Recognition of double-stranded RNA by human toll-like receptor 3 and downstream receptor signaling requires multimerization and an acidic pH.
Studies involving Toll-like receptor 3 (TLR3)-deficient mice suggest that this receptor binds double-stranded RNA. In the present study, we analyzed ligand/receptor interactions and receptor-proximal events leading to TLR3 activation. The mutagenesis approach showed that certain cysteine residues and glycosylation in TLR3 amino-terminal leucine-rich repeats were necessary for ligand-induced signaling. Furthermore, inactive mutants had a dominant negative effect, suggesting that the signaling module is a multimer. We constructed a chimeric molecule fusing the amino-terminal ectodomain of TLR3 to the transmembrane and carboxyl terminal domains of CD32a containing an immunoreceptor tyrosine-based motif. Expression of TLR3-CD32 in HEK293T cells and the myeloid cell line U937 resulted in surface localization of the receptor, whereas the nonrecombinant molecule was intracellularly localized. The synthetic double-stranded RNAs poly(I-C) and poly(A-U) induced calcium mobilization in a TLR3-CD32 stably transfected U937 clone but not in control cells transfected with other constructs. An anti-TLR3 antibody also induced Ca(2+) flux but only when cross-linked by a secondary anti-immunoglobulin antibody, confirming that multimerization by the ligand is a requirement for signaling. The inhibitors of lysosome maturation, bafilomycin and chloroquine, inhibited the poly(I-C)-induced biological response in immune cells, showing that TLR3 interacted with its ligand in acidic subcellular compartments. Furthermore, TLR3-CD32 activation with poly(I-C) was only observed within a narrow pH window (pH 5.7-6.7), whereas anti-TLR3-mediated Ca(2+) flux was pH-insensitive. The importance of an acidic pH for TLR3-ligand interaction becomes critical when using oligomeric poly(I-C) (15-40-mers). These observations demonstrate that engagement of TLR3 by poly(I-C) at an acidic pH, probably in early phagolysosomes or endosomes, induces receptor aggregation leading to signaling.


Bio-objectifying European bodies: standardisation of biobanks in the Biobanking and Biomolecular Resources Research Infrastructure.
The article traces the genealogy of the Minimum Information About Biobank Data Sharing model, created in the European Biobanking and Biomolecular Resources Research Infrastructure to facilitate collaboration among biobanks and to foster the exchange of biological samples and data. This information model is aimed at the identification of biobanks; unification of databases; and objectification of the information, samples, and related studies - to create a completely new 'bio-object infrastructure' within the EU. The paper discusses key challenges in creating a 'universal' information model of such a kind, the most important technical translations of European research policy needed for a standardised model for biobank information, and how this model creates new bio-objects. The author claims that this amounts to redefinition of biobanks and technical governance over virtually bio-objectified European populations. It is argued here that old governance models based on the nation-state need radical reconsideration so that we are prepared for a new and changing situation wherein bodies of information that lack organs flow from one database to another with a click of a mouse.


Physical fitness before and during the COVID-19 pandemic: Results of annual national physical fitness surveillance among 16,647,699 Japanese children and adolescents between 2013 and 2021.
Limited nationally representative evidence is available on temporal trends in physical fitness (PF) for children and adolescents during the coronavirus disease 2019 (COVID-19) pandemic. The primary aim was to examine the temporal trends in PF for Japanese children and adolescents before and during the COVID-19 pandemic. The secondary aim was to estimate the concurrent trends in body size (measured as body mass and height) and movement behaviors (exercise, screen, and sleep time). Census PF data for children in Grade 5 (aged 10-11 years) and adolescents in Grade 8 (aged 13-14 years) were obtained for the years 2013-2021 from the National Survey of Physical Fitness, Athletic Performance, and Exercise Habits in Japan (n = 16,647,699). PF and body size were objectively measured, and movement behaviors were self-reported. Using sample-weighted linear regression, temporal trends in mean PF were calculated before the pandemic (2013-2019) and during the pandemic (2019-2021) with adjustments for age, sex, body size, and exercise time. When adjusted for age, sex, body size, and exercise time, there were significant declines in PF during the pandemic, with the largest declines observed in 20-m shuttle run (standardized (Cohen's) effect size (ES) = -0.109 per annum (p.a.)) and sit-ups performance (ES = -0.133 p.a.). The magnitude of the declines in 20-m shuttle run and sit-ups performances were 18- and 15-fold larger, respectively, than the improvements seen before the pandemic (2013-2019), after adjusting for age, sex, body size, and exercise time. During the pandemic, both body mass and screen time significantly increased, and exercise time decreased. Declines in 20-m shuttle run and sit-ups performances suggest corresponding declines in population health during the COVID-19 pandemic.


Bone Quality in Paget's Disease of Bone.
Paget's disease of bone is produced by a localized increase in osteoclastic and osteoblastic activity which can progress slowly to involve an entire bone if untreated. A common feature is enlarged bones which are deformed, particularly in weight-bearing regions of the skeleton such as the lower extremity. Pathologic fractures may be a consequence, and nonunion of femoral fractures is not uncommon. Analyses of bone biopsies from patients with Paget's disease indicate that there is a lower, heterogeneous degree of bone mineralization and a younger tissue age than that found in control bone. Pagetic bone also has less resistance to plastic deformation and a straighter crack path than control bone.


Clinical Effects of Lactobacillus rhamnosus in Non-Surgical Treatment of Chronic Periodontitis: A Randomized Placebo-Controlled Trial With 1-Year Follow-Up.
Probiotics are living microorganisms that provide beneficial effects for the host when administered in proper quantities. The aim of this double-masked placebo-controlled parallel-arm randomized clinical trial is to evaluate the clinical effects of a Lactobacillus rhamnosus SP1-containing probiotic sachet as an adjunct to non-surgical therapy. Twenty-eight systemically healthy volunteers with chronic periodontitis were recruited and monitored clinically at baseline and 3, 6, 9, and 12 months after therapy. Clinical parameters measured included plaque accumulation, bleeding on probing, probing depths (PDs), and clinical attachment loss. Patients received non-surgical therapy, including scaling and root planing (SRP), and were assigned randomly to a test (SRP + probiotic, n = 14) or control (SRP + placebo, n = 14) group. The intake, once a day for 3 months, of an L. rhamnosus SP1 probiotic sachet commenced after the last session of SRP. Both test and control groups showed improvements in clinical parameters at all time points evaluated. However, the test group showed greater reductions in PD than the control. Also, at initial visits and after 1-year follow-up, the test group showed a statistically significant reduction in the number of participants with PD ≥6 mm, indicating a reduced need for surgery, in contrast to the placebo group. The results of this trial indicate that oral administration of L. rhamnosus SP1 resulted in similar clinical improvements compared with SRP alone.


Regioselective Hydroalkylation of Vinylarenes by Cooperative Cu and Ni Catalysis.
Disclosed here is a dual copper and nickel catalytic system with a silyl hydride source for promoting the linear selective hydroalkylation of vinylarenes. This carbon-carbon bond-forming protocol is applied to couple a variety of functionalized vinylarenes with alkyl halides applying a nickel(II) NNN pincer complex in the presence of an NHC-ligated copper catalyst. This combination allows for a 1 mol % loading of the nickel catalyst leading to turnover numbers of up to 72. Over 40 examples are presented, including applications for pharmaceutical diversification. Labeling experiments demonstrated the regioselectivity of the reaction and revealed that the copper catalyst plays a crucial role in enhancing the rate for formation of the reactive linear alkyl nickel complex. Overall, the presented work provides a complimentary approach for hydroalkylation reactions, whilst providing a preliminary mechanistic understanding of the cooperativity between the copper and nickel complexes.


A novel COOH-GO-COOH-MWNT/pDA/AuNPs based electrochemical aptasensor for detection of AFB1.
Aflatoxin B1 (AFB1), one of the most common mycotoxins in food matrixes, has been identified as the most toxic contaminant with mutagenic, teratogenic, immunosuppressive, and carcinogenic effects. In this study, an electrochemical aptamer sensor was developed for the on-site detection of AFB1. Carboxylated graphene oxide (COOH-GO) and carboxylated multi-walled carbon nanotubes (COOH-MWNT) nanocomposites, dopamine polymers (pDA) and gold nanoparticles (AuNPs) were used to enhance the electrochemical activity and the biocompatibility of the screen-printed electrodes (SPE). Once AFB1 was captured by the aptamer immobilized on the electrode surface, the redox current of [Fe(CN)6]3-/4- decreased. Therefore, the binding of aptamer (Apt) and AFB1 can be reflected by the change of the peak current. The as-prepared sensor showed a wide detection range of 0.1 fg ml-1-100 pg ml-1 and a low detection limit of 15.16 ag ml-1. It is also simple and low-cost, which shows great potential in practical application.


[Search of Chlamydia trachomatis in 159 women from private consultation].
Chlamydia trachomatis infection is very common and is associated with recurring cervicovaginitis, pelvic inflammatory disease, abortion and ectopic pregnancy. It may cause sterility. To determine Chlamydia trachomatis frequency at outpatient consultation. One hundred fifty-nine patients were retrospectively evaluated at the gynecology consultation, to whom an endocervical specimen was taken with a cotton swab to corroborate the presence of Chlamydia trachomatis immunofluorescent antibodies. Forty-nine out of 159 cases (28.9%) were positive to Chlamydia trachomatis. After dividing by subgroups, 26 out of 100 women with cervicovaginitis were positive (26%). Eleven out of 41 cases with pelvic inflammatory disease were positive (26.8%). In 14 patients with recent abortion, 7 were positive (50%) and 2 out of 4 with ectopic pregnancy were positive (50%). Chlamydia trachomatis detection in patients with cervicovaginitis, pelvic inflammatory disease, ectopic pregnancy and abortion will allow an appropriate treatment to avoid progressive damage and its sequelae to the female genital tract, reproductive and economic levels.


Unusual metastatic patterns of prostate adenocarcinoma.
While prostatic adenocarcinoma is associated classically with osseous or lymphatic metastasis, it can present clinically with protean manifestations. We describe 4 cases that demonstrate unusual patterns of metastatic spread, including isolated supraclavicular mass, pleural effusion, suprapubic and genital skin metastasis, and priapism. Newer immunohistochemical staining techniques for prostate specific antigen and prostatic acid phosphatase can assist in localization of the metastatic adenocarcinoma to a prostatic origin.


