to validate potential drugs' effects on lung cancer cell lines. A ferroptosis-related prognostic model involving 14 genes in NSCLC patients was established. The risk score model was developed in training set GSE31210 and validated in the test set TCGA. The low-risk score group showed a better prognosis than the high-risk score group. The single-cell analysis revealed that the risk score genes were mainly derived from lung tumor cells. Most risk score genes were more highly expressed in tumor tissue than in normal tissue, according to the HPA database. Besides, these genes were associated with 106 drugs in CellMiner database. Finally, the drug effects on NSCLC cell growth were evaluated by cck8 and colony formation. We identified an effective ferroptosis-related prognostic model based on single-cell sequencing. The potential prediction model is devoted to exploring clinical therapeutic targets for NSCLC.


Negative staining method with nigrosin for the detection of cryptosporidial oocysts: a comparative study.
A comparison was made between two methods for the detection of cryptosporidial oocysts in human faecal sediments of formalin-ether concentrates: a cover-slipped, wet method (containing Gram's iodine) and an air-dried, negative staining method using nigrosin solution. A modified Ziehl-Neelsen technique was used as a reference method. The negative staining method with nigrosin gave a positive diagnosis more often than the cover-slipped wet method. A comparison of the nigrosin method with the modified Ziehl-Neelsen method gave almost identical results. Restaining the nigrosin slides with the modified Ziehl-Neelsen technique showed that the round, refractile bodies were cryptosporidial occysts.


Intramitochondrial transfer of phospholipids in the yeast, Saccharomyces cerevisiae.
Translocation of phosphatidylinositol, which is synthesized on the outer aspect of the outer membrane of isolated yeast mitochondria, to the inner membrane is linked to phosphatidylinositol synthesis and is therefore a vectorial process. Phosphatidylinositol once integrated into the inner mitochondrial membrane is not transferred back to the mitochondrial surface. Phosphatidylserine is also translocated from the outer to the inner mitochondrial membrane, where it is decarboxylated to phosphatidylethanolamine. We made use of this metabolic modification to characterize the intramitochondrial transfer of phosphatidylserine and phosphatidylethanolamine. Intramitochondrial phosphatidylserine transfer is insensitive to the uncoupler carbonyl cyanide m-chlorophenylhydrazone and to valinomycin and is thus independent of an electrochemical gradient across the inner membrane. Transfer of phosphatidylserine from the outer to the inner mitochondrial membrane occurs not only in intact mitochondria but also in mitoplasts which are devoid of intermembrane space proteins but have the outer membrane still adherent to the inner membrane. This result suggests that specific contact sites are involved in the intramitochondrial translocation of phospholipids. 3H-Labeled phosphatidylethanolamine synthesized from [3H]serine in isolated mitochondria is readily exported from the inner to the outer mitochondrial membrane without prior mixing with the pool of phosphatidylethanolamine of the inner membrane.


O(6)-methylguanine DNA-methyltransferase (MGMT) overexpression in melanoma cells induces resistance to nitrosoureas and temozolomide but sensitizes to mitomycin C.
Alkylating agents play an important role in the chemotherapy of malignant melanomas. The activity of alkylating agents depends on their capacity to form alkyl adducts with DNA, in some cases causing cross-linking of DNA strands. However, the use of these agents is limited by cellular resistance induced by the DNA repair enzyme O(6)-methylguanine DNA-methyltransferase (MGMT) which removes alkyl groups from alkylated DNA strands. To determine to what extent the expression of MGMT in melanoma cells induces resistance to alkylating agents, the human cell line CAL77 Mer- (i.e., MGMT deficient) were transfected with pcMGMT vector containing human MGMT cDNA. Several clones expressing MGMT at a high level were selected to determine their sensitivity to chemotherapeutic drugs. Melanoma-transfected cells were found to be significantly less sensitive to nitrosoureas (carmustine, fotemustine, streptozotocin) and temozolomide with an increase of IC(50) values between 3 and 14 when compared to parent cells. No difference in cell survival rates between MGMT-proficient and -deficient cells was observed for melphalan, chlorambucil, busulphan, thiotepa and cisplatin which preferentially induce N(7) guanine lesions. Surprisingly, MGMT overexpression increased the sensitivity of CAL77 cells to mitomycin C by approximately 10-fold. Treatment of clonal cell lines with buthionine-[S,R]-sulfoximine (BSO), an inhibitor of gamma-glutamylcysteine synthetase which depletes cellular glutathione, completely reversed this unexpected increase in sensitivity to mitomycin C. This observation suggests that glutathione is involved in the sensitivity of MGMT-transfected cells to mitomycin C and may act synergistically with MGMT via an unknown mechanism.


Socioeconomic and demographic determinants of familial social capital inequalities: a cross-sectional study of young people in sub-Saharan African context.
Social capital is broadly acknowledged as a vital 'health asset' that promotes young people's health and wellbeing and has the potential to prevent social- and health-related risk behaviours in the life-course. However, limited research has investigated the determinants of social capital for young people in sub-Saharan Africa. This study examines the role of socioeconomic and demographic factors in establishing inequalities in familial social capital among young people in Ghana. The study utilised a cross-sectional survey data involving 2068 in-school adolescents (13-18 years) in the Upper West Region, Ghana. Familial social capital was assessed by 'family sense of belonging', 'family autonomy support' and 'family control'. Multinomial logistic regressions established the relationships between socioeconomic and demographic factors and the measures of familial social capital. Adolescents from low affluence households had about 63 and 61% lower odds of attaining a high family sense of belonging (FSB) (OR = 0.373; 95%CI: 0.27-0.513) and high family autonomy support (FAS) (OR = 0.387; 95%CI: 0.270-0.556) respectively but had 67% higher odds of reporting high family control (FC) (OR = 1.673; 95%CI: 1.187-2.359) than their counterparts. Males had about 55 and 71% higher odds to possess high FSB (OR = 1.549; 95%CI: 1.210-1.983) and high FAS (OR = 1.705; 95%CI: 1.272-2.284) respectively but had 38% lower odds to report high family control (OR = 0.624; 95%CI: 0.474-0.822) than females. The odd of young adolescents to attain high FSB than older adolescents were about 66% higher (OR = 1.662; 95%CI: 1.168-2.367). Religion, educational level, ethnicity, family structure, and marital status were also significant predictors of adolescents' family sense of belonging, family autonomy support and, family control. Socioeconomic and demographic factors influence inequalities in the amount of familial social capital possessed by young people which suggests possible risks of social inequality. The family context is possibly failing some cohorts of young people with particular reference to female and poor adolescents regarding familial cognitive social capital. Public health strategies should include families in addressing socioeconomic and demographic differences in social capital with a key focus on the cohorts of young people at risk of social capital inequality.


Solvational tuning of the unfolding, aggregation and amyloidogenesis of insulin.
Solvational perturbations, accomplished by the addition of the three model cosolvents glycerol, ethanol and trifluoroethanol, exert pronounced and diversified effects on the unfolding, non-native assembly and fibril formation of the amyloidogenic protein insulin. Fluorescence, CD and UV-spectroscopic methods as well as atomic force microscopy imaging have been employed to reveal distinct structural and kinetic features upon the aggregation of insulin under different solvational perturbations, which ultimately manifest in morphological variations of mature aggregates and fibrils. In particular, fluorescence anisotropy studies proved to be very valuable in characterizing the corresponding aggregation nuclei. Glycerol stabilizes, through enhanced hydration, native oligomerization and retards fibrillar aggregation at all concentrations studied (up to 40% (w/w)). In contrast, both monoalcohols facilitate the formation of aggregation-prone intermediates by destabilization of the native assembly. The reversal from a kosmotropic to a merely chaotropic solvational behaviour can explain the accelerating effect on ordered fibrillation of low concentrations and the inhibitory nature of high concentrations of ethanol and trifluoroethanol, ultimately leading to amorphous aggregate structures. Mechanistically, dimer dissociation under stabilizing and nucleation under destabilizing conditions have been identified to be the rate-limiting steps that account for the non-monotonic concentration effects of the monoalcohols on the aggregation kinetics. A rationale as to how solvational constraints can tune the stability of the species on the native self-assembly and non-native aggregation pathway, and the energetic barriers that need to be overcome for the required structural interconversions has been put forward. We may propose that the concept of perturbed solvation is generally applicable to phenomena that are related to pathogenic amyloidogenesis of proteins and, in general, solvational effects, besides other aspects of the cellular environment, may play a significant role in a reshaping of the folding/aggregation funnel of proteins.


