TLR2 and TLR4 in cutaneous leishmaniasis caused by Leishmania major.
Cutaneous leishmaniasis (CL) is a self-healing skin disease which rarely for unknown reason(s) the lesion develops to a non-healing form. It seems that the initial contact of Leishmania parasites with the host innate immune system is an important step in the outcome of the disease. Recent studies suggested that toll-like receptors (TLRs) play a role in Leishmania recognition. In this study, the level of TLR2 and TLR4 was checked in patients with healing form of lesion and compared with that of patients with non-healing form of lesion caused by Leishmania major. Gene expression of TLR2 and TLR4 in peripheral blood-derived macrophages, before and after stimulation with live L. major promastigotes, was evaluated using quantitative real-time reverse transcription PCR and flow cytometry. The results showed that the mean relative gene expression and difference membrane expression of TLR2 in macrophages of patients with healing form of lesion were significantly higher than patients with non-healing form of lesion (P < 0.0001 and P = 0.0034), respectively, and the mean relative gene expression and difference in protein expression of TLR4 in macrophages of patients with healing form of lesion were significantly higher than that of patients with non-healing form of lesion (P = 0.021 and P = 0.002), respectively. The data suggested a possible role for TLR2 and TLR4 in the outcome of CL lesion. Further studies are needed to understand more about the detail role of the immune factors in leishmaniasis.


Acoustic-electric interactions in the guinea pig auditory nerve: simultaneous and forward masking of the electrically evoked compound action potential.
The study investigated the time course of the effects of acoustic and electric stimulation on the electrically evoked compound action potential (ECAP). Adult guinea pigs were used in acute experimental sessions. Bursts of acoustic noise and high-rate (5000 pulses/s) electric pulse trains were used as maskers. Biphasic electric pulses were used as probes. ECAPs were recorded from the auditory nerve trunk. Simultaneous masking of the ECAP with acoustic noise featured an onset effect and a decrease in the amount of masking to a steady state. It was characterized by a two-component exponential function. The amount of masking increased with masker level and decreased with probe level. Post-stimulatory ECAP recovery often featured a non-monotonic time course, described by a three-component exponent. Electric maskers produced similar post-stimulatory effects in hearing and acutely deafened subjects. Acoustic stimulation affects the ECAP in a level- and time-dependent manner. Simultaneous masking follows a time course comparable to that of adaptation to an acoustic stimulus. Refractoriness, spontaneous activity, and adaptation are suggested to play a role in ECAP recovery. Post-stimulatory changes in synchrony, possibly due to recovery of spontaneous activity and an additional hair-cell independent mechanism, are hypothesized to contribute to the observed non-monotonicity of recovery.


The coping strategies of community pharmacists and pharmaceutical services provided during COVID-19 in Malaysia.
The coronavirus disease 2019 (COVID-19) pandemic has resulted in profound health challenges across the globe. Pharmacists' readiness to cope with the pandemic is critical in supporting and sustaining the healthcare workforce to meet the challenges. This study aims to examine community pharmacists' views on their work environment, policies and preparedness for safe retail patronage to prevent the transmission of COVID-19 and assess the variables influencing coping strategies during the pandemic. An online survey of pharmacists practising in community pharmacy setting in Malaysia. A questionnaire-based, cross-sectional study was conducted in Malaysia to evaluate coping strategies of community pharmacists and pharmaceutical services provided during COVID-19 pandemic. Between May 1 and July 31, 2020, the questionnaire was distributed to pharmacists working in community setting nationwide utilising a snowball sampling method. Community pharmacists' perceptions on safety, resilience, organisational support, and pharmaceutical services offered during COVID-19 pandemic. A total of 217 pharmacists participated in the study. The vast majority of community pharmacists reported a positive outlook and were able to balance working with self-care during this period. Most reported to have access to personal protective equipment such as gloves and hand sanitisers. A large proportion of community pharmacies also installed physical barriers in doorways or in front of the counter, put markings on the ground to section areas to ensure physical distancing, and controlled the number of customers who could access the pharmacy during this period. Innovations reported to be implemented included teleconsultations and providing curb-side or drive-through delivery and pickup services. The findings suggest the frontline and essential roles of community pharmacists in delivering pandemic responses, creating the opportunity to determine areas where community pharmacy services can be incorporated to strengthen the public health system and improve patient health outcomes.


Associations of Multimorbidity With Stroke Severity, Subtype, Premorbid Disability, and Early Mortality: Oxford Vascular Study.
Patients with multimorbidity are underrepresented in clinical trials. Inclusion in stroke trials is often limited by exclusion based on premorbid disability, concerns about worse poststroke outcomes in acute treatment trials, and a possibly increased proportion of hemorrhagic vs ischemic stroke in prevention trials. Multimorbidity is associated with an increased mortality after stroke, but it is unclear whether this is driven by an increased stroke severity or is confounded by particular stroke subtypes or premorbid disability. We aimed to determine the independent association of multimorbidity with stroke severity taking account of these main potential confounders. In a population-based incidence study (Oxford Vascular Study; 2002-2017), prestroke multimorbidity (Charlson Comorbidity Index [CCI]; unweighted/weighted) in all first-in-study strokes was related to postacute severity (≈24 hours; NIH Stroke Scale [NIHSS]), stroke subtype (hemorrhagic vs ischemic; Trial of Org 10172 in Acute Stroke Treatment [TOAST]), and premorbid disability (modified Rankin scale [mRS] score ≥2) using age-adjusted/sex-adjusted logistic and linear regression models and to 90-day mortality using Cox proportional hazard models. Among 2,492 patients (mean/SD age = 74.5/13.9 years; 1,216/48.8% male; 2,160/86.7% ischemic strokes; mean/SD NIHSS = 5.7/7.1), 1,402 (56.2%) had at least 1 CCI comorbidity, and 700 (28.1%) had multimorbidity. Although multimorbidity was strongly related to premorbid mRS ≥2 (adjusted odds ratio [aOR] per CCI comorbidity 1.42, 1.31-1.54, p < 0.001), and comorbidity burden was crudely associated with an increased severity of ischemic stroke (OR per comorbidity 1.12, 1.01-1.23 for NIHSS 5-9, p = 0.027; 1.15, 1.06-1.26 for NIHSS ≥10; p = 0.001), no association with severity remained after stratification by TOAST subtype (aOR 1.02, 0.90-1.14, p = 0.78 for NIHSS 5-9 vs 0-4; 0.99, 0.91-1.07, p = 0.75 for NIHSS ≥10 vs 0-4), or within any individual subtype. The proportion of intracerebral hemorrhage vs ischemic stroke was lower in patients with multimorbidity (aOR per comorbidity 0.80, 0.70-0.92, p < 0.001), and multimorbidity was only weakly associated with 90-day mortality after adjustment for age, sex, severity, and premorbid disability (adjusted hazard ratio per comorbidity 1.09, 1.04-1.14, p < 0.001). Results were unchanged using the weighted CCI. Multimorbidity is common in patients with stroke and is strongly related to premorbid disability but is not independently associated with an increased ischemic stroke severity. Greater inclusion of patients with multimorbidity is unlikely therefore to undermine the effectiveness of interventions in clinical trials but would increase external validity.


Low psoas muscle area is associated with postoperative complications in Crohn's disease.
Crohn's disease (CD) is associated with increased postoperative morbidity. Sarcopenia correlates with increased morbidity and mortality in various medical conditions. We assessed correlations of the lean body mass marker and psoas muscle area (PMA), with postoperative outcomes in CD patients undergoing gastrointestinal surgery. We included patients with CD who underwent gastrointestinal surgery between June 2009 and October 2018 and had CT/MRI scans within 8 weeks preoperatively. PMA was measured bilaterally on perioperative imaging. Of 121 patients, the mean age was 35.98 ± 15.07 years; 51.2% were male. The mean BMI was 21.56 ± 4 kg/m2. The mean PMA was 95.12 ± 263.2cm2. Patients with postoperative complications (N = 31, 26%) had significantly lower PMA compared with patients with a normal postoperative recovery (8.5 ± 2.26 cm2 vs. 9.85 ± 2.68 cm2, P = 0.02). A similar finding was noted comparing patients with anastomotic leaks to those without anastomotic leaks (7.48 ± 0.1 cm2 vs. 9.6 ± 2.51 cm2, P = 0.04). PMA correlated with the maximum degree of complications per patient, according to the Clavien-Dindo classification (Spearman's coefficient = -0.26, P = 0.004). Patients with major postoperative complications (Clavien-Dindo ≥ 3) had lower mean PMA (8.12 ± 2.75 cm2 vs. 9.71 ± 2.57 cm2, P = 0.03). Associations were similar when stratifying by gender and operation urgency. On multivariate analysis, PMA (HR = 0.72/cm2, P = 0.02), operation urgency (HR = 3.84, P < 0.01), and higher white blood cell count (HR = 1.14, P = 0.02) were independent predictive factors for postoperative complications. PMA is an easily measured radiographic parameter associated with postoperative complications in patients with CD undergoing bowel resection.


Enantioselectivity of alcohol-modified polymeric surfactants in micellar electrokinetic chromatography.
A novel method of modifying sodium undecanoyl-L-leucinate (SUL) micelles employed in chiral separation of analytes in micellar electrokinetic chromatography (MEKC) to enhance selectivity toward specific analytes is discussed. The current study aimed at modifying the SUL micelles by introducing different alcohols into the mono-SUL micelles. The micellar solutions were then polymerized in the presence of alcohols followed by postpolymerization extraction of the alcohols to yield alcohol-free polymeric surfactants (poly-L-SUL). The effects of hexanol (C(6)OH) and undecylenyl alcohol (C(11)OH) on micellar properties of this surfactant were investigated by use of surface tensiometry, fluorescence spectroscopy, pulsed field gradient-nuclear magnetic resonance (PFG-NMR), and MEKC. The surface tension and PFG-NMR studies indicated an increase in the critical micelle concentration (cmc) and micellar size upon increasing the alcohol concentration. Fluorescence measurements suggested that alcohols induce closely packed micellar structures. Coumarinic and benzoin derivatives, as well as (+/-)-1, 1'-binaphthyl-2,2'-dihydrogen phosphate (BNP) were used as test analytes for MEKC experiments. Examination of MEKC data showed remarkable resolutions and capacity factors of coumarinic derivatives obtained with modified poly-L-SUL as compared to the unmodified poly-L-SUL. Evaluation of fluorescence, PFG-NMR, and MEKC data suggest a strong correlation between the polarity and hydrodynamic radii of alcohol-modified micelles and the resolution of the test analytes.


