Natural antisense RNA inhibits the expression of BCMA, a tumour necrosis factor receptor homologue.
BCMA (B-cell maturation) belongs to the tumour necrosis factor receptor gene family, and is specifically expressed in mature B lymphocytes. Antisense BCMA RNA is produced by transcription from the same locus and has typical mRNA features, e.g, polyadenylation, splicing, Kozak consensus sequence and an ORF (p12). To investigate the function of antisense BCMA RNA, we expressed BCMA in cell lines, in the presence of antisense p12 or a mutant lacking the initiation ATG codon (p12-ATG). Overexpression of both p12 and p12-ATG antisense BCMA resulted in a large decrease in the amount of BCMA protein produced, with no change in BCMA RNA levels, indicating that BCMA expression is regulated by antisense BCMA RNA at the translational level. We have also observed slight adenosine modifications, suggestive of the activity of a double-stranded RNA-specific adenosine deaminase. These data suggest that antisense BCMA may operate under physiological conditions using similar antisense-mediated control mechanisms, to inhibit the expression of the BCMA gene.


In vivo biological activity of rocket extracts (Eruca vesicaria subsp. sativa (Miller) Thell) and sulforaphane.
Eruca is thought to be an excellent source of antioxidants like phenolic compounds, carotenoids, glucosinolates and their degradation products, such as isothiocyanates. Sulforaphane is one of the most potent indirect antioxidants of Eruca isolated until the date. In this work we investigate: (i) the safety and DNA protective activity of Eruca extracts and sulforaphane (under and without oxidative stress) in Drosophila melanogaster; and (ii) the influence on D. melanogaster life span treated with Eruca extracts and sulforaphane. Our results showed that among the four concentrations of Eruca extracts tested (from 0.625 to 5mg/ml), intermediate concentrations of the Es2 accession (1.25 and 2.5mg/ml) exhibited no genotoxic activity, as well as antigenotoxic activity (inhibition rate of 0.2-0.6) and the lowest concentration of Es2 and Es4 accessions (0.625 mg/ml) also enhanced the health span portion of the live span curves. Sulforaphane presented a high antigenotoxic activity in the SMART test of D. melanogaster and intermediate concentrations of this compound (3.75 μM) enhanced average healthspan. The results of this study indicate the presence of potent antigenotoxic factors in rocket, which are being explored further for their mechanism of action.


New players in the neurovascular unit: insights from experimental and clinical epilepsy.
The conventional notion that neurons are exclusively responsible for brain signaling is increasingly challenged by the idea that brain function in fact depends on a complex interplay between neurons, glial cells, vascular endothelium, and immune-related blood cells. Recent data demonstrates that neuronal activity is profoundly affected by an entire cellular and extracellular 'orchestra', the so-called neurovascular unit (NVU). Among the 'musical instruments' of this orchestra, there may be molecules long-known in biomedicine as important mediators of inflammatory and immune responses in the organism, as well as non-neuronal cells, e.g., leukocytes. We here review recent evidence on the structure and function of the NVU, both in the healthy brain and in pathological conditions, such as the abnormal NVU activation observed in epilepsy. We will argue that a better understanding of NVU function will require the addition of new players to the 'orchestra'.


Ion-specific effect on the gelation kinetics of concentrated colloidal silica suspensions.
The comparative gelation kinetics of concentrated colloidal silica suspensions at pH 9.8 has been studied in the presence of two different counterions, ammonium as a representative structure-breaker ion and sodium as a representative structure-maker ion. For the same ionic strength of 0.45 M, the experimental results have indicated a pronounced effect of the ion-specificity on the aggregation kinetics and on the microstructure of the gelified networks at the sol-gel transition. This was ascribed to the significant difference of the critical coagulation concentration value in the presence of the two electrolytes.


Sex differences in emotional memory consolidation: the effect of stress-induced salivary alpha-amylase and cortisol.
This study examined sex differences in the emotional memory consolidation, and the impact of stress-induced cortisol and salivary alpha amylase responses on emotional memory recall. Following baseline salivary measures, 39 healthy women and 41 men viewed 20 neutral and 20 negative arousing images, and then underwent either a cold pressor stress test or control condition, followed by further salivary measures. Participants returned two days later for a free recall test. The stress condition induced greater cortisol response, and negative images were better recalled than neutral. Whereas women displayed greater recall of negative images under stress than men, they recalled fewer negative images in the control condition. Stress-induced cortisol predicted recall of negative images in women, and neutral images in men. This suggests there is an enhanced consolidation of negative images under stress in women that may be a potential mechanism for the greater female prevalence for developing anxiety disorders.


Management of Central Diabetes Insipidus in Disabled Children with Diluted Oral Desmopressin Lyophilisate Formulation Administered Through Nasogastric Tube: A Retrospective Case Series.
Experience with nasogastric administration of oral DDAVP [desamino-D-arginine-8-vasopressin] lyophilisate (ODL) for central diabetes insipidus (CDI) in disabled children with swallowing coordination difficulties is limited. We aimed to assess the safety and efficacy of nasogastric use of ODL in disabled children with CDI. Time to serum sodium normalisation was compared with that of children with normal intellect and CDI treated with sublingual DDAVP. Clinical, laboratory and neuroimaging characteristics were evaluated for 12 disabled children with CDI treated with ODL through nasogastric tube at Dr Behcet Uz Children's Hospital, Turkey, between 2012 and 2022. Six boys and six girls with a mean (±SD) age of 43 (± 40) months were evaluated. These children (mean [±SD] weight standard deviation score [SDS] - 1.2 ± 1.7; mean [±SD] height SDS - 1.3 ± 1.4) presented with failure to thrive, irritability, prolonged fever, polyuria and hypernatraemia (mean serum sodium 162 [±3.6] mEq/L). At diagnosis, mean serum and urine osmolality were 321 (± 14) mOsm/kg and 105 (± 7.8) mOsm/kg, respectively. Arginine vasopressin (AVP) levels were undetectable (< 0.5 pmol/L) at diagnosis in all patients. Nasogastric tube administration of DDAVP lyophilisate (120 µg/tablet) dissolved in water (10 mL) was commenced at a dose of 1-5 µg/kg/day in two divided doses together with controlled water intake to avoid hyponatraemia. The frequency and dose of DDAVP were titrated based on urine output and serum sodium concentration. Serum sodium declined at a rate of 0.11 ± 0.03 mEq/L/h and reached normal range in a mean duration of 174 ± 46.5 h. Serum sodium declined faster in children with normal intellect and CDI treated with sublingual DDAVP (1.28 ± 0.39 mEq/L/h; p = 0.0003). Three disabled children needed rehospitalisation because of hypernatraemia due to unintentional DDAVP omission by caregivers. No episode of hyponatraemia was observed. Weight gain and growth were normal during the median (± interquartile range) follow-up duration of 32 ± 67 months. Nasogastric administration of oral DDAVP lyophilised formulation was safe and effective in the treatment of CDI in disabled children in this small retrospective series.


Androgen receptor knockdown enhances prostate cancer chemosensitivity by down-regulating FEN1 through the ERK/ELK1 signalling pathway.
Flap endonuclease 1 (FEN1) is highly upregulated in prostate cancer and promotes the growth of prostate cancer cells. Androgen receptor (AR) is the most critical determinant of the occurrence, progression, metastasis, and treatment of prostate cancer. However, the effect of FEN1 on docetaxel (DTX) sensitivity and the regulatory mechanisms of AR on FEN1 expression in prostate cancer need to be further studied. Bioinformatics analyses were performed using data from the Cancer Genome Atlas and the Gene Expression Omnibus. Prostate cancer cell lines 22Rv1 and LNCaP were used. FEN1 siRNA, FEN1 overexpression plasmid, and AR siRNA were transfected into cells. Biomarker expression was evaluated by immunohistochemistry and Western blotting. Apoptosis and the cell cycle were explored using flow cytometry analysis. Luciferase reporter assay was performed to verify the target relationship. Xenograft assays were conducted using 22Rv1 cells to evaluate the in vivo conclusions. Overexpression of FEN1 inhibited cell apoptosis and cell cycle arrest in the S phase induced by DTX. AR knockdown enhanced DTX-induced cell apoptosis and cell cycle arrest at the S phase in prostate cancer cells, which was attenuated by FEN1 overexpression. In vivo experiments showed that overexpression of FEN1 significantly increased tumour growth and weakened the inhibitory effect of DTX on prostate tumour growth, while AR knockdown enhance the sensitivity of DTX to prostate tumour. AR knockdown resulted in FEN1, pho-ERK1/2, and pho-ELK1 downregulation, and the luciferase reporter assay confirmed that ELK1 can regulate the transcription of FEN1. Collectively, our studies demonstrate that AR knockdown improves the DTX sensitivity of prostate cancer cells by downregulating FEN1 through the ERK/ELK1 signalling pathway.


The effect of humic acids on the cytotoxicity of silver nanoparticles to a natural aquatic bacterial assemblage.
The effect of a terrestrial humic acid (HA) and a river HA on the cytotoxicity of silver nanoparticles (AgNPs) to natural aquatic bacterial assemblages (0 μM, 2.5 μM and 5 μM) was measured with spread plate counting. The effect of HA (20 and 40 ppm) on the cytotoxicity of AgNPs ranging in size between 15 and 25 nm was tested in the presence and in the absence of natural sunlight. The experiment was a full factorial, completely randomized design and the results were analyzed using the General Linear Model in SAS. LSMEANS was used to separate the means or combinations of means. Significant main effects of all independent variables, plus interaction effects in all cases except HA/LI and HA/AgNPs/LI were observed. The toxicity of AgNPs to natural aquatic bacterial assemblages appears to be concentration dependent for concentrations between 0 μM and 5 μM. The data indicate that the light exposure inhibited viability more than the darkness exposure. The HA treatment groups in the presence of light showed greater reduced viability count compared to darkness exposure groups. The inhibition of bacterial viability counts by AgNPs exposure was less in the light treatment groups containing a terrestrial HA compared to that with a river HA. Difference in the extent of reactive oxygen species formation and adsorption/binding of AgNPs was speculated to account for the observed phenomenon.


[Overweight, obesity and risk factors for atrial fibrillation or flutter--secondary publication. The cohort study Diet, Cancer and Health].
We prospectively examined the association between body mass index and risk of atrial fibrillation or flutter among 47,589 participants (mean age 56 years) in the Danish Diet, Cancer and Health Study. During follow-up (mean 5.7 years), atrial fibrillation or flutter developed in 553 subjects (372 men and 181 women). We found that the risk of atrial fibrillation increased substantially as body mass index increased in both men and women.