Effects of Alcohol Administered With Flibanserin in Healthy Premenopausal Women: A Randomized, Double-Blind, Single-Dose Crossover Study.
Flibanserin is approved in the United States and Canada for the treatment of hypoactive sexual desire disorder in premenopausal women. The purpose of this trial was to evaluate the safety of concomitant administration of flibanserin with alcohol. In this single-center, randomized, double-blind, single-dose, crossover study, participants were randomly assigned to 1 of 12 sequences to receive each of 7 treatments: flibanserin 100 mg or placebo with ethanol 0.2 g/kg, 0.4 g/kg, or 0.6 g/kg, or flibanserin 100 mg only. Treatments were administered using a worst-case approach that included morning dosing and consumption of alcohol within 10 minutes. The primary end point was the proportion of participants who experienced dizziness, syncope, or hypotension. Safety end points included orthostatic vital signs. The study included 96 premenopausal women (mean age 31 ± 8 years). The incidence of dizziness for ethanol + flibanserin was 39.8% for ethanol 0.6 g/kg, 34.1% for 0.4 g/kg, and 27.4% for 0.2 g/kg compared with 31.1% for flibanserin without ethanol. Based on the available vital signs data, there was no effect of ethanol concentration on orthostatic blood pressure, vertigo, or hypotension; no instances of syncope were observed. The overall incidence of adverse events (AEs) was similar when flibanserin was administered alone (96.7%) or with ethanol (90.5-97.6%). Consumption of the tested amounts of alcohol (0.2-0.6 g/kg) does not have an additive effect on the AE profile of flibanserin 100 mg in healthy premenopausal women. Strengths include the study population (premenopausal women, as indicated for flibanserin) and range of ethanol doses. Limitations include the morning dosing of study medication, which is inconsistent with the bedtime dosing recommended for flibanserin, and the method of handling missing vital sign measurements. Co-administration of flibanserin 100 mg with varying doses of ethanol resulted in few AEs of special interest, with no notable alcohol dose response. However, a significantly greater percentage of participants administered flibanserin with 0.6 g/kg and 0.4 g/kg of alcohol were characterized as "Participants in Whom Standing Blood Pressure Was Not Obtained" compared with participants administered flibanserin alone. Simon JA, Clayton AH, Parish SJ, et al. Effects of Alcohol Administered With Flibanserin in Healthy Premenopausal Women: A Randomized, Double-Blind, Single-Dose Crossover Study. J Sex Med 2020;17:83-93.


Crystal structure of an anti-Ang2 CrossFab demonstrates complete structural and functional integrity of the variable domain.
Bispecific antibodies are considered as a promising class of future biotherapeutic molecules. They comprise binding specificities for two different antigens, which may provide additive or synergistic modes of action. There is a wide variety of design alternatives for such bispecific antibodies, including the "CrossMab" format. CrossMabs contain a domain crossover in one of the antigen-binding (Fab) parts, together with the "knobs-and-holes" approach, to enforce the correct assembly of four different polypeptide chains into an IgG-like bispecific antibody. We determined the crystal structure of a hAng-2-binding Fab in its crossed and uncrossed form and show that CH1-CL-domain crossover does not induce significant perturbations of the structure and has no detectable influence on target binding.


Women reconstruct more details than men for a complex five-minute narrative: implications for right-hemispheric factors in the serial memory effect.
A total of 20 men and 20 women listened to a 5-min. ambiguous narrative (the "Billy" story). When the subjects were asked (unexpectedly) to reconstruct the details of the story as a written narrative 30 min. later, women employed about 50% more words and comparably more accurate detail than the men. Covariance for pre-experimental experiences suggested that the women's proficiency could be related to the greater representation of linguistic processes within the right (non-dominant) hemisphere. The hypothesis that the classic deficit of reconstructional accuracy for events during the middle of a story is related to right-hemispheric inhibition was also supported. Subjects who recalled the fewest details reported more vivid dreaming during the subsequent two days.


Demographics, measures of professional achievement, and gender differences for diplomates of the American College of Veterinary Surgeons in 2015.
To characterize elements of employment, professional success, and personal life for American College of Veterinary Surgeons (ACVS) diplomates and identify elements of practice that may serve as barriers to work-life balance or affect men and women differently. 836 ACVS diplomates. An 81-item questionnaire was sent to 1,450 ACVS diplomates in 2015 via email and conducted by means of an online platform. Responses were analyzed to identify associations among selected variables. The survey response rate was 58% (836/1,450). The median age category among respondents was 41 to 45 years. The ratio of male to female diplomates was equivalent among those < 40 years old. Respondents in small animal private practice worked the fewest number of hours; those in equine or large animal private practice worked the most and had the most on-call responsibility. Women were more likely than men to be employed in academia. In both private practice and academia, respondents in small animal practice earned more than did those in equine or large animal practice, and women earned less than did men, even after adjustment for relevant covariates. Women were less likely than men to be practice owners or to hold a prestigious academic title and rank. Perceptions about the effect of gender in the workplace differed between men and women. Findings suggested that the veterinary surgical profession is demanding for both genders, although increased flexibility in certain areas may improve work-life balance.


Reconstruction of Paprosky Type III Acetabular Defects by Three-Dimensional Printed Porous Augment: Techniques and Clinical Outcomes of 18 Consecutive Cases.
To introduce the surgical technique of reconstruction of Paprosky type III acetabular defects by 3D printed porous augments. First, CT scans of pelvis were obtained to establish the 3D reconstruction model of 3D printed porous augment. Then, a nylon pelvis model was printed to simulate operation with the surgeons. At this time, the augment was designed and modified according to the surgeon's suggestions and the 3D printing principles. Eighteen patients with Paprosky type III acetabular defects receiving reconstructive surgery by 3D printed porous augments were included in current study. Their data, including general information, intra-operative findings, imaging results, functional scores, and complications were retrospectively analyzed. The mean follow-up time lasted 33.3 ± 2.0 (24-56) months. The average limb-length discrepancy (LLD) was 31.7 ± 4.2 (3-59) mm preoperatively, 7.7 ± 1.4 (1-21) mm postoperatively (P < 0.0001), and 7.5 ± 1.2 (0-18) mm at the latest follow-up. The mean vertical position of hip center of rotation (HCOR) from the interteardrop line changed from preoperative 50.7 ± 3.9 (23.3-75.3) mm to postoperative 22.9 ± 1.9 (10.1-40.3) mm (P < 0.0001), with the latest follow-up revealing an HCOR of 22.3 ± 1.7 (11.0-40.5) mm. Follow-up study showed that no hip had radiolucencies and radiological loosening of the acetabular components and augment. The average Harris hip score (HHS) improved from 40.3 ± 4.5 (10.5-71) before operation to 88.4 ± 1.9 (75-97) at the last follow-up (P < 0.0001). Moreover, follow-up exhibited that no periprosthetic joint infection, hip dislocation, fracture, and re-revision occurred. Surgical treatment of Paprosky type III acetabular defect with 3D printed porous augment was simple, achieved good match between porous augment and the defect bone surface and the acetabular component, ideally restored LLD and HCOR after operation, significantly improved HHS and attained good early clinical outcomes. It is a promising personalized solution for patients with severe acetabular bone defect.


Determination of polyphenols in three Capsicum annuum L. (bell pepper) varieties using high-performance liquid chromatography-tandem mass spectrometry: their contribution to overall antioxidant and anticancer activity.
A mixture of polyphenol components was isolated from the fruits of C. annuum L. cv. Cupra, C. annuum L. cv. Orange glory, and C. annuum L. cv. ST4712 (CLST), via 70% methanol extraction followed by column chromatography over silica gel. The polyphenol components of the mixture were analyzed via HPLC-MS/MS and compared with the reported data. Three cinnamic acid derivatives and five flavonoid components in the fruits of the three varieties were identified for the first time in this study. The antioxidant activity and anticancer effect of the polyphenol mixtures of the three fruits were determined. The antioxidant and anticancer activities of CLST were substantially higher than those of C. annuum L. cv. Cupra and C. annuum L. cv. Orange glory. The high activities of CLST were attributed to the much higher concentration of quercetin derivatives in CLST.


