Artemisinin resistance containment project in Thailand. (I): Implementation of electronic-based malaria information system for early case detection and individual case management in provinces along the Thai-Cambodian border.
The Bureau of Vector-borne Diseases, Ministry of Public Health, Thailand, has implemented an electronic Malaria Information System (eMIS) as part of a strategy to contain artemisinin resistance. The attempt corresponds to the WHO initiative, funded by the Bill & Melinda Gates Foundation, to contain anti-malarial drug resistance in Southeast Asia. The main objective of this study was to demonstrate the eMIS' functionality and outputs after implementation for use in the Thailand artemisinin-resistance containment project. The eMIS had been functioning since 2009 in seven Thai-Cambodian border provinces. The eMIS has covered 61 malaria posts/clinics, 27 Vector-borne Disease Units covering 12,508 hamlets at risk of malaria infections. The eMIS was designed as an evidence-based and near real-time system to capture data for early case detection, intensive case investigation, monitoring drug compliance and on/off-site tracking of malarial patients, as well as collecting data indicating potential drug resistance among patients. Data captured by the eMIS in 2008-2011 were extracted and presented. The core functionalities of the eMIS have been utilized by malaria staff at all levels, from local operational units to ministerial management. The eMIS case detection module suggested decreasing trends during 2009-2011; the number of malaria cases detected in the project areas over the years studied were 3818, 2695, and 2566, with sero-positive rates of 1.24, 0.98, and 1.16%, respectively. The eMIS case investigation module revealed different trends in weekly Plasmodium falciparum case numbers, when classified by responsible operational unit, local and migrant status, and case-detection type. It was shown that most Thai patients were infected within their own residential district, while migrants were infected either at their working village or from across the border. The data mapped in the system suggested that P. falciparum-infected cases and potential drug-resistant cases were scattered mostly along the border villages. The mobile technology application has detected different follow-up rates, with particularly low rates among seasonal and cross-border migrants. The eMIS demonstrated that it could capture essential data from individual malaria cases at local operational units, while effectively being used for situation and trend analysis at upper-management levels. The system provides evidence-based information that could contribute to the control and containment of resistant parasites. Currently, the eMIS is expanding beyond the Thai-Cambodian project areas to the provinces that lie along the Thai-Myanmar border.


RNA splicing process analysis for identifying antisense oligonucleotide inhibitors with padlock probe-based isothermal amplification.
RNA splicing, which mainly involves two transesterification steps, is a fundamental process of gene expression and its abnormal regulation contributes to serious genetic diseases. Antisense oligonucleotides (ASOs) are genetic control tools that can be used to specifically control genes through alteration of the RNA splicing pathway. Despite intensive research, how ASOs or various other factors influence the multiple processes of RNA splicing still remains obscure. This is largely due to an inability to analyze the splicing efficiency of each step in the RNA splicing process with high sensitivity. We addressed this limitation by introducing a padlock probe-based isothermal amplification assay to achieve quantification of the specific products in different splicing steps. With this amplified assay, the roles that ASOs play in RNA splicing inhibition in the first and second steps could be distinguished. We identified that 5'-ASO could block RNA splicing by inhibiting the first step, while 3'-ASO could block RNA splicing by inhibiting the second step. This method provides a versatile tool for assisting efficient ASO design and discovering new splicing modulators and therapeutic drugs.


Role of vasopressin in neurocardiogenic responses to hemorrhage in conscious rats.
Vasovagal reflexes, such as hypotension and bradycardia, are induced by rapid hemorrhage and mimic neurocardiogenic reflexes in mammals. We examined the role of vasopressin in the neurocardiogenic responses to mild, rapid hemorrhage (1 mL/100 g for 30 seconds) and severe hemorrhage (1 mL/100 g body wt for 30 seconds repeated three times at 11-minute intervals) in homozygous Brattleboro and Long-Evans rats. Mild, rapid hemorrhage induced severe bradycardia and hypotension only in Long-Evans rats. Exogenous vasopressin (1.85 pmol/kg per minute for 1 hour) restored both the bradycardic and hypotensive responses in Brattleboro rats. DDAVP, a vasopressin V2-receptor agonist (0.19 pmol/kg per minute for 24 hours), did not affect the cardiovascular responses to hemorrhage in Brattleboro rats, although it maintained urine production within normal limits. However, OPC-31260 (21.6 mumol/kg IV), a vasopressin V2-receptor antagonist, attenuated both the hypotensive and bradycardic responses to hemorrhage in Long-Evans rats. A vasopressin V1-receptor antagonist attenuated bradycardia and delayed the recovery of arterial pressure after hemorrhage but did not affect the hypotension that occurred immediately after hemorrhage in Long-Evans rats. Methylatropine also attenuated both the bradycardic and hypotensive responses induced by hemorrhage, but propranolol had no effect on the cardiovascular responses to hemorrhage in Long-Evans rats. The recovery of arterial pressure after repeated hemorrhage was less adequate in Brattleboro rats than in Long-Evans rats. Our results suggest that the neurocardiogenic responses to hemorrhage, especially hypotension, may be related to vasodilation induced by a V2-receptor-mediated mechanism and by the vagal reflex, both of which are substantiated by the existence of vasopressin. The coexistence of V1- and V2-receptor mechanisms may be necessary for the hypotensive response to hemorrhage. We found that a V2-receptor antagonist attenuated the hypotension mediated by the so-called neurocardiogenic reflex.


