Computed tomography-derived myocardial extracellular volume: an early biomarker of cardiotoxicity in esophageal cancer patients undergoing radiation therapy.
We aimed to assess extracellular volume (ECV) through non-gated, contrast-enhanced computed tomography (CT) before and after radiation therapy (RT) in patients with esophageal cancer (EC). EC patients who had undergone CT before and after RT were retrospectively assessed. Patients with preexisting cardiovascular disease or with heavily artifacted CT were excluded. ECV was calculated using density values for the myocardial septum and blood pool. Data were reported as mean and standard deviation or median and interquartile range according to their distribution; t test or Wilcoxon and Pearson r or Spearman ρ were subsequently used. Twenty-one patients with stage ≥ IB EC, aged 64 ± 18 years, were included. Mean and maximum RT doses were 21.2 Gy (16.9-24.1) and 42.5 Gy (41.8-49.2), respectively. At baseline (n = 21), hematocrit was 39% ± 4%, ECV 27.9% ± 3.5%; 35 days (30-38) after RT (n = 20), hematocrit was 36% ± 4%, lower than at baseline (p = 0.002), ECV 30.3% ± 8.3%, higher than at baseline (p = 0.081); at follow-up 420 days (244-624) after RT (n = 13), hematocrit was 36% ± 5%, lower than at baseline (p = 0.030), ECV 31.4% ± 4.5%, higher than at baseline (p = 0.011). No patients showed signs of overt cardiotoxicity. ECV early after RT was moderately positively correlated with maximum RT dose (ρ = 0.50, p = 0.036). In EC patients, CT-derived myocardial ECV was increased after RT and may thus appear as a potential early biomarker of cardiotoxicity.


Anaplastic lymphoma kinase (ALK) activates Stat3 and protects hematopoietic cells from cell death.
The anaplastic lymphoma kinase (ALK) gene is characteristically translocated in Anaplastic Large Cell Lymphomas (ALCL) and the juxtaposition of the ALK gene to multiple partners results in its constitutive protein tyrosine kinase activity. We show here that expression of activated ALK induces the constitutive phosphorylation of Stat3 in transfected cells as well as in primary human ALCLs. Furthermore, immunohistochemical studies demonstrate that among distinct human B and T cell lymphomas, activation of Stat3 nuclear translocation is uniquely associated with ALK expression. NPM-ALK also binds and activates Jak3; however, Jak3 is not required for Stat3 activation or for cell transformation in vitro. Moreover, src family kinases are not necessary for NPM-ALK-mediated Stat3 activation or transformation, suggesting that Stat3 may be phosphorylated directly by ALK. To evaluate relevant targets of ALK-activated Stat3, we investigated the regulation of the anti-apoptotic protein Bcl-x(L) and its role in cell survival in NPM-ALK positive cells. NPM-ALK expression caused enhanced Bcl-x(L) transcription, largely mediated by Stat3. Increased expression of Bcl-x(L) provided sufficient anti-apoptotic signals to protect cells from treatment with specific inhibitors of the Jaks/Stat pathway or the Brc-Abl kinase. These studies support a pathogenic mechanism whereby stimulation of anti-apoptotic signals through activation of Stat3 contributes to the successful outgrowth of ALK positive tumor cells.


Double-barrel pipette system for microinjection.
A method for constructing double-barrel micropipettes is described. The pipettes are made from theta-tubing, which permits easy beveling of the tips. The two sides of the system operate independently and are suitable for simultaneous microinjection, microperfusion, or servo-null micropressure measurements.


Evaluation of the efficacy of platelet rich fibrin (I-PRF) intra-articular injections in the management of internal derangements of temporomandibular joints - a controlled preliminary prospective clinical study.
The temporomandibular joint (TMJ) is one of the most complex joints in the body. Temporomandibular disorders (TMD) are among the most troublesome disorders for patients, as they can cause pain, affect oral functions and disturb dentists on the level of diagnosis and treatment. The most common symptoms of temporomandibular joint disorders are articulated sounds (such as clicking or TMJ crepitation), joint pain in comfort and function situations (jaw movements), pain or strain in masseter and jaw muscles and or restricted mandibular movements. One of the most modern biocompatible substances used to treat joint disorders, including the TMJ, is platelet-rich plasma (PRP) and injectable platelet-rich fibrin (I-PRF). This study aims to evaluate the efficacy of platelet-rich fibrin (I-PRF) intra-articular injections in managing internal derangements of temporomandibular. Twenty patients suffering from a unilateral click due to temporomandibular disorders were individually injected with 1 mL of (I-PRF) twice 1 week apart into the superior joint space of the TMJ with the internal disorder. Data were recorded and evaluated by the Helkimo index. This evaluation was conducted three times; 1 week after the first injection, 1 week after the second injection, and 6 months after the first injection. The clicking disappeared entirely in 14 out of 20 patients after 1 week of getting the first injection, and in all patients after 1 week of getting the second injection, and returned to two of them after a six-month since the first injection. Preliminary results showed the efficacy of the Injectable Platelet-Rich Fibrin (I-PRF) in managing articular clicking in patients with internal disorders of the temporomandibular joint. Injectable Platelet Rich Fibrin has significant clinical efficacy in treating the articular clicking resulting from internal temporomandibular joint disorders.


Serum vaspin concentrations are closely related to insulin resistance, and rs77060950 at SERPINA12 genetically defines distinct group with higher serum levels in Japanese population.
Vaspin is an adipokine with insulin-sensitizing effects identified from visceral adipose tissues of genetically obese rats. We investigated genetic and nongenetic factors that define serum concentrations of vaspin. Vaspin levels were measured with RIA in Japanese subjects with normal fasting plasma glucose (NFG; n = 259) and type 2 diabetes patients (T2D; n = 275). Single nucleotide polymorphisms (SNP) at SERPINA12 (vaspin) gene locus were discovered, and five SNP were genotyped in the subjects with varied body mass index (n = 1138). The level of serum vaspin in 93% of the samples was found to vary from 0.2 to nearly 2 ng/ml in NFG subjects (n = 259) and from 0.2 to nearly 3 ng/ml in T2D patients (n = 275) (Vaspin(Low) group), whereas a significant subpopulation (7%) in both groups displayed much higher levels of 10-40 ng/ml (Vaspin(High) group). In the Vaspin(Low) group, serum vaspin levels in T2D were significantly higher than healthy subjects (0.99 ± 0.04 vs. 0.86 ± 0.02 ng/ml; P < 0.01). Both in T2D and genotyped Japanese population, serum vaspin levels closely correlated with homeostasis model of assessment for insulin resistance rather than anthropometric parameters. By genotyping, rs77060950 tightly linked to serum vaspin levels, i.e. CC (0.6 ± 0.4 ng/ml), CA (18.4 ± 9.6 ng/ml), and AA (30.5 ± 5.1 ng/ml) (P < 2 × 10(-16)). Putative GATA-2 and GATA-3 binding consensus site was found at rs77060950. Serum vaspin levels were related to insulin resistance, and higher levels of serum vaspin in 7% of the Japanese population are closely linked to minor allele sequence (A) of rs77060950.


Genetic heterogeneity and predictive biomarker for pulmonary sarcomatoid carcinomas.
The aim of this study is to investigate the genetic heterogeneity (carcinomatous vs. sarcomatous components) and predictive biomarkers in patients with pulmonary sarcomatoid carcinoma (PSC). Genetic alterations and biomarkers of immunotherapy were performed in a discovery set (n = 6) of PSC. Next-generation sequencing (NGS) on a pan-cancer gene panel was applied to detect the genetic alterations in each component, and the respective mutation profiling and tumor mutation burden (TMB) were compared as well. Immunohistochemistry (IHC) assay with SP263 antibody was used to detect the protein expression of programmed death-ligand 1 (PD-L1) in each component. Comparative genetic analysis revealed that the separate carcinomatous and sarcomatous components shared strikingly common mutations. TP53 (4/6, 66.7%) was the most common genetic alteration in 6 PSC patients. MET exon 14 skipping was detected in one case, accounting for 16.7%. An EZR-ROS1 fusion (EZR: intron10-ROS1: intron32) was identified in one case. The TMB of the two components was similar. Nevertheless, significantly higher PD-L1 expression was found in carcinomatous components compared to sarcomatous components. MDM2 amplification was detected in 2/6 (33.3%) of cases and STK11 mutation in 1/6 (16.7%) of cases. PSC containing carcinomatous and sarcomatous components had a mild heterogeneity; the two components may evolve from common ancestral cells. High PD-L1 expression suggests that immunotherapy could be used as a potential therapy for PSC patients, while patients with negative immune-responsive genes need to be screened out. Altogether, these findings further highlight that the detection of genetic alteration and PD-L1 expression plays an important role in treatment of patients with PSC.


Impact of Physiologically Based Pharmacokinetics, Population Pharmacokinetics and Pharmacokinetics/Pharmacodynamics in the Development of Antibody-Drug Conjugates.
Antibody-drug conjugates are important molecular entities in the treatment of cancer, with 8 antibody-drug conjugates approved by the US Food and Drug Administration since 2000 and many more in early- and late-stage clinical development. These conjugates combine the target specificity of monoclonal antibodies with the potent anticancer activity of small-molecule therapeutics. The complex structure of antibody-drug conjugates poses unique challenges to pharmacokinetic (PK) and pharmacodynamic (PD) characterization because it requires a quantitative understanding of the PK and PD properties of multiple different molecular species (eg, conjugate, total antibody, and unconjugated payload) in different tissues. Quantitative clinical pharmacology using mathematical modeling and simulation provides an excellent approach to overcome these challenges, as it can simultaneously integrate the disposition, PK, and PD of antibody-drug conjugates and their components in a quantitative manner. In this review, we highlight diverse quantitative clinical pharmacology approaches, ranging from system models (eg, physiologically based pharmacokinetic [PBPK] modeling) to mechanistic and empirical models (eg, population PK/PD modeling for single or multiple analytes, exposure-response modeling, platform modeling by pooling data across multiple antibody-drug conjugates). The impact of these PBPK and PK/PD models to provide insights into clinical dosing justification and inform drug development decisions is also highlighted.