Membrane cholesterol interferes with neuronal apoptosis induced by soluble oligomers but not fibrils of amyloid-beta peptide.
Neuronal cell death in Alzheimer's disease (AD) is partly induced by the interaction of the amyloid-beta peptide (Abeta) with the plasma membrane of target cells. Accordingly, recent studies have suggested that cholesterol, an important component of membranes that controls their physical properties and functions, plays a critical role in neurodegenerative diseases. We report here that the enrichment of the neuronal plasma membrane with cholesterol protects cortical neurons from apoptosis induced by soluble oligomers of the Abeta(1-40) peptide. Conversely, cholesterol depletion using cyclodextrin renders cells more vulnerable to the cytotoxic effects of the Abeta-soluble oligomers. Increasing the cholesterol content of small unilamellar liposomes also decreases Abeta-dependent liposome fusion. We clearly demonstrate that cholesterol protection is specific to the soluble conformation of Abeta, because we observed no protective effects on cortical neurons treated by amyloid fibrils of the Abeta(1-40) peptide. This may provide a new opportunity for the development of an effective AD therapy as well as elucidate the impact of the cholesterol level during AD development.


Flow cytometric isolation of endodermal progenitors from mouse salivary gland differentiate into hepatic and pancreatic lineages.
Experimental injury is useful to induce tissue stem cells, which may exist in small numbers under normal conditions. The salivary glands originate from the endoderm and consist of acinar and ductal epithelial cells, which have exocrine function. After salivary gland duct ligation, acinar cells disappear as a result of apoptosis, and duct epithelium subsequently proliferates. In this study, we analyzed the tissue stem cells induced by salivary gland duct ligation in mice using immunohistochemistry and flow cytometry. We sorted the Sca-1(+)/c-Kit(+) fraction from adult mice salivary glands by way of fluorescence-activated cell sorting. The sorted cells were apparently homogeneous and were designated mouse salivary gland-derived progenitors (mSGPs). mSGP cells differentiated into a hepatic lineage when cultured in matrigel. In spherical culture in the presence of glucagon-like peptide-1 (GLP-1), these cells differentiated into a pancreatic endocrine lineage. When spheroidal bodies of mSGP, 20 to 30 microm in diameter, were transplanted into liver via the portal vein, the cells integrated into hepatic cords and expressed albumin and alpha1-antitrypsin, suggesting that they had differentiated into hepatic-type cells. Moreover, ductlike structures formed by mSGP cells also appeared, epithelial cells of which were positive for cytokeratin 19. In conclusion, fluorescence-activated cell sorting (FACS) based on histologic evidence is efficient in isolating adult tissue stem cells of the salivary gland. Tissue stem cells of endodermal origin (e.g., hepatic oval cells, pancreatic epithelial progenitor cells, and salivary gland progenitor cells) have similarities in their molecular markers and tissue location. Our findings suggest the existence of common tissue stem cells in endoderm-derived organs.


Synthesis and antitumor activity of lapathoside D and its analogs.
Phenylpropanoid sucrose esters are important class of plant-derived natural products and have greater potential to be leads for new drugs because of their structural diversity and broad-array of pharmacological and biological activities. Regio- and chemo-selective acylation of 2,1':4,6-O-di-isopropylidene sucrose 4 with cinnamoyl chloride 5 and p-acetoxycinnamoyl chloride 6 afforded mono-, di-, tri- and tetra- variant PSEs in moderate yields. The first total synthesis of di-substituted PSE, lapathoside D 1' has been achieved successfully in short and simple synthetic steps from sucrose 3 as an inexpensive starting material. Lapathoside D 1 and a set of selected synthesized PSEs were tested for in vitro cytotoxicity against human cervical epithelioid carcinoma (HeLa) cell lines. Most of the compounds exhibited significant antitumor activity with their IC(50) values ranging from 0.05 to 7.63 μM. The primary screening results indicated that PSEs might be valuable source for new potent anticancer drug candidates.


A mathematical model of cerebral circulation and oxygen supply.
A compartment model of cerebral circulation and oxygen supply including an autoregulation mechanism is presented. The model is focused on the analysis of slow dynamical variations of long term neurophysiological parameters like the partial oxygen pressure of brain tissue or the cerebral blood flow. The circulatory part of the model is built up of seven compartments including arteries, capillaries, veins, brain tissue, cerebrospinal fluid, the sagittal sinus and an artificial compartment for the simulation of brain swelling. The description of oxygen supply is based on a Krogh model. Numerical calculations reproduce the experimentally well established connection between arterial blood pressure and the production of cerebrospinal fluid. Furthermore we found an approximately linear correlation of the partial oxygen pressure of brain tissue to the mean arterial blood pressure in the case of an impaired autoregulation mechanism. In a first test such a linear dependence could also be detected in clinical data from the neurosurgical intensive care monitoring.


Recreational runners with Achilles tendinopathy have clinically detectable impairments: A case-control study.
To confirm what impairments are present in runners with Achilles tendinopathy (AT) and explore the variance of AT severity in an adequately powered study. Case-control study. Two private physiotherapy clinics in Australia and Spain. Forty-four recreational male runners with AT and 44 healthy controls matched by age, height, and weight. Demographics, activity (IPAQ-SF), pain and function (VISA-A), pain during hopping (Hop pain VAS), hopping duration, psychological factors (TSK-11, PASS20), and physical tests regarding lower-limb maximal strength and endurance. Body mass index (BMI), activity, VISA-A, pain, and duration of hopping, TSK-11, PASS20, standing heel raise to failure, seated heel raise and leg extension 6RM, hip extension and abduction isometric torque were significantly different between groups (P < 0.05) with varied effect sizes (V = 0.22, d range = 0.05-4.18). 46% of AT severity variance was explained by higher BMI (β = -0.41; p = 0.001), weaker leg curl 6RM (β = 0.32; p = 0.009), and higher pain during hopping (β = -0.43; p = 0.001). Runners with AT had lower activity levels, lower soleus strength, and were less tall. BMI, pain during hopping, and leg curl strength explained condition severity. This information, identified with clinically applicable tools, may guide clinical assessment, and inform intervention development.


Impact of adjuvant treatment modalities on survival outcomes in curatively resected pancreatic and periampullary adenocarcinoma.
We examined the impact of adjuvant modalities on resected pancreatic and periampullary adenocarcinoma (PAC). A total of 563 patients who were curatively resected for PAC were retrospectively analyzed between 2003 and 2013. Of 563 patients, 472 received adjuvant chemotherapy (CT) alone, chemoradiotherapy (CRT) alone, and chemoradiotherapy plus chemotherapy (CRT-CT) were analyzed. Of the 472 patients, 231 were given CRT-CT, 26 were given CRT, and 215 were given CT. The median recurrence-free survival (RFS) and overall survival (OS) were 12 and 19 months, respectively. When CT and CRT-CT groups were compared, there was no significant difference with respect to both RFS and OS, and also there was no difference in RFS and OS among CRT-CT, CT and CRT groups. To further investigate the impact of radiation on subgroups, patients were stratified according to lymph node status and resection margins. In node-positive patients, both RFS and OS were significantly longer in CRT-CT than CT. In contrast, there was no significant difference between groups when patients with node-negative disease or patients with or without positive surgical margins were considered. Addition of radiation to CT has a survival benefit in patients with node-positive disease following pancreatic resection.


Correlation between secosteroid-induced vitamin D receptor activity in melanoma cells and computer-modeled receptor binding strength.
To define the interaction of novel secosteroids produced by the action of cytochrome P450scc with vitamin D receptor (VDR), we used a human melanoma line overexpressing VDR fused with enhanced green fluorescent protein (EGFP) and tested the ligand induced translocation of VDR from the cytoplasm to the nucleus. Hydroxyderivatives of vitamin D(3) with a full length (D(3)) side chain and hydroxy-secosteroids with a shortened side chain (pD) stimulated VDR translocation and inhibited proliferation, however, with different potencies. In general the D(3) were more potent than pD analogues. Molecular modeling of the binding of the secosteroids to the VDR genomic binding pocket (G-pocket) correlated well with the experimental data for VDR translocation. In contrast, docking scores for the non-genomic binding site of the VDR were poor. In conclusion, both the length of the side chain and the number and position of hydroxyl groups affect the activation of VDR by novel secosteroids.


Staged Revision Hip Arthroplasty With Femoral Impaction Bone Grafting for Prosthetic Joint Infections: Radiostereometric Analyses and Clinical Outcomes at Minimum 5-Year Follow-Up.
There are ongoing concerns regarding the use of bone graft following prosthetic joint infection and subsequent implant subsidence. The aim of this study was to determine whether the use of a cemented stem combined with femoral impaction bone grafting (FIBG) at second stage revision for infection results in stable femoral stem fixation, determined by accurate methods, and good clinical results. A prospective cohort of 29 patients underwent staged revision total hip arthroplasty for infection using an interval prosthesis followed by FIBG at the final reconstruction. The mean follow-up was 89 months (range, 8 to 167 months). Femoral implant subsidence was measured with radiostereometric analysis. Clinical outcomes included the Harris Hip Score, Harris Pain score and Société Internationale de Chirurgie Orthopédique et de Traumatologie activity scores. At 2-years follow-up the median stem subsidence relative to femur was -1.36 mm (range, -0.31 to -4.98), while the cement subsidence relative to femur was -0.05 mm (range, 0.36 to -0.73). At 5-years follow-up, the median stem subsidence relative to femur was -1.89 mm (range, -0.27 to -6.35), while the cement subsidence relative to femur was -0.06 mm (range, 0.44 to -0.55). There were 25 patients who were confirmed infection-free after the second stage revision with FIBG. The median Harris Hip Score improved from 51 pre-operatively to 79 at 5 years (P = .0130), and Harris Pain score from 20 to 40 (P = .0038). Stable femoral component fixation can be achieved with FIBG when reconstructing the femur after revision for infection without compromising infection cure rates and patient-reported outcomes.


Experimental osteonecrosis induced by a combination of low-dose lipopolysaccharide and high-dose methylprednisolone in rabbits.
The pathogenetic mechanisms involved in steroid-induced osteonecrosis are poorly understood. Appropriate experimental models of the human disease are indispensable to the understanding of successful treatment modalities for osteonecrosis. In the present experiment we devised a novel rabbit model of steroid-induced osteonecrosis by use of two low-dose LPS and three high-dose MPS to investigate the development of osteonecrosis. Thirty eight rabbits were used and tissue assessments were performed on proximal third and distal condyles of femora and humeri obtained 6 weeks after the administration of LPS and MPS. MRI of these regions and intraosseous pressure of proximal femur were obtained at 0 and 6 weeks. Other assessments included serum plasminogen activator/inhibitor ratio, cholesterol level, LDL/HDL ratio, and triglyceride levels at various time points. The study showed that with this osteonecrosis induction protocol there was low animal mortality (6.2%), high rate of osteonecrosis (90%), induction of thrombotic state, and hypercholesterol/lipidemia. On the whole, this is a novel modified animal model of steroid associated osteonecrosis and it would be useful for elucidating the pathogenesis of steroid associated osteonecrosis and developing preventive and therapeutic strategies.