Population toxicokinetic analysis of 2,3,7,8-tetrachlorodibenzo-p-dioxin using Bayesian techniques.
Understanding the kinetics of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) concentrations in humans is an important step for TCDD cancer risk assessment. In this paper longitudinal series of serum TCDD concentration measurements on U.S. veterans of the Vietnam war, who were exposed to dioxin during herbicide-spraying operations, are studied. The overall aim is to use these data to infer the dynamics of TCDD concentrations in humans. This is done by identifying a kinetic model describing the dioxin time course at the individual level. The individual toxicokinetic model is then expanded into a population model within a Bayesian hierarchical framework which allows residual variations across subjects that cannot be explained by observed covariates. Other complications in the data, such as unknown exposure histories, are also resolved implicitly through the hierarchical model. Moreover, the choice of a Bayesian approach enables the accumulation of external source of information in the form of prior distributions. The model is subjected to various diagnostic checks and analyses of sensitivity to distributional assumptions showing a good fit in terms of both the population and the kinetic features.


Use of specific mitochondrial complex inhibitors to investigate mitochondrial involvement on horse sperm motility and ROS production.
Equine spermatozoa highly rely on oxidative phosphorylation for their energy management. The present work aimed to characterize the role of mitochondria on horse sperm motility and ROS production by incubating spermatozoa with specific inhibitors of the different mitochondrial complexes. Equine spermatozoa were incubated 1 h and 3 h at 37 °C with: complex I inhibitor rotenone (5 μM, ROT), complex II inhibitor dimethyl-malonate (10 mM, DMM), complex III inhibitor antimycin A (1.8 μM, ANTI), the uncoupling agent carbonyl cyanide m-chlorophenyl hydrazine (5 μM, CCCP), ATP synthase inhibitor oligomycin (5 μM, OLIGO), and 2 μL vehicle DMSO (control, CTL). Samples were analyzed for sperm motility and for mitochondrial membrane potential (MMP), mitochondrial integrity, mitochondrial O2•- production, and cytoplasmic H2O2. A multivariate analysis was performed on the data. CCCP caused a pronounced MMP reduction at both time points while ROT and ANTI showed the same effect at 3 h. All treatments at 3 h incubation significantly reduced the percentage of sperm with early changes in membrane permeability with active mitochondria. The H2O2 production of live cells was low at 1 h incubation in all treatments; after 3 h a slight decrease in the percentage of low-H2O2 producing cells was recorded. All treatments, except DMM, induced a significant decline in sperm motility and kinematics and modified the pattern of sperm subpopulations. The effect of DMM was evident only after 3 h, increasing the percentage of slow sperm subpopulation. In conclusion, the disruption of mitochondrial integrity induces an increase of mitochondrial ROS production that could be detrimental for cell function and survivior.


miR-126 identifies a quiescent and chemo-resistant human B-ALL cell subset that correlates with minimal residual disease.
Complete elimination of B-cell acute lymphoblastic leukemia (B-ALL) by a risk-adapted primary treatment approach remains a clinical key objective, which fails in up to a third of patients. Recent evidence has implicated subpopulations of B-ALL cells with stem-like features in disease persistence. We hypothesized that microRNA-126, a core regulator of hematopoietic and leukemic stem cells, may resolve intratumor heterogeneity in B-ALL and uncover therapy-resistant subpopulations. We exploited patient-derived xenograft (PDX) models with B-ALL cells transduced with a miR-126 reporter allowing the prospective isolation of miR-126(high) cells for their functional and transcriptional characterization. Discrete miR-126(high) populations, often characterized by MIR126 locus demethylation, were identified in 8/9 PDX models and showed increased repopulation potential, in vivo chemotherapy resistance and hallmarks of quiescence, inflammation and stress-response pathway activation. Cells with a miR-126(high) transcriptional profile were identified as distinct disease subpopulations by single-cell RNA sequencing in diagnosis samples from adult and pediatric B-ALL. Expression of miR-126 and locus methylation were tested in several pediatric and adult B-ALL cohorts, which received standardized treatment. High microRNA-126 levels and locus demethylation at diagnosis associate with suboptimal response to induction chemotherapy (MRD > 0.05% at day +33 or MRD+ at day +78).


Metformin and β-cell function in insulin-treated patients with type 2 diabetes: A randomized placebo-controlled 4.3-year trial.
In this trial, 390 insulin-treated patients with type 2 diabetes were randomized to either placebo or metformin. Fasting levels of glucose, insulin and C peptide were determined at baseline, after 4 months and yearly thereafter for 4 years to assess fasting estimates of beta cell function. The primary endpoint was the fasting C peptide-to-glucose ratio (FCPGR) and secondary measures were the disposition index (DI) and the fasting C peptide (FCP). We analysed the results with a general linear mixed model. Baseline FCPGR was 5.27 (95% CI, 4.83 - 5.71). Compared to placebo, FCPGR increased in the metformin group with 1.48 (95% CI, 1.09 - 1.87, P < 0.001). The DI showed comparable results with a treatment effect of 1.50 (95% CI, 1.17 - 1.83; P < 0.001). FCP also increased in the metformin group but did not reach statistical significance vs placebo (0.034 nmol, 95% CI, -0.005 - 0.072; P = 0.085). Treatment with metformin vs placebo, added to insulin in patients with type 2 diabetes, improves long-term estimates of beta cell function in the fasting state.


Sagittal Alignment Outcomes in Lordotic Cervical Spine: Does Three-Level Anterior Cervical Discectomy and Fusion Outperform Laminoplasty?
A prospective cohort study. To compare the radiological outcomes between three-level anterior cervical discectomy and fusion (ACDF) and plate-only open-door laminoplasty (LAMP) in patients with lordotic cervical spine. Both three-level ACDF and LAMP are important surgical methods for multilevel cervical spondylotic myelopathy, but the cervical sagittal alignment outcomes and the lordosis preserving abilities between the two specific approaches have not been carefully compared. Sixty patients, all of whom had lordotic cervical spines and underwent three-level ACDF (n = 22) or LAMP (n = 38) for the treatment of multilevel cervical spondylotic myelopathy, were prospectively studied. Upright neutral cervical lateral radiographs were assessed preoperatively, at 3 days after surgery, and at the last follow-up (1.5 years). The primary radiological outcomes are C2-7 Cobb angle, and the secondary outcomes include C2-7 sagittal vertical axis , disc height, Harrison angle, and Ishihara index. The relationship between preoperative parameters and follow-up outcomes were assessed. Preoperative cervical sagittal alignment parameters were similar between the two groups. Patients in the ACDF group obtained larger C2-7 Cobb angles than those the LAMP group early after surgery. However, the lordosis in the ACDF group decreased significantly during follow-up, leading to similar outcomes between the two groups. The lordosis-preserving ability of ACDF is significantly poorer than that of LAMP. Secondary radiological parameters also showed similar trends. In both groups, the preoperated Ishihara index was significantly related to follow-up C2-7 Cobb angles. For patients with Ishihara index less than 20, ACDF is more likely to produce a larger C2-7 Cobb angled at the time of follow-up. In patients with lordotic cervical spine, the sagittal alignment outcomes of the two approaches were similar, and the lordosis-preserving ability was poorer in ACDF. Further research should focus on the factors related to the lordosis-preserving ability of multilevel ACDF. 3.


TSA restores hair follicle-inductive capacity of skin-derived precursors.
The genesis of the hair follicle relies on signals derived from mesenchymal cells in the dermis during skin morphogenesis and regeneration. Multipotent skin-derived precursors (SKPs), which exhibit long term proliferation potential when being cultured in spheroids, have been shown to induce hair genesis and hair follicle regeneration in mice, implying a therapeutic potential of SKPs in hair follicle regeneration and bioengineering. However, the hair-inductive property of SKPs declines progressively upon ex vivo culture expansion, suggesting that the expressions of the genes responsible for hair induction are epigenetically unstable. In this study, we found that TSA markedly alleviated culture expansion induced SKP senescence, increased the expression and activity of alkaline phosphatase (AP) in the cells and importantly restored the hair inductive capacity of SKPs. TSA increased the acetylation level of histone H3, including the K19/14 sites in the promoter regions of bone morphogenetic proteins (BMPs) genes, which were associated with elevated gene expression and BMP signaling activity, suggesting a potential attribution of BMP pathway in TSA induced recovery of the hair inductive capacity of SKPs.


Observation on the schizont stage of an unidentified Theileria sp. in experimentally infected sheep.
The schizont stage of an unidentified Theileria sp. infective for small ruminants was observed. Intact sheep were infected with adult Haemaphysalis qinghaiensis collected from fields where ovine theileriosis was prevalent. When the infested sheep developed theileriosis, tissue and organ smears were prepared. Theileria schizonts were demonstrated in liver, spleen, lung, kidney, lymph node and peripheral blood. Most of the schizonts were found outside the host cells, which was probably a smear artifact. This study adds our knowledge that lung and kidney can be parasitized by schizonts of Theileria sp.


Analysis of in vitro ADCC and clinical response to trastuzumab: possible relevance of FcγRIIIA/FcγRIIA gene polymorphisms and HER-2 expression levels on breast cancer cell lines.
Trastuzumab is a humanized monoclonal antibody (mAb) currently used for the treatment of breast cancer (BC) patients with HER-2 overexpressing tumor subtype. Previous data reported the involvement of FcγRIIIA/IIA gene polymorphisms and/or antibody-dependent cellular cytotoxicity (ADCC) in the therapeutic efficacy of trastuzumab, although results on these issues are still controversial. This study was aimed to evaluate in vitro the functional relationships among FcγRIIIA/IIA polymorphisms, ADCC intensity and HER-2 expression on tumor target cells and to correlate them with response to trastuzumab. Twenty-five patients with HER-2 overexpressing BC, receiving trastuzumab in a neoadjuvant (NEO) or metastatic (MTS) setting, were genotyped for the FcγRIIIA 158V>F and FcγRIIA 131H>R polymorphisms by a newly developed pyrosequencing assay and by multiplex Tetra-primer-ARMS PCR, respectively. Trastuzumab-mediated ADCC of patients' peripheral blood mononuclear cells (PBMCs) was evaluated prior to therapy and measured by (51)Chromium release using as targets three human BC cell lines showing different levels of reactivity with trastuzumab. We found that the FcγRIIIA 158F and/or the FcγRIIA 131R variants, commonly reported as unfavorable in BC, may actually behave as ADCC favorable genotypes, in both the NEO (P ranging from 0.009 to 0.039 and from 0.007 to 0.047, respectively) and MTS (P ranging from 0.009 to 0.032 and P = 0.034, respectively) patients. The ADCC intensity was affected by different levels of trastuzumab reactivity with BC target cells. In this context, the MCF-7 cell line, showing the lowest reactivity with trastuzumab, resulted the most suitable cell line for evaluating ADCC and response to trastuzumab. Indeed, we found a statistically significant correlation between an increased frequency of patients showing ADCC of MCF-7 and complete response to trastuzumab in the NEO setting (P = 0.006). Although this study was performed in a limited number of patients, it would indicate a correlation of FcγR gene polymorphisms to the ADCC extent in combination with the HER-2 expression levels on tumor target cells in BC patients. However, to confirm our findings further experimental evidences obtained from a larger cohort of BC patients are mandatory.