Loss of Axdnd1 causes sterility due to impaired spermatid differentiation in mice.
Spermiogenesis, the process of deformation of sperm head morphology and flagella formation, is a phenomenon unique to sperm. Axonemal dynein light chain proteins are localized to sperm flagella and are known to be involved in sperm motility. Here, we focused on the gene axonemal dynein light chain domain containing 1 (Axdnd1) with the aim to determine the function of its protein product AXDND1. To elucidate the role of AXDND1 in spermatogenesis, we generated Axdnd1 knockout (KO) mice using the CRISPR/Cas9 system. The generated mice were subjected to fertility tests and analyzed by immunohistochemistry. The Axdnd1 KO mouse exhibited sterility caused by impaired spermiogenesis during the elongation step as well as abnormal nuclear shaping and manchette, which are essential for spermiogenesis. Moreover, AXDND1 showed enriched testicular expression and was localized from the mid-pachytene spermatocytes to the early spermatids. Axdnd1 is essential for spermatogenesis in the mouse testes. These findings improve our understanding of spermiogenesis and related defects. According to a recent report, deleterious heterozygous mutations in AXDND1 were found in non-obstructive azoospermia (NOA) patients. Therefore, Axdnd1 KO mice could be used as a model system for NOA, which will greatly contribute to future NOA treatment studies.


Influence of a buffered solution on the adsorption isotherm and overloaded band profiles of an ionizable compound.
The overloaded band profiles and the adsorption isotherms of propranolol were acquired at 23 degrees C, on the endcapped C18-Kromasil stationary phase, using two aqueous solutions of methanol as the mobile phase. The first solution contained 40% methanol and no buffer. The second contained an aqueous acetate buffer at C(buffer) = 0.20 M and pH = 5.9. In both cases, 33 isotherm data points were acquired by frontal analysis (FA), to achieve an accurate description of the isotherms in the concentration range between 1.54 x 10(-3) and 1.54 x 10(-1) mol/l of propranolol. The isotherms obtained were best described by a bi-Langmuir and a bi-Moreau isotherm model, depending on whether the mobile phase was buffered or not. This shows that the adsorption of propranolol takes place on two different types of sites, a behavior similar to the one already observed with phenol and caffeine on the same column. The presence of the buffer in the mobile phase drastically changes the adsorption mechanism of propranolol. Weak adsorbate-adsorbate interactions (two and three times RT on the low- and the high-energy sites, respectively) take place in the absence of buffer but vanish when the mobile phase is buffered. As expected, the adsorption constants on the abundant low-energy sites with or without buffer are comparable because the mobile phase composition was adjusted to give similar retention times in the two cases. On the other hand, the adsorption of propranolol on the high-energy sites is stronger in presence of the buffer. The difference probably comes from ion-pair formation in the adsorbed phase between the propranolol cation and the acetate anion. The change in total saturation capacity of the adsorbent (22%) compared to that for phenol is explained by the difference in methanol content of the mobile phase.


Updates of lifetime costs of care and quality-of-life estimates for HIV-infected persons in the United States: late versus early diagnosis and entry into care.
Lifetime costs of care and quality-of-life estimates for HIV-infected persons depend on the disease stage at which these persons are diagnosed, enter care, and start antiretroviral therapy. Updated estimates were used to analyze the effects of late versus early diagnosis/entry on US lifetime care costs, quality-of-life estimates, and HIV transmissions. The Progression and Transmission of HIV/AIDS model was used to estimate discounted (3%) lifetime treatment costs ($US 2011) and quality-of-life variables from time of infection for cohorts of 10,000 HIV-infected index patients in 4 categories of CD4 count at diagnosis: (I) ≤200 cells/μL, (II) 201-350 cells/μL, (III) 351-500 cells/μL, and (IV) 501-900 cells/μL. It is assumed that index patient diagnoses were uniformly distributed across the CD4 count range in each category and that patients entered care at the time of diagnosis, remained in care, and were eligible to initiate antiretroviral therapy at a CD4 count of 500 cells/μL. Lifetime transmissions of the index patients were also estimated. Discounted average lifetime costs varied from $253,000 for category I index patients to $402,000 for category IV patients. Discounted quality-adjusted life years lost decreased from 7.95 to 4.45 across these categories, additional years of life expectancy increased from 30.8 to 38.1, and lifetime transmissions decreased from 1.40 to 0.72. Early diagnosis and treatment of HIV infection increases lifetime costs but improves length and quality of life and reduces the number of new infections transmitted by nearly 50%.


Effect of oxime ether incorporation in acyl indole derivatives on PPAR subtype selectivity.
Compounds that simultaneously activate peroxisome proliferator-activated receptor (PPAR) subtypes α and γ have the potential to effectively treat dyslipidemia and type 2 diabetes (T2D) in a single pharmaceutically active molecule. The frequently observed side effects of selective PPARγ agonists, such as edema and weight gain, were expected to be overcome by using additive PPARα activity, leading to dual PPARα/γ agonists with balanced activity for both subtypes. Herein we report the discovery, synthesis, and optimization of a new series of α-ethoxyphenylpropionic acid bearing 5- or 6-substituted indoles. The incorporation of oxime ethers on the carbonyl portion of the benzoyl group can bring the PPARα/γ potency ratio equal to or slightly greater than one, as is the case for compounds 20 c and 21 a. Compound 20 c shows high efficacy in an ob/ob mouse model of T2D and dyslipidemia, similar to that of rosiglitazone and tesaglitazar, but with a significant increase in body weight gain. In contrast, compound 21 a, less potent as a dual PPARα/γ activator than 20 c, showed an interesting pharmacological profile, as it elicits a decrease in body weight relative to reference compounds.


Paraneoplastic erythrocytosis in a young adult with an erythropoietin-producing Wilms' tumor.
Paraneoplastic erythrocytosis in patients with Wilms' tumors is exceedingly rare, with only three reported cases in the literature. We report a case of a young man with Wilms' tumor with a significant erythrocytosis but a normal serum erythropoietin level and a tumor that elaborated erythropoietin.


Dyadic digital health interventions: Their rationale and implementation.
While most psychosocial and behavioral digital health interventions have been designed to be consumed by an individual, intervening at the level of a dyad - two interdependent individuals - can more comprehensively address the needs of both individuals and their relationship. The clinical utility of the dyadic digital health intervention approach, as well as the practical implementation of this design, will be demonstrated via three examples: eSCCIP, FAMS, and OurRelationship.


Identification of a novel calcium antagonist binding site in rat brain by SR 33557.
1. In K(+)-depolarized taenia preparations from guinea-pig caecum SR 33557 was a potent antagonist of Ca(2+)-induced contractions and antagonized the effect of the calcium channel activator Bay K 8644. 2. SR 33557 displayed high affinity (pKi 9.54 +/- 0.04, nH 1.01) for the [3H]-(+/-)-PN 200-110 binding site in rat cerebral cortex membranes. In the presence of 5 mM Ca2+ this affinity was reduced (pKi 8.82 +/- 0.01, nH 1.05) whilst the affinity of nitrendipine was unaffected by this concentration of Ca2+. 3. Saturation binding experiments in rat cerebral cortex carried out in the absence and presence of SR 33557 (0.1-1.0 nM) indicated an apparently competitive interaction at the dihydropyridine site, in that SR 33557 reduced the KD of [3H]-(+/-)-PN 200-110 binding without any effect on Bmax. In kinetic experiments, the rate of dissociation of [3H]-(+/-)-PN 200-110 from rat cerebral cortex was unchanged in the presence of SR 33557 (5 nM). 4. D-cis-diltiazem fully reversed the inhibition [3H]-nitrendipine binding to rat cerebral cortex produced by SR 33557 indicating the site of action of SR 33557 to be distinct from the dihydropyridine (DHP) binding site. 5. Saturation analysis indicated that [3H]-SR 33557 (0.01-0.8 nM) labelled a single class of binding sites in rat cerebral cortex membranes with high affinity (KD 0.12 +/- 0.01, Bmax 222 +/- 20 fmol mg-1 protein), although kinetic data indicated the existence of negative cooperativity between the binding sites. 6.In competition studies, a variety of different calcium antagonists displayed similar affinity for [3H]-SR 33557 and [3H]-(+/-)-PN 200-110 sites. The [3H]-SR 33557 site was sensitive to the inhibitory effect of divalent cations. The affinity of Cd2+ was 0.026 +/- 0.015 mM and the rank order of affinity was Cd2+ >Ca2+ >Mn2+ >Mg2+ >Na+.7. We propose that SR 33557 labels a distinct site in rat cerebral cortex. The coupling between the SR 33557 and DHP site appears to be very close, resulting in apparently competitive interactions in some experimental protocols but can be revealed as negatively allosteric in other circumstances.