Factors Predicting Progression to Severe COVID-19: A Competing Risk Survival Analysis of 1753 Patients in Community Isolation in Wuhan, China.
Most studies of coronavirus disease 2019 (COVID-19) progression have focused on the transfer of patients within secondary or tertiary care hospitals from regular wards to intensive care units. Little is known about the risk factors predicting the progression to severe COVID-19 among patients in community isolation, who are either asymptomatic or suffer from only mild to moderate symptoms. Using a multivariable competing risk survival analysis, we identify several important predictors of progression to severe COVID-19-rather than to recovery-among patients in the largest community isolation center in Wuhan, China from 6 February 2020 (when the center opened) to 9 March 2020 (when it closed). All patients in community isolation in Wuhan were either asymptomatic or suffered from mild to moderate COVID-19 symptoms. We performed competing risk survival analysis on time-to-event data from a cohort study of all COVID-19 patients (n = 1753) in the isolation center. The potential predictors we investigated were the routine patient data collected upon admission to the isolation center: age, sex, respiratory symptoms, gastrointestinal symptoms, general symptoms, and computed tomography (CT) scan signs. The main outcomes were time to severe COVID-19 or recovery. The factors predicting progression to severe COVID-19 were: male sex (hazard ratio (HR) = 1.29, 95% confidence interval (CI) 1.04-1.58, p = 0.018), young and old age, dyspnea (HR = 1.58, 95% CI 1.24-2.01, p < 0.001), and CT signs of ground-glass opacity (HR = 1.39, 95% CI 1.04-1.86, p = 0.024) and infiltrating shadows (HR = 1.84, 95% CI 1.22-2.78, p = 0.004). The risk of progression was found to be lower among patients with nausea or vomiting (HR = 0.53, 95% CI 0.30-0.96, p = 0.036) and headaches (HR = 0.54, 95% CI 0.29-0.99, p = 0.046). Our results suggest that several factors that can be easily measured even in resource-poor settings (dyspnea, sex, and age) can be used to identify mild COVID-19 patients who are at increased risk of disease progression. Looking for CT signs of ground-glass opacity and infiltrating shadows may be an affordable option to support triage decisions in resource-rich settings. Common and unspecific symptoms (headaches, nausea, and vomiting) are likely to have led to the identification and subsequent community isolation of COVID-19 patients who were relatively unlikely to deteriorate. Future public health and clinical guidelines should build on this evidence to improve the screening, triage, and monitoring of COVID-19 patients who are asymtomatic or suffer from mild to moderate symptoms.


Oncomodulin regulates spontaneous calcium signalling and maturation of afferent innervation in cochlear outer hair cells.
Cochlear outer hair cells (OHCs) are responsible for the exquisite frequency selectivity and sensitivity of mammalian hearing. During development, the maturation of OHC afferent connectivity is refined by coordinated spontaneous Ca2+ activity in both sensory and non-sensory cells. Calcium signalling in neonatal OHCs can be modulated by oncomodulin (OCM, β-parvalbumin), an EF-hand calcium-binding protein. Here, we investigated whether OCM regulates OHC spontaneous Ca2+ activity and afferent connectivity during development. Using a genetically encoded Ca2+ sensor (GCaMP6s) expressed in OHCs in wild-type (Ocm+/+ ) and Ocm knockout (Ocm-/- ) littermates, we found increased spontaneous Ca2+ activity and upregulation of purinergic receptors in OHCs from Ocm-/- cochlea immediately following birth. The afferent synaptic maturation of OHCs was delayed in the absence of OCM, leading to an increased number of ribbon synapses and afferent fibres on Ocm-/- OHCs before hearing onset. We propose that OCM regulates the spontaneous Ca2+ signalling in the developing cochlea and the maturation of OHC afferent innervation. KEY POINTS: Cochlear outer hair cells (OHCs) exhibit spontaneous Ca2+ activity during a narrow period of neonatal development. OHC afferent maturation and connectivity requires spontaneous Ca2+ activity. Oncomodulin (OCM, β-parvalbumin), an EF-hand calcium-binding protein, modulates Ca2+ signals in immature OHCs. Using transgenic mice that endogenously expressed a Ca2+ sensor, GCaMP6s, we found increased spontaneous Ca2+ activity and upregulated purinergic receptors in Ocm-/- OHCs. The maturation of afferent synapses in Ocm-/- OHCs was also delayed, leading to an upregulation of ribbon synapses and afferent fibres in Ocm-/- OHCs before hearing onset. We propose that OCM plays an important role in modulating Ca2+ activity, expression of Ca2+ channels and afferent innervation in developing OHCs.


Automated function imaging: a new operator-independent strain method for assessing left ventricular function.
Speckle tracking is a new technique based on pure 2D grayscale ultrasound acquisition allowing calculation of segmental strains. To facilitate clinical application, speckle tracking has been integrated into the most recent echocardiographic systems for quick, automated evaluation of left ventricular function (Automated Function Imaging, AFI). To evaluate the feasibility, calculation time, accuracy and reproducibility of global longitudinal strain (GLS) from AFI in determining LV function in comparison to reference echocardiographic and angiographic methods-whatever the operator's experience. Echocardiography was performed in 65 patients scheduled for cardiac catheterization using a Vivid 7 system. They were divided into 3 groups according to EF (>55%, 35< or =EF< or =55%,<35%). Image quality, global LV function parameters (ejection fraction, aortic flow, dp/dt) and segmental contraction were analyzed by one experienced operator and one beginner. GLS was obtained from apical 2, 3 and 4 chamber views. GLS was compared to both echocardiographic and angiographic EF, as well as to other echocardiographic parameters. GLS was obtained successfully in 97% of patients. Mean calculation time including correction of endocardial detection was less than 60 seconds. GLS was significantly different between the 3 groups, respectively -18.1+/-2.5%, -11.5+/-2.1% and -6.0+/-1.2% (p<0.01). Strong correlations were observed between GLS and LV function (r from 0.68 to 0.87) with a high level of reproducibility. No difference was observed between experienced and inexperienced operators. AFI is clinically applicable and an effective means of assessing LV function due to its short acquisition time, feasibility and accuracy, whatever the experience of the operator.


In situ evaluation of the functional state of chromatin by means of Quinacrine Mustard staining and time-resolved fluorescence microscopy.
A pulsed laser microfluorometer with high spatial and temporal resolution was employed to study the functional state of chromatin, using Quinacrine Mustard as the fluorescent probe. Corresponding segments of polytene chromosomes of embryo suspensor cells of Phaseolus coccineus and parenchymal cells of Helianthus tuberosus, in phases not involving DNA synthesis, were selected as models. The fluorescence decay time turned out to be a discriminating parameter for the chromatin fractions of differing functional engagement. The results were interpreted on the basis of a different accessibility of the DNA to the intercalating agent, as a result of the different structural situation of the chromatin. This fact determines modifications of the energy transfer rates between dye molecules in different quantum efficiency conditions, which results in variations of fluorescence decay times.


Recombination-ready Sindbis replicon expression vectors for transgene expression.
Sindbis viruses have been widely used as tools to study gene function in cells. Despite the utility of these systems, the construction and production of alphavirus replicons is time consuming and inefficient due to potential additional restriction sites within the insert region and lack of directionality for insert ligation. In this report, we present a system useful for producing recombinant Sindbis replicons that uses lambda phage recombination technology to rapidly and specifically construct replicon expression plasmids that contain insert regions in the desired orientation. Recombination of the gene of interest with the replicon plasmid resulted in nearly 100% recombinants, each of which contained a correctly orientated insert. Replicons were easily produced in cell culture and packaged into pseudo-infectious viral particles. Insect and mammalian cells infected with pseudo-infectious viral particles expressed various transgenes at high levels. Finally, inserts from persistently replicating replicon RNA were easily isolated and recombined back into entry plasmids for sequencing and subsequent analysis. Replication-ready replicon expression plasmids make the use of alphavirus replicons fast and easy as compared to traditional replicon production methods. This system represents a significant step forward in the utility and ease of use of alphavirus replicons in the study of gene function.


Risk Factors for Adjacent Segment Disease in Short Segment Lumbar Interbody Fusion-A Case Series.
Adjacent segment disease (ASD) is a common problem after lumbar spinal fusions. Ways to reduce the rates of ASD are highly sought after to reduce the need for reoperation. To find predisposing factors of ASD after lumbar interbody fusions, especially in mismatch of pelvic incidence and lumbar lordosis (PI-LL). We conducted a retrospective cohort study of all patients undergoing lumbar interbody fusions of less than 4 levels from June 2015 to July 2020 with at least 1 year of follow-up and in those who had obtained postoperative standing X-rays. We found 243 patients who fit inclusion and exclusion criteria. Fourteen patients (5.8%) developed ASD, at a median of 24 months. Postoperative lumbar lordosis was significantly higher in the non-ASD cohort (median 46.4° ± 1.4° vs 36.9° ± 3.6°, P < .001), pelvic tilt was significantly lower in the non-ASD cohort (16.0° ± 0.66° vs 20.3° ± 2.4°, P = .002), PI-LL mismatch was significantly lower in the non-ASD cohort (5.28° ± 1.0° vs 17.1° ± 2.0°, P < .001), and age-appropriate PI-LL mismatch was less common in the non-ASD cohort (34 patients [14.8%] vs 13 [92.9%] of patients with high mismatch, P < .001). Using multivariate analysis, greater PI-LL mismatch was predictive of ASD (95% odds ratio CI = 1.393-2.458, P < .001) and age-appropriate PI-LL mismatch was predictive of ASD (95% odds ratio CI = 10.8-970.4, P < .001). Higher PI-LL mismatch, both age-independent and when adjusted for age, after lumbar interbody fusion was predictive for developing ASD. In lumbar degenerative disease, correction of spinopelvic parameters should be a main goal of surgical correction.


Entomopathogenic fungus disrupts the phloem-probing behavior of Diaphorina citri and may be an important biological control tool in citrus.
Citrus is among the most important fruit crops worldwide; however, numerous pests and diseases affect the orchards, increasing production costs. The psyllid Diaphorina citri, is a vector of the phloem-limited bacteria 'Candidatus Liberibacter spp.', the causal agent of Huanglongbing (HLB) disease. The lack of a cure for HLB requires management of the vector, mainly by intensive use of chemical insecticides, leading to the selection of resistant populations. Our study determined the effects of the entomopathogenic fungus Cordyceps fumosorosea on the probing behavior of D. citri at different time points after the fungus was applied by spraying. The electrical penetration graph technique was used to monitor the stylet activities of D. citri after application of the microbiological product. The effects were more pronounced between 30 and 96 h after the insects were sprayed, with significant disruption of the stylet activities related to the phloem and directly associated with the transmission of HLB. Our study indicated that the microbiological product Challenger®, with the active ingredient C. fumosorosea fungus, can significantly change the probing behavior of D. citri, may be helpful in more-sustainable management of the vector, and can be used to reduce the spread of HLB.