Determinants of infant feeding practices among mothers living with HIV attending prevention of mother to child transmission Clinic at Kiambu Level 4 hospital, Kenya: a cross-sectional study.
Human immunodeficiency virus infection and acquired immune deficiency syndrome is global pandemic with around 150,000 children infected with HIV in 2015. In Kenya, it is estimated that 11,000 children who are under 15 years old were infected. Safe infant feeding practices are the major important determinants of the prevention of mother to child transmission. The decision to breastfeed or not is considered a very tough choice for mothers living with HIV. This study assessed the infant feeding practices and its determinants among mothers living with HIV with infants 0-12 months old. This was a mixed methods cross-sectional study adopting qualitative and quantitative data collection procedures. A sample of 180 systematically selected mothers living with HIV with infants 0-12 months old attending prevention of mother to child transmission clinic participated in the study. Exclusive breastfeeding rate was 71.4%, mixed feeding (18.2%) and replacement feeding (10.4%). Complementary feeding with continued breastfeeding rate was 63.1%. Similarly, qualitative results showed that exclusive breastfeeding was the most preferred feeding method by mothers living with HIV. Age (Adjusted Odds Ratio (AOR) 0.19; (95% Confidence Interval (CI) 0.41, 0.85; p = 0.030) and infant feeding practice knowledge (AOR 0.20; 95% CI 0.06, 0.64; p = 0.007) were determinants of exclusive breastfeeding. Education AOR 0.17; 95% CI 0.03, 0.85; p = 0.002) and occupation (AOR 3.91; 95% CI 1.24, 12.32; p = 0.020) were determinants of complementary feeding with continued breastfeeding. Exclusive breastfeeding is attainable in this population. However, poor infant feeding practice knowledge led to non-adherence to safe infant feeding practices such as exclusive breastfeeding. Socio-demographic factors such as age, education and occupation were established as determinants of infant feeding practices among mothers living with HIV. Ministry of Health should come up with strategies on infant feeding counseling that are aligned to a local context, to allow mothers to understand the importance of recommended infant feeding options for HIV-exposed infants.


The role of heparin-binding EGF-like growth factor and amphiregulin in the epidermal proliferation of psoriasis in cooperation with TNFalpha.
Heparin-binding EGF-like growth factor (HB-EGF) and amphiregulin (AREG) are the members of EGF family that bind to common EGF receptor (EGFR) in the epidermis. However, the role of these two growth factors in epidermal hyperplasia of psoriasis has not been established. On the other hand, CD4+ T cells are responsible for the development of the psoriatic plaques. However, inflammatory cytokines, such as TNFalpha, IL-1beta and IFNgamma, inhibit the growth of human keratinocytes in vitro. The expression of HB-EGF, AREG and EGFR proteins in normal (n = 22) and psoriatic (n = 34) skin tissues was examined by immunohistochemistry. Then, the effects of HB-EGF and AREG on the growth of cultured adult normal human epidermal keratinocytes (NHEK-AD) with or without TH1 cytokines, such as TNFalpha, IL-1beta and IFNgamma, were examined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, and the effects of these cytokines on the expression of EGFR mRNA in NHEK-AD were examined by real-time reverse transcriptase-polymerase chain reaction. The expression of HB-EGF and AREG in the epidermis was not specific to psoriatic plaques, but the distribution of positive cells throughout the epidermis was different between normal skins and psoriatic plaques. On the other hand, in the dermis and the papillary dermis, most of vascular endothelial cells and infiltrating mononuclear cells expressed both HB-EGF and AREG in normal skins and psoriatic plaques, and these positive cells were more frequent in psoriasis compared to normal skin. In the in vitro growth assay, HB-EGF, not AREG, stimulated the proliferation of NHEK-AD at the optimal concentration of 1 ng/ml. Furthermore, HB-EGF compensated the growth-suppressing effects of TNFalpha, IL-1beta and IFNgamma on NHEK-AD, and TNFalpha promoted the growth of NHEK-AD at the concentration of 2 and 20 U/ml in combination with HB-EGF and, in lesser extent, with AREG. However, TNFalpha did not affect the expression of EGFR mRNA in NHEK-AD. Growth factors and inflammatory cytokines produced in the dermis would be important for the epidermal proliferation in psoriatic plaques and TNFalpha may play a key role in cooperation with HB-EGF and AREG in the proliferation of epidermal keratinocytes at the psoriatic skin lesions.


Local radiotherapy and E7 RNA-LPX vaccination show enhanced therapeutic efficacy in preclinical models of HPV16+ cancer.
Human papilloma virus (HPV) infection is a causative agent for several cancers types (genital, anal and head and neck region). The HPV E6 and E7 proteins are oncogenic drivers and thus are ideal candidates for therapeutic vaccination. We recently reported that a novel ribonucleic acid lipoplex (RNA-LPX)-based HPV16 vaccine, E7 RNA-LPX, mediates regression of mouse HPV16+ tumors and establishes protective T cell memory. An HPV16 E6/E7 RNA-LPX vaccine is currently being investigated in two phase I and II clinical trials in various HPV-driven cancer types; however, it remains a high unmet medical need for treatments for patients with radiosensitive HPV16+ tumors. Therefore, we set out to investigate the therapeutic efficacy of E7 RNA-LPX vaccine combined with standard-of-care local radiotherapy (LRT). We demonstrate that E7 RNA-LPX synergizes with LRT in HPV16+ mouse tumors, with potent therapeutic effects exceeding those of either monotherapy. Mode of action studies revealed that the E7 RNA-LPX vaccine induced high numbers of intratumoral-E7-specific CD8+ T cells, rendering cold tumors immunologically hot, whereas LRT primarily acted as a cytotoxic therapy, reducing tumor mass and intratumor hypoxia by predisposing tumor cells to antigen-specific T cell-mediated killing. Overall, LRT enhanced the effector function of E7 RNA-LPX-primed T cell responses. The therapeutic synergy was dependent on total radiation dose, rather than radiation dose-fractionation. Together, these results show that LRT synergizes with E7 RNA-LPX and enhances its anti-tumor activity against HPV16+ cancer models. This work paves into a new translational therapy for HPV16+ cancer patients.


Weak inhibitors protect cholinesterases from strong inhibitors (paraoxon): in vitro effect of tiapride.
Weak and reversible inhibitors of cholinesterases, when administered before potent organophosphorus inhibitors (pretreatment), have the ability, to a certain extent, to protect enzymes from inhibition. Such a protective effect was demonstrated in vitro for metoclopramide and ranitidine. The putative mode of protective action of these substances is, when administered in excess, competition for the active site of the enzyme with the more potent organophosphate. The present paper presents results using another benzamide with weak cholinesterase inhibitory properties: tiapride (TIA). The purpose of the study was to quantify in vitro the extent that TIA conferred protection, using paraoxon (POX) as an inhibitor, and to compare the results with existing data obtained using TIA as a protective agent against dichlorvos (DDVP). POX is a highly toxic non-neuropathic organophosphate. While the use of parathion (the inactive prodrug which is metabolically converted to POX) has been restricted in most countries, the organophosphate is still responsible for a large number of accidental or suicidal exposures. DDVP is a moderately toxic, non-neuropathic organophosphate. Red blood cell (RBC) acetylcholinesterase (AChE) activities in whole blood and butyrylcholinesterase (BChE) activities in human plasma were measured photometrically in the presence of different POX and TIA concentrations and the IC(50) was calculated. Determinations were repeated in the presence of increasing TIA concentrations. The IC(50) of POX increases with the TIA concentration in a linear manner. The protective effect of tiapride on cholinesterase could be of practical relevance in the pretreatment of organophosphate poisoning. It is concluded that in vivo testing of TIA as an organophosphate protective agent is warranted.


Platelet-derived growth factor from astrocytes drives the clock that times oligodendrocyte development in culture.
The various cell types in a multicellular animal differentiate on a predictable schedule but the mechanisms responsible for timing cell differentiation are largely unknown. We have studied a population of bipotential glial (O-2A) progenitor cells in the developing rat optic nerve that gives rise to oligodendrocytes beginning at birth and to type-2 astrocytes beginning in the second postnatal week. Whereas, in vivo, these O-2A progenitor cells proliferate and give rise to postimitotic oligodendrocytes over several weeks, in serum-free (or low-serum) culture they stop dividing prematurely and differentiate into oligodendrocytes within two or three days. The normal timing of oligodendrocyte development can be restored if embryonic optic-nerve cells are cultured in medium conditioned by type-1 astrocytes, the first glial cells to differentiate in the nerve: in this case the progenitor cells continue to proliferate, the first oligodendrocytes appear on the equivalent of the day of birth, and new oligodendrocytes continue to develop over several weeks, just as in vivo. Here we show that platelet-derived growth factor (PDGF) can replace type-1-astrocyte-conditioned medium in restoring the normal timing of oligodendrocyte differentiation in vitro and that anti-PDGF antibodies inhibit this property of the appropriately conditioned medium. We also show that PDGF is present in the developing optic nerve. These findings suggest that type-1-astrocyte-derived PDGF drives the clock that times oligodendrocyte development.


Surveillance of multiple subtype specific antibodies against avian influenza viruses among egg yolk in wild ducks from northeast China, 2017-2019.
Over the past decade, various avian influenza viruses have been isolated from wild ducks found in the northeast of China. To monitor the prevalence of multiple subtype specific AIVs antibodies, 1705 wild ducks' eggs from six wetlands of northeast China were analyzed for surveillance of H1, H3, H5, and H7 AIVs antibodies by c-ELISA and HI test from Jan 2017 to Dec 2019. The results show that the combined frequency of multiple subtype specific AIVs antibodies were H1 (12.32%), H3 (8.15%), H5 (2.05%), and H7 (3.46%) respectively. Therefore, a detailed analysis of the geographical distribution of AIVs in China, and the risk factors for human infection is of vital importance. This study provides basic data for other researchers to deeply study AIVs distribution characteristics, and for governments to develop detailed measures to control the spread of AIVs.


Canine angiostrongylosis in Italy: occurrence of Angiostrongylus vasorum in dogs with compatible clinical pictures.
This study evaluated the perception of veterinarians in including the emerging metastrongyloid Angiostrongylus vasorum in the differential diagnosis for dogs referred with appropriate signs. Veterinarians at teaching hospitals, private practices or kennels collected faecal samples from 606 symptomatic dogs in six different areas of Italy. The samples were examined with the Baermann technique. Fourteen dogs scored positive for A. vasorum, all showing different clinical pictures, with the most common reason for inclusion in the study being a combination of respiratory signs along with general signs. The most reported sign was cough, followed by aspecific distresses. Other respiratory signs were dyspnoea, tachypnoea and breathing sounds, while bleeding diathesis occurred in one dog. Gastrointestinal disorders were diarrhoea and vomitus, while no neurological signs were detected. Four key cases are herein described, with a special focus on their clinical description, findings at the collateral diagnostic imaging and therapeutic options with moxidectin. This survey provides new data on distribution of A. vasorum in Europe and on the clinical impact of the disease. The results confirm that clinical angiostrongylosis overlaps a series of other conditions of dogs. Awareness among vet practitioners is discussed along with the importance of including A. vasorum on the list of differential diagnoses in canine clinical practice and of performing appropriate diagnostic methods.