Investigation of the oral microbiome of children associated with dental caries: A systematic study.
The present study aims to investigate the variations in dental caries (DC) related microbiome abnormality and metabolomics shift in children. The patients were divided into two groups healthy control (C) and highly affected DC children based on inclusion and exclusion criteria. Saliva samples were collected and used for the taxonomic and functional characterization of oral microbiota. Metatranscriptomics analysis revealed the alterations and composition of oral microbiota in the C and DC groups. Relative abundance in the C group was associated with Firmicutes, Actinobacteria, and Bacteroidetes. Whereas, the microbial composition in the DC group was found to be considerably altered with increases in the abundance of the Proteobacteria (25%), Fusobacteria (15%), and Cyanobacteria (8%) while decreases in the abundance of Firmicutes (10%) and Bacteroidetes (23%). Alterations in the phylum composition were positively and negatively correlated with several metabolites of sugars (such as fructose, sorbose, ribose, allose, and mannose) and amino acids (such as arginine, lysine, tryptophan, and proline). Moreover, in comparison with the C group, the metabolic shift of the DC group was different with an increase in certain tricarboxylic acid cycle intermediates levels, and a decrease in fatty acid. Such alterations can enhance the growth of oral pathogens and contribute to DC development. The findings of this study suggest that an altered abundance of Actinobacillus, Fusobacterium, and Shuttleworthia can serve as biomarkers of DC in children.


Evaluation of the Sensitivity of the Nipple-Areola Complex in Patients Undergoing Breast Ptosis Correction with Periareolar Dermis Release: A Randomized Controlled Trial.
In the surgical treatment of breast ptosis, the plastic surgeon's main challenge is to mobilize the nipple-areola complex (NAC) over long distances, preserving its sensitivity. Herein, we aimed to evaluate the NAC sensitivity following a periareolar dermis release in patients undergoing surgical correction of breast ptosis. This is a prospective, double-blinded, randomized study in which 39 patients (78 breasts) were operated on for the treatment of breast ptosis. Patients' breasts were divided into two groups, each breast in a group. In breasts of the experiment group, the periareolar dermis release was used after decortication. This maneuver was not used in the control group's breasts. To analyze the sensitivity in relation with the distance that the NAC should be mobilized to correct breast ptosis, the value of 6 cm was used as boundary. Sensitivity of the NAC was assessed with monofilaments preoperatively and postoperatively-at three and six weeks and one year. Regarding the sensitivity of the areola, a significant difference was observed in the comparison between the control and experiment groups at the NAC mobility ≤ 6.0 cm in the preoperative. Regarding the nipple, a significant difference was observed in the comparison between the control and experiment groups at three weeks postoperatively for a NAC mobility ≤ 6.0 cm. After one year, there was no difference in the comparison between the groups in either areola or nipple. The periareolar dermis release does not compromise NAC vascularization and pressure sensitivity in patients undergoing breast ptosis correction. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .


Determination of propofol glucuronide from hair sample by using mixed mode anion exchange cartridge and liquid chromatography tandem mass spectrometry.
The main objective of this study was to develop and validate a simpler and less time consuming analytical method for determination of propofol glucuronide from hair sample, by using mixed mode anion exchange cartridge and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The study uses propofol glucuronide, a major metabolite of propofol, as a marker for propofol abuse. The hair sample was digested in sodium hydroxide solution and loaded in mixed-mode anion cartridge for solid phase extraction. Water and ethyl acetate were used as washing solvents to remove interfering substances from the hair sample. Consequently, 2% formic acid in ethyl acetate was employed to elute propofol glucuronide from the sorbent of mixed-mode anion cartridge, and analyzed by LC-MS/MS. The method validation parameters such as selectivity, specificity, LOD, LLOQ, accuracy, precision, recovery, and matrix effect were also tested. The linearity of calibration curves showed good correlation, with correlation coefficient 0.998. The LOD and LLOQ of the propofol glucuronide were 0.2 pg/mg and 0.5 pg/mg, respectively. The intra and inter-day precision and accuracy were acceptable within 15%. The mean values of recovery and matrix effect were in the range of 91.7-98.7% and 87.5-90.3%, respectively, signifying that the sample preparation, washing and extraction procedure were efficient, and there was low significant hair matrix effect for the extraction of propofol glucuronide from hair sample on the mixed mode anion cartridge. To evaluate the suitability of method, the hair of propofol administered rat was successfully analyzed with this method.


Obesity Increases Disease Activity of Norwegian Patients with Axial Spondyloarthritis: Results from the European Map of Axial Spondyloarthritis Survey.
To investigate the prevalence of overweight and obesity, as well as the association between body mass index (BMI) and disease activity in patients with axial spondyloarthritis (axSpA). Norwegian axSpA patients from the European Map of Axial Spondyloarthritis (EMAS) survey were included in this analysis. Sociodemographic, anthropomorphic, and disease-related variables (HLA-B27, comorbidities, BASDAI, and self-reported spinal stiffness) were reported. Patients were categorized into under/normal weight (BMI < 25 kg/m2), overweight (BMI ≥ 25 to < 30 kg/m2), and obese (≥ 30 kg/m2). Of the 509 participants in the EMAS survey, 35% were categorized as under/normal weight, 39% overweight, and 26% obese. Compared to under/normal-weight patients, overweight patients had significantly higher degree of spinal stiffness (mean (SD) 7.91 ± 2.02 vs 7.48 (2.15) and number of comorbidities (2.45 ± 2.11, vs 1.94), both p < 0.001. Obese patients had significantly higher disease activity (BASDAI mean (SD) 5.87 ± 1.78 vs 4.99 ± 2.08, p < 0.001), degree of spinal stiffness (8.18 ± 2.03 vs 7.48 ± 2.15, p = 0.006), and number of comorbidities (3.43 ± 2.43 vs 1.94. ± .38, p < 0.001) than under/normal weight patients. After adjusting for gender and age, obesity proved to be independently associated with disease activity. Obesity was associated with higher reported BASDAI score, and being overweight or obese was associated with a higher degree of spinal stiffness and number of comorbidities compared to under/normal weight respondents. The results highlight the serious impact of obesity on health status, and obesity should therefore be considered as a modifiable risk factor for disease activity within the disease management of axSpA.


Prognostic significance of metabolic tumour volume and maximum standard uptake value of fluor-18-fluorodeoxyglucose positron emission tomography with computed tomography in nasopharyngeal carcinoma.
To evaluate the prognostic role of markers of fluor-18-fluorodeoxyglucose positron emission tomography with computed tomography (18F-FDG-PET-CT), such as maximum standard uptake value (SUVmax) and metabolic tumour volume (MTV) measured at primary and nodal disease, and their clinical significance in terms of predicting treatment outcomes and survival. Between January 2017 and January 2020, 20 case records of nasopharyngeal carcinoma patients who underwent 18F-FDG-PET-CT as part of staging workup before radiotherapy and as a part of response evaluation after radiotherapy were retrospectively reviewed. At a median follow-up of 34.7 months, the 2-year progression-free survival (PFS) was 70% and 2-year overall survival (OS) was 79%. Patients with a lower nodal SUVmax (SUVmax-N) had a better 2-year PFS (91% vs. 46%; p = 0.035) and 2-year OS (95% vs. 58%; p = 0.015). A high SUVmax-N of > 10.58 was a negative predictor of OS (95% confidence interval [CI]: 0.93-1; p = 0.003) as well as PFS (95% CI: 0.64-1; p = 0.017). Also, a high MTV > 25.8 cm3 was a negative predictor of PFS (95% CI: 0.58-0.98; p = 0.048). MTV was an independent predictor of PFS and OS on univariate analysis, whereas it was not significant in the Cox regression multivariate analysis. High values of MTV and SUVmax-N can be considered as independent prognostic factors of OS and PFS in nasopharyngeal cancer patients treated with concurrent chemoradiation, highlighting the need for more intensified treatment.


Antitumor and immunomodulatory activity of a polysaccharide from fungus Coprinus comatus (Mull.:Fr.) Gray.
In our study, a water-soluble polysaccharide (CCPa-1) was successfully purified from the fruiting bodies of Coprinus comatus by DEAE-cellulose and Sepharose CL-6B column chromatography. The molecular weight was evaluated to be 53.6 kDa as determined by high performance size exclusion chromatography (HPSEC). Sugar composition analysis revealed that CCPa-1 consisted primarily of galactose, glucose and arabinose in a molar ratio of 6.6:1.2:2.2. CCPa-1 could not only inhibit the growth of sarcoma 180 (S180) tumor transplanted in mice, but also increase the relative spleen/thymus indexes and body weight of tumor bearing mice. Moreover, Con A- or LPS-induced splenocyte proliferation was also enhanced after CCPa-1 administration in tumor-bearing mice. Furthermore, CCPa-1 significantly enhanced the Con A- or LPS-induced splenocyte proliferation and increased the production of TNF-α and IL-2. All the data demonstrated that CCPa-1 had a potential application as natural antitumor agent with immunomodulatory activity.


Establishment and application of suspension static method in blood group screening of automated blood group analyzer.
The accuracy of blood group identification is the basis of blood transfusion safety. In order to increase the detection rate of weak agglutination, unexpected antibodies (UAb) and blood subtypes for pre-transfusion testing, the blood group screening process of automated blood group analyzer (ABGA) is ameliorated by introducing one static step and establishing a suspension static method (SSM). One static step was introduced in the blood group screening process of ABGA and three static time conditions were designed: 300 s, 400 s and 500 s, from which the optimal static time was selected and SSM was established; By comparing the detection of weak agglutination and UAb before and after the application of SSM, the feasibility and effect of suspension static method were verified and evaluated. The last two steps of the automatic blood group screening process were replaced with static, light centrifugation and imaging. The optimal static time parameter was selected as 400 s and SSM was established; After the application of SSM, it was verified that: (1) The detection level of weak antibodies (anti-A and anti-B) and weak antigens (weak D phenotype) could be improved by SSM, including antibodies in plasma of known type O samples with 0, 2, 4, 8, 16 and 32 times serial dilutions(simulating weak anti-A and weak anti-B), weak antibodies (anti-B) in plasma of one normal A-type sample and weak antigens on red blood cells (RBC) of 5 weak D phenotype samples (weak D antigen); (2) Three blood donor samples (type A, O and B) with known UAb were detected by SSM. The results showed that SSM could detect both weak antibodies (anti-A and anti-B) and UAb; (3) SSM was applied to detect the samples of 3 A2B and 3 subtype B blood donors and the blood subtypes could be clearly detected; (4) The number of screening samples was 95,314 and 106,814 before SSM (2018) and after (2020) the application of SSM and the positive rate of UAb (63/95,314 and 187/106,814) increased after SSM, discrepancy of which was statistically significant (χ2 = 48.42, P < 0.01). The above results demonstrate that SSM of ABGA is conducive to the detection of weak agglutination, UAb and blood subtypes in blood samples, which can improve the sensitivity of blood group detection and ensure the safety of clinical blood transfusion to a certain extent.