Leakage of internal markers from erythrocytes and lipid vesicles induced by melittin, gramicidin S and alamethicin: a comparative study.
The membrane-disruptive capacities of melittin, derivatised melittins, alamethicin and gramicidin S have been compared for the human erythrocyte membrane and lipid vesicles of three different compositions (phosphatidylcholine, 85% phosphatidylcholine/15% phosphatidylserine, and a lipid analogue of the outer leaflet of the human erythrocyte membrane). The sensitivity to ionic strength, divalent metal ions and polylysine of release of fluorescent markers from liposomes and of haemoglobin from intact erythrocytes has been assayed. Acetyl melittin was found to he more effective than melittin in lysing phosphatidylcholine and phosphatidylcholine/phosphatidylserine vesicles, somewhat less effective in the lipid analogue and markedly less effective in lysing erythrocytes. Succinyl melittin was non-haemolytic, but was able to lyse lipid vesicles at a high concentration. Ca2+ inhibited melittin haemolysis at high ionic strength (150 mM NaCl), but produced a more complex response of stimulation followed by inhibition at low ionic strength. In lipid vesicles, Ca2+ either stimulated melittin lysis or was ineffective. Zn2+ exerted effects similar to Ca2+ with lipid vesicles at approx. 10-fold lower concentration except that a weak inhibition was observed for the erythrocyte membrane lipid analogue at high ionic strength. Polylysine strongly inhibited haemolysis by melittin at low ionic strength, but was ineffective or stimulatory in lipid vesicle lysis. High phosphate concentration also inhibited melittin haemolysis, but again no corresponding effect could he found in any of the lipid vesicle systems. These disparities between effects of melittin on erythrocytes and lipid vesicles support the proposal that melittin-protein interactions are of consequence to its haemolytic action. Similar experiments were performed with gramicidin S and alamethicin in order to compare their lytic properties with those of melittin. It was found that each lysin exhibited its own individual pattern of sensitivity to lipid composition, ionic strength and inhibition by cations. It thus appears likely that the detailed molecular interactions responsible for lysis are significantly different for each of these three agents.


Exploring the photosynthetic production capacity of sucrose by cyanobacteria.
Because cyanobacteria are photosynthetic, fast-growing microorganisms that can accumulate sucrose under salt stress, they have a potential application as a sugar source for the biomass-derived production of renewable fuels and chemicals. In the present study, the production of sucrose by the cyanobacteria Synechocystis sp. PCC6803, Synechococcus elongatus PCC7942, and Anabaena sp. PCC7120 was examined. The three species displayed different growth curves and intracellular sucrose accumulation rates in response to NaCl. Synechocystis sp. PCC6803 was used to examine the impact of modifying the metabolic pathway on the levels of sucrose production. The co-overexpression of sps (slr0045), spp (slr0953), and ugp (slr0207) lead to a 2-fold increase in intracellular sucrose accumulation, whereas knockout of ggpS (sll1566) resulted in a 1.5-fold increase in the production of this sugar. When combined, these genetic modifications resulted in a fourfold increase in intracellular sucrose accumulation. To explore methods for optimizing the transport of the intracellular sucrose to the growth medium, the acid-wash technique and the CscB (sucrose permease)-dependent export method were evaluated using Synechocystis sp. PCC6803. Whereas the acid-wash technique proved to be effective, the CscB-dependent export method was not effective. Taken together, these results suggest that using genetic engineering, photosynthetic cyanobacteria can be optimized for efficient sucrose production.


Automated detection of neutralizing SARS-CoV-2 antibodies in minutes using a competitive chemiluminescence immunoassay.
The SARS-CoV-2 pandemic has shown the importance of rapid and comprehensive diagnostic tools. While there are numerous rapid antigen tests available, rapid serological assays for the detection of neutralizing antibodies are and will be needed to determine not only the amount of antibodies formed after infection or vaccination but also their neutralizing potential, preventing the cell entry of SARS-CoV-2. Current active-virus neutralization assays require biosafety level 3 facilities, while virus-free surrogate assays are more versatile in applications, but still take typically several hours until results are available. To overcome these disadvantages, we developed a competitive chemiluminescence immunoassay that enables the detection of neutralizing SARS-CoV-2 antibodies within 7 min. The neutralizing antibodies bind to the viral receptor binding domain (RBD) and inhibit the binding to the human angiotensin-converting enzyme 2 (ACE2) receptor. This competitive binding inhibition test was characterized with a set of 80 samples, which could all be classified correctly. The assay results favorably compare to those obtained with a more time-intensive ELISA-based neutralization test and a commercial surrogate neutralization assay. Our test could further be used to detect individuals with a high total IgG antibody titer, but only a low neutralizing titer, as well as for monitoring neutralizing antibodies after vaccinations. This effective performance in SARS-CoV-2 seromonitoring delineates the potential for the test to be adapted to other diseases in the future.


The effectiveness of the random dot E stereotest as a preschool vision screening instrument.
Sixty nursery-school children (ages 39--76 months) were screened with a modified-clinical method (visual acuity, retinoscopy, cover test, NPC) and with a relatively new vectographic test for stereopsis--the Random Dot E. Screening outcomes indicate that the RDE was as effective in identifying children for referral as was the modified clinical battery. Implications are discussed.


Debonding orthodontic ceramic brackets by ultrasonic instrumentation.
Breakage of ceramic brackets because of brittleness may cause such problems as aspiration of fragments and injury by flying debris. Portions remaining on the tooth must be laboriously ground off with a handpiece. This study investigated a potential method of lowering the force required to remove ceramic brackets, i.e., use of an ultrasonic chisel. Ceramic brackets were bonded to extracted incisors and canines. The degree of cure of a light-activated cement was systematically varied with different exposure times to a curing light. The brackets were sheared from the teeth with a universal testing machine and a chisel tip on a Cavitron. The ultrasonic chisel markedly reduced the force required to debond the brackets; however, the time of application averaged 16.6 seconds. Reducing the degree of cure of the cement had only a slight effect on the time required and the forces generated. This method of debonding is not recommended without further development.


Development of an analytical method for the simultaneous determination of 50 semi-volatile organic contaminants in wastewaters.
This work describes the development of a robust analytical methodology for the simultaneous determination of 50 semi-volatile organic compounds (SVOCs) in wastewater effluent samples by solid-phase extraction (SPE) followed by gas chromatography coupled to mass spectrometry (GC-MS) analysis. In this work, we studied extensively whether the validated SPE method used for the analysis of polar compounds in wastewaters could be extended to the analysis of non-polar compounds in the same analytical run. To that aim, the effect of different organic solvents in the SPE process (i.e., sample conditioning prior to SPE, elution solvent and evaporation steps) was evaluated. In this sense, the addition of methanol to wastewater samples before the extraction, the use of hexane:toluene (4:1, v/v) mixture for the quantitative elution of target compounds, and the addition of isooctane during the evaporation were required to minimize analyte losses during SPE and enhance extraction yields. Overall, the developed methodology showed a good performance for the determination of 50 SVOCs, and was further applied to the analysis of real wastewater effluent samples.•A validated SPE method for polar compounds was extended to the analysis of non-polar compounds.•Elution with hex:tol (4:1, v/v) and the addition of isooctane during the evaporation yield good recoveries.•The developed methodology was suitable for the determination of 50 SVOCs in aqueous samples.


Triethylene glycol dimethacrylate induces large deletions in the hprt gene of V79 cells.
Acrylate esters are applied in industrial and consumer products often associated with polymers and resins. The difunctional methacrylate, triethylene glycol dimethacrylate (TEGDMA), is also frequently included in dental composite materials. Recently, mutagenicity testing of the compound revealed the induction of gene mutations at the hprt locus in V79 cell [H. Schweikl, G. Schmalz, K. Rackebrandt, The mutagenic activity of unpolymerized resin monomers in Salmonella typhimurium and V79 cells, Mutat. Res. 415 (1998) 119-130]. In the present study, TEGDMA caused a dose dependent increase of the number of micronuclei in V79 cells. Furthermore, the mutation spectra induced in exon sequences of the hprt gene in HPRT-deficient V79 cell clones were analyzed by the polymerase chain reaction (PCR). No DNA sequence deletions were observed in spontaneously occurring HPRT-deficient cell clones at the molecular level after PCR analysis, indicating that all spontaneous mutations were caused by point mutations. However, TEGDMA treated V79 cell cultures exhibited different mutation spectra. Only one cell clone among a total of 25 contained all exon sequences of the hprt gene. Large DNA sequences were deleted in 24 cell clones. Partial gene deletions occurred in four clones from exon 5 through 9, and exon 1 was not amplified in one cell clone. Exon sequences of the hprt gene were totally deleted in 19 HPRT-deficient clones. The induction of mostly large deletions in the genome of mammalian cells, like the mutation spectra induced by TEGDMA in V79 cells here, is probably typical for crosslinking agents, including anticancer drugs. Identical types of mutations including chromosomal aberrations and the formation of micronuclei in vitro were observed for acrylates and methacrylates tested so far in various mutation assays. Therefore, we conclude by analogy that the induction of large DNA sequence deletions as shown here with the reactive dimethacrylate, triethylene glycol dimethacrylate, is probably common for acrylates and methacrylates.