S1 kills MCF-7/ADR cells more than MCF-7 cells: A protective mechanism of endoplasmic reticulum stress.
Drug resistance in chemotherapy for breast cancer is associated with high levels of P-glycoprotein (P-gp) as well as endoplasmic reticulum (ER) stress. In this paper, we aimed to evaluate the efficacy of a pan-BH3 mimetic S1 on drug-resistant MCF-7/ADR cells, and the roles of S1-induced ER stress in cell death. S1 exhibited greater and faster mitochondrial apoptosis in MCF-7/ADR cells than in MCF-7 cells. We demonstrated by Bax/Bak activation and cyrochrome c release that the p-glycprotein had little influence on S1 entering cells and hitting its targets in MCF-7/ADR cells. An IRE1-mediated ER stress response followed by c-Jun N-terminal kinase (JNK) and extracellular signal-regulated protein kinase (ERK) activation was specifically induced by S1 in MCF-7 cells, but not in MCF-7/ADR cells. Coimmunoprecipitation and western blotting analysis determined that ER stress played a protective role in S1-induced apoptosis by triggering Bcl-2 phosphorylation, which grabbed more pro-apoptotic proteins. The synergism effect of ERK inhibitor PD98059 with S1 confirmed the protective role of ER stress. Defective ER stress in MCF-7/ADR cells confers the more sensitivity toward S1.


Long-term progression-free survival of hepatocellular carcinoma with synchronous diffuse peritoneal metastasis treated by CRS+HIPEC: A case report and literature review.
Hepatocellular carcinoma (HCC) with peritoneal metastasis (PM) is rare. There has been no standard treatment for this severe disease, and the conventional palliative therapy could only reach an overall survival of 6 to 14 months PATIENT CONCERNS:: A 38-year-old male with a chief complain of "abdominal distension and diagnosis of HCCPM for 3 months", was suffering from severe diarrhea and moderate anemia. Diagnostic laparoscopic exploration with biopsy and the following pathology confirmed the diagnosis of HCC with PM. The patient was treated with cytoreductive surgery (CRS) plus hyperthermic intraperitoneal chemotherapy (HIPEC) followed by 6 cycles of intraperitoneal chemotherapy and 6 cycles of intravenous chemotherapy. Till January 15, 2019, the patient has progression-free survival for over 22 months. CRS plus HIPEC combined adjuvant intraperitoneal and intravenous chemotherapy may improve progression-free survival for some HCC with PM patients and should be considered as an option for such patients.


Accumulation of organotin compounds in the deep-sea environment of Nankai Trough, Japan.
The concentration of butyltin (BT) and phenyltin (PT) compounds was measured in sediment, gastropods (Colliloconcha nankaiensis), sea cucumbers (Psychropotes verrucosa), galatheid crabs (Munidopsis albatrossae and Munidopsis subsquamosa), and bivalves (Clyptogena tsubasa and Clyptogena nautilei) collected from the Nankai Trough (water depth about 3000 m). Sediment at depths of up to 1 cm was taken by a core sampler and at depths of up to 15 cm was taken by rake. The concentration of BTs in sediment core from a depth of 0-1 cm (0.041 mg kg(-1) dry) was higher than in sediment core from 0-15 cm (0.021 mg kg(-1) dry). The relative proportion of different BTs in the 0-1-cm sediment core was similar to that in the 0-15-cm sediment core, but the concentration of PTs in the former (0.028 mg kg(-1) dry) was lower than in the latter 0-15 cm (0.052 mg kg(-1) dry). Organotin (OT) compounds were also detected in deep-sea organisms. The means of BT concentrations in C. nankaiensis, P. verrucosa, M. albatrossae, M. subsquamosa, Cl. tsubasa, and Cl. nautilei were, respectively, 0.089, 0.057, 0.018, 0.016, 0.019, and 0.026 mg kg(-1). The corresponding concentrations of PTs were 0.212, 0.363, 0.166, 0.186, 0.030, and 0.025 mg kg(-1). High concentrations of BTs and PTs were observed in gastropods and sea cucumbers. The species of deep-sea organism can be classified by delta(13)C value into two groups (A and B). The organisms in group A use organic matter chemosynthesized by symbiotic bacteria while those in group B depend on photosynthesis carried out near the surface by phytoplankton. No difference in BT or TBT concentration is observed between the two groups, but PT and TPT concentrations are higher in group B. Trophic levels in the food chain are often estimated using delta(15)N values. Group B showed a higher trophic level than group A. Although no change in BT and TBT concentration was observed to accompany increases of delta(15)N values, PT and TPT concentrations generally increased with increasing delta(15)N values. The compositions of BTs in deep-sea organisms were calculated. An increasing proportion of MBT and a declining proportion of DBT were observed at higher trophic levels. No correlation between the shell length of Cl. nautilei and BT or PT concentration was observed. The average partition coefficients of TBT for C. nankaiensis, P. verrucosa, M. albatrossae, M. subsquamosa, Cl. Tsubasa, and Cl. nautilei were 2.6, 0.72, 0.63, 0.19, 0.43, 0.30, and 0.46.


Prognostic value of different scoring models in patients with multiple organ dysfunction syndrome associated with acute COPD exacerbation.
Chronic obstructive pulmonary disease (COPD) represents an increasing healthcare concern as a leading cause of morbidity and mortality worldwide. Our objective was to predict the outcome of COPD patients associated with multiple organ dysfunction syndrome (MODS) by scoring models. A retrospective study was performed on severe COPD patients within 24 hours of the onset of MODS. The Acute Physiology and Chronic Health Evaluation (APACHE) II, APACHE III, Multiple Organ Dysfunction Score (MODS), Simplified Acute Physiology Score II (SAPS II), and Sepsis-related Organ Failure Assessment (SOFA) scores were calculated for patients. A total of 153 elderly patients were recruited. Compared to 30-day survivors, the number of failing organs and all of the scoring models were significantly higher in 30-day non-survivors. The SOFA showed the highest sensitivity and area under the curve (AUC) for predicting the prognosis of patients with MODS induced by acute exacerbation of COPD. The results of logistic regression indicated that factors that were correlated with the prognosis of COPD included the exacerbation history, SOFA score, number of failing organs, and duration of ICU stay. The value of exacerbation frequency for predicting the outcome of COPD was excellent (AUC: 0.892), with a sensitivity of 0.851 and a specificity of 0.797. The SOFA score, determined at the onset of MODS in elderly patients with COPD, was a reliable predictor of the prognosis. The exacerbation frequency, number of failing organs, and the SOFA score were risk factors of a poor prognosis, and the exacerbation frequency could also effectively predict the outcome of COPD.


Absence of a protective effect of corticosterone on 0-0-diisopropyl phosphorofluoridate (DFP) induced delayed neurotoxicity in chickens.
Chickens given 15 to 45 ppm dietary corticosterone 1 day prior to and 5 days following subcutaneous administration of 1 mg/kg 0,0-diisopropyl phosphorofluoridate (DFP) were not protected from either clinical delayed neuropathy, depression of neurotoxic esterase activity or advanced degenerative peripheral nerve myelinated fiber damage.


Mitigating the toxic effects of CdSe quantum dots towards freshwater alga Scenedesmus obliquus: Role of eco-corona.
The extensive use of semiconducting nanoparticles such as quantum dots in biomedical and industrial products can lead to their inadvertent release into the freshwater system. Natural exudates in the aquatic system comprising extracellular polymeric substance (EPS) and protein-rich metabolites can eventually adsorb onto the quantum dots (QDs) surface and form an eco-corona. The alterations in the physio-chemical and toxicological behavior of CdSe/ZnS QDs under the influence of eco-corona in the freshwater system have not been explored yet. In the present study, lake water medium conditioned with exudate secreted by Scenedesmus obliquus was utilized as an eco-corona forming matrix. The time-based evolution of the eco-corona on the differently charged CdSe/ZnS QDs was analyzed using transmission electron microscopy and dynamic light scattering. Aging of amine-QDs in algal exudate for 72 h showed enhanced aggregation (Mean Hydrodynamic Diameter- 1969 nm) as compared to carboxyl-QDs (1543 nm). Further, eco-coronation tends to impart an overall negative charge to the QDs. The fluorescence intensity of amine-QDs was quenched by 84% due to the accumulation of higher eco-corona. An integrative effect of surface charge and accumulated eco-corona layer influenced the Cd2+ ion leaching from the QDs. An enhancement in the algal cell viability treated with carboxyl - CdSe/ZnS (90%) and amine- CdSe/ZnS QDs (94%) aged for 72 h suggested that eco-corona can effectively mitigate the inherent toxicity of the QDs. The oxidative stress markers in the algal cells (LPO, SOD, and CAT) were in correlation with the cytotoxicity results. The algal photosynthetic efficiency depended on the deposition of eco-coronated QDs on the cell surface. Cellular uptake results indicated low Cd2+ concentration of nearly 13.9 and 11.5% for carboxyl- and amine- CdSe/ZnS QDs respectively. This suggests that eco-coronation directly influences the bioavailability of engineered nanoparticles.


Design of a fluorogenic PNA probe capable of simultaneous recognition of 3'-overhang and double-stranded sequences of small interfering RNAs.
We developed a new fluorescent peptide nucleic acid (PNA) probe, COT probe, capable of simultaneous recognition of 3'-overhang and double stranded sequences of target small interfering RNA (siRNA).


Incorporation of hydroxyproline in bacterial collagen from Streptococcus pyogenes.
Bacterial collagen-like proteins differ from vertebrate collagens in that they do not contain hydroxyproline, which is seen as a characteristic of the vertebrate collagens, and which provides a significant contribution to the stability of the collagen triple-helix at body temperature. Despite this difference, the bacterial collagens are stable at around body temperature through inclusion of other stabilising sequence elements. Another difference is the lack of aggregation, and certain vertebrate collagen binding domains that can be introduced into the bacterial sequence lack full function when hydroxyproline is absent. In the present study we have demonstrated that a simple method utilising co-translational incorporation during fermentation can be used to incorporate hydroxyproline into the recombinant bacterial collagen. The presence and amount of hydroxyproline incorporation was shown by amino acid analysis and by mass spectrometry. A small increase in thermal stability was observed using circular dichroism spectroscopy. STATEMENT OF SIGNIFICANCE: Recombinant bacterial collagens provide a new opportunity for biomedical materials as they are readily produced in large quantity in E. coli. Unlike animal collagens, they are stable without the need for inclusion of a secondary modification system for hydroxyproline incorporation. In animal collagens, however, introduction of hydroxyproline is essential for stability and is also important for functional molecular interactions within the mammalian extracellular matrix. The present study has shown that hydroxyproline can be readily introduced into recombinant S. pyogenes bacterial collagen through direct co-translational incorporation of this modified imino acid during expression using the codons for proline in the introduced gene construct. This hydroxylation further improves the stability of the collagen and is available to enhance any introduced molecular functions.