Novel immunomodulator FTY720 is phosphorylated in rats and humans to form a single stereoisomer. Identification, chemical proof, and biological characterization of the biologically active species and its enantiomer.
In vivo phosphorylation of FTY720 (1) in rats and humans resulted exclusively in the biologically active (S)-configured enantiomer, which was proven by an ex vivo o-phthaldialdehyde derivatization protocol especially elaborated for phosphates of 1. Starting from the prochiral amino alcohol 1, racemic and enantiomerically pure phosphates of 1 were synthesized. Pure enantiomers were obtained after purification of a partially protected key intermediate on an enantioselective support. The absolute stereochemistry was determined by X-ray diffraction.


Non-cholinesterase effects of anticholinesterases.
The presence in the nervous tissue of extremely potent peptides arises from the enzymatic degradation of larger molecular weight proteins. These smaller units are thought to exert modulatory actions on known transmitters. Their short half-life when injected is due to the degradative action of specific EXO and ENDOPEPTIDASES. It is suggested from the meager evidence available that organophosphates may adversely affect the processes of formation and/or degradation to account for many of the adverse effects of organophosphate anticholinesterases which cannot be attributed to acetylcholine accumulation.


Assessment of anterior capsule opacification: photographic technique and quantification.
To develop and evaluate a standardized slitlamp photographic technique to document and quantify anterior capsule opacification (ACO) in the pseudophakic eye. University of Vienna, Medical School, Department of Ophthalmology, Vienna, Austria. In this prospective study, slitlamp photographs were taken in 29 eyes of 19 patients with ACO of varying degrees and intraocular lenses of different materials. Digital slitlamp photographs were taken by 2 examiners using a standardized protocol. The intensity of anterior fibrosis was evaluated using subjective grading by 2 masked examiners and by image analysis. A standardized protocol was developed for documentation of ACO. The intraexaminer (or short-term) reproducibility (r = 0.96, P < .01) and interexaminer reproducibility (r = 0.93, P < .01) were excellent. The results of image analysis used to objectively quantify ACO correlated with the subjective grading (r = 0.95, P < .01). This standardized technique of slitlamp photography and image analysis provided reproducible documentation and can therefore serve as a basis for the quantification of ACO.


Changes in the structure and function of the testes and epididymides in vasectomized rams.
Vasectomized rams were studied with regard to the structure and function of their testes and epididymides at different periods up to 3 years and 9 months after the operation, and compared with intact control rams. Control rams showed a marked seasonal pattern with higher values for size and weight of testes, diameter of seminiferous tubules, Sertoli cell ratios for germ cells, and size of cauda epididymidis immediately prior to and during the breeding season than at other times of the year. In vasectomized animals the seasonal pattern was ill-defined, the testicular parameters were lower, and the cauda epididymidis was larger than in intact rams. Spermatogenesis was qualitatively and quantitatively affected after vasectomy, with a sequential ebb-and-flow pattern of spermatogenic arrest and hypospermatogenesis. Epididymal changes included the development of multiple spermatoceles surrounded by a granulomatous reaction, and disturbances in the process of sperm maturation.


Convergent prefrontal and nigral projections to the striatum of the rat.
Projections to the striatum from the prefrontal cortex, the substantia nigra's pars compacta (SNC), and the ventral tegmental area (AVT) were examined autoradiographically. These projections converge on the striatum in an organized manner such that the prefrontal areas innervated by AVT project to the same part of the striatum as AVT, and prefrontal areas innervated by SNC project to the same part of the striatum as SNC.


Pre-excitation of the ventricle associated with total intra His bundle block.
A case with total intra-His bundle block and intermittent pre-excitation syndrome is presented. During A-V conduction there was a P-delta interval of 130 msec. with a P-A interval of 20 msec., an A-H interval of 60 msec. and an H-V interval of 50 msec. During rapid atrial pacing the P-delta interval increased primarily due to an A-H1 prolongation and a Mobitz type 2 block and total A-V block occured at increasing rates showing H1 following every A spike. The escape beats showed a normal width of the QRS complexes with preceding H2 spikes. After administration of Ajmaline the bypass tract was blocked and constant total A-V block occurred. It was concluded that there was a constant total intra-His bundle block and a nodoventricular or fasciculoventricular bypass tract with prolonged conduction to the ventricle. This bypass tract blocked sometimes spontaneously and could also be blocked by rapid atrial pacing and administration of drugs. The close anatomic proximity of the His bundle and Mahaim fibers is responsible for the simultaneous block resulting in total atrioventricular block.


Australian sonographers' perceptions of patient safety in ultrasound imaging: Part two - translation into practice.
A lack of patient safety research hampers capacity to improve safety in healthcare.Ultrasound is often considered 'safe' as it does not use ionising radiation, a simplistic view of patient safety. Understanding sonographers' actions towards patient safety is crucial; however, self-reported measures cannot always predict behaviour. This study is part of a PhD exploring patient safety in medical diagnostic ultrasound. The aim of this paper is to explore sonographers' responses to the patient safety concerns identified in Part one of this study. The ultimate aim of the study is to inform the final phase of the doctoral study which will consider the next steps in improving the quality and safety of healthcare experienced by patients. A qualitative study using semi-structured, one-on-one interviews. The Theory of Planned Behaviour (TPB) explained how sonographers respond to perceived patient safety risks in practice. Thirty-one sonographers were interviewed. Based on the seven themes identified in Part one of the study, results showed that incongruences exist between identifying patient safety risks and the actions taken in practice to manage these risks. The TPB showed that behavioural, normative and control beliefs impact sonographers' responses to perceived patient safety risks in practice and can lead to risk avoidance. Lack of regulation in ultrasound creates a challenge in dealing with Fitness to Practice issues. Collective actions are required to support sonographers in taking appropriate actions to enhance patient safety from multiple stakeholders including accreditation bodies, regulatory authorities, educational institutions and employers.


Arterial spin labeling MRI for assessment of perfusion in native and transplanted kidneys.
To apply a magnetic resonance arterial spin labeling (ASL) technique to evaluate kidney perfusion in native and transplanted kidneys. This study was compliant with the Health Insurance Portability and Accountability Act and approved by the institutional review board. Informed consent was obtained from all subjects. Renal perfusion exams were performed at 1.5 T in a total of 25 subjects: 10 with native and 15 with transplanted kidneys. A flow-sensitive alternating inversion recovery (FAIR) ASL sequence was performed with respiratory triggering in all subjects and under free-breathing conditions in five transplant subjects. Thirty-two control/tag pairs were acquired and processed using a single-compartment model. Perfusion in native and transplanted kidneys was compared above and below an estimated glomerular filtration rate (eGFR) threshold of 60 ml/min per 1.73 m² and correlations with eGFR were determined. In many of the transplanted kidneys, major feeding vessels in the coronal plane required a slice orientation sagittal to the kidney. Renal motion during the examination was observed in native and transplant subjects and was corrected with registration. Cortical perfusion correlated with eGFR in native (r=0.85, P=.002) and transplant subjects (r=0.61, P=.02). For subjects with eGFR >60 ml/min per 1.73 m², native kidneys demonstrated greater cortical (P=.01) and medullary (P=.04) perfusion than transplanted kidneys. For subjects with eGFR <60 ml/min per 1.73 m², native kidneys demonstrated greater medullary perfusion (P=.04) compared to transplanted kidneys. Free-breathing acquisitions provided renal perfusion measurements that were slightly lower compared to the coached/triggered technique, although no statistical differences were observed. In conclusion, FAIR-ASL was able to measure renal perfusion in subjects with native and transplanted kidneys, potentially providing a clinically viable technique for monitoring kidney function.


Basal insulin delivery reduction for exercise in type 1 diabetes: finding the sweet spot.
Exercise poses significant challenges to glucose management in type 1 diabetes. In spite of careful planning and manipulation of subcutaneous insulin administration, increased risk of hypoglycaemia and glycaemic variability during and after exercise may occur as a result of inherent delays in insulin action and impaired counter-regulatory hormone responses. Various strategies to mitigate this issue have been advocated in clinical practice, including ingestion of supplementary carbohydrate prior to exercise, reducing background and pre-meal insulin bolus and performing bouts of resistance/high intensity exercise before aerobic exercise. Insulin pump therapy, considered the most physiological form of insulin replacement for type 1 diabetes allows modulation of basal insulin delivery before, during and after exercise. However uncertainty remains regarding the optimal strategy to reduce basal insulin delivery and its efficacy. In this issue of Diabetologia, McAuley and colleagues (DOI: 10.1007/s00125-016-3981-9 ) report on the impact of a 50% reduction of basal insulin delivery before, during and after moderate-intensity aerobic exercise. Results from this study may contribute to a better understanding of the effects of basal insulin delivery manipulation and may aid in devising therapeutic approaches for glucose management during exercise.


Distribution of galanin-binding sites in the monkey and human telencephalon: preliminary observations.
Using X-ray film autoradiography the distribution of 125I-galanin binding sites was studied in the forebrain of monkey and man. In the monkey a high density was found in all areas of the neocortex, especially layer 4, and in certain subfields in the hippocampal region. Also in the human brain high activity was seen in neocortex, mainly layer 6 and in hippocampal areas, as well as in amygdala, piriform cortex and hypothalamus. These results suggest that the 29-amino acid peptide galanin may be involved in the regulation of higher cortical functions in primates.


Importance of Right Marginal Branch Protection During Primary Angioplasty in Inferior STEMI With Right Ventricular Involvement.
We document the high effectiveness of an extra guidewire insertion aimed at mechanical lumen protection of the right marginal branch from displaced thrombus, which frequently occurs during proximal right coronary artery stenting.


Legal requirements for digestate as fertilizer in Austria and the European Union compared to actual technical parameters.
Biogas is mainly discussed in the context of renewable energy, digestate is playing a minor role. However, in the upcoming EU Fertilizer Product Regulation digestate as eligible component material is in line with the circular economy action plan. This article discusses the quality of raw digestate based on analysis data from Austrian biogas plants regarding to the current Austrian as well as the European fertilizer regulatory framework. Therefore, a survey among Austrian biogas plant operators for results of digestate analysis and substrate input and energy output is used. The results show, that the legislative framework applies to a minimum of 1.5 million tonnes of digestate. Austrian digestate is of high quality with regards to the nutrient content, required limit values for heavy metals and is also safe with respect to their hygienic parameters. The following European fertilizer product function categories are available for international trade of digestate: "organic soil improver", "growing medium", and "organic, non-microbial plant biostimulant". Establishing legal criteria determining End-of-Waste and subsequently the product status for digestate is another important step in the European circular economy.