Inhibition of DNA replication in Saccharomyces cerevisiae by araCMP.
Cytosine arabinoside (araC), a potent inhibitor of DNA replication in mammalian cells, was found to be completely ineffective in Saccharomyces cerevisiae. The 5' monophosphate derivative, araCMP, is toxic and effectively inhibits both nuclear and mitochondrial DNA synthesis in this organism. Although wild-type strains can be inhibited by araCMP, dTMP permeable (tup-) strains were found to be much more sensitive to the analogue. In vivo labelling experiments indicate that araC enters yeast cells; however, it is extensively catabolized by deamination and breakage of the glycosidic bond. In addition, the analogue is not efficiently phosphorylated in S. cerevisiae owing to an apparent lack of deoxynucleoside kinase activity. These results provide further evidence that deoxyribonucleotides can be synthesized only through de novo pathways in this organism. Finally, araCMP was found to be recombinagenic in S. cerevisiae which suggests, together with other previous studies, that, in general, inhibition of DNA synthesis in yeast promotes mitotic recombination events.


Genomic-based classification of Catenulispora pinisilvae sp. nov., novel actinobacteria isolated from a pine forest soil in Poland and emended description of Catenulispora rubra.
Two actinobacteria, strains NF3 and NH11T, isolated from a pine forest soil, near Torun, Poland were examined for diverse chemotaxonomic and morphological properties that placed them in the genus Catenulispora. They produced an extensively branched stable mycelium, contained LL-diaminopimelic acid as the diamino acid of the peptidoglycan, arabinose as the diagnostic whole-organism sugar, tetra-, hexa- and octa-hydrogenated menaquinones with nine isoprenoid units as the predominant isoprenologues, iso-C16:0 and anteiso-C17:0 as major fatty acids, and formed a well supported clade within the Catenulispora 16S rRNA gene tree together with Catenulispora acidiphila DSM 44928T and Catenulispora rubra DSM 44948T sharing sequence similarities with the latter of 98.8 and 99.0%, respectively. The sizes of whole genome sequences generated for the isolates and the C. rubra strain ranged from 11.20 to 12.80 Mbp with corresponding in silico DNA G+C values of 69.9-70.0%. The isolates and the C. acidiphila and C. rubra strains formed a well supported branch in the actinobacterial phylogenomic tree. Isolates NF3 and NH11T belong to the same species as they have identical 16S rRNA gene sequences, share many chemotaxonomic, cultural and phenotypic features and show very high average nucleotide identity (ANI) and digital DNA:DNA relatedness (dDDH) similarities. They can be distinguished from their closest phylogenomic neighbours by using a combination of chemotaxonomic and phenotypic properties and by ANI and dDDH values well below the thresholds of these metrics used to assign closely related strains to different species. Consequently, we propose that the isolates be classified as a new Catenulispora species, Catenulispora pinisilvae sp. nov., the type strain is NH11T (=DSM 111109T =PCM 3046T). An emended description is given for C. rubra based on data acquired in the present study. Analyses of the draft genomes of the isolates and the C. acidiphila and C. rubra strains revealed the presence of many biosynthetic gene clusters with the potential to synthesize novel drug-like metabolites. In vitro screens showed that the isolates inhibited the growth of Gram-positive bacteria and wheat pathogens belonging to the genus Fusarium.


Organ culture of adult mouse intestine. I. Morphological results after 24 and 48 hours of culture.
Explants of adult mouse intestine have been maintained in organ culture for 24 to 48 hr. The best results have been obtained with a mixture of DMEM-HEPES medium and NCTC-135 enriched with 10% fetal bovine serum. The morphology of the mucosa is well preserved at the light and electron microscopic level: absorbing cells exhibit an increase in secondary lysosomes; goblet cells and Paneth cells remain active; numerous mitoses are observed in the crypts; and vigorous re-epithelization takes place on the margin of the explants.


An activity-maintaining sequential protein extraction method for bioactive assay and proteome analysis of velvet antlers.
The exceptional growth rate of velvet antler makes it a valuable model for studying the development of tissues, such as blood vessels, cartilage and bone. Meanwhile, investigating the activities of extracted proteins from velvet antlers promisingly leads to the discovery of new active factors which regulate the development of above-mentioned tissue types. In this study, a novel sequential protein extraction method was developed for proteome profiling and bioactivity study of velvet antlers. Herein, four antler growing tips were pooled to create a proportional pooled sample, and three aliquots of which were extracted in parallel using the developed extraction method. For each sample, proteins were extracted sequentially by saline solvent (0.15M sodium chloride, pH 7.0), mild acid buffer (0.15M acetate buffer, pH 4.0) and mild alkaline buffer (0.15M glycine-sodium hydroxide buffer, pH 10.0) with good bio-compatibility to prevent proteins denaturation. Then STD lysis buffer, containing 4% SDS, 0.1M Tris-HCl and 0.1M DTT, was used to extract hydrophobic proteins. The tryptic digest of each fraction was analyzed by nanoRPLC-ESI-MS/MS in triplicates, with false discovery rate for peptide identification adjusted to 1% to create filtered protein group list. In total, 1423 protein groups were identified, which expanded up to 3 times of the previous published dataset. The relative standard deviation of identified peptide and protein group number for all analyses indicated the good reproducibility of the developed sequential protein extraction method. Additionally, proteins extracted by acid buffer and alkaline buffer showed obvious promoting effect on the proliferation of human umbilical vein endothelial cells. All these results demonstrate that the developed sequential extraction method is efficient for the comprehensive proteome analysis and activity investigation of velvet antlers.


Seismicity in Romania--evidence for the sinking lithosphere.
The revision of Romanian earthquakes shows a distribution suggesting a sinking lithosphere under the Carpathian arc. Thermal and gravitational anomalies, as well as petrological and tectonic features, provide further evidence on the cause and character of intermediate earthquakes of Romania. This is consistent with the theory of plate tectonics in south-east Europe.


Kainic acid-induced lesion of dopaminergic target cells in the striatum: consequences on the dynamics of cerebellar cGMP.
The bilateral intrastriatal injection of kainic acid (2 microgram/caudate) caused a marked decrease in the activity of striatal dopamine-sensitive adenylate cyclase. This probable loss of target cells for dopamine was associated with a parallel decrease of the cGMP level in the cerebellar cortex, which was maximal (80% loss) by 24 h and prevented apomorphine from raising cerebellar cGMP levels. On the contrary harmaline and isoniazid both increased the levels of cerebellar cGMP in kainic-lesioned rats to the same extent as in control rats. The results indicate that dopamine mechanisms in the striatum are involved in the regulation of cerebellar cortex cGMP.


Comparative analysis of microbial communities associated with bacteriomes, reproductive organs and eggs of the cicada Subpsaltria yangi.
Plant sap-feeding insects of Hemiptera often form intimate symbioses with microbes to obtain nutrients. The cicada Subpsaltria yangi is the only species of the subfamily Tettigadinae known from China. Using high-throughput sequencing combined with fluorescence in situ hybridization analysis, we characterize the bacterial composition of the bacteriomes, testes, ovaries and eggs of two representative populations of this species which occur in different habitats and feed on different plant hosts. In both populations, the bacterial community diversity in the testes was significantly higher than that in other tissues. The obligate endosymbiont Candidatus Sulcia muelleri was observed in all samples and was dominant in the bacteriomes, ovaries and eggs. The usual co-resident endosymbiont Candidatus Hodgkinia cicadicola found in some other cicadas was not detected. Instead, a novel Rhizobiales bacterium which shows a ~ 81% 16S rDNA similarity to Ca. Hodgkinia cicadicola was detected. Given that the genome of Ca. Hodgkinia cicadicola exhibits rapid evolution, it is possible that this novel Rhizobiales bacterium is a related endosymbiont with beneficial trophic functions similar to that of Ca. Hodgkinia cicadicola hosted by several certain other cicadas. The presence of the novel Rhizobiales species in other cicadas and its involvement with the adaptive evolution of related cicada hosts require further investigation. Discrepancy of bacterial communities associated with testes between the two populations may be closely related to the geographic isolation and divergence of habitats and host plants. Our results are informative for further studies of evolutionary divergence of related endosymbionts hosted in cicadas.