Scoring and Interpretation of the FSFI: What can be Learned From 20 Years of use?
Over the past 20 years, the Female Sexual Function Index (FSFI) has been considered the gold standard for the measurement of sexual function in women, with over 1,000 published manuscripts citing the article. Despite the measure's widespread usage and excellent psychometric properties, there has been some confusion over how to best implement and score the measure and interpret corresponding findings. The aim of the current article is to provide guidance, drawing from 20 years of use, on how to best implement the FSFI in research settings and interpret results based on the validation studies that have been conducted to date. The overview of scoring and interpretation procedures found in this article is drawn from a review of the published literature on the psychometric properties of the FSFI. The measure of interest for the present review is the FSFI. This review article provides information about implementing, scoring, and interpreting the full-scale FSFI. Domain-level scoring and interpretation procedures are also discussed across the 5 domains of the FSFI: arousal, satisfaction, desire, pain, and lubrication. Additionally, guidance is provided for evaluating translated versions of the FSFI and using the measure to examine sexual function in culturally diverse populations. Guidance on appropriately scoring and interpretating the FSFI has the potential to strengthen our empirical understanding of sexual function, and consequently, to guide theory-driven treatment development and clinical practice. The present review provides applied guidance for the appropriate use of the FSFI specifically, but does not cover other common measures of sexual function or adaptations of the original measure. It is our hope that the guidance found in this review will ultimately lead to more rigorous and appropriate usage of the FSFI in research settings. Meston CM, Freihart BK, Handy AB, et al. Scoring and Interpretation of the FSFI: What can be Learned From 20 Years of use? J Sex Med 2020;17:17-25.


Commentary: Concurrent administration of PTH and antiresorptives: Additive effects or DXA cosmetics.
Osteoanabolic therapy with parathyroid hormone (PTH(1-84)) or the PTH analogues teriparatide (PTH(1-34), TPTD) and abaloparatide induces a positive remodeling balance and increases modeling and remodeling activity on bone surfaces. As the anabolic action of PTH is primarily remodeling based increased bone turnover maximizes bone accrual. Increased remodeling, however, also increases cortical porosity and reduces mineralization of newly formed bone, which may cause initial reductions in BMD, particularly at sites rich in cortical bone. Increased cortical porosity may also have negative consequences for bone strength. Consequently, an interest developed in concurrent therapies offsetting the potential early negative cortical bone effects developed, and several studies using varying concurrent combinations of TPTD or PTH(1-84) with various antiresorptive (anti-catabolic) agents (estrogen, SERMs, bisphosphonates and denosumab) have been published. This commentary addresses the discrepancy between changes in areal bone mineral density (BMD) and bone turnover markers (BTM) in concurrent therapy studies leading to possible misinterpretations of the results. In studies of concurrent therapies increases in BMD are generally accompanied by decreases in biochemical markers of bone turnover. This includes Procollagen Type I N-Terminal Propetide (PINP), which has emerged as a reliable marker of bone formation during osteoanabolic therapy. We therefore want to submit, that the larger increases in BMD seen initially in patients on concurrent therapy mask the potential for later reduced osteoanabolic action of PTH. This notion is corroborated by: 1) the lesser impairment of bone anabolism seen with milder antiresorptive modalities like hormone replacement therapy (HRT) or Selective Estrogen Receptor Modulators (SERMs); 2) the changes in BMD seen in extension studies where treatment naïve patients previously treated with PTH alone are crossed over to antiresorptive drugs. We therefore advise against a general use of concurrent therapy with PTH and antiresorptive agents, as it entails blunting of osteoanabolic action of PTH in the long run.


Studies on wound healing potential of topical herbal formulations- do we need to strengthen study protocol?
Healing of wound is a normal biological process that occurs naturally as long as it is not obstructed by infection. Many monoherbal and polyherbal formulations have been reported to hasten/accelerate wound healing activity in freshly prepared incisional and excisional experimental wound models. In the present review, an attempt has been made to throw light on importance of microbial infection in the process of wound healing and antimicrobial activity of herbal formulations. Different herbal formulations have been reported to hasten/accelerate the process of wound healing by enhancing epitheliazation, neovascularization, formation of granulation tissue, collagen synthesis, wound contraction, tensile strength, etc. As these studies have been conducted in freshly prepared non-infected wounds, it is difficult to ascertain the wound healing potential of these formulations in absence of microbial colonization/infection and results are not justifiable because the healing is limited to non-infected wounds. It would be more appropriate to ascertain the wound healing potential and not hastening/accelerating the wound healing property of newer herbal formulations on wound healing in experimental animals in presence of colonization/infection. Hence, it is recommended to strengthen these study protocols further using suitable controls to find out the antimicrobial activities of herbal formulations and their effect on wounds colonized/infected with pathogenic microbes in significant numbers to achieve more meaningful and concrete conclusions.


Effects of amino acids enriched tears substitutes on the cornea of patients with dysfunctional tear syndrome.
To evaluate the effect of aminoacid enriched artificial tears on the ocular surface of patients with dysfunctional tear syndrome (DTS). Forty patients were divided into two groups: group 1 treated for 90 days with sodium hyaluronate (SH) 0.15% 1 drop × 5 times/day; group 2 treated for 90 days with SH 0.15% + aminoacids mixture 1 drop × 5 times/day. Symptom score questionnaire, tear break-up time (TBUT), corneal fluorescein stain, Shirmer's I test and confocal microscopy were performed at baseline and after 30 and 90 days. Confocal images underwent morphometric analysis. Both treatments improved symptoms after 1 month. Group 2 patients showed at 1 month an improvement of TBUT and corneal stain, maintained throughout the study. Also Shirmer's I test improved after 3 months. In group 1, an improvement of TBUT and corneal stain was observed after 3 months. The morphometric analysis of confocal images demonstrated at month 1 an improvement of nerve tortuosity in group 2; after 3 months both groups showed a significant improvement versus baseline. The epithelium showed, in both groups, a reduction in hyperreflective large cells starting from 1 month; the area of the cells was significantly reduced after 3 months, with a significant higher reduction in group 2. The perineural stromal opacity was significantly increased after 3 months, particularly in group 2. This is the first study addressing corneal changes after amino acids administration in a DTS population. The treatment with amino acids enriched SH can be considered a useful tool in the treatment of DTS.


Prevalence of monoclonal gammopathy of undetermined significance in Thailand.
Individuals with monoclonal gammopathy of undetermined significance (MGUS) develop multiple myeloma and related malignancies at the rate of 1% per year. Given differences in ethnicity, data on prevalence and risk factors of MGUS in Thai population will be helpful in understanding the pathogenesis of plasma cell disorders and designing an early cancer detection strategy. Subjects of 50 years or older were included. Demographic data and suspected risk factors were collected. Monoclonal proteins were detected using serum protein electrophoresis. Serum was obtained from 3,260 participants; 1,104 males (33.9%) and 2,156 females (66.1%). The median age was 57 years (range 50-93 years). Monoclonal proteins were detectable in 2.3% (95% confidence interval [CI] 1.8-2.8). M spikes were found in gamma- and beta-globulin regions in 50 (1.5%) and 25 (0.8%) subjects, respectively. The prevalence of MGUS in subjects 50-59, 60-69, and 70 years or older was 2.0% (41/1,975), 2.6% (22/851), and 2.8% (12/434), respectively. By multivariate analysis, MGUS was associated with living outside Bangkok (odds ratio 2.25, 95% CI 1.11-4.58). The overall prevalence of MGUS in the Thai population was 2.3%, which was lower than that in Western countries, but comparable to that in Japan.


An ultrastructural study of cytodifferentiation in the developing human root-tip.
Transmission electron microscopy showed that in growing human teeth, the root sheath consisted of inner and outer epithelial cells. The inner epithelial cells formed a basal lamina associated filamentous layer which increased in density in coronal direction. Extensions of developing odontoblasts were in contact with the basal lamina. Direct contact with the epithelial plasmalemma was not observed. The odontoblasts obtained their fully-developed cylindrical appearance after making contact with the basal lamina of the inner epithelial cells. But, they possessed already abundant RER before these contacts were present, indicating that the differentiation of dental papillary cells into collagen-producing cells did not require heterotypic epithelio-mesenchymal contacts. After deposition of dentine, the odontoblast processes were withdrawn from the epithelium and the outer layer of root dentine. Differentiation of cementoblasts was observed in the dental follicle in which mesenchymal cells develop into RER-containing cells, migrating through apical root-sheath fenestrations to their final position between the continuous root sheath and outer dentinal layer. The findings suggest that all differentiation steps of odontogenic epithelium and odontogenic papillary and follicular mesenchyme proceed without actual contact between epithelium and mesenchyme.


Establishment of a controlled slow freezing-based approach for experimental clinical cryopreservation of human prepubertal testicular tissues.
To develop an efficient, clinical-grade, freezing protocol toward experimental clinical cryopreservation of testicular tissues in prepubertal boys suffering from cancer. Experimental cryopreservation of testicular tissue. University Medical Center. Adult patients undergoing orchiectomy for various tumors and prepubertal boys scheduled for gonadotoxic treatment. None. Histopathological analysis of tissue architecture, structural integrity, and cellular morphology was performed for control and frozen-thawed cryopreserved tissues.The number of seminiferous tubules per testicular section was calculated. The survival of spermatogonial stem cells (SSCs) and Sertoli cells of the control and frozen-thawed cryopreserved tissues was analyzed by immunofluorescence staining. Uncontrolled Slow Freezing, Controlled slow freezing, and vitrification similarly preserved the integrity of the adult testicular tissues and the survival of SSCs and Sertoli cells. Controlled slow freezing of prepubertal testicular tissues effectively preserved their architecture, the number of tubules, SSCs, and Sertoli cells. In addition, we observed SSC loss after chemotherapy in prepubertal boys, reemphasizing the importance of fertility preservation before gonadotoxic treatment. Future fertility restoration for male survivors of pediatric cancers depends on the development of an optimal prepubertal testicular tissue cryopreservation method. Our findings demonstrate the effectiveness of controlled slow freezing for cryopreservation of human prepubertal testicular tissues and may contribute to more effective banking of these tissues and potential fertility restoration. NIH research clinical trials number: NCT02529826.


Improving biogas upgrading and liquid chemicals production simultaneously by a membrane biofilm reactor.
In this study, a novel membrane biofilm reactor (MBfR) was developed for simultaneously biogas upgrading and liquid chemicals production. With external hydrogen supplied from inside of the gas permeable hollow fiber of the MBfR, CO2 in biogas could be captured via a biological process as liquid chemicals and simultaneously producing high-purity methane. Continuous operation of MBfR further confirmed that higher solubilized hydrogen was favorably affecting acetate and ethanol titer and rate, and methane purity. Moreover, by retaining biomass on the outer surface of hollow fiber, the highest biogas purity (96.7%) and acetate and ethanol production rates (37.8 and 13.5 mmol L-1d-1) were achieved at a hydraulic retention time of 2.0 d. Meanwhile, the CO2 and hydrogen conversion efficiency reached to the maximum of 93.8% and 98.1%, respectively. The findings obtained can pave a new way for efficient liquid chemical production and biogas upgrading with both economic and environmental benefits.