Snake venomics of Macrovipera mauritanica from Morocco, and assessment of the para-specific immunoreactivity of an experimental monospecific and a commercial antivenoms.
Proteomic analysis of the venom of the medically relevant snake Macrovipera mauritanica from Morocco revealed a complex proteome composed of at least 45 toxins from 9 protein families targeting the hemostatic system of the prey or victim. The toxin profile of Moroccan M. mauritanica displays great similarity, but also worth noting departures, with the previously reported venom proteome of M. lebetina from Tunisia. Despite fine compositional differences between these Macrovipera taxa, their overall venom phenotypes explain the clinical picture observed in M. mauritanica and M. lebetina envenomings. However, M. mauritanica venom also contains significant amounts of orphan molecules whose presence in the venom seems to be difficult to rationalize in the context of a predator-prey arms race. The paraspecific immunoreactivity of an experimental monospecific (M. mauritanica) antivenom and a commercial bivalent antivenom, anti-C. cerastes and anti-M. lebetina, against the venoms of Moroccan M. mauritanica and Tunisian M. lebetina, was also investigated through an affinity chromatography-based antivenomics approach. Both antivenoms very efficiently immunodepleted homologous venom toxins and displayed a high degree of paraspecificity, suggesting the clinical utility of the two antivenoms for treating bites of both M. mauritanica or M. lebetina.


An improved acidified glycerol lysis test (AGLT) used to detect spherocytes in pregnancy.
The acidified glycerol lysis test (Zanella et al, 1980) had proved unreliable in our hands. We now report the test to be stable and reliable when conducted at 37 degrees C rather than at ambient laboratory temperature as used by earlier workers. With this modification, blood samples were tested from 167 pregnant subjects. Forty-eight were less than 26 weeks pregnant and gave normal AGLT values (T50 greater than 1800 s) while of 119 more than 26 weeks pregnant 57 (48%) gave shortened T50 values indicative of the presence of spherocytes.


Novel Graphene oxide-Polyethylene Glycol mono-4-nonylphenyl Ether adsorbent for solid phase extraction of Pb2+ in blood and water samples.
A novel and efficient Graphene Oxide-Polyethylene Glycol mono-4-nonylphenyl Ether (GO-PEGPE) nanocomposite was synthesized and used for solid phase extraction of trace levels of Pb2+ in different water and blood samples. The synthesized adsorbent was then characterized by the Fourier Transform-Infrared spectrophotometry (FT-IR), Field Emission-Scanning Electron Microscopy (FE-SEM), Energy dispersive X-ray spectroscopy (EDX) and X-ray diffraction analysis (XRD). To optimize the critical parameters including pH of samples solution, amounts of adsorbent and extraction time, the response surface methodology based on the central composite design (RSM-CCD) was used and based on the results, pH = 6.0, extraction time = 22 min and amounts of adsorbent = 15 mg were selected as the optimum conditions. The relative standard deviation based on seven replicate analysis of 2 µg L-1 Pb2+ was 5.2% and the limit of detection was 0.023 µg L-1 (n = 8). The results of adsorption isotherm investigation show that the adsorption of Pb2+ onto the GO-PEGPE nanocomposite obeyed by the Langmuir isotherm with the maximum adsorption capacity of 69.44 mg g-1. Also, based on the Temkin and Dubinin-Radushkevich (DR) isotherms, the adsorption of Pb2+ onto the GO-PEGPE nanocomposite is a physisorption phenomenon and the consequences of the kinetic models illustrated that the adsorption of Pb2+ followed by the pseudo second order adsorption kinetic model. Finally, the proposed method was successfully applied for preconcentration of Pb2+ in different water and blood samples of turning industry workers.


Small molecules combined with collagen hydrogel direct neurogenesis and migration of neural stem cells after spinal cord injury.
Complete spinal cord injury (SCI) leads to cell death, interruption of axonal connections and permanent functional impairments. In the development of SCI treatments, cell transplantation combined with biomaterial-growth factor-based therapies have been widely studied. Another avenue worth exploring is the generation of neurons from endogenous neural stem cells (NSCs) or reactive astrocytes activated by SCI. Here, we screened a combination of four small molecules, LDN193189, SB431542, CHIR99021 and P7C3-A20, that can increase neuronal differentiation of mouse and rat spinal cord NSCs. Moreover, the small molecules loaded in an injectable collagen hydrogel induced neurogenesis and inhibited astrogliogenesis of endogenous NSCs in the injury site, which usually differentiate into astrocytes under pathological conditions. Meanwhile, induced neurons migrated into the non-neural lesion core, and genetic fate mapping showed that neurons mainly originated from NSCs in the parenchyma, but not from the central canal of the spinal cord. The neuronal regeneration in the lesion sites resulted in some recovery of locomotion. Our findings indicate that the combined treatment of small molecules and collagen hydrogel is a potential therapeutic strategy for SCI by inducing in situ endogenous NSCs to form neurons and restore damaged functions.


Vitamin D receptor, oestrogen receptor-alpha and calcium-sensing receptor genotypes, bone mineral density and biochemical markers in Paget's disease of bone.
The significance of genetic polymorphisms in the development of Paget's disease of bone is unclear at present. We analysed the BsmI polymorphism of the vitamin D receptor (VDR) gene, the PvuII and XbaI polymorphisms of the oestrogen receptor-alpha (ER alpha) gene, and the A986S polymorphism of the calcium-sensing receptor (CaSR) gene in 69 pagetic patients and 120 healthy subjects. We also examined the relationship of these polymorphisms with lumbar spine and femoral neck BMD as well as with biochemical parameters (serum alkaline phosphatase, osteocalcin and parathyroid hormone) in Paget's disease. The XbaI and PvuII genotype distributions of the ER alpha gene were significantly different between patients with Paget's disease and control subjects (P<0.001). Also, the CaSR A986S genotype frequency was significantly different between pagetic patients and controls (P<0.01). No significant effect of gene polymorphisms on BMD or biochemical parameters of bone turnover was observed. Our results suggest that the ER alpha PvuII/XbaI and CaSR A986S polymorphisms may contribute to genetic susceptibility to Paget's disease. However, further studies are required to investigate the underlying pathomechanism and to replicate the associations.


Quantification of mRNA for endothelial NO synthase in mouse blood vessels by real-time polymerase chain reaction.
The mouse is useful in studies of vascular biology because of its well-defined genetics and because the mouse genome can be manipulated. However, because only small amounts of mRNA can be extracted from blood vessels, the quantification of gene expression in individual mice is difficult. Endothelial NO synthase (eNOS) plays a major role in the regulation of vascular tone and growth. In addition, there appear to be sex differences in the production of NO under basal conditions in mouse aortas. The goals of this study were to develop a real-time polymerase chain reaction (PCR) method to quantify eNOS mRNA in blood vessels from mice and to examine eNOS mRNA levels in vessels from male and female mice. Blood vessels were isolated from C57BL/6 mice. Total RNA from individual mice was isolated and reverse-transcribed. The number of molecules of eNOS mRNA (after reverse transcription) was determined against cDNA standards, with 18S rRNA used as a control for RNA input and reverse-transcription efficiency. When expressed as copy numbers per nanogram of total RNA or as the ratio of eNOS to 18S rRNA, eNOS mRNA was lower in the aortas of female mice than in those of male mice at 7 to 9 months of age. In contrast, no difference in eNOS mRNA was found in the aortas of 2-month-old mice. In addition, eNOS mRNA levels were similar in the carotid, cerebral, and coronary arteries. These findings provide the first quantitative measurements of eNOS mRNA by using real-time PCR in the vessels of mice and suggest age- and sex-related differences in the basal levels of eNOS mRNA in mice. In addition, the eNOS region that was used for real-time PCR was amplified and sequenced for monkeys and other species. With modifications, this region may be used to design real-time PCR for eNOS in other species.


Reproductive defects in the male diabetic athymic nude mouse.
We have previously reported spontaneous hyperglycemia and impaired glucose tolerance in male athymic nude Balb/c mice. This colony also demonstrates impaired fertility. Previous studies in both athymic nude and diabetic obese (ob/ob) mice have shown reproductive abnormalities. It was the purpose of this investigation to further elucidate the endocrine abnormalities which might contribute to the reduced fertility observed in this model. Fasting plasma glucose was measured on each nude mouse to ensure that a diabetic state existed. Testes were removed and intratesticular testosterone and LH receptors were measured. Testosterone levels in diabetic, athymic nude Balb/c (DAN) mice were significantly decreased to 46.5% of control Balb/c mice. LH receptors in DAN mice were significantly elevated by 23.6% when compared to control animals. These results demonstrate that the impaired fertility observed in the DAN mouse may result from decreased testosterone production. The increased LH receptor levels seen in the DAN mouse might reflect a regulatory event in response to the decreased intratesticular testosterone levels.


Real-time fluorescence polymerase chain reaction (PCR) identification of Streptococcus pneumoniae from pleural fluid and tissue.
A diagnosis of pneumococcal pneumonia and empyema was established by real-time fluorescence polymerase chain reaction (PCR) performed in pleural fluid and tissue samples from a child in whom cultures were negative. This case demonstrates that the use of solid-phase nucleic acid purification technology and real-time fluorescence PCR have an application in the rapid diagnosis of S. pneumoniae pneumonia.


The responses of neurons in the cortex in the superior temporal sulcus of the monkey to band-pass spatial frequency filtered faces.
There are neurons in the cortex in the anterior part of the superior temporal sulcus of the macaque monkey with visual responses which would be useful for face recognition (Rolls, 1984; Baylis et al., 1985). To analyze further the information which leads them to respond, their responses were measured to parametrically filtered stimuli. The responses of 48 such single neurons were measured to faces which were digitized and were bandpass spatial frequency filtered. The octave width bands were 2-4, 4-8, 8-16, 16-32, 32-64 and 64-128 cycles per image. It was found that the neurons could respond well to single octaves of the spatial frequencies normally present in faces, that the most effective bands were 4-8, 8-16 and 16-32 cycles per face (cpf), and that the bands 2-4 and 32-64 cpf were partly effective. In investigations of whether the responses of the neurons to an unfiltered face, and to low-pass and high-pass filtered images could be predicted by linear addition of their responses to each of the octave bands shown separately, it was found that the majority of the neurons were non-linear, and responded much less than predicted. It was also shown that this occurred even when the contrast was reduced to 0.25 of that normally present in a face, so that the result was not due just to a ceiling effect of the maximum firing rate. These results help to define parametrically the aspects of the information normally present in a face which are sufficient to produce responses of these neurons to them, and show that linear operations cannot account for information processing in this part of the visual system.