Evaluation of a social determinants of health screening questionnaire and workflow pilot within an adult ambulatory clinic.
There is increased recognition in clinical settings of the importance of documenting, understanding, and addressing patients' social determinants of health (SDOH) to improve health and address health inequities. This study evaluated a pilot of a standardized SDOH screening questionnaire and workflow in an ambulatory clinic within a large integrated health network in Northern California. The pilot screened for SDOH needs using an 11-question Epic-compatible paper questionnaire assessing eight SDOH and health behavior domains: financial resource, transportation, stress, depression, intimate partner violence, social connections, physical activity, and alcohol consumption. Eligible patients for the pilot receiving a Medicare wellness, adult annual, or new patient visits during a five-week period (February-March, 2020), and a comparison group from the same time period in 2019 were identified. Sociodemographic data (age, sex, race/ethnicity, and payment type), visit type, length of visit, and responses to SDOH questions were extracted from electronic health records, and a staff experience survey was administered. The evaluation was guided by the Reach, Effectiveness, Adoption, Implementation, and Maintenance (RE-AIM) framework. Two-hundred eighty-nine patients were eligible for SDOH screening. Responsiveness by domain ranged from 55 to 67%, except for depression. Half of patients had at least one identified social need, the most common being stress (33%), physical activity (22%), alcohol (12%), and social connections (6%). Physical activity needs were identified more in females (81% vs. 19% in males, p < .01) and at new patient/transfer visits (48% vs. 13% at Medicare wellness and 38% at adult wellness visits, p < .05). Average length of visit was 39.8 min, which was 1.7 min longer than that in 2019. Visit lengths were longer among patients 65+ (43.4 min) and patients having public insurance (43.6 min). Most staff agreed that collecting SDOH data was relevant and accepted the SDOH questionnaire and workflow but highlighted opportunities for improvement in training and connecting patients to resources. Use of evidence-based SDOH screening questions and associated workflow was effective in gathering patient SDOH information and identifying social needs in an ambulatory setting. Future studies should use qualitative data to understand patient and staff experiences with collecting SDOH information in healthcare settings.


Fixation of platelet-rich plasma and fibrin gels on knee cartilage defects after microfracture with arthroscopy.
An investigation of arthroscopic surgery combined with coverage of the microfractured wound surface with platelet-rich plasma (PRP) and fibrin gels (FG) to treat knee cartilage defects. Between February 2017 and February 2020, 145 patients with knee cartilage defects were treated. Only isolated full-thickness cartilage defects were included, and 28 patients (12 men and 16 women) were included in this study. They were all treated with arthroscopic surgery on subchondral bones, filled with PRP and thrombin, and sealed with FG. The knee pain visual analogue scale (VAS) scores were measured after the patients climbed ten stairs up and down, and the Western Ontario and McMaster Universities osteoarthritis index and the area of cartilage defects were measured through the pre-operative and post-operative follow-up. The complication incidences were also observed. All patients were followed up for ten to 15 months (median 12 months). The knee pain VAS scores decreased from 6.57 ± 1.07 pre-operatively to 2.09 ± 1.35 at the last follow-up. The WOMAC osteoarthritis index decreased from 44.32 ± 3.95 (mean ± sd) pre-operatively to 16.57 ± 2.20 by the last follow-up. The cartilage defect decreased from 2.93 ± 0.65 cm2 pre-operatively to 1.09 ± 0.69 cm2 at the last follow-up. All scores showed statistically significant improvements after surgery (p < 0.05). No complications were observed. The combination therapy of arthroscopic surgery and covering the microfractured wound surface with PRP and FG can repair knee cartilage defects, relieve pain, and improve function, and is a safe and effective treatment.


Environmentally adaptive MOF-based device enables continuous self-optimizing atmospheric water harvesting.
Harvesting water vapor from desert, arid environments by metal-organic framework (MOF) based devices to deliver clean liquid water is critically dependent on environment and climate conditions. However, reported devices have yet been developed to adapt in real-time to such conditions during their operation, which severely limits water production efficiency and unnecessarily increases power consumption. Herein, we report and detail a mode of water harvesting operation, termed 'adaptive water harvesting', from which a MOF-based device is proven capable of adapting the adsorption and desorption phases of its water harvesting cycle to weather fluctuations throughout a given day, week, and month such that its water production efficiency is continuously optimized. In performance evaluation experiments in a desert, arid climate (17-32% relative humidity), the adaptive water harvesting device achieves a 169% increase in water production (3.5 LH2O kgMOF-1 d-1) when compared to the best-performing, reported active device (0.7-1.3 LH2O kgMOF-1 d-1 at 10-32% relative humidity), a lower power consumption (1.67-5.25 kWh LH2O-1), and saves time by requiring nearly 1.5 cycles less than a counterpart active device. Furthermore, the produced water meets the national drinking standards of a potential technology-adopting country.


Genome-Wide Single-Nucleotide Polymorphism-Based Genomic Diversity and Runs of Homozygosity for Selection Signatures in Equine Breeds.
The horse, one of the most domesticated animals, has been used for several purposes, like transportation, hunting, in sport, or for agriculture-related works. Kathiawari, Marwari, Manipuri, Zanskari, Bhutia, Spiti, and Thoroughbred are the main breeds of horses, particularly due to their agroclimatic adaptation and role in any kind of strong physical activity, and these characteristics are majorly governed by genetic factors. The genetic diversity and phylogenetic relationship of these Indian equine breeds using microsatellite markers have been reported, but further studies exploring the SNP diversity and runs of homozygosity revealing the selection signature of breeds are still warranted. In our study, the identification of genes that play a vital role in muscle development is performed through SNP detection via the whole-genome sequencing approach. A total of 96 samples, categorized under seven breeds, and 620,721 SNPs were considered to ascertain the ROH patterns amongst all the seven breeds. Over 5444 ROH islands were mined, and the maximum number of ROHs was found to be present in Zanskari, while Thoroughbred was confined to the lowest number of ROHs. Gene enrichment of these ROH islands produced 6757 functional genes, with AGPAT1, CLEC4, and CFAP20 as important gene families. However, QTL annotation revealed that the maximum QTLs were associated with Wither's height trait ontology that falls under the growth trait in all seven breeds. An Equine SNP marker database (EqSNPDb) was developed to catalogue ROHs for all these equine breeds for the flexible and easy chromosome-wise retrieval of ROH along with the genotype details of all the SNPs. Such a study can reveal breed divergence in different climatic and ecological conditions.


Micromonospora haikouensis sp. nov., isolated from mangrove soil.
An actinomycete strain 232617(T) was isolated from a composite mangrove sediment sample collected in Haikou, China. Phylogenetic analysis of the 16S rRNA gene sequence of strain 232617(T) indicated the highest similarity with Micromonospora siamensis TT2-4(T) (99.05%), Micromonospora krabiensis A-2(T) (98.99%) and Micromonospora carbonacea DSM 43815(T) (98.91%). The gyrB gene sequence analysis also indicated that 232617(T) should be assigned to the genus Micromonospora. The cell wall contains meso-DAP and glycine. The major menaquinones were MK-10(H(4)) and MK-10(H(6)), with MK-9(H(4)) as minor components. The characteristic whole-cell sugars are xylose, arabinose and glucose. The phospholipid profile comprises phosphatidylethanolamine, diphosphatidlglycerol and phosphatidylinositol mannoside. The DNA G+C content is 71.5 mol%. Furthermore, a combination of DNA-DNA relatedness and some physiological and biochemical properties indicated that the novel strain could be readily distinguished from the closest related species. On the basis of these phenotypic and genotypic data, strain 232617(T) represents a novel species of the genus Micromonospora, for which the name Micromonospora haikouensis sp. nov. is proposed. The type strain is 232617(T) (= CCTCC AA 201112 (T) = DSM 45626 (T)).


A Case of Complete Resolution of Severe Plantar Dyshidrotic Eczema With Dupilumab
A 44-year-old woman with a history of asthma, hypercholesterolemia, and impaired glucose tolerance presented with severely painful and intensely pruritic plantar dermatitis for more than two years that impaired her ability to walk.


Testicular suture: a significant risk factor for infertility among formerly cryptorchid men.
Although fertility is decreased after cryptorchidism, the importance of risk factors, including parenchymal testicular suture, is unknown. The aim of this study was to examine the relationship between parenchymal testicular suture and failure to conceive a child for 1 year or longer among formerly cryptorchid men. Men who underwent orchidopexy between 1955 and 1972 at the Children's Hospital of Pittsburgh (n = 619) were surveyed by questionnaire and their medical records reviewed. Only the men who attempted to conceive a child (n = 387) are included. Logistic regression analysis determined significant risk factors for infertility. Testicular suture was strongly related to infertility (RR, 7.56; 95% CI, 1.66, 34.39) as were bilateral cryptorchidism (RR, 5.51; 95% CI, 1.58, 19.24), varicocele (RR, 4.72; 95% CI, 1.42, 15.75), hormone treatment before surgery (RR, 3.69; 95% CI, 1.22, 11.11), and partner conception problem (RR, 3.32; 95% CI, 1.11, 9.90). Testicular suture was a potent independent determinant of infertility among formerly cryptorchid men who have orchidopexy. Bilateral cryptorchidism, hormone treatment, varicocele, and partner conception problems also were associated with increased infertility.