[Association between telomere length and cognitive impairment in older adults].
Cognitive impairment is a transition stage between normal aging and dementia, the prevalence of last one increases with age; the damage of the functions and physical integrity, places the older adult in a greater susceptibility to get sick. Telomere length is a hallmark of aging to characterize this phenotype, as well as a biomarker that reflects the underlying state of the cell. In this work, the relative length of telomeres in older adults with cognitive impairment was correlated. Observational-analytical study, in samples of adult patients older than 65 years with and without cognitive impairment, in whom the relative length of telomeres was measured. Ninety samples of older adults were included in the study and in the association analysis according to multivariate logistic models, cognitive impairment showed almost five times more risk for telomere shortening in relation to the presence of the diagnosis of cognitive impairment (Odds ratio 4.88, p=0.027). When correlating the relative length of telomeres in older adults diagnosed with cognitive impairment, this association was confirmed for shorter.


Clinical efficacy of novel unidirectional buccoadhesive vs. vaginoadhesive bromocriptine mesylate discs for treating pathologic hyperprolactinemia.
To test the clinical effectiveness of new bioadhesive unidirectional buccal and vaginal bromocriptine methylate discs in hyperprolactinemic patients. A preliminary randomized comparative study. A pharmaceutical phase at the departments of Pharmaceutics, Faculties of Pharmacy, Assiut and El-Minea universities and a clinical phase at the Infertility Out-patient Clinic of Women's Health University Center, Assiut University, Assiut, Egypt. A total of 42 patients with pathologic hyperprolactinemia. Patients were randomly divided into two groups. Group A comprised 21 patients who used unidirectional buccoadhesive bromocriptine methylate discs once daily for 1 month. Group B included 21 patients who used vaginoadhesive bromocriptine methylate discs once daily for 1 month. Serum prolactin (PRL) was measured before and after therapy in all cases. Decline of serum PRL level after 1 month of therapy. Pharmaceutically, tests for swelling, surface pH, in vitro and in vivo bioadhesion and in vitro release expressed satisfactory results. The in vitro release of vaginal bromocriptine from the discs is increased in pH 4.5 media. Both groups showed a highly statistically significant reduction of serum PRL levels after 1 month of therapy without any significant difference between both groups. The decline of serum PRL was not correlated with age, parity, or indication of entering into this study. Both buccoadhesive and vaginoadhesive discs containing bromocriptine are of equal efficacy for treating pathologic hyperprolactinemia. Buccoadhesive discs have the advantages of being gender nonspecific (i.e., could be used by men), avoiding manipulating the vagina, which could be inconvenient to some patients, such as virgins; not being dependent on cyclic estrogen (E) levels; and could be easily used during menstruation.


Effects of gamma irradiation on osteoinduction associated with demineralized bone matrix.
Gamma irradiation is frequently used to sterilize implanted devices but has limitations when used on biologically active materials and composites. In this study, we have evaluated the changes of biological activity of demineralized bone matrix (DBM) in the dry state and in the presence of aqueous and non-aqueous carriers while exposed to various levels of ionizing radiation. The activity of DBM in the dry state remains relatively stable with only a small loss of activity. Composites of DBM with a carrier such as lecithin, to which no water has been added, lose activity at approximately the same rate as DBM in the anhydrous form. In composites that contain water, the loss of activity occurs even at much lower levels of radiation exposure. Gamma irradiation does not change cell attachment to the DBM matrix but has an influence on both stem cell and osteoprecursor cell proliferation rates. Because of the limitations imposed by radiation, it seems most practical to handle DBM aseptically throughout the procedures of compositing pastes, putties, or suspensions, and only if necessary exposing the inert excipients to radiation sterilization prior to mixing.


Assessing the fertility of two mares cloned from the same founder animal.
Two cloned mares, produced from the same sample of skin fibroblasts, were bred during four breeding seasons from their second year of age, as embryo donors, in exactly the same conditions, using the same stallions for both cloned mares. The aim of this study was to test the embryo donor potential of cloned mares and to compare the results obtained from two cloned mares of the same mare with other embryo donor mares (n = 31-39 per breeding season) at the same stud. For both cloned mares, 19 embryos were recovered by 43 collection attempts (44%) (7/22 for one; 12/21 for the other), 16 (84%) pregnancies (5/7 for one, 11/12 for the other) were obtained at day 14 post-ovulation (D14 ), and 12 (3/7 for one; 9/12 for the other) foals were born. One cloned mare was a less efficient donor mare than the other (p < .05), In control donor mares, 623 embryo collections were performed, with a recovery rate (80%-496/623) significantly higher than for cloned mares. The recovery rate in the subpopulation of 2-5-year-old control donor mares (same age of cloned mares) (89%-127/143) and The recovery rate in the subpopulation of 12 control mares bred with the seven same stallions as clones (55%-17/31), were both higher than for cloned mare (p < .05). The success rate of transfer was not different between embryos produced by cloned mares (84%-16/19) and those produced by control donor mares (79%-392/496). However, the foaling rate per embryo collection was significantly lower for cloned mares (28%-12/43) than for control donor mares (52% - 325/623) (p < .05).


The prognostic role of loss of insulin-like growth factor-binding protein-3 expression in head and neck carcinogenesis.
The activities of insulin-like growth factors (IGFs) in regulating cell proliferation, differentiation, and apoptosis are modulated by a family of high-affinity specific IGF-binding proteins (IGFBPs), IGFBP-3 being the most abundant in circulation. Down-regulation of IGFBP-3 has been shown to be associated with a shorter disease-specific survival probability in early non-small-cell lung cancer (NSCLC). We examined the prognostic role of IGFBP-3 protein expression loss in 34 squamous cell carcinomas (SCC) of the tongue (stages II-IV) and 30 premalignant lesions of the oral cavity and the larynx. Reduced IGFBP-3 expression was found in 11 (32%) of 34 tongue SCC cases, and was associated with significantly shorter disease-specific and disease-free survival (P=0.0002 and <0.0001 by Log-rank test). In premalignant lesions, IGFBP3 loss was found in 8 (27%) of 30 cases without significant association with cancer-free survival. Therefore, down-regulation of IGFBP-3 is an early event during head and neck carcinogenesis, that bears an adverse prognostic significance in tongue cancer and could serve as both a marker of aggressive disease and target for intervention.


[Analysis of the Neuraminidase Amino Acid Sequences of Influenza A/H1N1pdm09, A/H3N2, and B Viruses Isolated from Influenza Patients in the 2013/14 Japanese Influenza Season].
Neuraminidase (NA) is an essential surface protein for influenza virus replication. NA inhibitors are commonly used for the treatment of influenza patients in Japan. Several mutations that reduce the effect of NA inhibitors have been reported. We sequenced the whole NA segment of isolated virus from influenza patients and investigated the relation between the NA amino acid sequence and the 50% inhibitory concentration (IC50) of four NA inhibitors. A total of 20 viruses that showed high or low IC50 of NA inhibitors were selected from A/H1N1pdm09, A/H3N2, and B isolates from the viruses isolated from patients in the 2013-14 influenza season. Viral RNA was extracted and RT-PCR was done. The amplified genome was sequenced using a next generation sequencer", and the deduced amino acid sequences were analyzed. Two A/H1N1pdm09 viruses that showed very high IC50 for oseltamivir (150 nM and 130 nM) contained the H275Y mutation. Otherwise, no significant relation was found between the NA amino acids and the IC50 of the four NA inhibitors. There was no significant relation between the NA amino acids and the IC50 of the four NA inhibitors for A/H3N2 viruses. The B viruses that showed a high IC50 for oseltamivir and laninamivir shared some amino acids. The B viruses that showed a high IC50 of zanamivir and peramivir also shared some amino acids. They were different from the shared amino acids found for oseltamivir and laninamivir. The previously reported H275Y mutation that causes oseltamivir resistance was found in the two A/H1N1pdm09 viruses that showed a very high IC50 for oseltamivir. No additional NA amino acid sequences related to the IC50 of the four NA inhibitors was found. The meaning of the shared amino acids among B viruses that showed a high IC50 would be an interesting target for further investigation.


Cybernetic model of psychophysiologic pathways: III. Clinical impairment of tension and kinesthesia.
It is unclear whether peripheral, subcortical or intracortical loops are directly involved between receptors in muscles and tendons and the cerebral cortex in signaling movement magnitude and muscular tension information. Previous experiments have indicated that this information does reach consciousness. Data from voluntary compression of springs and strain-gauge were analyzed in patients with unilateral focal lesions of the cerebral hemispheres. It was found that the perception of signals of muscular tension is abolished by lesions of the contralateral cortex near the central sulcus. It was concluded that the possibility exists of separate cortical projection areas for kinesthetic signals from muscles and from joints.


Adaptive UNet-based Lung Segmentation and Ensemble Learning with CNN-based Deep Features for Automated COVID-19 Diagnosis.
COVID-19 disease is a major health calamity in twentieth century, in which the infection is spreading at the global level. Developing countries like Bangladesh, India, and others are still facing a delay in recognizing COVID-19 cases. Hence, there is a need for immediate recognition with perfect identification of infection. This clear visualization helps to save the life of suspected COVID-19 patients. With the help of traditional RT-PCR testing, the combination of medical images and deep learning classifiers delivers more hopeful results with high accuracy in the prediction and recognition of COVID-19 cases. COVID-19 disease is recently researched through sample chest X-ray images, which have already proven its efficiency in lung diseases. To emphasize corona virus testing methods and to control the community spreading, the automatic detection process of COVID-19 is processed through the detailed medication reports from medical images. Although there are numerous challenges in the manual understanding of traces in COVID-19 infection from X-ray, the subtle differences among normal and infected X-rays can be traced by the data patterns of Convolutional Neural Network (CNN). To improve the detection performance of CNN, this paper plans to develop an Ensemble Learning with CNN-based Deep Features (EL-CNN-DF). In the initial phase, image scaling and median filtering perform the pre-processing of the chest X-ray images gathered from the benchmark source. The second phase is lung segmentation, which is the significant step for COVID detection. It is accomplished by the Adaptive Activation Function-based U-Net (AAF-U-Net). Once the lungs are segmented, it is subjected to novel EL-CNN-DF, in which the deep features are extracted from the pooling layer of CNN, and the fully connected layer of CNN are replaced with the three classifiers termed "Support Vector Machine (SVM), Autoencoder, Naive Bayes (NB)". The final detection of COVID-19 is done by these classifiers, in which high ranking strategy is utilized. As a modification, a Self Adaptive-Tunicate Swarm Algorithm (SA-TSA) is adopted as a boosting algorithm to enhance the performance of segmentation and detection. The overall analysis has shown that the precision of the enhanced CNN by using SA-TSA was 1.02%, 4.63%, 3.38%, 1.62%, 1.51% and 1.04% better than SVM, autoencoder, NB, Ensemble, RNN and LSTM respectively. The comparative performance analysis on existing model proves that the proposed algorithm is better than other algorithms in terms of segmentation and classification of COVID-19 detection.