EIF4A3-induced circular RNA PRKAR1B promotes osteosarcoma progression by miR-361-3p-mediated induction of FZD4 expression.
Emerging evidence indicates that circRNAs are broadly expressed in osteosarcoma (OS) cells and play a crucial role in OS progression. Recently, cancer-specific circRNA circPRKAR1B has been identified by high-throughput sequencing and is recorded in publicly available databases. Nevertheless, the detailed functions and underlying mechanisms of circPRKAR1B in OS remains poorly understood. By functional experiments, we found that circPRKAR1B enhanced OS cell proliferation, migration, and promotes OS epithelial-mesenchymal transition (EMT). Mechanistic investigations suggested that circPRKAR1B promotes OS progression through sponging miR-361-3p to modulate the expression of FZD4. Subsequently, we identified that EIF4A3 promoted cirPRKAR1B formation through binding to the downstream target of circPRKAR1B on PRKAR1B mRNA. Further rescue study revealed that overexpression of the Wnt signalling could impair the onco-suppressor activities of the silencing of circPRKAR1B. Interestingly, further experiments indicated that circPRKAR1B is involved in the sensitivity of chemoresistance in OS. On the whole, our results demonstrated that circPRKAR1B exerted oncogenic roles in OS and suggested the circPRKAR1B/miR-361-3p/FZD4 axis plays an important role in OS progression and might be a potential therapeutic target.


Companion Diagnostics and Predictive Biomarkers for MET-Targeted Therapy in NSCLC.
Dysregulation of the MET tyrosine kinase receptor is a known oncogenic driver, and multiple genetic alterations can lead to a clinically relevant oncogenesis. Currently, a number of drugs targeting MET are under development as potential therapeutics for different cancer indications, including non-small cell lung cancer (NSCLC). However, relatively few of these drugs have shown sufficient clinical activity and obtained regulatory approval. One of the reasons for this could be the lack of effective predictive biomarkers to select the right patient populations for treatment. So far, capmatinib is the only MET-targeted drug approved with a companion diagnostic (CDx) assay, which is indicated for the treatment of metastatic NSCLC in patients having a mutation resulting in MET exon 14 skipping. An alternative predictive biomarker for MET therapy is MET amplification, which has been identified as a resistance mechanism in patients with EGFR-mutated NSCLC. Results obtained from different clinical trials seem to indicate that the MET/CEP7 ratio detected by FISH possesses the best predictive properties, likely because this method excludes MET amplification caused by polysomy. In this article, the concept of CDx assays will be discussed, with a focus on the currently FDA-approved MET targeted therapies for the treatment of NSCLC.


Continuous fluorescence anisotropy-based assay of BOCILLIN FL penicillin reaction with penicillin binding protein 3.
We report a simple, rapid, and reproducible fluorescence anisotropy-based method for measuring rate constants for acylation and deacylation of soluble penicillin binding protein (PBP) constructs by compounds in microtiter plates by means of competition with time-dependent acylation by BOCILLIN FL. The method is demonstrated by measuring the acylation rate constants of the PBP3 periplasmic domains from Pseudomonas aeruginosa and Acinetobacter baumannii by BOCILLIN FL, aztreonam, meropenem, and ceftazidime. The new method requires very little protein and can be completed in approximately 1h per compound. A set of BOCILLIN FL acylation progress curves collected over a range of competitor concentrations is fit globally to a kinetic model by numerical integration. First-order deacylation rate constants could also be measured, as demonstrated with a catalytically impaired mutant OXA-10 β-lactamase.


The Role of Probiotics in Chronic Rhinosinusitis Treatment: An Update of the Current Literature.
Chronic rhinosinusitis (CRS) is a significant health problem. It affects 5-12% of the general population. The causes that underlie the onset of CRS are not yet well known. However, many factors may contribute to its onset, such as environmental factors and the host's general condition. Medical treatment mainly uses local corticosteroids, nasal irrigation, and antibiotics. In recent years, a new therapeutic approach that employs the use of probiotics emerged. Probiotics have been extensively studied as a therapy for dysbiosis and inflammatory pathologies of various parts of the body. We aimed to examine the studies in vivo and in vitro and clinicals reports in the existing literature to update probiotics' role in rhinosinusitis chronic medical treatment.


Radiosynthesis, in vitro cellular uptake and in vivo biodistribution of 3'-O-(3-benzenesulfonylfuroxan-4-yl)-5-[125I]iodo-2'-deoxyuridine, a nucleoside-based nitric oxide donor.
3'-O-(3-Benzenesulfonylfuroxan-4-yl)-5-iodo-2'-deoxyuridine (1) is a cytotoxic nitric oxide (NO) donor-nucleoside dual action prodrug designed to exploit both NO and an antimetabolite nucleoside for cancer therapy. 1 was radiolabeled by radioiodide exchange and purified by HPLC in 16% overall radiochemical yield. The specific activity of [(125)I]1 was 31.8 microCi/mug (680 MBq/microM). Protein binding, deiodination, cellular uptake and incorporation of 1 into cellular nucleic acids were measured by standard methods, and its in vivo biodistribution was determined in Balb/c mice bearing implanted EMT-6 tumors following intravenous injection. [(125)I]1 degraded rapidly during the in vitro tests, thus impeding unequivocal assessment but indicating that it was only weakly protein bound and that it was resistant to deiodination under test conditions. Uptake of [(125)I]1 by murine tumor cells (KBALB and KBALB-STK) in vitro was low (approximately 17 fmol/microg protein over 2 h) with only approximately 0.3% (0.04-0.06 fmol/microg protein) of total uptake present in the DNA fraction. In the murine tumor model, liver, kidney, intestine and tumor showed the greatest uptake, with liver, intestine and blood all containing >5 injected dose per gram of tissue (%ID/g) during the 15-min to 2-h postinjection period. Maximum tissue/blood level ratios were 3.6 (2 h) for tumor and 6.4 (24 h) for liver. Low uptake in thyroid and stomach was indicative of minimal in vivo deiodination. [(125)I]1 undergoes only minimal deoiodination under both in vitro and in vivo conditions. Under conditions of the in vitro NO release assay, 1 reacts to produce a single, major, unstable adduct that decomposes upon workup. Protein binding of [(125)I]1 could not be assessed because of similar chemical reaction with albumin. Incorporation of radioactivity into the cellular nucleic acid fraction was low, and in vivo distribution of [(125)I]1 was consistent with nonspecific reactivity towards tissue nucleophiles. The chemical reactivity of [(125)I]1 mitigates against its use as a NO donor and as a tracer for this class of compounds.


Use of alpha-tocopherol esters for topical vitamin E treatment: evaluation of their skin permeation and metabolism.
The aim of this work was to investigate new pro-vitamins based on α-tocopherol (α-Toc) and fatty acids, and to compare their properties with those of α-tocopherol acetate (α-TAc). Skin levels of α-Toc-fatty acid ester conjugates, total α-Toc and endogenous α-Toc were measured in skin samples taken from separate groups of treated and untreated rats. Multiple and extensive treatment with α-Toc oleate and α-TAc was also carried out to assess the skin accumulation and safety of these esters. The in-vivo studies revealed that α-Toc-fatty acid conjugates penetrated into the skin quantitatively while being comparable with the permeation of α-TAc. Differences were found between the levels of total α-Toc and endogenous α-Toc after application of α-TAc, α-Toc oleate, α-Toc linoleate, α-Toc-α linolenate and α-Toc palmitate, indicating that α-Toc conjugates of these fatty acids, but not α-Toc γ-linolenate or α-Toc stearate, were hydrolysed to free α-Toc. In long-term and extensive treatment, α-TAc was found to be lethal to rats treated with 1.15 mg/kg of this agent, which had been spread over 16 cm(2) of skin. Similar treatment with α-Toc oleate did not produce any side effects. This study suggests that α-Toc conjugates with unsaturated fatty acids may be a good alternative as stable vitamin E derivatives, rather than the α-TAc ester.


Evaluation of a series of phenyl-type stationary phases in supercritical fluid chromatography with the linear solvation energy relationship model and its application to the separation of phenolic compounds.
In recent years, supercritical fluid chromatography (SFC) has become a powerful tool in modern analytical chemistry, and the diversity of stationary phases in SFC promotes phenyl-type phases to confront with a significant resurgence of interest. In this paper, a series of phenyl-type stationary phases with different substituted benzenes involving N-propylbenzamide (PB), 4-fluoro-N-propylbenzamide (PB-F), and 4-ethyl-N-propylbenzamide (PB-ET) were synthesized. Retention mechanism of these phases in SFC was investigated using a linear solvation energy relationship (LSER) model. The phenyl-type stationary phases with all positive parameters can provide all types of interaction, typically involving hydrogen bonding, dipole-dipole and dispersive interactions. The different benzene's substituents of the stationary phases mainly affected their hydrogen bonding and dipole-dipole interactions, which could be reflected by the angle between the solvation vectors to some extent. The k-k plot showed that the selectivity difference of phenyl-type stationary phases was closely related to the type of solute. Thus, based on twenty-five natural phenolic compounds, two systems with high orthogonality (63.49%) were constructed using three columns, namely phenyl column (PHE) × PB-F and PB × PB-F. Finally, after investigating the influence of chromatographic conditions, ten flavonoids could be separated by using PB, PB-F and PHE columns in SFC.


Performance and microbial diversity of palm oil mill effluent microbial fuel cell.
To evaluate the bioenergy generation and the microbial community structure from palm oil mill effluent using microbial fuel cell. Microbial fuel cells enriched with palm oil mill effluent (POME) were employed to harvest bioenergy from both artificial wastewater containing acetate and complex POME. The microbial fuel cell (MFC) showed maximum power density of 3004 mW m(-2) after continuous feeding with artificial wastewater containing acetate substrate. Subsequent replacement of the acetate substrate with complex substrate of POME recorded maximum power density of 622 mW m(-2). Based on 16S rDNA analyses, relatively higher abundance of Deltaproteobacteria (88.5%) was detected in the MFCs fed with acetate artificial wastewater as compared to POME. Meanwhile, members of Gammaproteobacteria, Epsilonproteobacteria and Betaproteobacteria codominated the microbial consortium of the MFC fed with POME with 21, 20 and 18.5% abundances, respectively. Enriched electrochemically active bacteria originated from POME demonstrated potential to generate bioenergy from both acetate and complex POME substrates. Further improvements including the development of MFC systems that are able to utilize both fermentative and nonfermentative substrates in POME are needed to maximize the bioenergy generation. A better understanding of microbial structure is critical for bioenergy generation from POME using MFC. Data obtained in this study improve our understanding of microbial community structure in conversion of POME to electricity.