Disrupting effects of antibiotic sulfathiazole on developmental process during sensitive life-cycle stage of Chironomus riparius.
Antibiotics in the environment are a concern due to their potential to harm humans and interrupt ecosystems. Sulfathiazole (STZ), a sulfonamide antibiotic, is commonly used in aquaculture and is typically found in aquatic ecosystems. We evaluated the ecological risk of STZ by examining biological, molecular and biochemical response in Chironomus riparius. Samples were exposed to STZ for 12, 24 and 96 h, and effects of STZ were evaluated at the molecular level by analyzing changes in gene expression related to the endocrine system, cellular stress response and enzyme activity of genes on antioxidant and detoxification pathways. STZ exposure induced significant effects on survival, growth and sex ratio of emergent adults and mouthpart deformity in C. riparius. STZ caused concentration and time-dependent toxicity in most of the selected biomarkers. STZ exposure leads to significant heat-shock response of protein genes (HSP70, HSP40, HSP90 and HSP27) and to disruption by up-regulating selected genes, including the ecdysone receptor gene, estrogen-related receptors, ultraspiracle and E74 early ecdysone-responsive gene. Furthermore, STZ induced alteration of enzyme activities on antioxidant and detoxification responses (catalase, superoxide dismutase, glutathione peroxidase and peroxidase) in C. riparius. By inducing oxidative stress, antibiotic STZ disturbs the endocrine system and produces adverse effects in growth processes of invertebrates.


Candidate target genes for the Saccharomyces cerevisiae transcription factor, Yap2.
In Saccharomyces cerevisiae, the Yap family of basic leucine zipper (bZip) proteins contains eight members. The Yap family proteins are implicated in a variety of stress responses; among these proteins, Yap1 acts as a major regulator of oxidative stress responses. However, the functional roles of the remaining Yap family members are poorly understood. To elucidate the function of Yap2, we mined candidate target genes of Yap2 by proteomic analysis. Among the identified genes, FRM2 was previously identified as a target gene of Yap2, which confirmed the validity of our screening method. YNL134C and YDL124W were also identified as candidate Yap2 target genes. These genes were upregulated in strains overexpressing Yap2 and possess Yap2 target sequences in their promoter regions. Furthermore, chromatin immunoprecipitation assays showed that YNL134C and YDL124W have Yap2 binding motif. These data will help to elucidate the functional role of Yap2.


Elementary students' perceptions of 3Dmetric: A cross-sectional study.
Rapid changes in the 21st century demand the use of technology in learning geometry in elementary schools. One such technology is augmented reality (AR). 3Dmetric (3D and Geometric) is a geometry learning medium on AR-based 3D space material. Students' perceptions, which refer to their interpretation, are a key factor in studying the changes in their interpretations of a particular phenomenon. The purpose of the current study was to investigate the perceptions of elementary school students after using 3Dmetric to learn geometric shapes. The differences and the relationship between the students' level of perception and level of spatial ability were also investigated. This study applied a cross-sectional approach with quantitative and qualitative designs. A total of 36 students in one elementary school in Indonesia participated in this study. The instruments used were the Perception Scale for Using 3Dmetric in Geometry Teaching, Spatial Ability Scale, and In-Depth Interview Form. Results showed that the positive perception of elementary school students regarding the use of 3Dmetric does not depend on the level of their spatial ability. Moreover, the difference in their perceptions is not caused by the level of their spatial ability. The positive findings in this cross-sectional study can contribute to the success of AR-based learning and teaching in the 21st century, especially with regard to learning materials for 3D geometry. They can also lead to the formation of the spatial abilities and improvement in the academic performance of elementary school students.


Subdural effusion in the acute stage of Kawasaki disease (Mucocutaneous lymph node syndrome).
A 6-month-old girl with complications of subdural effusions at the acute stage of Kawasaki disease is reported. Based on the pathology of Kawasaki disease and considering the possibility of systemic vasculitis, the subdural effusions were assumed to be attributable to vasculitis involving the dura mater.


Letters.
COMMUNITY CLINICS FOR LEG ULCERS COMMUNITY CLINICS IMPROVE HEALING RATES AND QUALITY OF LIFE ABSORBENCY OF PERFORATED FILM WOUND DRESSINGS CORRECT TECHNIQUE FOR ABPI MEASUREMENT.


Evaluation of a rebound tonometer for measuring intraocular pressure in dogs and horses.
To compare intraocular pressure (IOP) measurements obtained with a rebound tonometer in dogs and horses with values obtained by means of applanation tonometry and direct manometry. Prospective study. 100 dogs and 35 horses with clinically normal eyes, 10 enucleated eyes from 5 dogs, and 6 enucleated eyes from 3 horses. In the enucleated eyes, IOP measured by means of direct manometry was sequentially increased from 5 to 80 mm Hg, and IOP was measured with the rebound tonometer. In the dogs and horses, results of rebound tonometry were compared with results of applanation tonometry. For the enucleated dog and horse eyes, there was a strong (r2 = 0.99) linear relationship between pressures obtained by means of direct manometry and those obtained by means of rebound tonometry. Mean +/- SD IOPs obtained with the rebound tonometer were 10.8 +/- 3.1 mm Hg (range, 5 to 17 mm Hg) and 22.1 +/- 5.9 mm Hg (range, 10 to 34 mm Hg) for the dogs and horses, respectively. Mean IOPs obtained with the applanation tonometer were 12.9 +/- 2.7 mm Hg (range, 8 to 18 mm Hg) and 21.0 +/- 5.9 mm Hg (range, 9 to 33 mm Hg), respectively. Values obtained with the rebound tonometer were, on average, 2 mm Hg lower in the dogs and 1 mm Hg higher in the horses, compared with values obtained with the applanation tonometer. Results suggest that the rebound tonometer provides accurate estimates of IOP in clinically normal eyes in dogs and horses.


Heterologous expression of the membrane proteins that control cellular excitability.
Versatile and potent expression systems are needed to decipher the structure and functions of the many excitability proteins that have been identified through molecular cloning. This article reviews the use of recombinant vaccinia viruses (VV), which have been recently explored for the heterologous expression of eukaryotic proteins. Vaccinia viruses feature a series of favourable properties, most of all a broad host range and high efficiency of infection, that make them uniquely suited as flexible expression vectors. In one type of experiment, the recombinant virus simply harbors the cDNA for the foreign protein; in a second type the virus harbors the cDNA for the specific and efficient RNA polymerase of bacteriophage T7, which in turn generates RNA from a separate introduced plasmid or virus. Both variations have been successfully applied to the expression and analysis of voltage-dependent ion channels, neurotransmitter receptors and other excitability proteins in many cell lines and postmitotic cells in culture. VV vectors promise to be particularly useful to study membrane proteins that require posttranslational processing, association with cell-specific subunits or coupling to endogenous second messengers pathways.


Author Correction: Sustained perfusion of revascularized bioengineered livers heterotopically transplanted into immunosuppressed pigs.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Age-dependent decline of copper clearance at the blood-cerebrospinal fluid barrier.
The homeostasis of copper (Cu) in the central nervous system is regulated by the blood-brain barrier and blood-cerebrospinal (CSF) barrier (BCB) in the choroid plexus. While proteins responsible for Cu uptake, release, storage and intracellular trafficking exist in the choroid plexus, the influence of age on Cu clearance from the CSF via the choroid plexus and how Cu transporting proteins contribute to the process are unelucidated. This study was designed to test the hypothesis that the aging process diminishes Cu clearance from the CSF of rats by disrupting Cu transporting proteins in the choroid plexus. Data from ventriculo-cisternal perfusion experiments demonstrated greater 64Cu radioactivity in the CSF effluents of older rats (18 months) compared to younger (1 month) and adult (2 months) rats, suggesting much slower removal of Cu by the choroid plexus in old animals. Studies utilizing qPCR and immunofluorescence revealed an age-specific expression pattern of Cu transporting proteins in the choroid plexus. Moreover, proteomic analyses unraveled age-specific proteomes in the choroid plexus with distinct pathway differences, particularly associated with extracellular matrix and neurodevelopment between young and old animals. Taken together, these findings support an age-dependent deterioration in CSF Cu clearance, which appears to be associated with altered subcellular distribution of Cu transporting proteins and proteomes in the choroid plexus.