Estrogen receptor purification by affinity chromatography using an orange triazine dye.
A rapid two-step procedure was devised for the purification of the estrogen receptor from the calf uterus. A 900- to 1700-fold purification of the estrogen receptor was obtained using ammonium sulfate precipitation followed by dye affinity chromatography with Reactive Orange 14 immobilized to Sepharose. The Reactive Orange 14-Sepharose was used to purify the estrogen receptor in the presence or absence of estradiol as well as to purify the progesterone receptor. The purified estrogen receptor retained its estradiol- and DNA-binding properties and sedimented into sucrose gradients as the 5 S receptor dimer. The Reactive Orange 14-Sepharose is easily prepared and offers a higher yield and purity of the estrogen receptor than that afforded by estrogen- or heparin-Sepharose chromatography.


The interplay of ligand binding and quaternary structure in the diverse interactions of dynein light chain LC8.
Dynein light chain LC8 is a small, dimeric, and very highly conserved globular protein that is an integral part of the dynein and myosin molecular motors but appears to have a broader role in multiple protein complexes unrelated to molecular motors. LC8 binds to two families of targets: those having a KXTQT sequence fingerprint and those having a GIQVD fingerprint. All known LC8 binding partners containing these fingerprints share a common binding site on LC8 that raises the question of what determines binding specificity. Here, we present the crystal structure of apo-LC8 at 1.7-A resolution, which, when compared with the crystal structures of several LC8 complexes, gives insight into the mechanism underlying the binding diversity of LC8. Peptide binding is associated with a shift in quaternary structure that expands the hydrophobic binding surface available to the ligand, in addition to changes in tertiary structure and ordering of LC8 around the binding groove. The observed quaternary shift suggests a mechanism by which binding at one of the two identical sites can influence binding at the other. NMR spectra of titrations with peptides from each fingerprint family show evidence of allosteric interaction between the two binding sites, to a differing degree in the two ligand families. Allosteric interaction between the binding sites may be a mechanism to promote simultaneous binding of ligands from the same family, providing a physiological role for the two fingerprints.


Occurrence of antifouling biocides in sediment and green mussels from Thailand.
Various antifouling biocides were surveyed in sediment and green mussels (Perna viridis) from the coastal area of Thailand. The concentrations of butyltin (MBT), dibutyltin (DBT), and tiributyltin (TBT) in sediment from Thailand were in the range of 1-293 microg kg(-1) dry wt., 1-368 microg kg(-1) dry wt., and 2-1246 microg kg(-1) dry wt., respectively. A higher concentration of TBT was observed in industrial areas, where many tankers and cargo ships sail and moor. Phenyltin (PT) compounds were not detected in most of the sampling sites. The concentrations of MBT, DBT, and TBT in green mussels from Thailand were in the range of 8-20 microg kg(-1) wet wt., 4-9 microg kg(-1) wet wt., and 4-45 microg kg(-1) wet wt., respectively. The concentration of TBT was high in an area where aquaculture practice is common. The detection frequencies of TPT were low. Representative booster biocides were surveyed in sediment. The detection frequencies of Sea Nine 211 were low (2/13). Furthermore, Sea Nine 211 concentrations in the detected samples were at values near the detection limit (0.051-0.094 microg kg(-1) dry wt.). Diuron was detected at the highest level among the booster biocides in the range of 0.07-25 microg kg(-1) dry wt. Irgarol 1051 was detected in the range of 0.03-3.2 microg kg(-1) dry wt., and concentrations of the degradation product's M1 were 0.03-4.9 microg kg(-1) dry wt. Diuron and Irgarol 1051 showed higher concentrations in industrial areas and fishery sites, respectively. Diuron was only detected among green mussels in the range of <0.64-9.6 microg kg(-1) wet wt. The lower ratio of Diuron and Irgarol 1051 for TBT concentration suggests that TBT is still being used in Thailand.


Neurodevelopmental disorder associated with de novo SCN3A pathogenic variants: two new cases and review of the literature.
SCN3A was recently recognized as a gene associated with neurodevelopmental disorder and epilepsy. We present two additional patients with a novel de novo SCN3A pathogenic variant, and a review of all published cases of de novo variants. In one of our patients brain magnetic resonance imaging (MRI) disclosed a severe polymicrogyria and in the other it was normal. The clinical phenotype was characterized by a severe developmental delay and refractory epilepsy in the patient with polymicrogyria and intellectual disability with autistic features and pharmacoresponsive epilepsy in the subject with normal MRI. Polymicrogyria, a disorder of progenitor cells proliferation and migration, is an unanticipated finding for an ion channel dysfunction.


A critical period for acoustic trauma in the hamster and its relation to cochlear development.
Young hamsters pass through a developmental stage during which they are unusually susceptible to acoustic trauma. This sensitive period occurs after apparent structural and functional maturation of the ear and appears to be dependent on unidentified developmental changes within the cochlea.


Gene expression of male pathway genes sox9 and amh during early sex differentiation in a reptile departs from the classical amniote model.
Sex determination is the process whereby the bipotential embryonic gonads become committed to differentiate into testes or ovaries. In genetic sex determination (GSD), the sex determining trigger is encoded by a gene on the sex chromosomes, which activates a network of downstream genes; in mammals these include SOX9, AMH and DMRT1 in the male pathway, and FOXL2 in the female pathway. Although mammalian and avian GSD systems have been well studied, few data are available for reptilian GSD systems. We conducted an unbiased transcriptome-wide analysis of gonad development throughout differentiation in central bearded dragon (Pogona vitticeps) embryos with GSD. We found that sex differentiation of transcriptomic profiles occurs at a very early stage, before the gonad consolidates as a body distinct from the gonad-kidney complex. The male pathway genes dmrt1 and amh and the female pathway gene foxl2 play a key role in early sex differentiation in P. vitticeps, but the central player of the mammalian male trajectory, sox9, is not differentially expressed in P. vitticeps at the bipotential stage. The most striking difference from GSD systems of other amniotes is the high expression of the male pathway genes amh and sox9 in female gonads during development. We propose that a default male trajectory progresses if not repressed by a W-linked dominant gene that tips the balance of gene expression towards the female trajectory. Further, weighted gene expression correlation network analysis revealed novel candidates for male and female sex differentiation. Our data reveal that interpretation of putative mechanisms of GSD in reptiles cannot solely depend on lessons drawn from mammals.


Roles of vaccinia virus in the development of new vaccines.
Vaccinia virus is an efficient expression vector with broad host range infectivity and large DNA capacity. This vector has been particularly useful for identifying target antigens for humoral and cell-mediated immunity. With increased levels of gene expression, obtained either with stronger vaccinia promoters or through incorporation of the bacteriophage T7 RNA polymerase gene into the vaccinia genome, proteins may be synthesized in mammalian cells for use as subunit vaccines. For use as a live recombinant vaccine, efforts are being made to attenuate vaccinia virus further, either by inactivating genes contributing to virulence or by introducing human lymphokine genes into the vaccinia genome.


Understanding practitioners' characteristics and perspectives prior to the dissemination of an evidence-based intervention.
To describe the characteristics and repertoires of community practitioners serving families involved in child physical abuse that may inform training and treatment dissemination efforts. The aims are to: (a) describe the background characteristics of these clinicians; (b) document their most common intervention techniques; (c) examine clinicians' attitudes toward manualized treatment; and (d) understand the organizational climate of community agencies. Practitioners from nine different agencies who have worked with cases in which there was child physical abuse (n=77) completed focus group discussions and three self-report questionnaires. Qualitative and quantitative analyses were conducted in tandem. Practitioners reported that they most often focused on family issues and employed family therapy techniques. Although treatment manuals were important, it was difficult to determine how practitioners used or would use these materials with families. An almost equal number of concerns and benefits were raised regarding using manuals in treatment. There was moderate agreement in how practitioners perceived their work environments. Age and attainment of a graduate degree were positively related to greater caring and job satisfaction, respectively, while length of time at a given agency was positively related to greater reports of emotional exhaustion on a measure of organizational climate. Given the promising benefits of evidence-based treatments for child abuse, it is important that researchers begin to form partnerships with community agencies that will lead to the dissemination and evaluation of effective treatment strategies.


Short-term effect of percutaneous recanalization of chronic total occlusions on QT dispersion and heart rate variability parameters.
QT dispersion (QTd), which is a measure of inhomogeneity of myocardial repolarization, increases following impaired myocardial perfusion. Its prolongation may provide a suitable substrate for life-threatening ventricular arrhythmias. We investigated the changes in QTd and heart rate variability (HRV) parameters after successful coronary artery revascularization in a patient with chronic total occlusions (CTO). This study included 139 successfully revascularized CTO patients (118 men, 21 women, mean age 58.3±9.6 years). QTd was measured from a 12-lead electrocardiogram and was defined as the difference between maximum and minimum QT interval. HRV analyses of all subjects were obtained. Frequency domain (LF: HF) and time domain (SDNN, pNN50, and rMSSD) parameters were analyzed. QT intervals were also corrected for heart rate using Bazett's formula, and the corrected QT interval dispersion (QTcd) was then calculated. All measurements were made before and after percutaneous coronary intervention (PCI). Both QTd and QTcd showed significant improvement following successful revascularization of CTO (55.83±14.79 to 38.87±11.69; p<0.001 and 61.02±16.28 to 42.92±13.41; p<0.001). The revascularization of LAD (n=38), Cx (n=28) and RCA (n=73) resulted in decrease in HRV indices, including SDDN, rMSSD, and pNN50, but none of the variables reached statistical significance. Successful revascularization of CTO may result in improvement in regional heterogeneity of myocardial repolarization, evidenced as decreased QTcd after the PCI. The revascularization in CTO lesions does not seem to have a significant impact on HRV.


Exploring substance use and mental health for minority transgender youth: Implications for advanced practice nurses.
Transgender youth, particularly if they are transgender female and of a minority racial or ethnic group (i.e., minority transgender youth [MTY]), are known to have higher incidence of new HIV infections, substance abuse, and suicidality when compared with cisgender youth. Mental health disparities and substance use among MTY, between the ages of 13-24 years, have been underexplored in the United States, although they have serious public health concerns contributing to morbidity and mortality. To examine the US literature to discover what is known about the intersection of substance use, mental health disorders, and associated health risks for MTY. An integrative review exploring experimental and nonexperimental research was conducted from 2010 to 2018 using Cumulative Index for Nursing and Allied Health (CINAHL), PsycINFO, and PubMed. Articles were selected if the research identified MTY, mental health and/or substance use risks, data-driven outcomes, or considerations that could potentially influence MTY outcomes. Ten articles were included in the final selection. All studies addressed mental health, substance use, and/or health risks, although many of the articles explored all transgender youth outcomes without a particular focus on MTY. Identified factors linked to poor health outcomes included socioeconomic vulnerability, substance use, mood disorders, self-harm, and risky sexual behaviors. The authors have provided recommendations regarding culturally appropriate, compassionate, evidence-informed practices for engaging MTY and their families to promote greater well-being for all youth, regardless of gender identity.