Transcriptomic and metabolomic perspectives for the growth of alfalfa (Medicago sativa L.) seedlings with the effect of vanadium exposure.
Hitherto, the effect of vanadium on higher plant growth remains an open topic. Therefore, nontargeted metabolomic and RNA-Seq profiling were implemented to unravel the possible alteration in alfalfa seedlings subjected to 0.1 mg L-1 (B group) and 0.5 mg L-1 (C group) pentavalent vanadium [(V(V)] versus control (A group) in this study. Results revealed that vanadium exposure significantly altered some pivotal transcripts and metabolites. The number of differentially expressed genes (DEGs) markedly up- and down-regulated was 21 and 23 in B_vs_A, 27 and 33 in C_vs_A, and 24 and 43 in C_vs_B, respectively. The number for significantly up- and down-regulated differential metabolites was 17 and 15 in B_vs_A, 43 and 20 in C_vs_A, and 24 and 16 in C_vs_B, respectively. Metabolomics and transcriptomics co-analysis characterized three significantly enriched metabolic pathways in C_vs_A comparing group, viz., α-linolenic acid metabolism, flavonoid biosynthesis, and phenylpropanoid biosynthesis, from which some differentially expressed genes and differential metabolites participated. The metabolite of traumatic acid in α-linolenic acid metabolism and apigenin in flavonoid biosynthesis were markedly upregulated, while phenylalanine in phenylpropanoid biosynthesis was remarkably downregulated. The genes of allene oxide cyclase (AOC) and acetyl-CoA acyltransferase (fadA) in α-linolenic acid metabolism, and chalcone synthase (CHS), flavonoid 3'-monooxygenase (CYP75B1), and flavonol synthase (FLS) in flavonoid biosynthesis, and caffeoyl-CoA O-methyltransferase (CCoAOMT) in phenylpropanoid biosynthesis were significantly downregulated. While shikimate O-hydroxycinnamoyltransferase (HCT) in flavanoid and phenylpropanoid biosynthesis were conspicuously upregulated. Briefly, vanadium exposure induces a readjustment yielding in metabolite and the correlative synthetic precursors (transcripts/unigenes) in some branched metabolic pathways. This study provides a practical and in-depth perspective from transcriptomics and metabolomics in investigating the effects conferred by vanadium on plant growth and development.


Antioxidant properties of pioglitazone limit nicotinamide adenine dinucleotide phosphate hydrogen oxidase and augment superoxide dismutase activity in cardiac allotransplantation.
Many non-immunologic factors contribute to the development of cardiac allograft vasculopathy (CAV), chief among them being ischemia-reperfusion injury associated with oxidative stress. We hypothesized that pioglitazone, a peroxisome proliferator-activated receptor (PPAR)-γ agonist, could attenuate graft oxidant stress in cardiac transplantation. Experiments were performed using a murine heterotopic cardiac allotransplantation model. Pioglitazone was administered to recipients once daily, beginning 1 day before transplantation. At 4 hours after transplantation, pioglitazone significantly reduced the expression of endothelial cell adhesion receptors and infiltration of polymorphonuclear leukocytes (PMNs). The anti-oxidant balance in pioglitazone-treated cardiac allografts was significantly bolstered by reduced nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) oxidase (Nox1 and p22(phox) sub-units) activity and preservation of manganese superoxide dismutase (SOD) activity, resulting in the mitigation of oxidative damage at the level of lipids, proteins, and DNA. At 7 days after transplantation, PPAR-γ was significantly up-regulated by pioglitazone, but nuclear factor-κB and inducible nitric oxide synthase were significantly down-regulated. A concomitant reduction of inflammatory cytokines and chemokines and graft leukosequestration was noted. Pioglitazone consequently prolonged cardiac allograft survival and attenuated CAV development. In vitro experiments demonstrated that pioglitazone decreased transendothelial PMN migration, NADPH oxidase activity, and loss of SOD activity in PMNs and endothelial cells. Pioglitazone can suppress the oxidative stress and damage and can stimulate antioxidant capacity in cardiac allografts after transplantation. Mitigation of graft oxidant stress could be an important mechanism through which pioglitazone confers benefit after cardiac transplantation.


Nucleotide sequence of the gene coding for ovine corticotropin-releasing factor and regulation of its mRNA levels by glucocorticoids.
The ovine gene CRF, coding for corticotropin-releasing factor, has been isolated and the nucleotide sequence determined. The degree of nucleotide sequence homology between the ovine and human CRF genes is unusual, in that the 5' flanking regions are more highly conserved than the protein-coding regions. This striking degree of homology would indicate that a strong selective pressure is being exerted over an extensive area of the 5' flanking region, which could include transcriptional control elements. The 5' flanking region of the ovine CRF gene contains five elements which share homology with the glucocorticoid receptor DNA binding sequence. Also Northern blot analysis indicates that hypothalamic CRF mRNA levels are negatively regulated by glucocorticoids. Dexamethasone treatment halves the CRF mRNA content of the hypothalamus, whereas adrenalectomy causes a three- to four-fold increase in CRF mRNA levels.


Glomerular charge alterations in human minimal change nephropathy.
A theoretical model of charge and size selectivity for the glomerulus has been applied to human data. Using previously published values for GFR, renal plasma flow, systemic oncotic pressure, and fractional clearances of neutral dextrans, albumin, salivary amylase, and transferrin, membrane parameters describing the glomerular barrier were determined for normal individuals under control conditions and during lysine infusion (which retards tubule protein reabsorption), and for patients with minimal change nephropathy (MCN). To permit the estimation of membrane charge from fractional clearances, molecular charge values for human transferrin (-9.4 Eq/mole) and human salivary amylase (-4.1) were determined by measuring electrophoretic mobilities of these proteins in polyacrylamide gels. Assuming no large changes in the transmural hydraulic pressure difference (delta P), the glomerular ultrafiltration coefficient (Kf, the product of hydraulic permeability and capillary surface area) was calculated to be reduced by greater than 50% in MCN. The effective pore radius (approximately 55 A) is virtually unaltered in MCN, suggesting that the decline in Kf is due to a reduced number of pores. The degree of albuminuria observed in MCN is attributable to an approximately 50% reduction in the concentration of fixed negative charges in the glomerular capillary wall. The concentrations of fixed charges calculated from albumin data in normal individuals (140 to 160 mEq/liter) and in patients with MCN (60 to 90 mEq/liter) are insensitive to the assumed values of delta P.


Melanocytes in the nasal cavity and paranasal sinus. Incidence and distribution in Japan.
Random, nonselected tissue specimens from 99 Japanese-20 cylindrically cut nasal blocks removed during autopsy (A.C., Autopsy Cases) and 79 cases removed during surgery, consisting of 32 chronic sinusitis cases (C.S.) and 47 nasal polyp cases (N.P.)-were examined histopathologically and electronmicroscopically with respect to distribution and frequency of melanocytes in the nasal cavity and paranasal sinus. Malignant melanoma cases were excluded. The overall incidence of positive cases for melanocytes revealed 21.2% (21 of 99 cases), with an incidence ratio of male to female of 0.9:1.0. Melanocytes were found beginning in the under 19 age group with incidence increasing proportionately with age. Peak incidence was in the 50-year age group at 50%. Melanocytes and melanotic cell foci were distributed in the stroma of the propria mucosa beneath the pseudostratified columnar epithelium and focused around the nasal and paranasal glands and sinuses. In 2 of the 21 cases positive for stromal melanocytes, intraepithelial melanocytes with dendritic processes were found. The histogenesis of malignant melanoma arising from the nasal cavity and paranasal sinus are also discussed in this study.


Jobelyn® improves motor dysfunctions induced by haloperidol in mice via neuroprotective mechanisms relating to modulation of cAMP response-element binding protein and mitogen-activated protein kinase.
The clinical efficacy of haloperidol in the treatment of psychosis has been limited by its tendency to cause parkinsonian-like motor disturbances such as bradykinesia, muscle rigidity and postural instability. Oxidative stress-evoked neuroinflammation has been implicated as the key neuropathological mechanism by which haloperidol induces loss of dopaminergic neurons and motor dysfunctions. This study was therefore designed to evaluate the effect of Jobelyn® (JB), an antioxidant supplement, on haloperidol-induced motor dysfunctions and underlying molecular mechanisms in male Swiss mice. The animals were distributed into 5 groups (n = 8), and treated orally with distilled water (control), haloperidol (1 mg/kg) alone or in combination with each dose of JB (10, 20 and 40 mg/kg), daily for 14 days. Thereafter, changes in motor functions were evaluated on day 14. Brain biomarkers of oxidative stress, proinflammatory cytokines (tumor necrosis factor-alpha and interleukin-6), cAMP response-element binding protein (CREB), mitogen-activated protein kinase (MAPK) and histomorphological changes were also investigated. Haloperidol induces postural instability, catalepsy and impaired locomotor activity, which were ameliorated by JB. Jobelyn® attenuated haloperidol-induced elevated brain levels of MDA, nitrite, proinflammatory cytokines and also boosted neuronal antioxidant profiles (GSH and catalase) of mice. It also restored the deregulated brain activities of CREB and MAPK, and reduced the histomorphological distortions as well as loss of viable neuronal cells in the striatum and prefrontal cortex of haloperidol-treated mice. These findings suggest possible benefits of JB as adjunctive remedy in mitigating parkinsonian-like adverse effects of haloperidol through modulation of CREB/MAPK activities and oxidative/inflammatory pathways.


[Pharmacology of Janus kinase inhibitors].
Modern dermatotherapy is dominated by the development of various biologicals and small molecules. Janus kinase inhibitors (JAKi) form a novel class of small molecular synthetic compounds inhibiting the intracellular signal transduction of cytokine receptors. Cytokines are key mediators in the pathophysiology of numerous inflammatory skin diseases. Many cytokines use so-called type I and II cytokine receptors, which associate with the Janus kinases JAK1, JAK2, JAK3 or TYK2. JAKi are under clinical investigation for inflammatory skin disease, specifically in phase 3 trials for psoriasis or atopic dermatitis. Since JAKi are tested in oral as well as in topical formulations, they could become very popular in dermatotherapy. The mechanisms of JAKi, their selectivity, preliminary efficacy data, and their safety profile are discussed in this article.