Synthesis of a novel 3,5-diacrylamidobenzoic acid based hyper-cross-linked resin for the efficient adsorption of Congo Red and Rhodamine B.
A novel hyper cross-linked, 3,5-diacrylamidobenzoic acid based resin (APEADA) has been synthesized for the highly efficient removal of Congo red (CR) and Rhodamine B (RhB) from aqueous solution. The synthesized resin was extensively characterized by FT-IR, FESEM, PXRD, EDX, BET analysis, TGA and solid state 13C (CP-MAS) NMR. APEADA exhibits thermal stability (≈ 200 °C) and a considerable surface area (64.78 m2 g-1), as suggested by Thermogravimetric Analysis (TGA) and Brunauer-Emmett-Teller (BET) analysis, respectively. The experimental data of dyes adsorption onto resin was examined by a variety of isotherm models wherein Langmuir and Temkin models were found to be the best fit for explaining the adsorption of both RhB (R2 = 0.9899) and CR (R2 = 0.9919). The resin can efficiently remove CR (92.03%) at pH 8.14 and moderately adsorb RhB (45.3%) at pH 5.01 and exhibits maximum adsorption capacity (Qmax) for CR (280.0 mg g-1) and RhB (23.28 mg g-1) at 318.6 K. The thermodynamic analysis of the data revealed that the adsorption processes were exothermic and spontaneous. Moreover, APEADA showed efficient removal efficiency (80.13%) for dyes in simulated effluents which warrants its utility and effectiveness in industrial waste water treatment.


Microplastics pollution in agricultural farms soils: preliminary findings from tropical environment (Klang Valley, Malaysia).
The aim of this research is to investigate the occurrence, characteristics, and potential sources of microplastic pollution at four agricultural farms in Malaysia's tropical region of Klang Valley. The mean number of microplastic particles found in the agricultural soils were 2.1 ± 0.44 to 3.4 ± 1.2 particles/kg. Farms B and D had the lowest and highest total microplastic particle counts, 1.5 and 6.0 particles/kg, respectively, which was in line with the intensity of plastic consumption at these farms. Microplastics particle sizes ranged from 16.7 to 1.246 µm, attributed to their extensive breakdown processes. The microplastic particle shapes (film, fiber, and fragment) and colors (black, white, red, and blue) from the soil samples reflected the type of plastic products used and unmanaged plastic waste at these farms. Plastic nets, mulching films, and unmanaged plastics waste constituted the major microplastics sources at these farms. Our findings confirm microplastic pollution in tropical agricultural soils as well as the need to assess the negative effects of long-term plastic use on agricultural soils.


Enhanced susceptibility of trypsinized HeLa cells to Chlamydia trachomatis infection.
An improved cell culture method for the recovery of Chlamydia trachomatis from clinical samples was evaluated. Freshly trypsinized HeLa 229 cells were infected in suspension culture and compared to a standard monolayer culture method. Among 1085 specimens evaluated, 84 were positive by both methods, 978 were negative by both methods, 2 were positive by the monolayer method only, and 21 were positive by the suspension method only. Inclusion counts were two-fold higher in the suspension culture method than in the monolayer method (p less than .001). It is concluded that freshly trypsinized HeLa 229 cells infected in suspension culture are more susceptible to infection with Chlamydia trachomatis than standard monolayer cells.


Factors associated with different forms of alcohol use behaviors among college students in Bhutan: a cross-sectional study.
Alcohol use has impacts on several dimensions of health, including physical health and mental health, particularly in college-age populations. Therefore, this study aimed to estimate the prevalence and to determine factors associated with alcohol use behaviors among college students in Bhutan. A cross-sectional design was applied to collect data from the participants. A simple random method was used to select the participants from the lists of students who were attending the four selected colleges in Bhutan in the academic year of 2019. A questionnaire was developed, and validity and reliability were verified before use. Descriptive statistics were applied to describe the general characteristics of participants, while logistic regression was used to detect the associations between variables at the significance level of α = 0.05. A total of 432 college students were recruited into the study, of whom 62.0% were females. The average age was 20.0 years, 40.7% were the third-year students, and 79.9% lived in college dormitories. The prevalence of current drinking was 51.6% and that of binge drinking was 19.4%. After controlling for all potential confounder factors, three variables were found to be associated with current drinking: students who had low income were more likely to be current drinkers than those who had high income (AOR = 2.59, 95% CI = 1.29-5.21); students who did not use tobacco were more likely to be current drinkers than those students who used tobacco (AOR = 6.99, 95% CI = 2.90-16.81); and students who had close friends who did not use alcohol were more likely to be current drinkers than those who had close friends who used alcohol (AOR = 5.14, 95% CI = 3.04-8.69). Four factors were found to be associated with binge drinking after controlling for all possible confounder factors: students who had high income were more likely to be binge drinkers than those who had low income (AOR = 3.03, 95% CI = 1.72-5.31); students who used tobacco were more likely to be binge drinkers than those students who did not use tobacco (AOR = 2.28, 95% CI = 1.35-3.87); students whose parents used alcohol were more likely to be binge drinkers than those students whose parents did not use alcohol (AOR = 1.75, 95% CI = 1.02-3.01); and students whose close friends used alcohol were more likely to be binge drinkers than those who had close friends who did not use alcohol (AOR = 2.26, 95% CI = 1.05-4.87). There is a high prevalence in alcohol use among the college students in Bhutan. Effective health promotion programs should be implemented by focusing on reducing the alcohol consumption among college students.


Developing a prioritisation framework for patients in need of coronary artery angiography.
Effective waiting list management and comprehensive prioritisation can provide timely delivery of appropriate services to ensure that the patient needs are met and increase equity in the provision of health services. We developed a prioritisation framework for patients in need of coronary artery angiography (CAA). We used a multi-methods approach to elicit effective factors that affect CAA patient prioritisation. Qualitative data wase collected using semi-structured interviews with 15 experts. The final set of factors was selected using experts' consensus through modifed Delphi technique. The framework was finalised during expert panel meetings. 212 effective factors were identified based on the literature review, interviews, and expert panel discussion of them, 37 factors were selected for modifed Delphi study. Following two rounds of Delphi discussions, seven final factors were selected and weighed by ten experts using pair-wise comparisons. The following weights were given: the severity of pain and symptoms (0.22), stress testing (0.18), background diseases (0.15), number of myocardial infarctions (0.15), waiting time (0.10), reduction of economic and social performance (0.12), and special conditions (0.08). Clinical effective factors were important for CAA prioritisation framework. Using this framework can potentially lead to improved accountability and justice in the health system.


In vivo imaging of alphaherpesvirus infection reveals synchronized activity dependent on axonal sorting of viral proteins.
A clinical hallmark of human alphaherpesvirus infections is peripheral pain or itching. Pseudorabies virus (PRV), a broad host range alphaherpesvirus, causes violent pruritus in many different animals, but the mechanism is unknown. Previous in vitro studies have shown that infected, cultured peripheral nervous system (PNS) neurons exhibited aberrant electrical activity after PRV infection due to the action of viral membrane fusion proteins, yet it is unclear if such activity occurs in infected PNS ganglia in living animals and if it correlates with disease symptoms. Using two-photon microscopy, we imaged autonomic ganglia in living mice infected with PRV strains expressing GCaMP3, a genetically encoded calcium indicator, and used the changes in calcium flux to monitor the activity of many neurons simultaneously with single-cell resolution. Infection with virulent PRV caused these PNS neurons to fire synchronously and cyclically in highly correlated patterns among infected neurons. This activity persisted even when we severed the presynaptic axons, showing that infection-induced firing is independent of input from presynaptic brainstem neurons. This activity was not observed after infections with an attenuated PRV recombinant used for circuit tracing or with PRV mutants lacking either viral glycoprotein B, required for membrane fusion, or viral membrane protein Us9, required for sorting virions and viral glycoproteins into axons. We propose that the viral fusion proteins produced by virulent PRV infection induce electrical coupling in unmyelinated axons in vivo. This action would then give rise to the synchronous and cyclical activity in the ganglia and contribute to the characteristic peripheral neuropathy.


Therapeutic DNA vaccines against tuberculosis: a promising but arduous task.
To review recent developments in therapeutic DNA vaccines against tuberculosis. The data used in this review were obtained mainly from the studies of therapeutic DNA vaccines against tuberculosis reported from 2000 to 2006. Relevant articles about studies of therapeutic DNA vaccines against tuberculosis were selected. Data were mainly extracted from the 32 articles listed in the reference section of this review. Some DNA vaccines which previously showed to induce protective immunity against infection by Mycobacterium tuberculosis in a prophylactic manner are also surprisingly effective when used therapeutically, including persistent Mycobacterium tuberculosis and multidrug-resistant tuberculosis which are refractory to immune system and antibacterial chemotherapy alone. When used in combination with antibacterial drugs, therapeutic DNA vaccines could effectively eliminate residual bacteria in infected animals and shorten the therapy course of conventional chemotherapy. Detailed studies demonstrated that therapeutic effects of DNA vaccines may at least partly be due to the restoration of the Th(1)/Th(2) balance. Some problems have also emerged along with these exciting results. Therapeutic DNA vaccine is a promising strategy against tuberculosis, however developing an ideal DNA vaccine for therapy of tuberculosis will require further development.


Novel GRIA2 variant in a patient with atypical autism spectrum disorder and psychiatric symptoms: a case report.
As sequencing technology has advanced in recent years, a series of synapse-related gene variants have been reported to be associated with autism spectrum disorders (ASDs). The α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor is a subtype of the ionotropic glutamate receptor, whose number or composition changes can regulate the strength and plasticity of synapses. Here, we report a de novo GRIA2 variant (NM_001083619.3: c.2308G > A, p.Ala770Thr) in a patient with obvious behavior regression and psychiatric symptoms. It encodes GluA2, which is the crucial subunit of the AMPA receptor, and the missense variation is predicted to result in instability of the protein structure. The association between GRIA2 variants and onset of ASD symptoms is rare, and our study expands the spectrum of phenotypic variations. For patients with an unexplained etiology of ASD accompanied by psychiatric symptoms, genetic causes should be considered, and a complete genetic evaluation should be performed.