An essential care course for healthcare assistants.
There is growing consensus that healthcare assistants (HCAs) need access to professional training, education and support. The authors describe a short course designed to educate and support HCAs, focusing on the care of older people.


Rheumatic manifestations of hyperlipidemia and antihyperlipidemia drug therapy.
Conflicting data exist with respect to the existence and clinical manifestations of a hyperlipidemic arthropathy. Reasonable evidence supports the existence of a migratory polyarthritis similar to rheumatic fever in patients homozygous for type II hyperlipidemia. Although similar complaints have been described in patients heterozygous for this condition, findings have been inconsistent among various reports. It is possible that high lipid levels are required to induce rheumatic complaints, and these are found predominantly in homozygous patients. Even so, rheumatic syndromes appear to be more attributable to periarthritis because evidence of inflammatory arthritis is largely lacking. In contrast, Achilles tendinitis appears to be associated with heterozygous type II hyperlipidemia and presumably is based on lipid deposits within the tendon. Gout is an accepted association of type IV hyperlipidemia. In addition, oligoarticular symptoms have been described with type IV hyperlipidemia. However, no consistent clinical entity has emerged. Drugs used in the treatment of hyperlipidemia are associated with a variety of rheumatic problems, including proximal myopathy and lupus-like syndromes. The most commonly implicated drugs are the hydroxymethylglutaryl-coenzyme A reductase inhibitors and the fibric acid derivatives.


Induction of proliferative and cytotoxic responses in resting Lyt-2+ T cells with lectin and recombinant interleukin 2.
The lectin leucoagglutinin has been used to induce reactivity to interleukin 2 (IL2) in unseparated spleen cells and in highly purified Lyt-2+ lymph node T cells. Recombinant human IL2 and various other IL2-containing preparations, including concanavalin A-induced spleen cell supernatant, were compared for their capacity to support DNA synthesis and cytotoxic activity. In contrast to published reports, we found that the capacity of all preparations tested was identical in both functional assays, if they were adjusted to the same IL2 titer. Our inability to detect a requirement for an externally added cytotoxic T cell differentiation factor could either mean that IL2 is sufficient for the promotion of both proliferation and differentiation in leucoagglutinin-activated resting cytotoxic T cell precursors, or that under our experimental conditions, T cell differentiation factor is endogenously produced by Lyt-2+ T cells.


Lineage tracing of acute myeloid leukemia reveals the impact of hypomethylating agents on chemoresistance selection.
Chemotherapy-resistant cancer recurrence is a major cause of mortality. In acute myeloid leukemia (AML), chemorefractory relapses result from the complex interplay between altered genetic, epigenetic and transcriptional states in leukemic cells. Here, we develop an experimental model system using in vitro lineage tracing coupled with exome, transcriptome and in vivo functional readouts to assess the AML population dynamics and associated molecular determinants underpinning chemoresistance development. We find that combining standard chemotherapeutic regimens with low doses of DNA methyltransferase inhibitors (DNMTi, hypomethylating drugs) prevents chemoresistant relapses. Mechanistically, DNMTi suppresses the outgrowth of a pre-determined set of chemoresistant AML clones with stemness properties, instead favoring the expansion of rarer and unfit chemosensitive clones. Importantly, we confirm the capacity of DNMTi combination to suppress stemness-dependent chemoresistance development in xenotransplantation models and primary AML patient samples. Together, these results support the potential of DNMTi combination treatment to circumvent the development of chemorefractory AML relapses.


[Intrauterine blood transfusion as a method of treatment of the edematous form of hemolytic disease of newborn].
Results are reported with intrauterine donor blood transfusions in 7 fetuses with hydropic hemolytic disease. Eight intraabdominal and 8 intravascular sonographically guided fetal transfusions were given at 25-30 weeks' gestation with fetal heart rate monitoring. The criteria for efficacy were disappearance of ascites, reduction in fetal hepatic and splenic sizes, thinning of the placenta. Three fetuses died in utero from symptomatic hydropic hemolytic disease despite the therapy. Four other fetuses were delivered at 30-34 gestational weeks by cesarean section and showed no abdominal ascites. One of the fetuses died from birth trauma on the second postnatal day while the other three were discharged in a satisfactory condition.


Long-term renal function in cyclosporine-treated renal allograft recipients.
The purpose of this study was to analyze the factors affecting long-term renal function in cyclosporine-treated kidney transplants. The study population comprised 167 patients with more than 5 years of graft function on cyclosporine therapy. Patients were subdivided into those with a serum creatinine level 2.0 mg/dL or less (group I, n = 123) versus those with a level more than 2.0 mg/dL (group II, n = 44) at 5 years post-transplant. Patient survival, graft survival, rejection episodes, renal function, cyclosporine dose and trough level, and proteinuria were compared in these two groups. There was no significant difference between groups I and II in terms of race, sex, donor source, donor age, primary renal disease, retransplants, transfusions, presensitization, histocompatibility locus antigen match, or initial nonfunction. At 6 months post-transplantation, the mean serum creatinine level in group II was significantly higher than group I (P = 0.00001), and this difference increased at subsequent follow-up intervals. The incidence of proteinuria was significantly higher in group II compared with group I (P < 0.001). Renal allograft survival beyond 5 years post-transplant was significantly better in group I compared with group II (P = 0.005). There was no significant difference in the mean cyclosporine dose or the mean cyclosporine trough level between groups I and II at any time following transplantation. The most important difference between groups I and II was the finding of significantly more early (P < 0.001) and late (P < 0.001) rejection episodes in group II. These data suggest that long-term renal function in cyclosporine-treated kidney transplant patients is primarily influenced by the occurrence of early and late rejection episodes rather than by the dosage or duration of cyclosporine therapy.


Interaction between adenosine A1 and A2 receptor-mediated responses in the rat hippocampus in vitro.
Previous work has been carried out on the effects of adenosine on transmitter release and on the excitability of postsynaptic neurones, but little is known about the effects of adenosine on the coupling between the two. In this study, we examine the effects of specific adenosine receptor agonists and antagonists on the population excitatory postsynaptic potential (population EPSP) slope, the population spike amplitude, and the relationship between the two (E-S coupling) in the CA1 area of rat hippocampus. Activation of adenosine A1 receptors by adenosine or the selective agonist N6-cyclopentyladenosine resulted in a decrease of the population spike amplitude by a greater extent than could be accounted for by the decrease in population EPSP slope, resulting in a dissociation in the E-S relationship, reflected as a right-shift in the E-S curve. Activation of adenosine A2A receptors by the selective agonist 2-p-(2-carboxyethy)phenethylamino-5'-N-ethylcarboxamidoadeno sine (CGS 21680), or blockade by antagonists ZM 241385 and CP 66713 had no effect on evoked responses. However, when both adenosine A1 and A2A receptors were activated at the same time, a significant attenuation of the inhibitory effects of N6-cyclopentyladenosine on population spike amplitude was observed, resulting in a left-shift in the E-S curve. Intracellular recording indicated that N6-cyclopentyladenosine raised the threshold for spike induction by pulses of depolarising current, even at a concentration which did not produce hyperpolarisation of the neurone. At 30 nM, CGS 21680 prevented this effect of N6-cyclopentyladenosine, and this apparent antagonism was prevented by the A2A receptor antagonist ZM 241385. The results show that adenosine A1 receptors change the coupling between presynaptic transmitter release and postsynaptic cell firing, and that this effect is attenuated by A2A receptor activation.


A method for the study of lay conceptions of psychopathology: a free-response approach.
A free-response approach for identifying a lay group's conception of psycopathology is described in this paper and is illustrated with a college student population. An initial sample of individuals from the population is asked to give "reasons why someone might need to be examined by a professional person in the field of mental health." A second sample of individuals sort the problem statements on the basis of their judgements of the similarity among the statements. A measure of psychological distance for each pair of statements, based on the proportion of times the two statements are sorted together, is used as input for clustering and multidimensional scaling. A third sample contributes to the interpretation of the resulting structures by rating each problem on several psychological properties. The most relevant properties found for college students are: (a) degree of abnormality; (b) referral source--self versus others. Also disc1ssed are possible applications of the method to other lay groups in the interest of providing improved community mental health services to such groups.


Physical, morphological, and wound healing properties of a polyurethane foam-film dressing.
We investigated the physicochemical properties of Medifoam® N and its wound healing performance compared to other commercially available polyurethane (PU) foam dressing in vitro and in vivo to gain insight in their clinical performance. Wound contact layer and cross-section of eleven polyurethane foam dressings were assessed with field-emission scanning electron microscope. Thickness, density, tensile strength, elongation, moisture-vapor transmission rate (MVTR), retention and absorptivity were measured to compare physical properties. Phosphate-buffered saline (PBS) solution absorption patterns were compared. An animal model for wound-healing was applied to validate in vitro findings. Among eleven tested foam dressings, Medifoam® N has the smallest pore and cell sizes with excellent uniformity, i.e. it has 25 ~ 75 μm on the wound contact layer and 100 ~ 350 μm in the cross-section while other dressings have a larger pose size with larger variability. Compared to other PU foams, Medifoam® N also has moderate thickness, density, tensile strength, elongation and MVTR. Furthermore, it has excellent fluid absorption and retention capacity. These intrinsic properties of Medifoam® N contributed to improve fluid absorption patterns, i.e. other dressing material flawed out PBS solution on the dressings while Medifoam® N retained all the tested solutions. In animal wound-healing study, Medifoam® N treated animals showed excellent angiogenesis and collagen deposition even though epithelial recovery rate was not significantly different to other dressings. Medifoam® N has optimized physical properties and thus improved fluid absorption/retention capacity. Compared to other dressings, Medifoam® N showed excellent fluid absorption patterns and these characteristics contributed to improved wound healing and excellent angiogenic potential. We found that Medifoam® N showed the best results among the employed dressing samples.


Human brain aldehyde dehydrogenase: activity with DOPAL and isozyme distribution.
Aldehyde dehydrogenase (ALDH, EC 18.104.22.168), has been shown to be the most important enzyme for DOPAL metabolism in human brain (Agarwal et al.). In the present investigation cerebellum, corpus striatum and pons showed the highest ALDH activity. Most of the enzyme activity was found in the mitochondrial and microsomal fractions. Two activity bands on IEF gels and dual Km values indicate the presence of two distinct isozymes in all the fractions. Two cerebella from alcoholics yielded the same results as the control group regarding their total ALDH activity, subcellular distribution pattern and protein content. The presence of DOPAC (acid metabolite of DOPAL), pargyline, pyrazole or ethanol in the assay mixture did not alter the ALDH activity significantly.