Comparative power spectral analysis of simultaneous elecroencephalographic and magnetoencephalographic recordings in humans suggests non-resistive extracellular media.
The resistive or non-resistive nature of the extracellular space in the brain is still debated, and is an important issue for correctly modeling extracellular potentials. Here, we first show theoretically that if the medium is resistive, the frequency scaling should be the same for electroencephalogram (EEG) and magnetoencephalogram (MEG) signals at low frequencies (<10 Hz). To test this prediction, we analyzed the spectrum of simultaneous EEG and MEG measurements in four human subjects. The frequency scaling of EEG displays coherent variations across the brain, in general between 1/f and 1/f(2), and tends to be smaller in parietal/temporal regions. In a given region, although the variability of the frequency scaling exponent was higher for MEG compared to EEG, both signals consistently scale with a different exponent. In some cases, the scaling was similar, but only when the signal-to-noise ratio of the MEG was low. Several methods of noise correction for environmental and instrumental noise were tested, and they all increased the difference between EEG and MEG scaling. In conclusion, there is a significant difference in frequency scaling between EEG and MEG, which can be explained if the extracellular medium (including other layers such as dura matter and skull) is globally non-resistive.


Sulphated glycoproteins induced by herpes simplex virus.
BHK cells infected with strain 17 syn+ (HSV-1) or HG52 (HSC-2) incorporated inorganic sulphate into polypeptides which co-migrated on SDS-polyacrylamide gels with virus-induced glycoproteins. The major sulphated glycoprotein was glycoprotein E. In addition, less-intense sulphated bands co-migrated with glycoprotein D and HSV-1 glycoprotein A/B/C. Sulphate label co-migrating with HSV-2 glycoprotein A/B/C was occasionally observed. We have investigated which sulphated polypeptides are excreted from infected cells. Major ones of apparent mol. wt. 32000, 34000 and 35000 were excreted from cells infected with syn+. In addition, polypeptides which migrated in the vicinity of glycoprotein D were often excreted from cells infected with either 17 syn+ or HG52. The 32K, 34K and 35K polypeptides were antigenically related to glycoprotein D and over 95% of the total amount synthesized was excreted. Analysis of intracellular sulphated polypeptides using intertypic recombinants mapped glycoprotein E to between 0.832 and 0.950 units of the HSV genome.


Hemispheric differences in motor control.
Two lines of evidence are presented to suggest that the left hemisphere in human beings plays a special role in the organization of complex motor behaviour, an idea first put forward by Liepmann and extended more recently by Kimura. The results of one line of research suggest that the right-sided asymmetries observed in movements of the mouth during verbal and non-verbal tasks reflect the fact that mechanisms within the left hemisphere are particularly involved in selecting individual movements and facilitating the transition from one movement to another. The results of the second line of research extend this idea and suggest that the organization of eye and limb movements during visually guided reaching is also dependent on these left-hemisphere mechanisms. These findings, together with the work of a number of other workers, all point to the same conclusion: that speech is but one example of a great number of different motor patterns mediated in part by neural systems within the so-called 'dominant' hemisphere.


Bio-antioxidants - a chemical base of their antioxidant activity and beneficial effect on human health.
The paradox of aerobic life is that higher eukaryotic organisms cannot exist without oxygen, yet oxygen is inherently dangerous to their existence. Autoxidation of organic substances frequently occurs via free radical mechanism which generates different active radicals and peroxides OH(•), O2 (•-), LO2 (•), HOOH, LOOH, so called reactive oxygen species (ROS), which appear to be responsible for oxygen toxicity. To survive in such an unfriendly oxygen environment, living organisms generate - or obtain from food - a variety of water- and lipid-soluble antioxidant compounds. Biologically active compounds with antioxidant potential, i.e. bio-antioxidants (natural and their synthetic analogues) have a wide range of applications. They are important drugs, antibiotics, agrochemical substitutes, and food preservatives. Many of the drugs today are synthetic modifications of naturally obtained substances. This review presents information about the chemical base of antioxidant activities and beneficial effects on human health of known and new bio-antioxidants. There is abundant literature on the phenolic antioxidants and tocopherols in particular. In this review the following bio-antioxidants are considered: A) Carotenoids, B) Cathecholamines, C) Phospholipids, D) Chalcones, E) Coumarins, F) Phenolic acids, G) Flavonoids, H) Lignans, and I) Tannins.