Trends in cesarean delivery rates in primipara and the associated factors.
Few studies have focused on cesarean delivery (CD) trends among primipara under the one-child and the two-child policies. This study aimed to explore the trends in CD rates among primipara during 1995-2019 and the associated factors with CD risk. This study obtained clinical data on primiparous mothers and newborns from 1995 to 2019 at a large tertiary hospital in Wuhan, China. Trends in CD rates were calculated using the joinpoint regression analysis. The Chi-square tests and log-binomial regression analyses were used to evaluate the associations between primary variables and CD risk. CD rates showed a significant upward trend with an average annual percentage change (APC) of 2.2% (95% CI: 0.6, 3.8%) during the study period. In 1995-2006, the CD rates continued to increase with an APC of 7.8% (95% CI: 4.8, 10.9%). After 2006, the CD rates started to decline by an APC of - 4.1% (95% CI: - 5.5, - 2.6%). The CD rates non-significantly increased from 36.2% in 2016 to 43.2% in 2019. Moreover, the highest CD rate during 2015-2019 was observed on August 30 (59.2%) and the lowest on September 1 (29.7%). Primipara of older age and those with >3pregnancies had higher risks of CD. Furthermore, primipara who gave birth to newborns with low birth weight and macrosomia had higher risks of CD. Maternal and fetal as well as social and cultural factors may contribute to the rising trend of CD rates. Effective measures should be taken to control CD under the two-child policy, especially for primipara.


Heterogeneous grain-scale response in ferroic polycrystals under electric field.
Understanding coupling of ferroic properties over grain boundaries and within clusters of grains in polycrystalline materials is hindered due to a lack of direct experimental methods to probe the behaviour of individual grains in the bulk of a material. Here, a variant of three-dimensional X-ray diffraction (3D-XRD) is used to resolve the non-180° ferroelectric domain switching strain components of 191 grains from the bulk of a polycrystalline electro-ceramic that has undergone an electric-field-induced phase transformation. It is found that while the orientation of a given grain relative to the field direction has a significant influence on the phase and resultant domain texture, there are large deviations from the average behaviour at the grain scale. It is suggested that these deviations arise from local strain and electric field neighbourhoods being highly heterogeneous within the bulk polycrystal. Additionally, the minimisation of electrostatic potentials at the grain boundaries due to interacting ferroelectric domains must also be considered. It is found that the local grain-scale deviations average out over approximately 10-20 grains. These results provide unique insight into the grain-scale interactions of ferroic materials and will be of value for future efforts to comprehensively model these and related materials at that length-scale.


The value of thermography in the early diagnosis of postoperative sternal wound infections.
Peristernal skin temperatures were recorded postoperatively by infrared thermography in 150 patients. Persistent elevation of peristernal skin temperature during the 3rd and 4th post-operative week was found in 5 patients, all of whom developed sternal wound infection. A further group of 18 patients, all suspected to have occult wound infection, showed persistent temperature elevation in 7 patients, 6 of these patients were proven later to have manifest infection and needed treatment. Close thermographic scruting of the incision in patients with suspected but not proven infection appears to be useful in deleting early stages of deep seated infections.


Perceived quality of communication amongst outpatients in western and traditional Chinese medicine clinics in a Chinese population.
This study investigates differences in perceived communication by patients of the quality of outpatient episodes in an urban Chinese population. A representative, population based survey was conducted in Hong Kong, China. A random sample of respondents aged >15 years who had received outpatient care the 30 days previous to enumeration were invited to evaluate communication with their doctors and the overall quality of their latest consultation. The majority thought that their clinicians were listening carefully (93.5%), explaining clearly (93.1%), expressing appropriate respect (93.8%), and were allocating sufficient time for consultation (89.5%). 97.6% gave 5-10 points out of 10 for overall healthcare quality. Compared to users of private western medicine (WM) services, multivariate analysis showed that traditional Chinese medicine (TCM) patients were more likely to rate their clinicians highly for listening skills but low for respect. Users of WM public services were more likely to be dissatisfied with all evaluated aspects. Favourable ratings were more likely to be expressed by those who are older, less educated, and those who paid by indemnity insurance. We have highlighted three unique observations: first, patients' perception of clinician-patient communication in out patients differs between WM and TCM. Second, patients who used public WM services rate the quality of their visits lower than those in the private WM sector. Lastly, we have observed a negative association between higher education background and satisfaction level. Further researches are warranted.


From Individuals to Systems and Contributions to Creations: Novel Framework for Mapping the Efforts of Individuals by Convening The Center of Health and Health Care.
People with lived health care experiences (often referred to as "patients") are increasingly contributing to health care and are most effective when they are involved as partners who can contribute complementary knowledge alongside other stakeholders in health care. Convening The Center aimed to bring together "people known as patients"-the center of health care-to address priorities as they defined them. According to the original project design, an in-person gathering was to be conducted; however, as a result of the COVID-19 pandemic, the in-person gathering was transformed into a series of digital gatherings, including an in-depth interview phase, small-group gatherings, and a collective convening of 25 participants (22 women and 3 men from the United States, India, Costa Rica, Sweden, and Pakistan). Each participant was interviewed on Zoom (Zoom Video Communications Inc), and the interview data were thematically analyzed to design a subsequent small group and then full cohort Zoom sessions. Visual note-taking was used to reinforce a shared understanding of each individual- and group-level conversation. The interviews and gatherings for Convening The Center offered unique perspectives on patient activities in research, health innovation, and problem-solving. This project further developed a novel, two-spectrum framework for assessing different experiences that patients may have or seek to gain, based on what patients actually do, and different levels of patients' involvement, ranging from individual to community to systemic involvement. The descriptors of patients in academic literature typically focus on what health care providers think patients "are" rather than on what patients "do." The primary result of this project is a framework for mapping what patients "do" and "where" they do their work along two spectra: from creating their own projects to contributing to work initiated by others and from working at levels ranging from individual to community to systems. A better understanding of these spectra may enable researchers to more effectively engage and leverage patient expertise in health care research and innovation.


Epidemiological aspects of "repetition strain injury" in Telecom Australia.
An epidemic of "repetition strain injury" ("RSI") in Telecom Australia is described. In the years 1981-1985, there have been 3976 reports of "RSI". The occupation that was most affected was that of telephonist, with 1886 reports and a rate of 343 per 1000 keyboard staff members over five years; among other occupations that were affected were clerical workers (1421 reports; rate, 284 per 1000 keyboard staff members); telegraphists (17 reports; rate, 34 per 1000 staff members); and process workers (235 reports; rate, 116 per 1000 staff members). Women accounted for 3330 (83%) of all reports; in the telephonist group, 27% of female and 20% of male staff members were affected (P less than 0.001); for women, younger staff members were more affected. There was a significant difference among states in the prevalence of "RSI" in telephonists. There was a high morbidity, and 644 (16%) subjects were affected for more than 26 weeks; the cost-effectiveness of treatment and rehabilitation varied appreciably among state administrations. The costs of the epidemic exceed $15 million, including $1.8 million in medical costs. Reasons for the rise and decline of the epidemic are discussed.


Cytomegalovirus-induced gingival hyperplasia.
We report a rare presentation of gingival hyperplasia associated with cytomegalovirus infection in a 33-year-old human immunodeficiency virus-infected man.


Inhibition of human choriotropin binding to receptor by human choriotropin alpha peptides. A comprehensive synthetic approach.
Synthetic overlapping peptides of the alpha-subunit of human chorionic gonadotropin (hCG) were made by solid-phase peptide synthesis employing a comprehensive synthetic approach. The entire primary structure of the alpha-subunit was synthesized as a series of nine consecutive peptides, each 15 residues in length, and overlapping with its two adjacent neighbors by 5 residues on each side. Receptor binding activity of each synthetic peptide was measured by the inhibition of binding of 125I-labeled hCG to rat ovarian receptor. Peptides alpha 21-35, alpha 31-45, alpha 71-85, and alpha 81-92 were shown to compete for binding with native hCG, thus demonstrating that at least two regions on the alpha-subunit may be part of the binding site(s) of the hormone. The low affinity of the peptides (10(-5)-10(-6) M) compared to native hormone (10(-10) M) for receptor is not unexpected due to the probability of discontinuous and multiple sites involved in receptor binding. An ultrapure preparation of hCG alpha-subunit also had low affinity (10(-5), suggesting that conformational changes upon combination with beta-subunit to form dimer or changes in conformation after binding are necessary for high affinity interaction. These results correlate with previous predictions of binding sites based on studies employing chemical and enzymatic modifications of intact hormone and show that synthetic peptide strategies are helpful in the elucidation of protein structure and function.


Rural age-friendly ecosystems for older adults: An international scoping review with recommendations to support age-friendly communities.
The population of older adults in rural areas is rising, and they experience higher rates of poverty and chronic illness, have poorer health behaviors, and experience different challenges than those in urban areas. This scoping review seeks to (1) map the state of the science of age-friendly systems in rural areas regarding structural characteristics, processes for delivering age-friendly practices, and outcomes of age-friendly systems, (2) analyze strengths, weakness, opportunities, and threats of age-friendly system implementation, and (3) make person, practice, and policy-level recommendations to support active aging and development of age-friendly communities. An international scoping review was conducted of articles that used age-friendly framing, had a sample age of 45 years of age or older, self-identified as rural, and reported empiric data. Searches were conducted in PubMed, CINAHL, AgeLine, PsychINFO, EMBASE, Scopus, and Academic Search Elite on October 26, 2021, and rerun March 10, 2023. Data were charted across three analytic layers: socioecological model, Donabedian's framework, and SWOT analysis. Results reveal limited data on outcomes relevant to organizations, such as return on investment or healthcare utilization. While the SWOT analysis revealed many strengths of age-friendly systems, including their impact on persons' outcomes, it also revealed several weaknesses, threats, and gaps. Namely, age-friendly systems have weaknesses due to reliance on trained volunteers and staff, communication, and teamwork. System-level threats include community and health system barriers, and challenges in poor/developing areas. While age-friendly systems in this review were heterogeneous, there is an opportunity to focus on unifying elements including the World Health Organization age-friendly cities framework or 4Ms framework for age-friendly care. Despite the many benefits of age-friendly systems, we must acknowledge limitations of the evidence base, pursue opportunities to examine organizational metrics to support implementation and sustainability of age-friendly systems, and leverage improvements in age-friendliness at a community level.