Early pharmacokinetics and clinical effects of oral D-amphetamine in normal subjects.
Seven normal subjects received 0.25 mg/kg D-amphetamine orally, both after an overnight fast and again after a standard breakfast. Plasma levels, subjective and cardiovascular effects, and observer-rated activation were assessed hourly for 5 hr. Food did not affect amphetamine levels. Plasma levels peaked at 2-3 hr. Maximum cardiovascular effects generally occurred at 1 hr, whereas maximum behavioral and subjective effects occurred at 2 hr. Subjective and behavioral effects declined thereafter, in spite of substantial amphetamine levels. A separate group of 8 subjects received 0.5 mg/kg D-amphetamine orally. Plasma levels, subjective and cardiovascular effects, and activation ratings were assessed hourly for 4 hr. Maximum plasma levels were approximately twice those seen in the first group. In this case, plasma levels peaked at 3-4 hr; blood pressure and subjective and behavioral effects were all maximal at 2-3 hr and were declining by 4 hr, in spite of stable or rising plasma levels.


An integrated plasma and urinary metabonomic study using UHPLC-MS: intervention effects of Epimedium koreanum on 'Kidney-Yang Deficiency syndrome' rats.
A metabonomic strategy based on UHPLC-MS with principal component analysis was developed to investigate the intervention effects of Epimedium koreanum on metabolism characters of 'Kidney-Yang Deficiency syndrome' rats. The rats were injected intraperitoneally hydrocortisone once daily for 15 days to simulate 'Kidney-Yang Deficiency syndrome' and then administered orally E. koreanum extract once daily for the following 15 days. Plasma and urine samples before hydrocortisone injection, on day 15 of hydrocortisone injection and on days 3, 6, 9, 12, 15 exposed to E. koreanum extract were collected. Significant metabolic disorders were observed in 'Kidney-Yang Deficiency syndrome' rats and sixteen potential biomarkers were identified. The disturbed plasma levels of phenylalanine, tryptophan, cholic acid, lysophosphatidylcholines and urinary levels of phenylalanine, hippurate, phenylacetylglycine, N(2)-succinyl-l-ornithine, creatinine, α-ketoglutarate, citrate, phenol sulfate, indoxyl sulfate, cresol sulfate in model rats were gradually restored to normal after administration of E. koreanum extract, which indicated that E. koreanum had time-dependent recovering effects via regulating oxidant-antioxidant balance, amino acid metabolism, lipid metabolism, energy metabolism, and gut microflora. This work highlights that metabonomics is a promising tool for studying the essence of Chinese medicine's syndrome theory and the action mechanism of traditional Chinese medicine, and provides scientific and reasonable information on safety and efficacy of traditional Chinese medicine.


Comparing transforming growth factor beta-2 and fibronectin as pleurodesing agents.
We have recently demonstrated that transforming growth factor beta-2 (TGF-beta2) can produce effective pleurodesis. Whether this effect can be reproduced by the use of its downstream proteins is not known. This study compared the effectiveness of TGF-beta2 and fibronectin in inducing pleurodesis in rabbits. New Zealand white rabbits (1.5-2.0 kg) were given 1.7 microg of TGF-beta2 (n=5) or 2.0 mg of cellular fibronectin (n=4) intrapleurally via a chest tube. The induced pleural fluid was collected and analyzed. The rabbits were sacrificed after 14 days. The pleurodesis was graded macroscopically from 1 (none) to 8 (symphysis > 50%). All rabbits in the TGF-beta2 group developed effective pleurodesis while none in the fibronectin group had scores > 2 (pleurodesis scores 7.0 +/- 0.6 vs 1.3 +/- 0.3, P < 0.001). Rabbits that received TGF-beta2 produced large amounts of pleural fluid initially (< 4 days). Microscopically, the pleura of rabbits in the TGF-beta2 group showed prominent spindle cell proliferation and collagen deposition, but no significant inflammation or mesothelial proliferation. Pleural tissues of rabbits in the fibronectin group had occasional thin collagen deposits only. The intrapleural administration of 2.0 mg of fibronectin, a downstream product of TGF-beta, did not induce effective pleurodesis, as did the intrapleural administration of TGF-beta2. The present study suggests that the mechanism by which TGF-beta2 induces pleurodesis is not predominantly dependent on the production of fibronectin.


CRH-stimulation of cyclic adenosine 5'-monophosphate pathway is partially inhibited by the coexpression of CRH-R1 and CRH-R2alpha.
Corticotropin-releasing hormone (CRH) is one of the major proteins responsible for brain stress regulation. Two well-known receptors have been described: type 1 and type 2alpha, both members of the receptor superfamily of G protein-coupled receptors (GPCR). We investigated receptor regulation when both CRH receptor subtypes are coexpressed in the same mammalian cell line. When both types of receptors are coexpressed, cAMP second messenger production is partially inhibited compared to when receptors are expressed separately. However, neither binding kinetics nor internalization rates are modified by coexpression of these receptors. To our knowledge this is the first demonstration of receptor interaction that results in the modification of CRH-mediated signal transduction pathway. Because CRH-R1 and CRH-R2alpha have overlapping mRNA expression patterns in the brain, these receptors may be coexpressed in neurons, suggesting that receptor interaction may play an important role in the effect evoked by CRH, contributing to the complexity of differential coupling of the CRH receptors in different endocrine and stress behavior responses.


From concepts to percepts in human and machine face recognition: A reply to Blauch, Behrmann & Plaut.
Intact recognition of familiar faces is critical for appropriate social interactions. Thus, the human face processing system should be optimized for familiar face recognition. Blauch et al. (2020) used face recognition deep convolutional neural networks (DCNNs) that are trained to maximize recognition of the trained (familiar) identities, to model human unfamiliar and familiar face recognition. In line with this model, we discuss behavioral, neuroimaging and computational findings that indicate that human face recognition develops from the generation of identity-specific concepts of familiar faces that are learned in a supervised manner, to the generation of view-invariant identity-general perceptual representations. Face-trained DCNNs seem to share some fundamental similarities with this framework.


Critical aortic stenosis. Survival and management.
The factors associated with survival in 40 neonates (age less than 28 days) with critical aortic stenosis undergoing either open (22 patients) or closed (18 patients) transventricular aortic valvotomy were reviewed. Significant adverse correlates with survival included evidence of poor perfusion preoperatively (low pH, greater than Grade 2/6 soft ejection systolic murmur) and marked congestive heart failure (hepatomegaly, cardiomegaly, elevated left atrial pressure). Congenital mitral stenosis (anulus less than 11 mm), a small aortic anulus (less than 6.5 mm), and failure to achieve an adequate aortic orifice (greater than 6 mm), at operation were identified as factors associated with increased mortality. Initial perioperative survival was better with closed aortic valvotomy. However, there was no significant difference in overall operative survival between closed (9/18, 50%) and open (8/22, 36%) aortic valvotomy (p = 0.26). The incidence of early reoperation (less than 1 year of age) was greater in perioperative survivors undergoing closed valvotomy (7/13, 54%) rather than open valvotomy (1/10, 10%) (p less than 0.05). In conclusion, long-term survival among patients with critical neonatal aortic valve stenosis remains disturbingly low (13/40, 32%) and has not significantly improved over the past 20 years.


A theoretical model of circulatory interstitial fluid flow and species transport within porous cortical bone.
A three-dimensional model of interstitial fluid flow and passive species transport within mineralized regions surrounding cross-cortical vessel canals is developed. In contrast to earlier studies, the present model applies to circulatory, non-stress-induced interstitial flow in porous cortical bone. Based on previous experimental observations, the canals are modeled as line sources that pass at an oblique angle through the cortex. Cross-cortical interstitial flow from the endosteal surface to the periosteal surface is also taken into account. It is found that model transport characteristics are qualitatively consistent with reported observations. In addition, parametric studies reveal the following: (1) Solute contact with the matrix is maximized when the ratio of canal radius to cortex thickness (R) is near physiological R values. (2) Solute-matrix contact falls to low levels when R falls below the physiological range. (3) Solute-matrix contact is maximized when the cross-cortical velocity is approximately an order of magnitude smaller than the canal outflow velocity. The first and second findings suggest that within porous bone physiological ranges of R promote near optimal species contact with the mineralized matrix. The third finding suggests that relatively impermeable layers of bone within the cortex can effectively promote solute-matrix contact by limiting cross-cortical flow. Finally, the model suggests that intra-canal resorption associated with reduced external loading may effectively compensate for reduced stress-induced interstitial flow by enhancing circulatory interstitial flow and species transport.


Cardiovascular autonomic control in paraplegic and quadriplegic.
Spinal cord injury (SCI) is commonly associated with devastating paralysis. This condition also results in cardiovascular autonomic dysfunction associated with increased mortality from cardiovascular disease. The purpose of this study was to explore the differences in cardiovascular autonomic modulation in individuals with and without SCI. The study included 60 individuals: 30 individuals without SCI, who formed the control group-CG and 30 individuals with SCI, who formed the SCI group-SCIG. The latter group was divided into two, one group of subjects with SCI above the spinal segment T6-SCIG (above T6) and a group of individuals with SCI below T6-SCIG (below T6). The subjects were evaluated by linear and nonlinear analysis of heart rate variability (HRV). The SCIG showed significantly lower square root of the mean squares differences of successive NN intervals (rMSSD), number of pairs of adjacent NN intervals differing by more than 50 ms (pNN50), standard deviation of short-term HRV (SD1), and high frequency power (HF). Their low frequency power (LF) in absolute units (ms(2)) was significantly lower and their normalized units (n.u.) were significantly higher. Their LF/HF ratio was significantly higher, and sample entropy (SampEn), which indicates the complexity and irregularity of the NN intervals time series, was significantly lower compared to the CG. The differences between the SCIG and CG were derived mainly from the SCIG (above T6). The correlation test revealed very low values between each of the parameters evaluated for CG and SCIG. The SCIG (above T6) showed greater cardiovascular autonomic impairment compared to SCIG (below T6) and CG. The SCIG (below T6) also presented some degree of autonomic dysfunction. All parameters, linear or nonlinear, are suitable to demonstrate the differences between the SCIG and CG.