Cell spinpods are a simple inexpensive suspension culture device to deliver fluid shear stress to renal proximal tubular cells.
Rotating forms of suspension culture allow cells to aggregate into spheroids, prevent the de-differentiating influence of 2D culture, and, perhaps most importantly of all, provide physiologically relevant, in vivo levels of shear stress. Rotating suspension culture technology has not been widely implemented, in large part because the vessels are prohibitively expensive, labor-intensive to use, and are difficult to scale for industrial applications. Our solution addresses each of these challenges in a new vessel called a cell spinpod. These small 3.5 mL capacity vessels are constructed from injection-molded thermoplastic polymer components. They contain self-sealing axial silicone rubber ports, and fluoropolymer, breathable membranes. Here we report the two-fluid modeling of the flow and stresses in cell spinpods. Cell spinpods were used to demonstrate the effect of fluid shear stress on renal cell gene expression and cellular functions, particularly membrane and xenobiotic transporters, mitochondrial function, and myeloma light chain, cisplatin and doxorubicin, toxicity. During exposure to myeloma immunoglobulin light chains, rotation increased release of clinically validated nephrotoxicity cytokine markers in a toxin-specific pattern. Addition of cisplatin or doxorubicin nephrotoxins reversed the enhanced glucose and albumin uptake induced by fluid shear stress in rotating cell spinpod cultures. Cell spinpods are a simple, inexpensive, easily automated culture device that enhances cellular functions for in vitro studies of nephrotoxicity.


The prostate cancer bone marrow niche: more than just 'fertile soil'.
The hematopoietic stem cell (HSC) niche in the bone marrow has been studied extensively over the past few decades, yet the bone marrow microenvironment that supports the growth of metastatic prostate cancer (PCa) has only been recently considered to be a specialized 'niche' as well. New evidence supports the fact that disseminated tumor cells (DTCs) of PCa actually target the HSC niche, displace the occupant HSCs and take up residence in the pre-existing niche space. This review describes some of the evidence and mechanisms by which DTCs act as molecular parasites of the HSC niche. Furthermore, the interactions between DTCs, HSCs and the niche may provide new targets for niche-directed therapy, as well as insight into the perplexing clinical manifestations of metastatic PCa disease.


Thermal Signature Measurements for Ammonium Nitrate/Fuel Mixtures by Laser Heating.
Measurements were carried out to obtain thermal signatures of several ammonium nitrate/fuel (ANF) mixtures, using a laser-heating technique referred to as the laser-driven thermal reactor (LDTR). The mixtures were ammonium nitrate (AN)/kerosene, AN/ethylene glycol, AN/paraffin wax, AN/petroleum jelly, AN/confectioner's sugar, AN/cellulose (tissue paper), nitromethane/cellulose, nitrobenzene/cellulose, AN/cellulose/nitromethane, AN/cellulose/nitrobenzene. These mixtures were also compared with AN/nitromethane and AN/diesel fuel oil, obtained from an earlier investigation. Thermograms for the mixtures, as well as individual constituents, were compared to better understand how the sample thermal signature changes with mixture composition. This is the first step in development of a thermal-signature database, to be used along with other signature databases, to improve identification of energetic substances of unknown composition. The results indicated that each individual thermal signature was associated unambiguously with a particular mixture composition. The signature features of a particular mixture were shaped by the individual constituent signatures. It was also uncovered that the baseline signature was modified after an experiment due to coating of unreacted residue on the substrate surface and a change in the reactor sphere oxide layer. Thus, care was required to pre-oxidize the sphere prior to an experiment. A minimum sample mass (which was dependent on composition) was required to detect the signature characteristics. Increased laser power served to magnify signal strength while preserving the signature features. For the mixtures examined, the thermal response of each ANF mixture was found to be different, which was based on the mixture composition and the thermal behavior of each mixture constituent.


Factors Associated With Measles Transmission in the United States During the Postelimination Era.
Measles cases and outbreaks continue to occur in the United States after the introduction of measles from endemic settings. To discern the factors associated with measles transmission in the United States after measles had been eliminated. This cross-sectional study was conducted from January 1, 2001, to December 31, 2017, in the United States among US residents and international visitors with confirmed measles. A maximum likelihood algorithm that uses the observed dates of rash onset and the known distribution of the serial interval (time between symptom onset in related consecutive cases) was applied to outbreak notification data to estimate the effective reproduction number (R), or the mean number of new infections generated per case. Transmissibility was assessed by comparing R based on the characteristics of primary and secondary cases of measles. Measles virus. Effective reproduction number (R), or the mean number of successful transmission events per case of measles (ie, the mean number of persons to whom each patient with measles spreads measles). A total of 2218 individuals with confirmed measles cases (1025 female, 1176 male, and 17 sex not reported; median age, 15 years [range, 0-89 years]) reported from 2001 to 2017 were evaluated. Among patients who received no doses of measles vaccine, R was 0.76 (95% CI, 0.71-0.81); among patients who received 1 dose of measles vaccine, R was 0.17 (95% CI, 0.11-0.26); among patients who received 2 doses or more of measles vaccine, R was 0.27 (95% CI, 0.17-0.39); and among patients with unknown vaccination status, R was 0.52 (95% CI, 0.44-0.60). Among patients born before 1957, R was 0.35 (95% CI, 0.20-0.58), and among those born on or after 1957, R was 0.64 (95% CI, 0.61-0.68). R was higher when primary and secondary cases of measles were patients aged 5 to 17 years (0.36 [95% CI, 0.31-0.42]) compared with assortative transmission in other age groups (<1 year, 0.14 [95% CI, 0.10-0.20]; 1-4 years, 0.25 [95% CI, 0.20-0.30]; 18-29 years, 0.19 [95% CI, 0.15-0.24]; 30-49 years, 0.15 [95% CI, 0.11-0.20]; ≥50 years, 0.04 [95% CI, 0.01-0.10]). The findings of this study support having high targets for 2-dose measles vaccine coverage, particularly among school-aged children in the United States.


Repair of rectal injury during robotic-assisted laparoscopic prostatectomy.
Rectal injury is a potential devastating complication in radical prostatectomy. We reviewed the incidence and management of rectal injuries with robotic-assisted laparoscopic radical prostatectomy performed by a single surgeon. Of the first 251 robotic-assisted laparoscopic radical prostatectomies performed, 2 (0.8%) were complicated by rectal injury. All patients underwent bowel preparation and received preoperative intravenous antibiotics. Both rectal injuries were detected intraoperatively and treated robotically. The rectotomy was closed in two layers of absorbable sutures and imbricated with a nonabsorbable suture. The rectum was then tacked to the levator fibers on one side with nonabsorbable suture to pull the injury away from the vesicourethral anastomosis. An omental flap was developed and tacked posterior to the urethra to cover the repair. A closed suction drain was placed. After surgery, the patients received broad-spectrum intravenous antibiotics and resumed oral intake on postoperative day 1. At 2 weeks, the Foley catheter was removed after voiding cystourethrography confirmed no extravasation. Rectal injury during robotic-assisted laparoscopic radical prostatectomy can be identified and managed intraoperatively without requiring open conversion. A three-layer closure tacked away from the vesicourethral anastomosis and reinforced by an omental flap resulted in an uneventful recovery.


[4 + 4] photodimerization of azaanthracenes in both solution and solid phase controlled by cation-π interactions.
Regio- and stereoselective [4 + 4] photodimerization reactions of 1- and 2-azaanthracenes were performed in both methanol solution and solid phases to give anti-HT dimers in high yields. In these reactions, intermolecular cation-π interactions between the pyridinium cation and the benzene ring play a key role in preorientation prior to the photodimerization reactions.


Whose consent is it anyway? A poststructuralist framing of the person in medical decision-making.
This paper explores the value of a Poststructuralist psychoanalytic model of persons, or "Subjects," as an expanded frame for the question "Whose consent is it anyway?" The elaboration of the need for this expanded frame, its tenets and its value form the substance of the paper. This frame incorporates the emotional, linguistic, and socio-cultural dimensions that help restore patients and physicians to their full status as persons from their restricted status, in the current dominant theory and model, as unidimensional, rationalistic, medico-legally constructed players; emphasizes their interconnectedness; and, focuses broadly on responsibility as bearing consequences, and not only accountability. This frame does not deny the role and importance of cognition or rationality, it supplements them. It does not supplant rationality, but rather includes it in a view of the person that also includes those other human capacities which are not based on an ideal of pure reason.


Origin of action potential recorded by fluid electrodes.
The concept that action potentials recorded by fluid electrodes may be generated at the point of the entrance to and the exit from the partition and that the action potential recorded by the fluid electrodes with a number of partitions between them may be equal to the algebraic sum of the action potentials recorded by the electrodes adjacent to each partition was directly verified in this study.