Prediabetes and risk of mortality, diabetes-related complications and comorbidities: umbrella review of meta-analyses of prospective studies.
The term prediabetes is used for individuals who have impaired glucose metabolism whose glucose or HbA1c levels are not yet high enough to be diagnosed as diabetes. Prediabetes may already be associated with an increased risk of chronic 'diabetes-related' complications. This umbrella review aimed to provide a systematic overview of the available evidence from meta-analyses of prospective observational studies on the associations between prediabetes and incident diabetes-related complications in adults and to evaluate their strength and certainty. For this umbrella review, systematic reviews with meta-analyses reporting summary risk estimates for the associations between prediabetes (based on fasting or 2 h postload glucose or on HbA1c) and incidence of diabetes-related complications, comorbidities and mortality risk were included. PubMed, Web of Science, the Cochrane Library and Epistemonikos were searched up to 17 June 2021. Summary risk estimates were recalculated using a random effects model. The certainty of evidence was evaluated by applying the GRADE tool. This study is registered with PROSPERO, CRD42020153227. Ninety-five meta-analyses from 16 publications were identified. In the general population, prediabetes was associated with a 6-101% increased risk for all-cause mortality and the incidence of cardiovascular outcomes, CHD, stroke, heart failure, atrial fibrillation and chronic kidney disease, as well as total cancer, total liver cancer, hepatocellular carcinoma, breast cancer and all-cause dementia with moderate certainty of evidence. No associations between prediabetes and incident depressive symptoms and cognitive impairment were observed (with low or very low certainty of evidence). The association with all-cause mortality was stronger for prediabetes defined by impaired glucose tolerance than for prediabetes defined by HbA1c. Prediabetes was positively associated with risk of all-cause mortality and the incidence of cardiovascular outcomes, CHD, stroke, chronic kidney disease, cancer and dementia. Further high-quality studies, particularly on HbA1c-defined prediabetes and other relevant health outcomes (e. g. neuropathy) are required to support the evidence.


Immunohistochemical application of D2-40 as basal cell marker in evaluating atypical small acinar proliferation of initial routine prostatic needle biopsy materials.
D2-40 has been recently discovered as a lymphatic endothelial cell marker, and some investigators have found that D2-40 is also expressed in myoepithelial cells of salivary gland or breast. In this study, we evaluated D2-40 expression of basal cells and applied D2-40 immunohistochemistry in the combination of P504S, cytokeratin 5, and p63 for ten lesions with atypical small acinar proliferation (ASAP) in initial prostatic needle biopsy. As a result, D2-40 was expressed in basal cells, lymphatic endothelial cells, and some stromal fibroblasts of normal prostatic tissue. Among ten ASAP lesions, the final diagnosis of seven lesions was resolved by combination immunohistochemistry. D2-40 was comparable to cytokeratin 5 and p63 as a basal cell marker, and there were no lesions that failed to provide an accurate final diagnosis using only D2-40 immunohistochemistry without cytokeratin 5 or p63. However, we found some D2-40-positive stromal fibroblasts or D2-40-positive lumen-collapsed lymphatic vessels neighboring atypical glands. Pathologists should pay attention to avoid recognizing these cells as basal cells. In conclusion, the combination of immunohistochemistry of P504S, cytokeratin 5, p63, and D2-40 may contribute to the accurate diagnosis of ASAP in the initial prostatic needle biopsy.


Plastic film mulching affects N2O emission and ammonia oxidizers in drip irrigated potato soil in northwest China.
In soil, ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) are regarded as key factors mediating nitrous oxide (N2O) production and emission. However, there are scarce reports about the effect of film mulching on ammonia oxidizers, and the biological nitrification process of N2O emission is unclear. This study was based on potato field experiments under different mulching films, including polyethylene mulching film (PM), transparent degradable mulching film (TDM), black degradable mulching film (BDM), and bare land (CK). AOB and AOA abundances were estimated using real-time PCR, and their diversity and community structure were measured using high-throughput sequencing. Result revealed that the total N2O emission from CK was 12.32%-41.03% higher than that from film mulching soil. Under film mulching, according to total N2O emission from soil and N2O concentration in soil treatments were ordered as PM > BDM > TDM, and N2O production was closely correlated with copy numbers of the amoA gene. BDM significantly increased the number of amoA-AOB gene copies (P < 0.01), and PM significantly increased the those of amoA-AOA (P < 0.01). BDM and TDM increased AOB operational taxonomic units (OTUs), Chao1, Simpson, and Shannon indices, while PM increased the AOA OTUs and Chao1 index. Variations in AOA abundance and diversity were closely related to soil mineral N and temperature changes induced by polyethylene film mulching (P < 0.05), whereas AOB showed no significant association with soil properties. Meanwhile, we did not find a distinct treatment effect on AOB community structure. Our findings indicate that (i) degradable film mulching increased AOB abundance and diversity and N2O concentration, but obviously reduced N2O emissions, and (ii) AOA were more sensitive than AOB to polyethylene mulching film.


Targeting brain angiotensin and corticotrophin-releasing hormone systems interaction for the treatment of mood and alcohol use disorders.
The brain renin-angiotensin system (RAS) participates importantly in the regulation of endocrine, autonomic, and behavioral response to stress. Recent data indicate that central action of AT(1) receptor antagonists can reduce anxiety symptoms in experimental animals. Furthermore, central inhibition of RAS activity decreases ethanol intake in an animal model of alcoholism. Pathological anxiety responses and the development of substance dependence are both critically mediated through corticotrophin-releasing hormone (CRH) systems, and the RAS is positioned to interact both with hypothalamic as well as extrahypothalamic CRH systems. The thesis of this paper is that the RAS is part of the neurochemical dysregulation underlying negative affective states, anxiety disorders, and ethanol dependence and that medications targeting the RAS should be considered to augment the treatment of these disorders.


MicroRNA let-7c-5p protects against cerebral ischemia injury via mechanisms involving the inhibition of microglia activation.
The present study was designed to reveal the potential role of let-7c-5p, a highly conserved miRNA in stroke. We found that the content of let-7c-5p was significantly decreased in the plasma of patients with ischemic stroke as well as in experimental animals. Moreover, we also observed a significant decrease of let-7c-5p in ipsilateral cortex and striatum in mice that were subjected to middle cerebral artery occlusion (MCAO) at 24h reperfusion. Overexpression of let-7c-5p via ICV injection decreased the infarction volume and attenuated the neurological deficits, and most interestingly, inhibited microglial activation. To further explore the mechanism, we checked let-7c-5p expression in BV2 cells and primary microglia in an OGD condition and in LPS-induced microglial activation. The results indicated that decreased let-7c-5p was evidenced in the activated microglia. Overexpression of let-7c-5p in BV2 cells remarkably inhibited the microglial activation. The inhibition of microglial activation by overexpression of let-7c-5p was also observed in mice with experimental stroke, which is in line with the decreased infarction volume and improved neurological deficits. We identified that let-7c-5p directly targeted to the 3'-untranslated region of the caspase 3 mRNA to reduce caspase 3 levels, which may underline the miRNA - modulated microglial activity. The present study revealed that suppression of microglia activation by let-7c-5p overexpression may be involved in the protection effects of ischemic damage. The mechanism may include the miRNA-mediated caspase 3 pathway.


Treatment of desmoid tumors in Gardner's syndrome. Report of a case.
Aggressive desmoid tumors present difficult problems among patients with Gardner's syndrome. Recently, attention has been directed toward metabolic or hormonal manipulation of these tumors. A 21-year-old woman with Gardner's syndrome was admitted because of recurrent abdominal wall tumors. She was treated with nonsteroidal anti-inflammatory drugs, tamoxifen, and ascorbate for seven months. During this therapy, CT scan showed a gradual increase in size of the tumors. Subsequent resection of the abdominal tumors and the colon was performed. Although these three types of drugs were administered to prevent postoperative recurrence, an abdominal wall desmoid tumor that invaded the mesentery developed within nine months. The known treatments, that is, chemotherapy, irradiation, and surgical resection are discussed in view of this experience.


Long-term follow-up after bilateral Artisan aphakia intraocular lens implantation in two children with Marfan syndrome.
We present two patients with Marfan syndrome and bilateral crystalline lens dislocation who underwent bilateral lens extraction and Artisan aphakia intraocular lens (IOL) implantation. With a follow-up of more than 12 years, we found a good visual outcome, no serious IOL-related complications, and endothelial cell densities within the expected range for eyes without cataract surgery.


Cre-inducible Adeno Associated Virus-mediated Expression of P301L Mutant Tau Causes Motor Deficits and Neuronal Degeneration in the Substantia Nigra.
Accumulation of microtubule associated protein tau in the substantia nigra is associated with several tauopathies including progressive supranuclear palsy (PSP). A number of studies have used mutant tau transgenic mouse model to mimic the neuropathology of tauopathies and disease phenotypes. However, tau expression in these transgenic mouse models is not specific to brain subregions, and may not recapitulate subcortical disease phenotypes of PSP. It is necessary to develop a new disease modeling system for cell and region-specific expression of pathogenic tau for modeling PSP in mouse brain. In this study, we developed a novel strategy to express P301L mutant tau to the dopaminergic neurons of substantia nigra by coupling tyrosine hydroxylase promoter Cre-driver mice with a Cre-inducible adeno-associated virus (iAAV). The results showed that P301L mutant tau was successfully transduced in the dopaminergic neurons of the substantia nigra at the presence of Cre recombinase and iAAV. Furthermore, the iAAV-tau-injected mice displayed severe motor deficits including impaired movement ability, motor balance, and motor coordination compared to the control groups over a short time-course. Immunochemical analysis revealed that tau gene transfer significantly resulted in loss of tyrosine hydroxylase-positive dopaminergic neurons and elevated phosphorylated tau in the substantia nigra. Our development of dopaminergic neuron-specific neurodegenerative mouse model with tauopathy will be helpful for studying the underlying mechanism of pathological protein propagation as well as development of new therapies.


Contaminants and impoSEX: transcriptomics of contaminant-induced sex change.
Reproduction is the goal of living organisms, and environmental conditions that influence sexual development are therefore critical to understanding adaptation in natural populations. It is not surprising that so much attention has been devoted to the impacts of the physical and chemical environment on this process (Vandenberg et al.2012). Chemicals of concern include a variety of endocrine disruptors (EDs) including oestrogen and oestrogen mimics that directly lead to malformation of the gonad. On the molecular side, the impact that EDs have on genes directly involved in the feminization or masculinization of the gonad such as Cyp 19A (or aromatase), foxl2,Sox9, Dmrt1 and NrOb1, has received considerable attention due to their direct involvement in the regulation of oestrogen and testosterone. In this issue of Molecular Ecology, Pascoal et al. (2013) examine the impact of a known endocrine disruptor (tributyltin or TBT) on the transcriptome of the dog whelk, Nucella lapillus (Fig. 1),in relation to the formation of imposex individuals (masculinized females). They conclude that TBT mimics the endogenous ligand of the nuclear retinoid X receptor (RXR) and/or peroxisome profilerator-activated receptor (PPAR) disrupting pathways.