The working memory capabilities of the spontaneously hypertensive rat.
This study addresses the working memory capabilities of the male spontaneously hypertensive rat (SHR) strain as compared to the normotensive inbred strain, Wistar Kyoto (WKY), and the out bred Sprague Dawley (SD) rat as a normal control. The objective was to use two working memory tasks in the water maze with different strategic demands: forced alternation (FA) which allows the use of either an allocentric ("place") or egocentric ("response") localisation strategy and delayed matching-to-place (DMP) which requires an allocentric strategy. In the FA task, SHR reached criterion at the same rate as WKY and SD controls and were impaired to the same extent as WKY at the long (1 h) delay. Furthermore, both SHR and WKY were impaired relative to SD when the memory load was increased through the use of massed trials. In the DMP task, the performance of SHR did not differ from that of either of the control strains, either during training or in response to delay. These findings do not provide evidence of short-term memory impairments in the SHR, which is a commonly-used animal model of Attention Deficit Hyperactivity Disorder (ADHD) in humans.


Structural-transformation-induced Drastic Luminescence Changes in an Organic-Inorganic Hybrid [ReN(CN)4 ]2- Salt Triggered by Chemical Stimuli.
We synthesized a (1-propylpyridinium)2 [ReN(CN)4 ]-type organic-inorganic hybrid exhibiting water-vapor-induced drastic structural changes of the [ReN(CN)4 ]2- assemblies. Specifically, upon exposure to water vapor, dehydrated nitrido-bridged chains were converted to hydrated cyanido-bridged tetranuclear clusters via rearrangements of large molecular building units in the crystals. These switchable assembly forms display substantially different photo-physical properties, although in both cases the emission is caused by a metal-centered d-d transition. The nitrido-bridged chain exhibited a near-infrared (749 nm) emission, which blue-shifted as the temperature increased, while a visible (561 nm) emission and its red shift was demonstrated by the cyanido-bridged cluster.


GIS-aided optimisation of faecal sludge management in developing countries: the case of the Greater Accra Metropolitan Area, Ghana.
This study employed GIS tools to help optimise faecal sludge (FS) management in the Greater Accra Metropolitan Area (GAMA) and its environs in Ghana. First, the rates of excreta generation, FS generation and FS collection were quantified based on literature, census and FS discharge data obtained from treatment plants in the study area. Next, we mapped the FS collection to the administrative areas in GAMA based on discharge records obtained from Lavender Hill, the main faecal treatment (FTP) and estimated the travel distance and travel time from the various FS desludging neighbourhoods to the plant. The results of the study show that the excreta and FS generation rates in GAMA are 604 L/cap/yr and 4,137 L/cap/yr, respectively. About 1 million m3 of FS was collected and treated in the study area in 2018, with a collection rate of 244 L/cap/yr. The private sector dominates this collection, haulage and treatment of FS in GAMA. The GIS analysis has provided fundamental data that will be useful in rationalising the FS emptying and transport cost in the study area. Moreover, it revealed that about 20-40% of the localities were outside the 15-25 km sustainable maximum transport distance recommended by some scholars. Finally, the findings highlight the importance of looking beyond administrative boundaries when planning for FS management logistics and infrastructure and also show that the most impoverished communities in the Accra metropolis may not necessarily be the least served when it comes to FS collection and haulage.


Crucial cytokine interactions in nitric oxide production induced by Mycoplasma arthritidis superantigen.
Mycoplasma arthritidis causes autoimmune arthritis in rodents. It produces a superantigen (MAM) that simultaneously activates antigen presenting cells and T cells inducing nitric oxide and cytokine release. Nitric oxide is a key inducer and regulator of the immune system activation. Here, we investigated nitric oxide and cytokine production and interactions of these molecules in MAM-stimulated co-cultures of macrophages (J774A.1 cell line) with spleen lymphocytes. We found that: a) MAM-induced nitric oxide, interferon-gamma, membrane-associated tumor necrosis factor and interleukin-2 production in co-cultures of macrophages with lymphocytes from BALB/c and C3H/HePas but not from C57Bl/6 mice; b) production of nitric oxide was dependent on interferon-gamma whereas that of interferon-gamma was dependent on interleukin-2 and membrane-associated tumor necrosis factor; c) these cytokines up regulated MAM-induced nitric oxide production. Unraveling the mechanisms of cell activation induced by MAM might be helpful to design strategies to prevent immune system activation by superantigens and therefore in seeking amelioration of associated immunopathologies.


Time to initiation or duration of S-1 adjuvant chemotherapy; which really impacts on survival in stage II and III gastric cancer?
Surgical resection with S-1 adjuvant chemotherapy (AC) is the standard of care for stage II-III gastric cancer (GC). However, it is unclear if time to initiation and duration of S-1 AC impact on survival. A multi-institutional GC database identified 498 patients who were treated with S-1 AC after D2 or more extended radical surgery for stage II-III gastric cancer. Patients were divided into four groups according to the interval between surgery and initiation of AC and the duration of AC as follows: group A (n = 226), who received AC earlier (≤6 weeks) and for longer (≥6 months) after surgery; group B (n = 160), who received AC later (>6 weeks) and for longer after surgery; group C (n = 46), who received AC earlier but for a shorter period (<6 months) after surgery; and group D (n = 66), who received AC later and for a shorter period after surgery. Prognostic factors for overall survival (OS) were investigated using multivariate analysis. The 5-year OS was 69.5%. Pathological stage II disease (hazard ratio (HR), 0.334; 95% confidence interval (CI), 0.215-0.499), with an OS of 85.8% versus 60.5% for stage III disease, as well as a longer duration (≥6 months) of S-1 (HR, 0.498; 95% CI, 0.355-0.706), with an OS of 74.3% versus 53.0% for a shorter duration (<6 months) of S-1, were identified as significant prognostic factors for long-term survival. Time to initiation was not associated with OS. A duration of S-1 AC of ≥6 months, but not time to initiation within 6 weeks, impacts on OS in stage II-III gastric cancer.


Comparing Data Collected on Amazon's Mechanical Turk to National Surveys.
In this study, we examined the impact of a range of methods to improve data quality on the demographic and health status representativeness of Amazon Mechanical Turk (MTurk) samples. We developed and field-tested a general survey of health on MTurk in 2017 among 5755 participants and 2021 among 6752 participants. We collected information on participant demographic characteristics and health status and implemented different quality checks in 2017 and 2021. Adding data quality checks generally improves the representativeness of the final MTurk sample, but there are persistent differences in mental health and pain conditions, age, education, and income between the MTurk population and the broader US population. We conclude that data quality checks improve the data quality and representativeness.


Compared promoting potential of D-galactosamine, carbon tetrachloride and partial hepatectomy in rapid induction of preneoplastic liver lesions in the rat.
Effectiveness of two different chemically induced stimuli for hepatocellular proliferation was compared with regard to that of commonly used partial hepatectomy (PH), for the purpose of developing short-term protocol for the assay of promoting agents of hepatocarcinogenesis. Enhancing effect of D-galactosamine (DGA) and carbon tetrachloride (CCl4) given during the promotion procedure by 2-acetylaminofluorene (2-AAF) was compared along with PH in rats initiated by diethylnitrosamine (DEN), using preneoplastic glutathione S-transferase positive (GST-P+) hepatocyte foci as an end-point marker lesion. The number of GST-P+ foci per cm2 was largest in the group given CCl4 followed by DGA, no treatment (2-AAF alone) and PH. In contrast, the area (mm2) per cm2 and mean diameter of the focus were largest in the PH group then DGA followed by CCl4 and no treatment. The results indicate that the number of GST-P+ foci were not clearly affected by 3 different treatments whereas area and size of foci which represented the result of promoting effect were clearly influenced by those treatments, indicating they caused differential proliferation of initiated cells. In this respect, even though PH is the most potent procedure, DGA is also efficient and preferred to CCl4 for the non-surgical enhancing method.


Timing of spike discharges in cat auditory cortex neurons: implications for encoding of stimulus periodicity.
Neurons at more central stations in the central auditory pathway show progressively poorer responses to high-frequency stimulus periodicities. This has been attributed to the relatively poorer spike timing in forebrain auditory neurons. This study directly examined the timing of spikes evoked by brief tone pulses which were varied in peak level and repetition rate. The experiments revealed that at tone repetition rates which produced progressively poorer entrainment, the timing of spike discharges remained sufficiently precise to support entrained responses. The fact that responses were poor indicates that imprecision in spike timing may not be the only factor limiting the encoding of temporal frequency. The data are discussed in relation to evidence on the temporal tuning of central neurons seen in studies using continuous amplitude modulations.


Warfarin therapy and incidence of cerebrovascular complications in left-sided native valve endocarditis.
Anticoagulant therapy has been anticipated to increase the risk of cerebrovascular complications (CVC) in native valve endocarditis (NVE). This study investigates the relationship between ongoing oral anticoagulant therapy and the incidence of symptomatic CVC in left-sided NVE. In a prospective cohort study, the CVC incidence was compared between NVE patients with and without ongoing warfarin. Among 587 NVE episodes, 48 (8%) occurred in patients on warfarin. A symptomatic CVC was seen in 144 (25%) patients, with only three on warfarin. CVC were significantly less frequent in patients on warfarin (6% vs. 26%, odds ratio [OR] 0.20, 95% confidence interval [CI] 0.06-0.6, p = 0.006). No increase in haemorrhagic lesions was detected in patients on warfarin. Staphylococcus aureus aetiology (adjusted OR [aOR] 6.3, 95% CI 3.8-10.4) and vegetation length (aOR 1.04, 96% CI 1.01-1.07) were risk factors for CVC, while warfarin on admission (aOR 0.26, 95% CI 0.07-0.94), history of congestive heart failure (adjusted OR 0.22, 95% CI 0.1-0.52) and previous endocarditis (aOR 0.1, 95% CI 0.01-0.79) correlated with lower CVC frequency.