Intraseasonal effects of clutch manipulation on parental provisioning and residual reproductive value of eastern phoebes (Sayornis phoebe).
First clutches of double-brooded eastern phoebes Sayornis phoebe were manipulated (up two eggs, down 2 eggs or no change) to test for intraseasonal reproductive tradeoffs and to test whether size of first brood influenced food delivery rates to nestlings and nestling quality in second broods.Considering all nests from both broods, rate of feeding nestlings increased linearly with brood size but nestling mass per nest decreased with increasing brood size. High nestling weights in small broods may have resulted from parents delivering better quality food, but we did not test this.Among treatment groups in first broods, nestlings from decreased broods weighed more than those in control or increased broods. Treatment did not influence the likelihood that second nests would be attempted after successful first nests nor did it alter the interval between nests. Nestlings of parents that renested weighed more than those of parents that did not, regardless of treatment, suggesting that post-fledging care may preclude renesting. Mass of individual females did not change between broods, regardless of brood size. Clutch sizes of second attempts were not affected by manipulations of first broods but increasing first broods reduced the number of nestlings parents were able to raise to day 11 in their second broods. However, manipulation of first broods did not affect mean nestling mass per nest of nestlings that survived to day 11.In phoebes, parents of small first broods are able to raise nestlings in better condition. We predict that in harsh years, parents of small first broods would be more likely to renest. Parents of enlarged first broods sacrificed quality of offspring in second broods, which seems a reasonable strategy if nestlings from second broods have lower reproductive value.


A two-step process for in silico screening to assess the performance of qRTPCR kits against variant strains of SARS-CoV-2.
Since inception of the COVID-19 pandemic, early detection and isolation of positive cases is one of the key strategies to restrict disease transmission. Real time reverse transcription polymerase chain reaction (qRTPCR) has been the mainstay of diagnosis. Most of the qRTPCR kits were designed against the target genes of original strain of SARS-CoV-2. However, with the emergence of variant strains of SARS-CoV-2, sensitivity of the qRTPCR assays has reportedly reduced. In view of this, it is critical to continuously monitor the performance of the qRTPCR kits in the backdrop of variant strains of SARS-CoV-2. Real world monitoring of assay performance is challenging. Therefore, we developed a two-step in-silico screening process for evaluating the performance of various qRTPCR kits used in India. We analysed 73 qRT-PCR kits marketed in India, against the two SARS-CoV-2 VoCs. Sequences of both Delta (B.1.617.2) and Omicron (B.1.1.529) VoCs submitted to GISAID within a specific timeframe were downloaded, clustered to identify unique sequences and aligned with primer and probe sequences. Results were analysed following a two-step screening process. Out of 73 kits analysed, seven were unsatisfactory for detection of both Delta and Omicron VoCs, 10 were unsatisfactory for Delta VoC whereas 2 were unsatisfactory for only Omicron VoC. Overall, we have developed a useful screening process for evaluating the performance of qRTPCR assays against Delta and Omicron VoCs of SARS-CoV-2 which can be used for detecting SARS-CoV-2 VoCs that may emerge in future and can also be redeployed for other evolving pathogens of public health importance.


Gut microbiota, a new frontier to understand traditional Chinese medicines.
As an important component of complementary and alternative medicines, traditional Chinese medicines (TCM) are gaining more and more attentions around the world because of the powerful therapeutic effects and less side effects. However, there are still some doubts about TCM because of the questionable TCM theories and unclear biological active compounds. In recent years, gut microbiota has emerged as an important frontier to understand the development and progress of diseases. Together with this trend, an increasing number of studies have indicated that drug molecules can interact with gut microbiota after oral administration. In this context, more and more studies pertaining to TCM have paid attention to gut microbiota and have yield rich information for understanding TCM. After oral administration, TCM can interact with gut microbiota: (1) TCM can modulate the composition of gut microbiota; (2) TCM can modulate the metabolism of gut microbiota; (3) gut microbiota can transform TCM compounds. During the interactions, two types of metabolites can be produced: gut microbiota metabolites (of food and host origin) and gut microbiota transformed TCM compounds. In this review, we summarized the interactions between TCM and gut microbiota, and the pharmacological effects and features of metabolites produced during interactions between TCM and gut microbiota. Then, focusing on gut microbiota and metabolites, we summarized the aspects in which gut microbiota has facilitated our understanding of TCM. At the end of this review, the outlooks for further research of TCM and gut microbiota were also discussed.


Portable sensor based on magnetic separation and enzyme-mediated immune nanomaterials for point-of-care testing of Listeria monocytogenes in food.
Listeria monocytogenes (L. monocytogenes), a typical foodborne pathogen, poses a serious threat to public health safety. This stimulates to develop a point-of-care testing (POCT) method to achieve rapid, sensitive detection of L. monocytogenes. In this study, polyethylene glycol (PEG) mediated ampicillin functionalized magnetic beads (Amp-PEG-MBs) was prepared successfully and it achieved high efficiency (>90%) and rapid (5 min) capture for L. monocytogenes at room temperature. The innovative combination of antibody (Ab), glucose oxidase (GOD) and graphene oxide (GO) prepared Ab@GO@GOD for the specific recognition of L. monocytogenes. Finally, Amp-PEG-MBs and Ab@GO@GOD were successfully assembled into Amp-PEG-MBs@L. monocytogenes-Ab@GO@GOD sandwich structure which could catalyze the glucose, and the final detection results were recorded by a blood glucose meter (BGM). Magnetic separation (MS) combined with enzyme-catalyzed sensor (MS-Ab@GO@GOD-BGM) was successfully established to achieve the detection of L. monocytogenes in artificially contaminated juice within 66 min with the limit of detection was 101 CFU/mL. This sensor has potential for other pathogens detection by modifying specific antibodies.


Survival of COPD patients using inhaled corticosteroids and long-acting beta agonists.
We conducted a historical cohort study to examine the relationship between survival and use of inhaled corticosteroids (ICS) and/or long-acting beta agonists (LABA) in patients with chronic obstructive pulmonary disease (COPD). All COPD patients aged 40 years who were enrolled in one of two regional managed care organizations during 1995-2000, and who had 90 days use of an ICS and/or LABA (N=1288) or of a short-acting bronchodilator (N=397), were identified. Of patients treated with ICS and/or LABA, 14.4% died during the follow-up period, as compared to 28.2% of comparison patients (P<0.01). In a Cox proportional hazards model that controlled for age, sex, comorbidities, COPD severity, and asthma status, a reduced risk of death was found for ICS treatment (HR 0.59 [95% CI 0.46-0.78]), LABA (HR 0.55 [0.34-0.89]), and ICS plus LABA treatment (HR 0.34 [0.21-0.56]). A second model that excluded any patient who also had an ICD-9 code for asthma (N=840) still found improved survival among those using the combination of ICS plus LABA (HR 0.35 [CI 0.17-0.71]). Additional analyses that varied the exposure criteria also found a consistent treatment benefit. Inclusion of ICS or bronchodilator treatment during the follow-up period as a time-dependent function appears to negate the survival benefit; however, the underlying assumptions for valid time-dependent modeling are clearly violated in this situation. In conclusion, we found that COPD patients who used ICS alone or in combination with LABA had substantially improved survival even after adjustment for asthma and other confounding factors.


Tumorigenic de-differentiation: the alternative splicing way.
The mechanism of acquisition of tumorigenic properties by somatic cells at the onset of cancer and later during relapse is a question of paramount importance in cancer biology. We have recently discovered a Muscleblind like-1 (MBNL1)-driven alternative-splicing mediated mechanism of tumorigenic de-differentiation that is associated with poor prognosis, relapse and metastasis in common cancer types.


The role of high dose chemotherapy and autologous stem-cell transplantation in peripheral T-cell lymphoma: a review of the literature and new perspectives.
Peripheral T-cell lymphoma (PTCL) is a heterogeneous group of non-Hodgkin's lymphoma that carries, except for ALK-positive anaplastic large cell lymphoma, a poor prognosis. Only a third of patients live 5years past diagnosis. The incidence of PTCL has been increasing during the last two decades. In recent years, there was a rising interest in PTCL manifested by the abundance of publications dedicated exclusively to this disease. The international T-cell lymphoma project was formed with an aim of unifying efforts towards a better understanding of the diagnosis and management of this disease. Given the poor outcomes of PTCL patients, high-dose chemotherapy and autologous stem-cell transplantation (HDT/ASCT) have been used in the up-front and salvage settings, with different success rates. However, there are no prospective randomized controlled trials addressing the role of HDT/ASCT in a PTCL-restricted population. This article critically reviews the data available from the retrospective and prospective studies addressing this topic. We will emphasize the favorable prognostic factors of HDT/ASCT such as a solid remission at the time of transplantation, a chemotherapy sensitive disease and a low prognostic index score. As novel agents and new therapeutic strategies are introduced, there is a continued need for prospective randomized trials to define the optimal use of HDT/ASCT in managing PTCL.


β-C(sp(3))-H Arylation of α-Hydroxy Acid Derivatives Utilizing Amino Acid as a Directing Group.
The Pd(II)-catalyzed arylation of unactivated β-C(sp(3))-H bonds in α-hydroxy aliphatic acid with a variety of aryl iodides was developed utilizing an amino acid auxiliary as a directing group. This protocol provides access to biologically active β-arylated-α-hydroxy acid derivatives.


Consequences of CK2 signaling to the nuclear matrix.
Protein kinase CK2 is recognized as one of the key cellular signals for cell growth and proliferation. Its nuclear targeting appears to be critical to its role in these functions. In the nucleus, nuclear matrix (NM) which plays a major role in growth-related activities is a primary locus for CK2 signaling. A variety of growth stimuli evoke a rapid translocation of the CK2 to the NM whereas removal of these factors has the opposite effect. These studies, employing various experimental models of cell growth (involving different growth-stimulatory factors), have suggested that rapid shuttling of CK2 to the NM is a key feature of early growth control. By contrast, removal of growth-stimulatory factors leading to the loss of cell viability is associated with early loss of CK2 from the NM (and chromatin). This indicates that absence of CK2 from the nuclear compartment is contributory to induction of cell death via apoptosis, implying a protective role for CK2 against cell death. Here, we review the evidence that suggests that CK2 signaling in the NM is not only involved in cell growth but also in cell survival.


Negative impact of noise on the principal component analysis of NMR data.
Principal component analysis (PCA) is routinely applied to the study of NMR based metabolomic data. PCA is used to simplify the examination of complex metabolite mixtures obtained from biological samples that may be composed of hundreds or thousands of chemical components. PCA is primarily used to identify relative changes in the concentration of metabolites to identify trends or characteristics within the NMR data that permits discrimination between various samples that differ in their source or treatment. A common concern with PCA of NMR data is the potential over emphasis of small changes in high concentration metabolites that would over-shadow significant and large changes in low-concentration components that may lead to a skewed or irrelevant clustering of the NMR data. We have identified an additional concern, very small and random fluctuations within the noise of the NMR spectrum can also result in large and irrelevant variations in the PCA clustering. Alleviation of this problem is obtained by simply excluding the noise region from the PCA by a judicious choice of a threshold above the spectral noise.