Addressing the gender gap in urinary continence care.
Urinary incontinence is a common condition that affects both men and women, and with profoundly negative effects. Prevalence figures do show that it is more common in younger women than men, but as people age the difference decreases, with research identifying that one-in-three older men have continence issues. However, even with this increase, there is little direct best practice guidance on addressing male urinary incontinence compared to that for women. Professionals seem to be unaware that men have known existing barriers to accessing health care and this would be especially true for incontinence care. There seems to be a lack of education in identifying and assessing symptoms, and little thought to appropriate management if required. This can also be true of manufacturers that provide continence management equipment. This article will look at some of these themes and highlight the gender gaps and give guidance on how professionals may address these.


Whole Genome Methylation Scanning Based on phi29 Polymerase Amplification.
Identifying differences in DNA methylation is critical to understanding how epigenetics influences gene expression during processes such as development. Here, we propose a method that employs a single, methylation-sensitive restriction endonuclease of choice, to produce discrete pools of methylated and unmethylated DNA from the same sample. A pool of restriction fragments representing unmethylated regions of the genome is first obtained by digestion with a methylation-sensitive endonuclease. The restriction-digested DNA is then concatamerized in the presence of stuffer-adaptor DNA, which prevents interference from originally unmethylated DNA by blocking the ends of the restriction fragments. The concatamerized DNA is amplified by phi29 polymerase to remove methylation marks, and again digested with the same endonuclease to produce a pool of DNA fragments representing methylated portions of the genome. The two pools of DNA fragments thus obtained can be analyzed by end-sequencing or hybridization to a genomic array. In this report we detail a proof of concept experiment that demonstrates the feasibility of our method.


One abutment one time vs. repeatable abutment disconnections in implants, restored with cemented / screw retained fixed partial dentures: Marginal bone level changes. A systematic review and meta-analysis.
The aim of this systematic review and meta-analysis is to evaluate the impact of abutment disconnections / reconnections on peri-implant marginal bone loss changes in partially edentulous patients. Clinical studies were selected via electronic and hand searches in English language journals until January 1, 2020. Only randomized clinical trials (RCGTs) and prospective controlled clinical trials (CCTs) showing direct comparison between the definitive implant abutments and multiple abutment replacements in the same patient or different patient groups in the partially edentulous patients were considered. The outcome measures were (1) the type of the abutment was used, (2) the time the abutment was placed, (3) marginal bone loss changes, (4) biological complications, (5) mechanical complications. After evaluation, 4 controlled clinical studies were included. Majority of the articles reveled protective marginal bone loss preservation for the implants with FAP (final abutment placement) at the time of implant placement compared with the implants with MAP (multiple abutment placements) in connected dental implants, in partially edentulous patients. Meta-analysis of the four studies with 280 implants reviled significantly greater bone loss in cases with multiple abutment disconnections/reconnections. The weighted mean difference in marginal bone loss was 0.4 mm (95% confidence interval, 0.16-0.63 mm), showing bone preservation in the FAP group. Within the limitations of this meta-analysis, multiple abutments disconnections significantly affected marginal bone loss changes in partially edentulous patients. The finding suggests to overview current prosthetic and surgical treatment planning protocols to prevent greater marginal bone loss.


Comparison of Ventilator Mechanics and Mortality Between COVID-19 Versus Non-COVID-19 Acute Respiratory Distress Syndrome Patients: A Cross-Sectional Study.
We aim to assess the differences in ventilator mechanics and mortality of acute respiratory distress syndrome (ARDS) between patients with and without COVID-19. It might serve as a milestone in reshaping management protocols by providing very preliminary evidence in this direction. It was a cross-sectional study that included adult patients aged 18 years or above admitted to the medical intensive care unit of our tertiary care hospital from January to December 2021 with the diagnosis of ARDS. Patients were divided into two groups. Group I were patients who had ARDS with COVID-19 infection while group II were those who had ARDS without COVID-19 infection. Both groups were compared in terms of clinical and respiratory mechanics of mechanical ventilators and mortality. The study included 135 patients, 68 of whom were in group I, and 67 were in group II. In the COVID-19 group, the median age was 60; while in the non-COVID-19 group, it was 64. There were 50% male patients in both groups. ARDS was more severe in COVID-19 (n = 44, 58%) than in the non-COVID group (n = 31, 41.3%, p-value = 0.030). The median PaO2/FiO2 ratio was 122.5 (interquartile range [IQR]: 93-160) in COVID-19 and was 180 (IQR: 127-248) in the non-COVID-19 group. Patient proning was higher (63% vs. 37%) in the COVID-19 group. In the COVID-19 group, 44 patients died compared to 32 in the non-COVID group (p-value = 0.060). COVID-19 patients had severe ARDS compared with non-COVID patients. Despite this, ventilator mechanics and mortality were not significantly different between both groups. It appears that more proning strategies were observed in the COVID-19 group and may have some positive effects.