Using CV-GLUE procedure in analysis of wetland model predictive uncertainty.
This study develops a procedure that is related to Generalized Likelihood Uncertainty Estimation (GLUE), called the CV-GLUE procedure, for assessing the predictive uncertainty that is associated with different model structures with varying degrees of complexity. The proposed procedure comprises model calibration, validation, and predictive uncertainty estimation in terms of a characteristic coefficient of variation (characteristic CV). The procedure first performed two-stage Monte-Carlo simulations to ensure predictive accuracy by obtaining behavior parameter sets, and then the estimation of CV-values of the model outcomes, which represent the predictive uncertainties for a model structure of interest with its associated behavior parameter sets. Three commonly used wetland models (the first-order K-C model, the plug flow with dispersion model, and the Wetland Water Quality Model; WWQM) were compared based on data that were collected from a free water surface constructed wetland with paddy cultivation in Taipei, Taiwan. The results show that the first-order K-C model, which is simpler than the other two models, has greater predictive uncertainty. This finding shows that predictive uncertainty does not necessarily increase with the complexity of the model structure because in this case, the more simplistic representation (first-order K-C model) of reality results in a higher uncertainty in the prediction made by the model. The CV-GLUE procedure is suggested to be a useful tool not only for designing constructed wetlands but also for other aspects of environmental management.


Predictors of Recurrent Stroke After Embolic Stroke of Undetermined Source in the RE-SPECT ESUS Trial.
Background We sought to determine recurrent stroke predictors among patients with embolic strokes of undetermined source (ESUS). Methods and Results We applied Cox proportional hazards models to identify clinical features associated with recurrent stroke among participants enrolled in RE-SPECT ESUS (Randomized, Double-Blind, Evaluation in Secondary Stroke Prevention Comparing the Efficacy and Safety of the Oral Thrombin Inhibitor Dabigatran Etexilate Versus Acetylsalicylic Acid in Patients With Embolic Stroke of Undetermined Source) trial, an international clinical trial evaluating dabigatran versus aspirin for patients with ESUS. During a median follow-up of 19 months, 384 of 5390 participants had recurrent stroke (annual rate, 4.5%). Multivariable models revealed that stroke or transient ischemic attack before the index event (hazard ratio [HR], 2.27 [95% CI, 1.83-2.82]), creatinine clearance <50 mL/min (HR, 1.69 [95% CI, 1.23-2.32]), male sex (HR, 1.60 [95% CI, 1.27-2.02]), and CHA2DS2-VASc ≥4 (HR, 1.55 [95% CI, 1.15-2.08] and HR, 1.66 [95% CI, 1.21-2.26] for scores of 4 and ≥5, respectively) versus CHA2DS2-VASc of 2 to 3, were independent predictors for recurrent stroke. Conclusions In RE-SPECT ESUS trial, expected risk factors previously linked to other common stroke causes were associated with stroke recurrence. These data help define high-risk groups for subsequent stroke that may be useful for clinicians and for researchers designing trials among patients with ESUS. Registration URL: https://www.clinicaltrials.gov; Unique identifier: NCT02239120.


In Vitro Analysis of the Cytocompatibility of a Novel Porcine Aortic Patch for Vascular Reconstruction.
Cardiovascular diseases are one of the most common causes of morbidity and mortality in the world. In the case of severe arteriosclerotic damage, surgical treatment is necessary. Although the use of autologous vessels is still considered to be the gold standard, sufficient autologous vessels for transplantation are lacking. In the present study, histological examination and in vitro cytotoxicity analysis according to DIN EN ISO 10993-5 were performed on a newly developed porcine vascular graft from a decellularized aorta. A conventional bovine graft was used as control. The ex vivo-histological analysis revealed the effectiveness of a new purification process on the microstructure and the removal of xenogeneic antigen-bearing structures in the new vessels. Furthermore, cell viability and cytotoxicity assays revealed full cytocompatibility. The novel graft shows no structural damage and gets completely decellularized by the purification process. Superior cytocompatibility, compared with the bovine-derived vascular graft, was demonstrated.


Effect of the reverse Trendelenburg position on the incidence of gastroesophageal reflux in dogs anesthetized for elective stifle surgery.
To determine if a 15° reverse Trendelenburg position decreases the incidence of gastroesophageal reflux (GER) compared with a horizontal position in dogs anesthetized for stifle surgery. Prospective, randomized parallel-arm study. A total of 44 healthy client-owned dogs were enrolled and data from 36 dogs were analyzed. Dogs requiring preoperative radiographs under anesthesia, or with a history of gastrointestinal signs or administered gastroprotectant therapy within 1 month of surgery were excluded. Anesthesia protocol was standardized to include hydromorphone, dexmedetomidine, ketamine, propofol and isoflurane. Dogs were randomly assigned at enrollment to be positioned in a 15° reverse Trendelenburg or a horizontal position for surgery. Continuous pH monitoring was documented throughout the procedure with a 6.4 Fr (2.13 mm) esophageal pH probe positioned in the distal esophagus via the oral cavity. GER was defined as pH < 4.0 (acidic) or > 7.5 (alkaline) for more than 30 seconds. The proportions of dogs developing GER were compared between groups using Fisher's exact test. Time to reflux was compared using survival curves and the Gehan-Breslow-Wilcoxon test. Statistical significance was set as p < 0.05. An episode of GER occurred in 11/36 (30%) dogs. Reflux was alkaline in two dogs and acidic in nine dogs. The proportion of dogs with GER was 5/18 (28%) and 6/18 (33%) for dogs in the reverse Trendelenburg position and horizontal position, respectively, and was not statistically significant (p > 0.99). Median (range) time until reflux was 44 (23-135) and 44.5 (9-56) minutes when dogs were positioned in reverse Trendelenburg position and horizontal position, respectively (p = 0.66; two-tailed Mann-Whitney U test). Positioning the surgery table in a 15° rostral elevation for dogs anesthetized for elective stifle surgical procedures did not decrease the incidence of GER.


Cutaneous Conundrums in Cardiac Sarcoidosis: A Series on Skin Disease in Immunosuppression.
Sarcoidosis is a complex systemic condition resulting in formation of non-caseating granulomas. Infiltrative disease in cardiac sarcoidosis can have significant ramifications on mortality and is one of the few indications for systemic immunosuppressive therapy. In the patient on immunosuppressive medication, resultant sequelae such as skin and soft tissue infections are common and must be differentiated from cutaneous forms of sarcoidosis and other skin pathologies. Patients with humoral or cellular immunodeficiencies may have cutaneous lesions secondary to endemic fungi, mycobacterium, viral diseases, parasites, or encapsulated organisms. We report a rare case of cardiac sarcoidosis on immunosuppressive therapy, with a series of cutaneous sequelae due to opportunistic infection.


Pneumoscrotum: A rare complication of ruptured emphysema bullae.
Subcutaneous emphysema and pneumoscrotum due to spontaneous pneumothorax is a rare clinical situation. We present the case of an 82 year old man who was admitted to the emergency department in state of respiratory distress and scrotum increased in size, painful and rapidly progressive installation. Clinical evaluation revealed scrotal emphysema associated with diffuse subcutaneous emphysema in the trunk, the face and both upper limbs. The treatment of this pathology is not yet standardized; antibiotic therapy and close monitoring are recommended.


The effect of torrefaction and ZSM-5 catalyst for hydrocarbon rich bio-oil production from co-pyrolysis of cellulose and low density polyethylene via microwave-assisted heating.
This study aims to investigate the effect of microwave torrefaction and ZSM-5 catalyst for hydrocarbon rich bio-oil production from microwave co-pyrolysis of cellulose and low density polyethylene (LDPE). FTIR analysis displayed remarkable reductions of active hydroxyl and ether groups in microwave torrefied cellulose (MTC), demonstrating that cellulose was less stable than MTC. GC/MS analysis indicated that the hydrocarbons content was ranged from 18.36% to 54.94% in the obtained bio-oils under different conditions, and the maximum hydrocarbons content (54.94%) which also contained the highest aromatic hydrocarbons (19.49%) was obtained from MTC catalytic co-pyrolysis. Microwave-assisted Thermogravimetric analyzer (MW-TGA) analysis showed that MTC catalytic co-pyrolysis apparently shifted the major thermal degradation to a lower temperature area, an evident synergistic effect was observed during MTC catalytic co-pyrolysis. Kinetics study revealed that the activation energy was significantly reduced from 97.87 kJ/mol to 63.86 kJ/mol for co-pyrolysis and MTC catalytic co-pyrolysis, respectively.


Treatment of primary hypertriglyceridemia states--General approach and the role of extracorporeal methods.
Hypertriglyceridemia (HTG) is a common metabolic disorder in which the concentration of very low density lipoproteins (VLDL) and of chylomicrons (CMs) is elevated in the plasma. HTG may be caused by primary and/or secondary causes and affected subjects may express HTG when children or in adulthood. In children and adults a genetic cause may underlie HTG which can be expressed as CMs a severe clinical picture known as Familial Hyperchylomicronemia due to lipoprotein lipase (LPL) or apolipoprotein (apo) CII deficiencies. Genetically determined HTG includes Familial Dysbetalipoproteinemia due to deficiency of apolipoprotein EIII of VLDL and Familial HTG. However, recent data suggest that classical Fredrickson phenotypes describing clinically HTG, which were once considered to be distinct based on biochemical features, have a shared genetic set up. The HTG has been classified according to a recent international paper: mild HTG: 2-10 mmol/L (176-882 mg/dL); severe HTG: > 10 mmol/L (>882 mg/dL) associated to CMs remnants, or Intermediate Density lipoprotein (IDL) like particles, and/or CMs. The treatment includes limitation of dietary content of saturated fat and alcohol, fibrates and omega3 fatty acids. When TG are severely elevated and associated to CMs the risk of acute pancreatitis suggests the use of more drastic therapeutic option such as therapeutic plasma exchange. This paper summarizes the experience with conventional plasmapheresis (Plasma-Exchange, PEX) and different Lipoprotein Apheresis methods with respect to acutely lowering TG levels in patients with normal TG, with mild and severe HTG. Upcoming promising therapies are gene therapy, novel apolipoprotein CIII inhibitors and lomitapide.