Prediction of inter-residue contacts map based on genetic algorithm optimized radial basis function neural network and binary input encoding scheme.
Inter-residue contacts map prediction is one of the most important intermediate steps to the protein folding problem. In this paper, we focus on the problem of protein inter-residue contacts map prediction based on neural network technique. Firstly, we use a genetic algorithm (GA) to optimize the radial basis function widths and hidden centers of a radial basis function neural network (RBFNN), then a novel binary encoding scheme is employed to train the network for the purpose of learning and predicting the inter-residue contacts patterns of protein sequences got from the protein data bank (PDB). The experimental evidence indicates the utility of our proposed encoding strategy and GA optimized RBFNN. Moreover, the simulation results demonstrate that the network got a better performance for these proteins, whose residue length falls into the area of (100, 300), and the predicted accuracy with a contact threshold of 7 Angstroms scores higher than the other 3 values with 5, 6, and 8 Angstroms.


Proteases are responsible for blister formation in recessive dystrophic epidermolysis bullosa and epidermolysis bullosa simplex.
The specific factors which induce blister formation in recessive dystrophic epidermolysis bullosa (RDEB) and epidermolysis bullosa simplex (EBS) were studied by culturing normal human skin with blister fluid from patients with RDEB and EBS. When skin from a healthy person was cultured with RDEB blister fluid, it developed a clean subepidermal blister with histology similar to that of a RDEB blister. The specific factor(s) which induced this subepidermal blister was inactivated by heat (60 degrees C, 30 min), trypsin digestion and by treating with EDTA, EGTA, alpha 2-macroglobulin, soybean trypsin inhibitor (SBTI) or N-ethylmaleimide (NEM), but was not affected by dialysis. These findings suggest that the active factor(s) in the blister fluid from patients with RDEB might include collagenase, neutral thiol protease and trypsin-like protease. By contrast, when normal skin was cultured with EBS blister fluid, this produced a clean intra-epidermal blister with histology similar to that of an EBS blister. The specific factor(s) inducing the intra-epidermal blister was inactivated by heat (60 degrees C, 30 min), trypsin digestion and by treating with NEM, but was not affected by dialysis, divalent cation chelators (EGTA, EDTA), alpha 2-macroglobulin, SBTI and pepstatin. These results suggest that the active factor(s) inducing the intra-epidermal blister in EBS might be a neutral SH-protease.


Excitatory effects of thyrotropin-releasing hormone (TRH) in hypoglossal motoneurons.
The effect of thyrotropin-releasing hormone (TRH) was studied in 30 hypoglossal motoneurons from brainstem slices of guinea pigs. Bath application of TRH resulted in an increase of the spontaneous excitatory synaptic activity, depolarization of the neurons, increase of the input resistance and change of the duration of the falling phase of excitatory postsynaptic potentials. The depolarizing response and membrane conductance change was the result of a direct postsynaptic action of TRH, possibly mediated by a reduction of a potassium conductance.


32Pi- and 45Ca-metabolism by matrix vesicle-enriched microsomes prepared from chicken epiphyseal cartilage by isosmotic Percoll density-gradient fractionation.
Matrix vesicle-enriched fractions were isolated from different zones of epiphyseal cartilage by nonenzymatic methods involving tissue homogenization, differential centrifugation, and isosmotic Percoll gradient fractionation. Uptakes of both 32Pi and 45Ca were studied concomitantly over periods from 20 min to 24 h. Percoll density gradients separated epiphyseal microsomes into two alkaline phosphatase-rich fractions: a low-density noncalcifiable fraction (P-I), and a higher-density fraction (P-II) which readily mineralized. The P-II fraction was found only in calcifying regions of the growth plate. Based on chemical and physical properties and enzyme activities, both fractions were similar except that P-II contained significantly higher levels of mineral ions than did P-I, and had lower levels of alkaline phosphatase. The mineral appeared to be primarily in a noncrystalline form. Metabolism of 32Pi and 45Ca by P-II followed a complex kinetic pattern in which accumulation of large amounts of both ions was preceded by an initial limited burst of uptake and a lag-phase of variable duration. During mineral ion loading, the density of the P-II fraction progressively increased as evidenced by co-migration of 45Ca, 32Pi, and alkaline phosphatase to increasingly higher densities. During the period of early mineral deposition (1-5 h), Ca/P uptake ratios were very low (1.0-1.2) and X-ray diffraction patterns showed a predominantly amorphous pattern. This suggests that the mineral accumulated in matrix vesicles is initially some form of noncrystalline calcium monohydrogenphosphate. L-tetramisole, a potent inhibitor of alkaline phosphatase, inhibited accumulation of both 45Ca and 32Pi in the absence of organic P substrates, 32Pi being preferentially inhibited over 45Ca. This finding, coupled with recent studies on the behavior of alkaline phosphatase at physiological pH, suggests that the protein is not acting as a phosphohydrolase, but rather as a Pi-binding or transport agent in vesicle-mediated calcification.


Democracy, economics, or health-Time to look to the east for leadership.
While the capabilities of the low and middle-income countries (LMIC) have always been defined as "doubtful" to deal with public health crises of Covid pandemic magnitude, a flip-flop and incoherent response by the high-income countries (HIC) was unexpected. In the unprecedented moment of the pandemic, it is that we are beginning to find among ourselves, across the globe a search for stronger, resilient, scalable, and economy-efficient foundations in health systems that are capable of delivering healthcare to all. The apparently underperforming international health bodies have only increased this willingness of individual nations to look towards multilateralism as the foundational principle for not just the exchange of ideas but to deliver on these. India, a vibrant and the largest democracy, has showed her willingness to overcome, for example, the limitation of the vaccines during COVID to a few by making availability and possible delivery of a safe and effective vaccine to populations across the world. This has not only been seen as a hope to surmount challenges through help from others but also as a show of human resilience in the face of adversity.


[Diagnosis and treatment of osteochondral lesions of the talus].
Delayed diagnosis of osteochondral lesions of the talus is common; acute osteochondral injuries to the talar dome frequently go unrecognized. While acute lesions may be repaired, typically the focus of operative treatment for osteochondral lesions is on: (a) débridement and drilling/microfracture (leading to filling of the defect with fibrocartilage) or (b) salvage procedures, including osteochondral autograft transfer system and autologous chondrocyte implantation (potentially leading to resurfacing of the defect with hyaline cartilage). In this article, we review current trends in preoperative assessment, defining indications and surgical management of osteochondral lesions of the talus.


COVID-19 and cutaneous manifestations: A review of the published literature.
COVID-19 is a highly contagious respiratory tract infection caused by severe acute respiratory syndrome coronavirus 2. COVID-19 outbreak, which caused thousands of deaths, has been declared a pandemic by the World Health Organization in March 2020. Skin manifestations related to SARS-CoV-2 infection can be divided mainly into five groups: chilblainlike lesions (CBLLs), maculopapular eruptions, urticarial eruptions, vesicular eruptions, and livedo or necrosis. Other skin findings reported are erythema multiforme (EM)-like lesions and skin findings associated with multisystem inflammatory syndrome in children (MIS-C) and rarely with multisystem inflammatory syndrome in adults (MIS-A). Other manifestations such as pityriasis rosea or shingles are also reported. A total of 60 articles including reviews, studies and case reports were selected for the evaluation in this review. The skin manifestations associated with COVID-19 infection are numerous and can vary widely. The major dermatological patterns of COVID-19 can be classified as inflammatory reactions (maculopapular/morbilliform, urticarial and vesicular rashes), or lesions of vascular origin (chilblain like rashes, petechiae/purpura, and livedo acemose-like pattern) CONCLUSION: We believe that the dermatologist could play an important role in the response to the SARS-CoV-2 pandemic through early recognition of skin lesions suggestive of COVID-19, particularly in paucisymptomatic infections where this recognition could direct toward an early diagnosis of infection that certainly leads to a better prognosis.


Trifluoromethylpyrimidine-based inhibitors of proline-rich tyrosine kinase 2 (PYK2): structure-activity relationships and strategies for the elimination of reactive metabolite formation.
The synthesis and SAR for a series of diaminopyrimidines as PYK2 inhibitors are described. Using a combination of library and traditional medicinal chemistry techniques, a FAK-selective chemical series was transformed into compounds possessing good PYK2 potency and 10- to 20-fold selectivity against FAK. Subsequent studies found that the majority of the compounds were positive in a reactive metabolite assay, an indicator for potential toxicological liabilities. Based on the proposed mechanism for bioactivation, as well as a combination of structure-based drug design and traditional medicinal chemistry techniques, a follow-up series of PYK2 inhibitors was identified that maintained PYK2 potency, FAK selectivity and HLM stability, yet were negative in the RM assay.


Thyroiditis due to Brucella melitensis--report of two cases.
Two cases of thyroiditis due to Brucella melitensis are reported. Both occurred in foreigners working in Switzerland and presented with characteristic symptoms of thyroiditis which masked those of brucellosis. Cultures were diagnostic. One patient had concomitant thyroid malignancy. Both showed normal thyroid function and were cured following antibiotic treatment.


Cascade associative memory storing hierarchically correlated patterns with various correlations.
In conventional models for storing hierarchically correlated patterns, correlations between ancestors (first-level patterns) and their descendants (second-level ones) are assumed to be uniform, so that the descendants are distributed around their ancestors with equal distances. However, this assumption might be unnatural. We believe that objects are encoded into patterns by preserving the similarity between them. In this case, descendants are distributed around their ancestors with various distances, so that the assumption is invalid and the conventional models become inapplicable. To overcome this, we propose a model CASM3 for storing hierarchically correlated patterns with various correlations. In CASM3, critical load levels vary with the descendants, and become higher with increasing correlations. Increase in load level successively destroys the memories of the descendants in descending order of their correlations. The size of the basins of attraction depends on the range of the correlations, and becomes larger as the correlation range is shifted toward lower levels.


Factors associated with health-related quality of life in renal transplant recipients: results of a national survey in France.
This study aims to identify factors associated with health related quality of life (HRQOL) through a comprehensive analysis of sociodemographic and clinical variables among a representative sample size of renal transplant recipients (RTR) in France. A cross-sectional multicenter study was carried out in 2008. All RTR over 18 years old with a functioning graft for at least one year were included. Data included socio-demographic, health status, and treatment characteristics. To evaluate HRQOL, the Short Form-36 Health Survey (SF-36) and a HRQOL instrument for RTR (ReTransQol) were administered. Multivariate linear regression models were performed. A total of 1061 RTR were included, with a return rate of 72.5%. The variance explained in regression models of SF-36 ranges from 20% to 40% and from 9% to 33% for ReTransQol. The originality of our study's findings was that novel variables, particularly treatment side effects and unemployment, have a negative effect on quality of life of RTR. The French Biomedicine Agency and the National Health Institute for Public Health Surveillance conduct specific actions for professional reintegration and therapeutic education programs in the national plan to improve the HRQOL of people living with chronic diseases.