Overactive EGF signaling suppresses a C. elegans pnc-1 egg-laying phenotype independent of known signaling mediators.
Nicotinamide recycling is critical to the development and function of Caenorhabditis elegans. Excess nicotinamide in a pnc-1 nicotinamidase mutant causes the necrosis of uv1 and OLQ cells and a highly penetrant egg laying defect. An EGF receptor (let-23) gain-of-function mutation suppresses the Egl phenotype in pnc-1 animals. However, gain-of-function mutations in either of the known downstream mediators, let-60/ Ras or itr-1, are not sufficient. Phosphatidylcholine synthesis is neither required nor sufficient, in contrast to its role in the let-23gf rescue of uv1 necrosis. The mechanism behind the let-23gf suppression of the pnc-1 Egl phenotype is unknown.


Modeling and simulation of cephalosporin C production in a fed-batch tower-type bioreactor.
Immobilized cell utilization in tower-type bioreactor is one of the main alternatives being studied to improve the industrial bioprocess. Other alternatives for the production of beta-lactam antibiotics, such as a cephalosporin C fed-batch process in an aerated stirred-tankbioreactor with free cells of Cephalosporium acremonium, or a tower-type bioreactor with immobilized cells of this fungus, have proven to be more efficient than the batch process. In the fed-batch process, it is possible to minimize the catabolite repression exerted by the rapidly utilization of carbon sources (such as glucose) in the synthesis of antibiotics by utilizing a suitable flow rate of supplementary medium. In this study, several runs for cephalosporin C production, each lasting 200 h, were conducted in a fed-batch tower-type bioreactor using different hydrolyzed sucrose concentrations. For this study's model, modifications were introduced to take into account the influence of supplementary medium flow rate. The balance equations considered the effect of oxygen limitation inside the bioparticles. In the Monod-type rate equations, cell concentrations, substrate concentrations, and dissolved oxygen were included as reactants affecting the bioreaction rate. The set of differential equations was solved by the numerical method, and the values of the parameters were estimated by the classic nonlinear regression method following Marquardt's procedure with a 95% confidence interval. The simulation results showed that the proposed model fit well with the experimental data, and based on the


Detection of two non-synonymous SNPs in SLC45A2 on BTA20 as candidate causal mutations for oculocutaneous albinism in Braunvieh cattle.
Cases of albinism have been reported in several species including cattle. So far, research has identified many genes that are involved in this eye-catching phenotype. Thus, when two paternal Braunvieh half-sibs with oculocutaneous albinism were detected on a private farm, we were interested in knowing whether their phenotype was caused by an already known gene/mutation. Analysis of genotyping data (50K) of the two albino individuals, their mothers and five other relatives identified a 47.61-Mb candidate haplotype on Bos taurus chromosome BTA20. Subsequent comparisons of the sequence of this haplotype with sequence data from four Braunvieh sires and the Aurochs genome identified two possible candidate causal mutations at positions 39,829,806 bp (G/A; R45Q) and 39,864,148 bp (C/T; T444I) that were absent in 1682 animals from various bovine breeds included in the 1000 bull genomes project. Both polymorphisms represent coding variants in the SLC45A2 gene, for which the human equivalent harbors numerous variants associated with oculocutaneous albinism type 4. We demonstrate an association of R45Q and T444I with the albino phenotype by targeted genotyping. Although the candidate gene SLC45A2 is known to be involved in albinism in different species, to date in cattle only mutations in the TYR and MITF genes were reported to be associated with albinism or albinism-like phenotypes. Thus, our study extends the list of genes that are associated with bovine albinism. However, further research and more samples from related animals are needed to elucidate if only one of these two single nucleotide polymorphisms or the combination of both is the actual causal variant.


Analytical Methods for the Quantification of Histamine and Histamine Metabolites.
The endogenous metabolite histamine (HA) is synthesized in various mammalian cells but can also be ingested from exogenous sources. It is involved in a plethora of physiological and pathophysiological processes. So far, four different HA receptors (H1R-H4R) have been described and numerous HAR antagonists have been developed. Contemporary investigations regarding the various roles of HA and its main metabolites have been hampered by the lack of highly specific and sensitive analytic methods for all of these analytes. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) is the method of choice for identification and sensitive quantification of many low-molecular weight endogenous metabolites. In this chapter, different methodological aspects of HA quantification as well as recommendations for LC-MS/MS methods suitable for analysis of HA and its main metabolites are summarized.


Analysis of the response of a pacemaker neuron model to transient inputs.
The response of a pacemaker neuron model to a train of transient inhibitory impulsive perturbations is examined. The model reproduces the heterogeneous discharge forms and abrupt switchings displayed by the crayfish slowly adapting stretch receptor organ (Segundo et al., 1994, Neuroscience 62(2), pp. 459-480). The non-monotonous aspect of the instantaneous firing rate of the model reflects the fact that in some regimes input and output rates are both increasing, despite the inhibitory effect of the former. We determine how such paradoxical acceleration takes place by analyzing the response of the model using its phase transition curve. We show that paradoxical acceleration results from the fact that the phase transition curve exhibits a large slowly increasing, almost linear section similar to that of living preparations.


Anti-epileptogenic and antioxidant effect of Lavandula officinalis aerial part extract against pentylenetetrazol-induced kindling in male mice.
Repeated application of Lavandula officinalis (L. officinalis) has been recommended for a long time in Iranian traditional medicine for some of nervous disorders like epilepsy and dementia. However, there is no available report for the effect of chronic administration of Lavandula extract in development (acquisition) of epilepsy. Therefore, this study was designed to investigate the anti-epileptogenic and antioxidant activity of repeated administration of Lavandula officinalis extract on pentylenetetrazol (PTZ) kindling seizures in mice model. Lavandula officinalis was tested for its ability (i) to suppress the seizure intensity and lethal effects of PTZ in kindled mice (anti-epileptogenic effect), (ii) to attenuate the PTZ-induced oxidative injury in the brain tissue (antioxidant effect) when given as a pretreatment prior to each PTZ injection during kindling development. Valproate (Val), a major antiepileptic drug, was also tested for comparison. Val and Lavandula officinalis extract showed anti-epileptogenic properties as they reduced seizure score of kindled mice and PTZ-induced mortality. In this regard, Lavandula officinalis was more effective than Val. Both Lavandula officinalis and Val suppressed brain nitric oxide (NO) level of kindled mice in comparison with the control and PTZ group. Meanwhile, Lavandula officinalis suppressed NO level more than Val and Lavandula officinalis also decreased brain MDA level relative to PTZ group. This is the first report to demonstrate NO suppressing and anti-epileptogenic effect of chronic administration of Lavandula officinalis extract on acquisition of epilepsy in PTZ kindling mice model. In this regard, Lavandula officinalis extract was more effective than Val, possibly and in part via brain NO suppression.


Mimicking B and T cell epitopes between Mycobacterium leprae and host as predictive biomarkers in type 1 reaction in leprosy.
Several Mycobacterial infections including leprosy and tuberculosis are known to evoke autoimmune responses by modulating homeostatic mechanism of the host. Presence of autoantibodies like, rheumatoid factor, anti-nuclear factor and antibodies to host, collagen, keratin, myelin basic protein (MBP) and myosin, have been earlier reported in leprosy patients. In the present study, we detected the role of mimicking epitopes between Mycobacterium leprae and host components in the induction of autoimmune response in leprosy. Based on our previous findings, we predicted and synthesized a total of 15 mimicking linear B cell epitopes (BCE) and 9 mimicking linear T cell epitopes (TCE) of keratin and MBP. Humoral and cell-mediated immune responses against these epitopes were investigated in Non-reaction (NR), Type 1 reaction (T1R) leprosy patients, and healthy controls. We observed significantly higher levels of antibodies against 8 BCE in T1R in comparison to NR leprosy patients. Further, we also found 5 TCE significantly associated with lymphocyte proliferation in the T1R group. Our results indicated that these epitopes play a key role in the induction of autoimmune response in leprosy and are also strongly associated with the inflammatory episodes of T1R. Conclusively, these molecules may be employed as a biomarker to predict the inflammatory episodes of T1R.


Skin sensitivity tests in Nigerians with bronchial asthma.
Skin sensitivity tests using the prick method were performed on 174 patients with bronchial asthma and 160 controls, using 20 common allergens. The results revealed a high incidence of positive skin test reactions to Dermatophagoides pteronyssinus, grass and flower pollens, feathers, and some ingesta such as cow's milk and fish. The difference in the reactions between the asthmatic patients and the controls was statistically significant. The results suggest that house-dust mite allergy is the most significant factor in the aetiology of bronchial asthma in Nigerians.


The Expression of HSPD1, SCUBE3, CXCL14 and Its Relations with the Prognosis in Osteosarcoma.
The aim of the study was to explore the expression of three genes, HSPD1, SCUBE3, and CXCL14, in osteosarcoma cells and tissue, as well as their association with the prognosis of patients with osteosarcoma. The expression of HSPD1, SCUBE3, and CXCL14 in osteosarcoma cells was detected by using Western blotting method. siRNA was used to knockdown the expression of the three genes. CCK8 cell proliferation assay was used to observe the effect of siRNA interference on U2OS cell proliferation. The expression of the three genes in osteosarcoma tissue was detected employing immunohistochemical method. Kaplan-Meier survival analysis was used to compare the relations between the expression of the three genes and prognosis. The Western blotting results showed that the expression of Hsp70, SCUBE3 protein, and CXCL14 chemotactic factor in osteosarcoma cells was significantly higher than that in normal osteocytes (p < 0.05). After the three genes were interfered by siRNA, the mRNA and protein expression levels of these genes in osteosarcoma cells were significantly decreased (p < 0.05). The growth rate of U2OS cell after the siRNA interference was significantly lower than that before interference and that in the control group transfected with negative control siRNA (p < 0.05). The result of immunohistochemistry demonstrated that the expression of Hsp70, SCUBE3 protein, and CXCL14 chemotactic factor in osteosarcoma tissues was significantly higher than that in adjacent muscle tissue (p < 0.05). Kaplan-Meier survival analysis indicated that the survival rate of the patients with high expression of those three kinds of genes was obviously lower than that of other patients (p < 0.05). There was no significant difference between the survival rates of patients with high or low expression of two genes (p > 0.05). The expression of HSPD1, SCUBE3, and CXCL14 was all high in osteosarcoma tissues and cells; moreover, the three kinds of genes had close correlations with the prognosis of the patients. Targeted inhibition of these three genes could inhibit the proliferation of the tumor, which may become a new therapeutic target.