Improved prediction of protein-protein interactions by a modified strategy using three conventional docking software in combination.
Proteins play a crucial role in many biological processes, where their interaction with other proteins are integral. Abnormal protein-protein interactions (PPIs) have been linked to various diseases including cancer, and thus targeting PPIs holds promise for drug development. However, experimental confirmation of the peculiarities of PPIs is challenging due to their dynamic and transient nature. As a complement to experimental technologies, multiple computational molecular docking (MD) methods have been developed to predict the structures of protein-protein complexes and their dynamics, still requiring further improvements in several issues. Here, we report an improved MD method, namely three-software docking (3SD), by employing three popular protein-peptide docking software (CABS-dock, HPEPDOCK, and HADDOCK) in combination to ensure constant quality for most targets. We validated our 3SD performance in known protein-peptide interactions (PpIs). We also enhanced MD performance in proteins having intrinsically disordered regions (IDRs) by applying the modified 3SD strategy, the three-software docking after removing random coiled IDR (3SD-RR), to the comparable crystal PpI structures. At the end, we applied 3SD-RR to the AlphaFold2-predicted receptors, yielding an efficient prediction of PpI pose with high relevance to the experimental data regardless of the presence of IDRs or the availability of receptor structures. Our study provides an improved solution to the challenges in studying PPIs through computational docking and has the potential to contribute to PPIs-targeted drug discovery. SIGNIFICANCE STATEMENT: Protein-protein interactions (PPIs) are integral to life, and abnormal PPIs are associated with diseases such as cancer. Studying protein-peptide interactions (PpIs) is challenging due to their dynamic and transient nature. Here we developed improved docking methods (3SD and 3SD-RR) to predict the PpI poses, ensuring constant quality in most targets and also addressing issues like intrinsically disordered regions (IDRs) and artificial intelligence-predicted structures. Our study provides an improved solution to the challenges in studying PpIs through computational docking and has the potential to contribute to PPIs-targeted drug discovery.


White matter tracts related to memory and emotion in very preterm children.
Very preterm (VP) children are at risk of memory and emotional impairments; however, the neural correlates remain incompletely defined. This study investigated the effect of VP birth on white matter tracts traditionally related to episodic memory and emotion. The cingulum, fornix, uncinate fasciculus, medial forebrain bundle and anterior thalamic radiation were reconstructed using tractography in 144 VP children and 33 full-term controls at age 7 years. Compared with controls, VP children had higher axial, radial, and mean diffusivities and neurite orientation dispersion, and lower volume and neurite density in the fornix, along with higher neurite orientation dispersion in the medial forebrain bundle. Support vector classification models based on tract measures significantly classified VP children and controls. Higher fractional anisotropy and lower diffusivities in the cingulum, uncinate fasciculus, medial forebrain bundle and anterior thalamic radiation were associated with better episodic memory, independent of key perinatal risk factors. Support vector regression models using tract measures did not predict episodic memory and emotional outcomes. Altered tract structure is related to adverse episodic memory outcomes in VP children, but further research is required to determine the ability of tract structure to predict outcomes of individual children. We studied white matter fibre tracts thought to be involved in episodic memory and emotion in VP and full-term children using diffusion magnetic resonance imaging and machine learning. VP children have altered fornix and medial forebrain bundle structure compared with full-term children. Altered tract structure can be detected using machine learning, which accurately classified VP and full-term children using tract data. Altered cingulum, uncinate fasciculus, medial forebrain bundle and anterior thalamic radiation structure was associated with poorer episodic memory skills using linear regression. The ability of tract structure to predict episodic memory and emotional outcomes of individual children based on support vector regression was limited.


A Pilot Comparison of High- Versus Low-Tech Palatability Assessment Tools in Young Children.
Medication refusal in children is largely driven by aversive taste profiles, which in turn influence adherence and therapeutic outcomes. However, there are no standardized methods for evaluating taste in young children. This study compares facial recognition technology with three hedonic visual scales in this population. Children, 3-7 years of age, were enrolled with informed parental permission into an institutional review board-approved, double-blind, randomized investigation. Each child received three test articles: prednisone (bitter), simple syrup (sweet), and filtered water (neutral), with an appropriate washout. Facial recognition software (Noldus FaceReader 7) recorded facial expression and intensity for 30-60 s after administration. Participants subsequently rated taste using three hedonic scales (5-point Sjövall and 5- and 3-point TASTY) and responded to simple questions on their perception of the test article. Repeated measures analysis of variance and multiple regression analysis were used to explore associations between palatability measures. Twelve children (seven males: ten white and two black) completed the study without adverse effects. There were no significant differences in participant characteristics by randomization sequence. The three hedonic scales tracked similarly for each test substance, with correlations between the 5-point scales (r = 0.899) comparable to those between the 3- and 5-point scales (r = 0.860-0.903). Hedonic scales appeared more reliable in assessing taste response than facial recognition, which did not effectively discriminate positive and negative responses. Our experience suggests that the TASTY scales appear to offer the greatest promise for assessing palatability in future clinical use.


Impact of conversion on short and long-term outcome in laparoscopic resection of curable colorectal cancer.
Long-term outcome of patients following conversion during laparoscopic surgery for colorectal cancer is not often reported. Recent data suggest a negative impact of conversion on long-term survival. This study aimed to evaluate the impact of conversion on the perioperative outcome and on long-term survival in patients who underwent laparoscopic resection for curable colorectal cancer. Evaluation of our prospective in-hospital collected data of patients who underwent laparoscopic surgery for curable colorectal cancer over a 5-year period. Long-term data were collected from our outpatient's clinic data and personal contact when necessary. During the study period, 175 patients were operated on laparoscopically for curable colon cancer (stage I-III). Mean follow-up was 33±18 months with a minimum follow-up of 12 months. For various reasons, 25 patients (14.4%) had to be converted to open surgery. Short-term outcome revealed a trend towards longer operations, a higher rate of surgical complications, and a longer hospital stay in the converted group. Five-year, Kaplan-Meier, disease-free analysis was worse for converted patients. Overall survival did not differ between the 2 groups. Cox proportional hazards regression analysis revealed that conversion and AJCC stage were independent risk factors for recurrence. Conversion in laparoscopic surgery for curable colorectal cancer is associated with a worse perioperative outcome and worse disease-free survival.


Improved Accuracy of Automated Estimation of Cardiac Output Using Circulation Time in Patients with Heart Failure.
Lung-to-finger circulation time of oxygenated blood during nocturnal periodic breathing in heart failure patients measured using polysomnography correlates negatively with cardiac function but possesses limited accuracy for cardiac output (CO) estimation. CO was recalculated from lung-to-finger circulation time using a multivariable linear model with information on age and average overnight heart rate in 25 patients who underwent evaluation of heart failure. The multivariable model decreased the percentage error to 22.3% relative to invasive CO measured during cardiac catheterization. This improved automated noninvasive CO estimation using multiple variables meets a recently proposed performance criterion for clinical acceptability of noninvasive CO estimation, and compares very favorably with other available methods.


Complications of Palatal Pillar Implants: An analysis of the MAUDE database and literature review.
The Pillar Palatal Implant System is a minimally invasive procedure for the treatment of obstructive sleep apnea and snoring. Prior studies have examined the effectiveness of this procedure, however no prior study has thoroughly evaluated its complications. We anticipate that this analysis will provide valuable insight into these procedures which can be used in patient education and post-operative follow-up. The Manufacturer and User Facility Device Experience (MAUDE) database was queried for cases reporting injury related to the pillar procedure. The case narratives were individually analyzed and categorized by type of complication. A comparison of extruding parts and select complications was also performed using chi-square analysis. Of the 261 cases reported, 73.6% reporting extruding parts, 47.5% described patient pain, and 38.7% included patients describing a foreign body sensation. Infection was reported in 7.7% of the cases with cellulitis and abscess formation specifically mentioned in one case. Cases describing pain and difficulty swallowing were significantly more likely to have reported extruding parts (p < 0.05). This study describes various complications associated with this procedure. The description of these complications and accompanying literature review may provide healthcare professionals and patients with an understanding of the scope of adverse events related to this procedure.


Analysis of the NCI-60 dataset for cancer-related microRNA and mRNA using expression profiles.
Recent studies have indicated that microRNA (miRNA) may play an oncogenic or tumor suppressor role in human cancer. To study the regulatory role of miRNAs in tumorigenesis, an integrated platform has been set up to provide a user friendly interface for query. The main advantage of the present platform is that all the miRNA target genes' information and disease records are drawn from experimentally verified or high confidence records. MiRNA target gene results are annotated with reference to the disease gene as well as the pathway database. The correlation strength between miRNA and target gene expression profile is quantified by computing the correlation coefficient using the NCI-60 expression profiling data. Comprehensive analysis of the NCI-60 data found that the cumulative percentage of negative correlation coefficients for cleavage regulation is slightly higher than its positive counterpart; which indicated that the mRNA degradation mechanism is slightly dominant. In addition, the RNAHybrid and TargetScans scores are computed which potentially served as quantitative estimators for miRNA-mRNA binding events. Three scores are defined for each miRNA-mRNA pair, which are based on the disease gene and pathway information. These three scores allow user to sort out high confidence cancer-related miRNA-mRNA pairs. Statistical tests were applied to investigate the relations of three chromosomal features, i.e., CpG island, fragile site, and miRNA cluster, with cancer-related miRNAs. A web-based interface has been set up for query, which can be accessed at: http://ppi.bioinfo.asia.edu.tw/mirna_target/ The main advantage of the present platform on miRNA-mRNA targeting information is that all the target genes' information and disease records are experimentally verified. Although this may limit the number of miRNA-mRNA relationships, the results provided here are more solid and have fewer false positive events. Certain novel cancer-related miRNA-mRNA pairs are identified and confirmed in the literature. Fisher's exact test suggests that CpG island and fragile site associated miRNAs tend to associate with cancer formation. In summary, the present platform provides an easy means of investigating cancer-related miRNAs.


Methane adsorption and methanol desorption of copper modified boron silicate.
Boron silicate (BS) with a chabazite framework structure was synthesised using a direct route and rigorously characterized before it was ion-exchanged with copper to form Cu-BS. Employing in situ infrared spectroscopy, we show that Cu-BS is capable of oxidising methane to methoxy species and methanol interacts with the boron sites without deprotonation.


Epiretinal membrane surgery: anatomic and functional outcomes.
To study the influence of anatomic preoperative characteristics (based on the parameter, foveal central thickness, measured by optical coherence tomography) and functional characteristics (based on the parameter, best corrected visual acuity, [BCVA]) on functional recovery after epiretinal membrane (ERM) surgery. A total of 88 eyes (of 86 patients), on whom a vitrectomy due to ERM was performed during a 3 years period were reviewed in this longitudinal, prospective study. An analysis was made of, ERM aetiology, BCVA, presence or absence of metamorphopsia, lens status, and central foveal thickness. Data relating to surgery and local complications, changes in BCVA, and changes in foveal central thickness were collected during the follow-up period. An improvement was in observed in BCVA in 82%, as well as a decrease in foveal thickness in 79% of the eyes which underwent surgery, both of these being statistically significant (P<.01). However, most of the patients showed different grades of oedema and/or macular thickening in the postoperative period. A significant correlation was found between preoperative and postoperative BCVA (P=.001), and also between preoperative and postoperative central foveal thickness (P=.004), but not between BCVA and foveal thickness. There is functional recovery in terms of BCVA in more than 80% of the patients after ERM surgery. Most of the eyes showed persistent macular thickening, but this did not seem to have influenced the final BCVA. The best determinant of postoperative functional recovery (postoperative visual acuity) is, in our experience, the preoperative BCVA, and not the macular thickness.