Physiotherapists' perceptions of implementing evidence-based practice for patients with low back pain through the Enhanced Transtheoretical Model Intervention: a qualitative study.
Physiotherapists have been urged to implement evidence-based practice for the management of low back pain (LBP). However, recent evidence suggests that many fail to do so, specifically in accordance with eliciting and addressing psychosocial factors that impact pain. The Enhanced Transtheoretical Model Intervention (ETMI) for the Treatment of patients with LBP was developed in Israel according to the clinical guidelines and demonstrated clinically and cost-effectiveness. This study's purpose is to explore physiotherapists' perceptions of implementing evidence-based practice through the ETMI approach. Qualitative study. Qualitative semi-structured interviews were conducted with 26 physiotherapists. The interviews were audio-recorded, transcribed, coded, and analyzed thematically to identify prevalent themes. Three main themes were identified from the transcripts, consisting of barriers and facilitators of implementing the intervention, and a meta-theme referring to shifts in the perceptions of professional role and identity. The findings suggest that physiotherapists believed that implementing ETMI and adopting a psychosocial approach to LBP can be achieved by overcoming issues around communication skills, patient reassurance, and inter-professional collaboration. Therapists also highlighted the need for physiotherapy training to gain skills in combined physical and psychological approaches.


Non-invasive diagnostic techniques in pigmentary skin disorders and skin cancer.
Diagnosis of pigmentary skin disorders, pre-cancerous and cancerous skin diseases is traditionally relied on visual assessment. The most widely applied invasive diagnostic technique is the skin biopsy. There have been significant technological advances in non-invasive diagnostic methods for skin disorders. The objective of this article is to discuss different non-invasive diagnostic modalities, used in the diagnosis of pigmentary skin disorders and cutaneous cancers. Comprehensive literature search was performed to screen articles related to non-invasive diagnostic techniques in pigmentary skin disorders and cutaneous cancers. Articles published in journals indexed in PubMed were searched along with those in Google Scholar. Clinical trials, review articles, case series, case reports and other relevant articles were considered for review. References of relevant articles were also considered for review. Dermoscopy and ultrasonography were the only non-invasive diagnostic and imaging techniques available to dermatologists for many years. The advent of computed tomography (CT) and magnetic resonance imaging (MRI) augmented the visualization of deeper structures. Confocal laser microscopy (CLM) and reflectance spectrophotometers have showed promising results in the non-invasive detection of pigmented lesions. Optical coherence tomography (OCT), electrical impedance spectroscopy (EIS), multispectral imaging, high frequency ultrasonography (HFUS) and adhesive patch biopsy aid in the accurate diagnosis of benign, as well as neoplastic skin diseases. There have been significant advancements in non-invasive methods for diagnosis of dermatological diseases. These techniques can be repeatedly used in a comfort manner for the patient, and may offer an objective way to follow the course of a disease.


Tyrosine hydroxylase- and/or aromatic L-amino acid decarboxylase-expressing neurons in the rat arcuate nucleus: ontogenesis and functional significance.
This study has evaluated in vivo, ex vivo and in vitro the ontogenesis and functional significance of the neurons of the arcuate nucleus (AN) expressing either individual enzymes of dopamine (DA) synthesis, tyrosine hydroxylase (TH) or aromatic L-amino acid decarboxylase (AADC) as well as both of them in rats from the 17th embryonic day (E) till adulthood. Immunocytochemistry, image analysis, confocal microscopy, high performance liquid chromatography with electrochemical detection and radioimmunoassay were used to solve this problem. Monoenzymatic TH-containing neurons were initially observed on E18 located in the ventrolateral AN whereas the neurons expressing only AADC or both AADC and TH first appeared on E20 in the dorsomedial AN. On E21, the monoenzymatic TH- or AADC-expressing neurons comprised more than 99% of the whole neuron population expressing the DA-synthesizing enzymes. In spite of an extremely small number (<1%) of the neurons expressing both enzymes (DArgic neurons), the dissected AN (ex vivo) and its primary cell culture (in vitro) contained a surprisingly high amount of DA and L-dihydroxyphenylalanine (L-DOPA) which were released in response to membrane depolarization. Furthermore, DA production in the AN of fetuses occurred to be sufficient to provide an inhibitory control of prolactin secretion, as in adults. The above data suggest that DA could be synthesized, at least in the AN of fetuses, by monoenzymatic neurons containing either TH or AADC, in co-operation. This hypothesis may be extended to adult animals as their AN contained the same populations of the neurons expressing DA-synthesizing enzymes as in fetuses though the proportion of true DArgic neurons increased up to 38%. During ontogenesis, the monoenzymatic TH- and AADC-containing neurons established axosomatic and axo-axonal junctions that might facilitate the L-DOPA transport from the former to the latter. Moreover, the monoenzymatic AADC-expressing neurons project their axons to the median eminence, thereby, providing the pathway for the DA transport toward the hypophysial portal circulation. Thus, DA appears to be synthesized in the AN not only by DArgic neurons but also by monoenzymatic TH- and AADC-expressing neurons in co-operation.


Heterogeneity and versatility of the extracellular matrix during the transition from pleomorphic adenoma to carcinoma ex pleomorphic adenoma: cumulative findings from basic research and new insights.
Pleomorphic adenoma (PA) is the most common salivary gland tumor, accounting for 50%-60% of these neoplasms. If untreated, 6.2% of PA may undergo malignant transformation to carcinoma ex-pleomorphic adenoma (CXPA). CXPA is a rare and aggressive malignant tumor, whose prevalence represents approximately 3%-6% of all salivary gland tumors. Although the pathogenesis of the PA-CXPA transition remains unclear, CXPA development requires the participation of cellular components and the tumor microenvironment for its progression. The extracellular matrix (ECM) comprises a heterogeneous and versatile network of macromolecules synthesized and secreted by embryonic cells. In the PA-CXPA sequence, ECM is formed by a variety of components including collagen, elastin, fibronectin, laminins, glycosaminoglycans, proteoglycans, and other glycoproteins, mainly secreted by epithelial cells, myoepithelial cells, cancer-associated fibroblasts, immune cells, and endothelial cells. Like in other tumors including breast cancer, ECM changes play an important role in the PA-CXPA sequence. This review summarizes what is currently known about the role of ECM during CXPA development.


Investing in what matters mostLife on the Rocks: Building a Future for Coral Reefs Juli Berwald Riverhead Books, 2022. 352 pp.
Faced with a family crisis, a marine scientist finds parallels with Earth's imperiled coral reefs.


A Novel Algorithm With Paired Predictive Indexes to Stratify the Risk Levels of Neonates With Invasive Bacterial Infections: A Multicenter Cohort Study.
Our aim was to develop a predictive model comprising clinical and laboratory parameters for early identification of full-term neonates with different risks of invasive bacterial infections (IBIs). We conducted a retrospective study including 1053 neonates presenting in 9 tertiary hospitals in China from January 2010 to August 2019. An algorithm with paired predictive indexes (PPIs) for risk stratification of neonatal IBIs was developed. Predictive performance was validated using k-fold cross-validation. Overall, 166 neonates were diagnosed with IBIs (15.8%). White blood cell count, C-reactive protein level, procalcitonin level, neutrophil percentage, age at admission, neurologic signs, and ill-appearances showed independent associations with IBIs from stepwise regression analysis and combined into 23 PPIs. Using 10-fold cross-validation, a combination of 7 PPIs with the highest predictive performance was picked out to construct an algorithm. Finally, 58.1% (612/1053) patients were classified as low-risk cases. The sensitivity and negative predictive value of the algorithm were 95.3% (95% confidence interval: 91.7-98.3) and 98.7% (95% confidence interval: 97.8-99.6), respectively. An online calculator based on this algorithm was developed for clinical use. The new algorithm constructed for this study was a valuable tool to screen neonates with suspected infection. It stratified risk levels of IBIs and had an excellent predictive performance.


Electrospray and atmospheric pressure chemical ionization tandem mass spectrometric behavior of eight anabolic steroid glucuronides.
Mass spectrometric and tandem mass spectrometric behavior of eight anabolic steroid glucuronides were examined using electrospray (ESI) and atmospheric pressure chemical ionization (APCI) in negative and positive ion mode. The objective was to elucidate the most suitable ionization method to produce intense structure specific product ions and to examine the possibilities of distinguishing between isomeric steroid glucuronides. The analytes were glucuronide conjugates of testosterone (TG), epitestosterone (ETG), nandrolone (NG), androsterone (AG), 5alpha-estran-3alpha-ol-17-one (5alpha-NG), 5beta-estran-3alpha-ol-17-one (5beta-NG), 17alpha-methyl-5alpha-androstane-3alpha,17beta-diol (5alpha-MTG), and 17alpha-methyl-5beta-androstane-3alpha,17beta-diol (5beta-MTG), the last four being new compounds synthesized with enzyme-assisted method in our laboratory. High proton affinity of the 4-ene-3-one system in the steroid structure favored the formation of protonated molecule [M + H]+ in positive ion mode mass spectrometry (MS), whereas the steroid glucuronides with lower proton affinities were detected mainly as ammonium adducts [M + NH4]+. The only ion produced in negative ion mode mass spectrometry was a very intense and stable deprotonated molecule [M - H]- . Positive ion ESI and APCI MS/MS spectra showed abundant and structure specific product ions [M + H - Glu]+, [M + H - Glu - H2O]+, and [M + H - Glu - 2H2O]+ of protonated molecules and corresponding ions of the ammonium adduct ions. The ratio of the relative abundances of these ions and the stability of the precursor ion provided distinction of 5alpha-NG and 5beta-NG isomers and TG and ETG isomers. Corresponding diagnostic ions were only minor peaks in negative ion MS/MS spectra. It was shown that positive ion ESI MS/MS is the most promising method for further development of LC-MS methods for anabolic steroid glucuronides.