Immunogenicity and antigenic relationships among spike proteins of porcine epidemic diarrhea virus subtypes G1 and G2.
Porcine epidemic diarrhea virus (PEDV) is a coronavirus that infects cells lining the small intestine of swine, resulting in vomiting, diarrhea, and dehydration. The amino acid sequence of the spike (S) protein, which is the principal target recognized by host immune cells, has multiple mutations that distinguish the two PEDV genotypes, G1 and G2. To determine whether these mutations lead to changes in antigenicity, as suggested by the failure of PEDV vaccines in China, we first optimized the codons of typical S genes of the CV777 vaccine strain (G1 subtype) and LNCT2 strain (G2 subtype) and expressed the recombinant full-length sequence of the S protein in a eukaryotic expression system. The IgG antibody levels of serum from mice immunized with purified S protein were markedly high. Antigenicity was compared by detection of polyclonal antibodies (PAbs) against the virus and S protein using an enzyme-linked immunosorbent assay (ELISA), an indirect immunofluorescence assay (IFA), and a serum cross-neutralization (SN) assay. Reactivity with the PAbs revealed significant cross-reactivity between the two PEDV subtypes, although there was a twofold difference in the antigenic responses based on PAb titers in the ELISA and IFA. Consistent with the variation in the S gene sequences, the SN titer suggested differences in the neutralization activity of the S protein between the two subtypes, which could explain the antigenic variation between the PEDV subtypes G1 and G2.


Barriers to microalgal mass cultivation.
Economically successful microalgal mass cultivation is dependent on overcoming several barriers that contribute to the cost of production. The severity of these barriers is dependent on the market value of the final product. These barriers prevent the commercially viable production of algal biofuels but are also faced by any producers of any algal product. General barriers include the cost of water and limits on recycling, costs and recycling of nutrients, CO2 utilization, energy costs associated with harvesting and biomass loss due to biocontamination and pond crashes. In this paper, recent advances in overcoming these barriers are discussed.


Association of serum total bilirubin and potential predictors with mortality in acute respiratory failure: A retrospective cohort study.
Total serum bilirubin (TBIL) levels are a risk factor in critically ill patients. However, the relationship between the dynamics of TBIL and the prognosis of acute respiratory failure (ARF) patients is unclear. This study aimed to investigate the impact of different levels of TBIL during hospitalization on mortality in ARF patients. This study used a retrospective cohort study. We extracted information on ARF patients from the Medical Information Bank for Intensive Care (MIMIC)-III (version 1.4). We used propensity score matching (PSM) to adjust for the level of potential baseline-level differences between groups. Cox regression was used to analyze mortality risk factors in patients with ARF. Subgroup analysis was used to explore special populations. 2673 patients were included in the study, and 19.7% developed hyperbilirubinemia (TBIL ≥ 2 mg/dL) during their hospitalization. After PSM, multivariate Cox regression showed a 50% and 135% increased risk of death for a maximum value of TBIL ≥ 5 mg/dL and minimum value of TBIL ≥ 2 mg/dL during hospitalization, respectively, compared to the control population. In addition, age ≥ 65 years, previous comorbid malignancies, respiratory rate ≥ 22 beats/min, SpO2 ≥ 95, BUN ≥ 20 mg/dL, lactate ≥ 5 mmol/L, platelet < 100 * 10 ^ 9/L were independent risk factors for 1-year mortality in ARF patients. Subgroup analysis showed that high bilirubin had a greater effect on patients aged less than 65 years (P for interaction < 0.05). Hyper TBIL (TBIL max ≥ 5 mg/dL or TBIL min ≥ 2 mg/dL) was an independent risk factor for 1-year mortality in patients with ARF. This study suggests that clinicians should be aware of TBIL levels and intervene early in these patients.


Effect of low pull headgear on head position.
To evaluate changes in head position following the use of low pull headgear (LHG) and compare these changes with an untreated control group. The test group comprised pre-treatment and post-treatment lateral cephalograms of 30 males, aged 11 ± 1.5 years, who were receiving LHG therapy for correction of Class II malocclusion. Pre-observation and post-observation lateral cephalograms of 25 untreated male subjects, aged 11 ± 1.6 years, served as controls. The average treatment time for the treatment group was 12 ± 2.02 months and the average observation period for the control group was 11 ± 1.03 months. Four postural variables (NSL/CVT, NSL/OPT, CVT/HOR, OPT/HOR) were measured to evaluate the head position in all subjects pre- and post-observations. There was no significant difference in all the measurements concerning the head position within each group (p > 0.05). The mean differences of pre- and post-observations of 4 postural variables in the LHG group were 1.43, 0.9, -1.13, and -1.08, while those of the control group were 1.56, -0.32, -0.24, and 0.04, respectively. There was no significant difference between the headgear and control groups for any of the postural variables measured (p = 0.924, 0.338, 0.448, and 0.398, respectively). Although postural variables showed considerable variability in both groups, head position exhibited no significant changes over a period of 11-12 months either in the control or headgear group.


Therapeutic potentials of fasudil in liver fibrosis.
Fasudil has the potential to prevent liver fibrosis by activating natural killer cells and inhibiting the proliferation of hepatic stellate cells. Fasudil may be a promising clinical therapeutic drug for the prevention and treatment of liver fibrosis.


Reduction of CT Artifacts Using Polyetheretherketone (PEEK), Polyetherketoneketone (PEKK), Polyphenylsulfone (PPSU), and Polyethylene (PE) Reconstruction Plates in Oral Oncology.
Surgical reconstruction after segmental mandibulectomy in patients with oral cancer commonly requires implantation of titanium plates, which cause metallic artifacts in CT examinations detrimental for image quality. Therefore, the aim of the present study was to evaluate CT artifacts and image quality of the polymer plate materials polyetheretherketone (PEEK), polyetherketoneketone (PEKK), polyphenylsulfone (PPSU), and polyethylene (PE) in mandible reconstruction. Five plates (titanium, PEEK, PEKK, PPSU, and PE) were successively surgically inserted in a human cadaveric specimen using eight (1) titanium and (2) zirconium oxide screws. Streak and blooming artifacts were assessed in 1 mm CT scans with and without metallic artifact reduction algorithms (MAR) in this cross-sectional study. The different plate materials were analyzed for their effect on Hounsfield unit (HU) values resulting in streak artifacts. Blooming artifacts were measured as virtual growth of plates compared to actual size. Subjective image quality was assessed by 3 independent observers. Statistics were conducted by multifactorial ANOVA and Bonferroni's post hoc test. Titanium (182.42 HU; SD ± 3.09) was associated with significantly more streak and blooming artifacts compared to the polymers PEEK (86.41 HU; SD ± 1.72; P < .001), PEKK (85.50 HU; SD ± 2.75; P < .001), PPSU (85.51 HU; SD ± 1.97; P < .001), and PE (85.52 HU; SD ± 3.33; P < .001) implants. No significant differences could be seen between the screw types (titanium vs zirconium oxide) (P = .75) or the activation or inactivation of MAR (P = .85). Image quality was best for PE and worst for titanium implants in combination with zirconium oxide screws. Polymers plate materials PEEK, PEKK, PPSU, and PE significantly reduce CT artifacts and improve image quality facilitating the radiological interpretation of implant surrounding tissues. This effect is beneficial for radiological tumor follow-up and CT-based planning of adjuvant radiotherapy.


Failure on voxilaprevir, velpatasvir, sofosbuvir and efficacy of rescue therapy.
There are limited data on patients with chronic HCV infection in whom combination voxilaprevir (VOX), velpatasvir (VEL), sofosbuvir (SOF) retreatment fails. Thus, we aimed to assess treatment failure and rescue treatment options in these patients. Samples from 40 patients with HCV genotypes (GT) 1-4 in whom VOX/VEL/SOF retreatment failed were collected within the European Resistance Study Group. Population-based resistance analyses were conducted and clinical parameters and retreatment efficacies were evaluated retrospectively in 22 patients. Most VOX/VEL/SOF failure patients were infected with HCV GT3a (n = 18, 45%) or GT1a (n = 11, 28%) and had cirrhosis (n = 28, 70%). Previous treatments included an NS3-inhibitor (30%), an NS5A-inhibitor (100%) and SOF (85%). Baseline RAS data from a subgroup of patients before VOX/VEL/SOF retreatment (78%) showed few NS3 RASs apart from Q80K in GT1a (40%), typical NS5A RAS patterns in most patients (74%) and no S282T in NS5B. Sequencing after VOX/VEL/SOF failure was available in 98% of patients and showed only minor changes for NS3 and NS5A RASs. In 22 patients, rescue treatment was initiated with glecaprevir, pibrentasvir alone (n = 2) or with SOF±ribavirin (n = 15), VOX/VEL/SOF±ribavirin (n = 4) or VEL/SOF and ribavirin (n = 1) for 12 to 24 weeks. Sustained virologic response was achieved in 17/21 (81%) patients with a final treatment outcome. Of these, 2 GT3a-infected patients had virologic failure after rescue treatment with VEL/SOF or glecaprevir/pibrentasvir+SOF+ribavirin, and 2 patients with cirrhosis died during treatment or before reaching SVR12. VOX/VEL/SOF failure was mainly observed in HCV GT3- and GT1a-infected patients with cirrhosis and was not associated with specific RAS patterns within NS3, NS5A or NS5B target regions. Rescue treatment with multiple targeted therapies was effective in most patients. The advent of direct-acting antivirals has enabled the effective cure of chronic hepatitis C in most patients. However, treatment failure occurs in some patients, who are often retreated with a combination regimen called VOX/VEL/SOF, which is associated with very high rates of cure. However, VOX/VEL/SOF retreatment also fails in some patients. Herein, we analysed samples from patients in whom VOX/VEL/SOF retreatment failed and we assessed the efficacy of different rescue therapies, showing that rescue treatment is effective in most patients (81%).


Aptamers for pharmaceuticals and their application in environmental analytics.
Aptamers are single-stranded DNA or RNA oligonucleotides, which are able to bind with high affinity and specificity to their target. This property is used for a multitude of applications, for instance as molecular recognition elements in biosensors and other assays. Biosensor application of aptamers offers the possibility for fast and easy detection of environmental relevant substances. Pharmaceutical residues, deriving from human or animal medical treatment, are found in surface, ground, and drinking water. At least the whole range of frequently administered drugs can be detected in noticeable concentrations. Biosensors and assays based on aptamers as specific recognition elements are very convenient for this application because aptamer development is possible for toxic targets. Commonly used biological receptors for biosensors like enzymes or antibodies are mostly unavailable for the detection of pharmaceuticals. This review describes the research activities of aptamer and sensor developments for pharmaceutical detection, with focus on environmental applications.