Effects of radio frequency heating on microbial populations and physicochemical properties of buckwheat.
Microbial contamination is a persistent problem for grain industry. Many studies have shown that radio frequency (RF) heating can effectively reduce pathogens populations in low moisture foods, but there is a lack on the efficacy to decontaminate natural microbiome. The main objectives of this study were to investigate the effects of different RF heating conditions on natural microbial populations and physicochemical properties of buckwheat. In this study, 30 buckwheat samples collected from 10 different Provinces in China were analyzed for their microbial loads, and the samples with the highest microbial populations were used for further study to select the suitable RF heating conditions. The results showed that microbial loads in tested buckwheat kernels were in the range of 3.4-6.2 log CFU/g. Samples from Shanxi (SX-3) had significantly higher microbial counts than other samples. The selected four temperature-time combinations: 75 °C-20 min, 80 °C-10 min, 85 °C-5 min, and 90 °C-0 min of RF heating could reduce microbial counts to <3.0 log CFU/g in buckwheat kernels at 16.5% w.b. moisture content. Furthermore, the reduction populations of the inoculated pathogens (Salmonella Typhimurium, Escherichia coli, Cronobacter sakazakii, and Bacillus cereus) reached 4.0 log CFU/g under the above conditions, and almost 5.0 log CFU/g especially at high temperature-short holding time combinations (85 °C-5 min and 90 °C-0 min). Besides, physicochemical properties evaluation also showed the insignificant color changes and nutrients loss after RF treatment at 90 °C-0 min. Therefore, the RF heating at 90 °C-0 min holds greater potential than the other lower temperature-longer holding time combinations for applications in buckwheat pasteurization.


Evidence that ghrelin may be associated with the food intake of gibel carp (Carassius auratus gibelio).
Ghrelin, a non-amidated peptide hormone, is a potent anorectic neuropeptide implicated in feeding regulation in mammals and non-mammalian vertebrates. However, the involvement of ghrelin in the feeding behavior of teleosts has not been well understood. To better understand the role of ghrelin in the regulation of appetite in fish, in this study, we cloned the cDNAs encoding ghrelin and investigated their mRNA distributions in gibel carp tissues. We also assessed the effects of different nutritional status on ghrelin mRNA abundance. Ghrelin mRNAs were ubiquitously expressed in ten tissues (intestine, liver, brain, mesonephron, head kidney, spleen, skin, heart, muscle, gill and pituitary gland), and relatively high expression levels were detected in the gut. Postprandial studies analysis revealed a significant postprandial decrease in ghrelin mRNA expression in the gut (1 and 3 h after the regular feeding time). In addition, ghrelin mRNA expression in the gut significantly increased at day 7 after fasting and declined sharply after refeeding, which suggested that ghrelin might be involved in the regulation of appetite in gibel carp. Overall, our result provides basis for further investigation into the regulation of feeding in gibel carp.


Development and validation of equation for cardiorespiratory fitness in patients with heart failure with preserved ejection fraction.
Cardiorespiratory fitness (CRF), as measured by maximal oxygen consumption (VO2max), is an important independent predictive factor of cardiovascular outcomes in patients with heart failure (HF). However, it is unclear whether conventional equations for estimating CRF are applicable to patients with HF with preserved ejection fraction (HFpEF). This study included 521 patients with HFpEF (EF ≥ 50%) whose CRF was directly measured by cardiopulmonary exercise test using a treadmill. We developed a new equation (Kor-HFpEF) for half of the patients in the HFpEF cohort (group A, n = 253) and validated it for the remaining half (group B, n = 268). The accuracy of the Kor-HFpEF equation was compared to that of the other equations in the validation group. In the total HFpEF cohort, the directly measured VO2max was significantly overestimated by the FRIEND and ACSM equations (p < 0.001) and underestimated by the FRIEND-HF equation (p <0.001) (direct 21.2 ± 5.9 mL/kg/min; FRIEND 29.1 ± 11.8 mL/kg/min; ACSM 32.5 ± 13.4 mL/kg/min; FRIEND-HF 14.1 ± 4.9 mL/kg/min). However, the VO2max estimated by the Kor-HFpEF equation (21.3 ± 4.6 mL/kg/min) was similar to the directly measured VO2max (21.7 ± 5.9 mL/kg/min, p = 0.124), whereas the VO2max estimated by the other three equations was still significantly different from the directly measured VO2max in group B (all p < 0.001). Traditional equations used to estimate VO2max were not applicable to patients with HFpEF. We developed and validated a new Kor-HFpEF equation for these patients, which had a high accuracy.


Insight into designing of 2-pyridone derivatives for COVID-19 drug discovery - A computational study.
Presently, the prime global focus is on SARS-CoV-2, as no fully established, licensed medicine has been found thus far, in spite of the existence of various reports and administration of partially proven certain class of natural products. However, in case of natural products, the extraction and purification limit their application. This situation drives researchers to explore synthetically viable drugs. In the present investigation, twenty-three 2-pyridone synthetic derivatives (P1-P23) have been theoretically tested for their suitability as potential inhibitors for COVID-19 main protease through DFT, molecular docking, and molecular dynamics simulations. DFT calculations offer insights into structure-property relationships, while ADMET studies indicate the pharmacological characteristics of these molecules. Molecular docking studies ascertain the nature and mode of interactions of these entities with COVID-19 main protease. Furthermore, covalent docking has been carried out to verify the feasibility of the formation of a covalent bond with the active site. The top protein-inhibitor complexes, such as P18, P11, and P12, were identified based on their glide score. These molecules, along with the covalent docked complexes, namely P18 and P16, were selected and subjected to molecular dynamics simulations. The 100 ns simulation process exhibited that the covalent docked ones, due to their stable form could serve as lead compounds against SARS-CoV-2. Hence, this study affirms the potential candidature of 2-pyridone-based inhibitors.


Analysis of DFS70 pattern and impact on ANA screening using a novel HEp-2 ELITE/DFS70 knockout substrate.
Indirect immunofluorescence (IIF) using human epithelial cell (HEp-2) substrate is a widely used and the recommended method for screening of antinuclear antibodies (ANA). Dense fine speckled (DFS70) pattern on HEp-2 has been widely reported in various healthy and disease groups. Interpretation of DFS70 pattern can be challenging on a conventional HEp-2 substrate due to its similarity to some of the disease associated patterns. The high prevalence of DFS70 autoantibodies in normal population, lack of association with a particular disease group and a general negative association with systemic and ANA associated autoimmune rheumatic diseases (SARD/AARD) necessitates the confirmation of DFS70 pattern. Results using available commercial assays for confirmation of DFS70 autoantibodies do not always agree with IIF screening results further complicating the lab work flow and ANA algorithms. In this review, we discuss the prevalence of DFS70 antibodies and factors affecting the performance of IIF and DFS70 specific confirmatory assays. Factors that contribute to disagreement between DFS70 suspicion by IIF and confirmatory assays will also be discussed. In addition, we also describe a novel IIF HEp-2 substrate, and its positive impact on DFS70 reporting and ANA screening-confirmation algorithm.


Microtubules Enhance Mesoscale Effective Diffusivity in the Crowded Metaphase Cytoplasm.
Mesoscale macromolecular complexes and organelles, tens to hundreds of nanometers in size, crowd the eukaryotic cytoplasm. It is therefore unclear how mesoscale particles remain sufficiently mobile to regulate dynamic processes such as cell division. Here, we study mobility across dividing cells that contain densely packed, dynamic microtubules, comprising the metaphase spindle. In dividing human cells, we tracked 40 nm genetically encoded multimeric nanoparticles (GEMs), whose sizes are commensurate with the inter-filament spacing in metaphase spindles. Unexpectedly, the effective diffusivity of GEMs was similar inside the dense metaphase spindle and the surrounding cytoplasm. Eliminating microtubules or perturbing their polymerization dynamics decreased diffusivity by ~30%, suggesting that microtubule polymerization enhances random displacements to amplify diffusive-like motion. Our results suggest that microtubules effectively fluidize the mitotic cytoplasm to equalize mesoscale mobility across a densely packed, dynamic, non-uniform environment, thus spatially maintaining a key biophysical parameter that impacts biochemistry, ranging from metabolism to the nucleation of cytoskeletal filaments.


A new multiplex assay allowing simultaneous detection of the inhibition of cell proliferation and induction of cell death.
The efficacy of distinct anti-cancer drugs used in the chemotherapy of human malignancies varies between tumor tissues and depends largely on the ability of the therapeutic agents to simultaneously inhibit cell proliferation and to eliminate malignant cells by apoptosis. Especially, detection of early apoptotic changes seems to be important because early stages of apoptosis differ from those of necrosis. Therefore, the development of a novel test allowing fast and concomitant screening of the anti-proliferative and pro-apoptotic action of a number of anti-cancer drugs is of great interest. For this purpose, we choose as an experimental model a well characterized anti-proliferative and pro-apoptotic effect of cisplatin (CP) on human cervical carcinoma HeLaS3 cells. As previously reported, exposure of HeLaS3 to CP resulted in a concomitant inhibition of cell proliferation and induction of apoptosis in a dose- and time-dependent manner. In the present study we performed two independent approaches. In the first approach, we examined the cell proliferation and activity of caspases-3/7 in two separate microtiter plates using the CellTiter-Glo Luminescent Cell Viability Assay and the Caspase-Glo 3/7 Assay, respectively. In the second approach, we determined the same parameters sequentially in one microtiter plate by a mutiplexing assay using CellTiter-Blue Cell Viability Assay and Caspase-Glo 3/7 Assay. The both approaches gave very similar results indicating that this new multiplexing assay offers an important advantage for simultaneous detection of cell number and activation of caspases-3/7. The new multiplexing assay offers a range of benefits over standard assays.


Synthesis and evaluation of paclitaxel C7 derivatives: solution phase synthesis of combinatorial libraries.
A novel and efficient two-step, automated solution phase synthesis of a 26-membered combinatorial chemistry library of paclitaxel C7 esters was accomplished using the HP 7686 Solution Phase Synthesizer. Results of combinatorial synthesis, purification, analysis, and biological evaluation are described.