Reconstructive burden and financial implications of wider excision margins for invasive primary cutaneous melanoma.
For invasive primary cutaneous melanoma, wider excision is advocated to reduce local recurrence risk and improve patient outcomes. Excision detail is controversial, especially in intermediate- and high-risk primary melanoma (AJCC pT2-pT4). Guidance varies from sizes 1 to 3 cm (translating into large defects of 2-6 cm). The aim of this study was to determine the reconstructive and resource burden of wider excision margins (EMs). Data analysis from our prospective database (2008-2017) included 1184 patients (563F:621 M) with cutaneous melanoma (pT1b-pT4b). Procedure tariff data were sourced from our financial services department. Two hundred twenty-nine patients had a narrower EM (1 cm) and 995 (80.7%) had a wider EM (2-3 cm). Reconstructive requirement significantly increased with a wider EM collectively (11.3% vs 29.3%, odds ratio (OR) = 3.2; p < 0.0001), in the extremities (15.2% vs 42.0%; p < 0.0001), and in the head and neck (H&N) (23.5 % vs 64.7%; p < 0.0001). Reconstruction significantly increased hospitalisation rates (26.6% vs 63.0%, OR = 4.7; p < 0.0001) collectively, in the H&N (26.8 % vs 53.9%), and in the upper (18.9 % vs 42.3%) and lower extremities (34.8% vs 77.3%). Narrower EMs significantly reduced hospitalisation rates in the upper and lower extremities (7.1% vs 28.5%; p = 0.004, 37.9% vs 58.5%; p = 0.005, respectively). Overall procedure cost significantly increased by £180 (mean, p < 0.0001) and £346 (median, p = 0.0004) per patient when reconstruction was required. Our data suggest substantial impact of wider EM on patients, which more than doubled in the functionally and cosmetically sensitive extremities and the H&N region. Reconstructions add significant financial and healthcare service burden. Without randomised controlled trial (RCT) evidence demonstrating increased efficacy of wider EM, narrower EM is advocated whilst awaiting future planned RCT results specifically investigating on this.


Arachidonic acid drives adaptive responses to chemotherapy-induced stress in malignant mesothelioma.
Background High resistance to therapy and poor prognosis characterizes malignant pleural mesothelioma (MPM). In fact, the current lines of treatment, based on platinum and pemetrexed, have limited impact on the survival of MPM patients. Adaptive response to therapy-induced stress involves complex rearrangements of the MPM secretome, mediated by the acquisition of a senescence-associated-secretory-phenotype (SASP). This fuels the emergence of chemoresistant cell subpopulations, with specific gene expression traits and protumorigenic features. The SASP-driven rearrangement of MPM secretome takes days to weeks to occur. Thus, we have searched for early mediators of such adaptive process and focused on metabolites differentially released in mesothelioma vs mesothelial cell culture media, after treatment with pemetrexed. Mass spectrometry-based (LC/MS and GC/MS) identification of extracellular metabolites and unbiased statistical analysis were performed on the spent media of mesothelial and mesothelioma cell lines, at steady state and after a pulse with pharmacologically relevant doses of the drug. ELISA based evaluation of arachidonic acid (AA) levels and enzyme inhibition assays were used to explore the role of cPLA2 in AA release and that of LOX/COX-mediated processing of AA. QRT-PCR, flow cytometry analysis of ALDH expressing cells and 3D spheroid growth assays were employed to assess the role of AA at mediating chemoresistance features of MPM. ELISA based detection of p65 and IkBalpha were used to interrogate the NFkB pathway activation in AA-treated cells. We first validated what is known or expected from the mechanism of action of the antifolate. Further, we found increased levels of PUFAs and, more specifically, arachidonic acid (AA), in the transformed cell lines treated with pemetrexed. We showed that pharmacologically relevant doses of AA tightly recapitulated the rearrangement of cell subpopulations and the gene expression changes happening in pemetrexed -treated cultures and related to chemoresistance. Further, we showed that release of AA following pemetrexed treatment was due to cPLA2 and that AA signaling impinged on NFkB activation and largely affected anchorage-independent, 3D growth and the resistance of the MPM 3D cultures to the drug. AA is an early mediator of the adaptive response to pem in chemoresistant MPM and, possibly, other malignancies.


Integrated genomic analysis of proteasome alterations across 11,057 patients with 33 cancer types: clinically relevant outcomes in framework of 3P medicine.
Proteasome, a cylindrical complex containing 19S regulatory particle lid, 19S regulatory particle base, and 20S core particle, acted as a major mechanism to regulate the levels of intracellular proteins and degrade misfolded proteins, which involved in many cellular processes, and played important roles in cancer biological processes. Elucidation of proteasome alterations across multiple cancer types will directly contribute to cancer medical services in the context of predictive, preventive, and personalized medicine (PPPM / 3P medicine). This study aimed to investigate proteasome gene alterations across 33 cancer types for discovery of effective biomarkers and therapeutic targets in the framework of PPPM practice in cancers. Proteasome gene data, including gene expression RNAseq, somatic mutation, tumor mutation burden (TMB), copy number variant (CNV), microsatellite instability (MSI) score, clinical characteristics, immune phenotype, 22 immune cells, cancer stemness index, drug sensitivity, and related pathways, were systematically analyzed with publically available database and bioinformatics across 11,057 patients with 33 cancer types. Differentially expressed proteasome genes were extensively found between tumor and control tissues. PSMB4 occurred the top mutation event among proteasome genes, and those proteasome genes were significantly associated with TMB and MSI score. Most of proteasome genes were positively related to CNV among single deletion, control copy number, and single gain. Kaplan-Meier curves and COX regression survival analysis showed proteasome genes were significantly associated with patient survival rate across 33 cancer types. Furthermore, the expressions of proteasome genes were significantly different among different clinical stages and immune subtypes. The expressions of proteasome genes were correlated with immune-related scores (ImmuneScore, StromalScore, and ESTIMATEScore), 22 immune cells, and cancer stemness. The sensitivities of multiple drugs were closely related to proteasome gene expressions. The identified proteasome and proteasome-interacted proteins were significantly enriched in various cancer-related pathways. This study provided the first landscape of proteasome alterations across 11,057 patients with 33 cancer types and revealed that proteasome played a significant and wide functional role in cancer biological processes. These findings are the precious scientific data to reveal the common and specific alterations of proteasome genes among 33 cancer types, which benefits the research and practice of PPPM in cancers. The online version contains supplementary material available at 10.1007/s13167-021-00256-z.


Chemokine CCL2 from proximal tubular epithelial cells contributes to sepsis-induced acute kidney injury.
Damage-associated molecular patterns secreted from activated kidney cells initiate the inflammatory response, a critical step in the development of sepsis-induced acute kidney injury (AKI). However, the underlying mechanism remains to be clarified. Here, we established a mouse model of sepsis-induced AKI through intraperitoneal injection of lipopolysaccharide (LPS) and demonstrated that LPS induced dramatical upregulation of C-C motif chemokine ligand 2 (CCL2) at both the mRNA and protein levels in the kidney, which was mainly expressed by tubular epithelial cells (TECs), especially by proximal TECs. Proximal tubule-specific ablation of CCL2 reduced LPS-induced macrophage infiltration, proinflammatory cytokine expression, and attenuated AKI. In vitro, using a Transwell migration assay, we found that deficiency of CCL2 in TECs decreased macrophage migration ability. However, myeloid-specific depletion of CCL2 could not protect the kidneys from the aforementioned effects. Mechanistically, LPS activated Toll-like receptor (TLR)2 signaling in TECs, which induced activation of its downstream effector NF-κB. Blockade of TLR2 signaling or inhibition of NF-κB activation in TECs significantly suppressed LPS-induced CCL2 expression. Furthermore, chromatin immunoprecipitation analyses confirmed a direct binding of NF-κB p65 in the CCL2 promoter region, and LPS increased the binding of NF-κB p65 to the CCL2 promoter, suggesting that TLR2/NF-κB p65 regulates CCL2 expression in TECs. Together, these results demonstrate that endogenous CCL2 released from proximal TECs, not from myeloid cells, was responsible for sepsis-induced kidney inflammation and AKI. Specifically targeting tubular TLR2/NF-κB/CCL2 signaling may be a potential therapeutic strategy for the prevention or attenuation of septic AKI.NEW & NOTEWORTHY This study provides a mechanistic insight into how C-C motif chemokine ligand 2 (CCL2) is upregulated in renal tubular epithelial cells (TECs) and contributes to kidney dysfunction during sepsis. The data reveal that lipopolysaccharide induces CCL2 expression through the Toll-like receptor 2/NF-κB signaling pathway in TECs. Endogenous CCL2 released from TECs, not from myeloid cells, is responsible for sepsis-induced kidney inflammation and acute kidney injury.


Absence of diabetes mellitus type 2 in obese children and adolescents in the north of Spain.
The worldwide epidemic of childhood obesity has been accompanied by an increase in the incidence of carbohydrate metabolism disorders. To determine the prevalence of type 2 diabetes mellitus (T2DM) and other carbohydrate metabolism disorders in obese young people in the Basque Country (Spain). Prospective observational study. We studied 136 obese Caucasian children and adolescents (body mass index ≥2 SDS above the mean). Their severity of obesity was classified as mild <3 SDS or moderate-to-severe ≥3 SDS. Data were collected on clinical and metabolic parameters; insulin resistance (IR) was calculated using the homeostasis model assessment, and an oral glucose tolerance test (OGTT) was carried out. T2DM was not found. Impaired glucose tolerance (IGT) was found in 9.6% of patients being higher in moderate-to-severe obesity (12.8% vs. 2.4%; p=0.048) and in patients with acanthosis nigricans (27.8% vs. 6.8%; p=0.016). No differences were detected by sex or pubertal development in metabolic results as a function of OGTT's response. IR (13.5%) was higher among those with moderate-to-severe obesity, in patients with acanthosis nigricans and was associated with other cardiovascular disease risk factors. We found no children with T2DM. The prevalence of IGT and IR was related to severity of obesity, to the association of acanthosis nigricans and was associated with cardiovascular risk.


The lattice arrangement of the collagen fibres in the submucosa of the rat small intestine: scanning electron microscopy.
The three-dimensional architecture of the submucosal collagen fibres of the rat (3 weeks old) small intestine was examined by scanning electron microscopy using a selective microdissection method. The main framework of the submucosa was composed of two arrays of collagen fibre bundles running diagonally around the intestinal wall, one set in a clockwise direction, the other counterclockwise. These fibre bundles were about 5 micron diameter and were oriented at a range of angle +/- 30 degrees -50 degrees to the longitudinal axis of the intestine. With the advantage of the SEM observation it was demonstrated that these fibres in different arrays did not constitute two separate layers but interwove to form a unified lattice sheet. An irregular network of fine collagen fibrils over the main framework was also seen. The significance of their arrangement is discussed with respect to the skeletal function of the submucosa in the intestine.


The psychophysiological nature of premature ejaculation.
The hypothesis that premature ejaculators (PEs) are less able than non-premature ejaculators (NPEs) to evaluate accurately their level of physiologically determined sexual arousal was tested. Twenty-six men (13 PEs and 13 NPEs) viewed a variety of videotaped vignettes, some of which were excerpts from sexually explicit films. Concurrent subjective (self-report) and objective (plethysmograph) ratings of sexual arousal were taken. Data revealed that both the PEs and NPEs were equally accurate in assessing their level of physiological sexual arousal. These results and those from a sexual history questionnaire were used to evaluate several hypotheses regarding the nature and etiology of premature ejaculation.