Non-invasive imaging of mouse hepatitis coronavirus infection reveals determinants of viral replication and spread in vivo.
Bioluminescence imaging (BLI) is a powerful new method to study virus dissemination in the live animal. Here we used this method to monitor the spatial and temporal progression of mouse hepatitis coronavirus (MHV) infection in mice using luciferase-expressing viruses. Upon intranasal inoculation, virus replication could initially be observed in the nasal cavity and the cervical lymph nodes, after which the infection spread to the brain and frequently to the eyes. The kinetics of virus spread to and clearance from the brain appeared to depend on the inoculation dose. After intraperitoneal inoculation, virus replication was predominantly observed in the liver and occasionally in the intestines, but interestingly also in the tail and paws. BLI thus elucidated new anatomic locations of virus replication. Furthermore, MHV dissemination was shown to be critically depended on the viral spike protein, but also on the mouse strain used. Widespread dissemination was observed in mice lacking a functional type I interferon response. The importance of the type I interferon system in limiting viral spread was also demonstrated by the administration of type I interferons to mice. Our results provide new insights in coronavirus pathogenesis and demonstrate the potential of BLI to study coronavirus-host interactions in vivo.


Evaluation of the cytotoxicity, genotoxicity and mutagenicity of diphenyl ditelluride in several biological models.
Diphenyl ditelluride (DPDT) is a potential prototype for the development of novel biologically active molecules. Thus, it is important to evaluate the toxic effects of this compound. In the present study, we evaluated the cytotoxic, genotoxic and mutagenic properties of DPDT in Chinese hamster fibroblast (V79) cells, in strains of the yeast Saccharomyces cerevisiae both proficient and deficient in several DNA repair pathways and in Salmonella typhimurium. DPDT induced frameshift mutations in both S.typhimurium and a haploid wild-type strain of S.cerevisiae. Mutants of S.cerevisiae defective in base excision repair and recombinational repair were more sensitive to DPDT. The results of a lactate dehydrogenase leakage assay suggest that DPDT is cytotoxic to V79 cells. At cytotoxic concentrations, this compound increased thiobarbituric reactive species levels and decreased the glutathione:GSSH ratio in yeast and V79 cells. DPDT generated single- and double-strand DNA breaks in V79 cells, both with and without metabolic activation, as revealed by alkaline and neutral comet assays. Moreover, an induction of oxidative DNA base damage was indicated by a modified comet assay using formamidopyrimidine DNA glycosylase and endonuclease III. Treatment with DPDT also induced micronucleus formation in V79 cells. Pre-incubation with N-acetylcysteine reduced DPDT's oxidative, genotoxic and mutagenic effects in yeast and V79 cells. Our results suggest that the toxic and mutagenic properties of DPDT may stem from its ability to disturb the redox balance of the cell, which leads to oxidative stress and the induction of DNA damage.


[Effects of Naomaitong and thrombolysis on injuries of lung and stomach in rats with cerebral ischemia].
To study the protective effects of Naomaitong, a compound traditional Chinese herbal medicine, used together with thrombolysis therapy, on injuries of the lung and stomach in rats with cerebral ischemia. Rats were randomly divided into sham-operated group, untreated group, urokinase group (thrombolysis group), Naomaitong group, thrombolysis plus Naomaitong group. Cerebral ischemia was induced in rats by autonomous blood blot and inserted nylon thread. Rats were administrated with thrombolysis therapy through artery 3, 6 and 9 h after cerebral ischemia respectively. Twenty-four hours after the administration, mortality of the rats, and the brain and stomach hemorrhage ratios, as well as the pathological changes of the brain, lung and stomach were observed, and then cerebral infarct size (CIS) and lung water ratio (LWR) were measured. Compared with the sham-operated group, the rat mortality, and the brain and stomach hemorrhage ratios increased, the CIS enlarged, pathological changes of the brain, lung and stomach appeared obvious, and the LWR increased. Naomaitong plus thrombolysis treatment improved the changes above significantly. In the untreated rats with cerebral ischemia, injuries of the brain, lung and stomach were aggravated, the CIS enlarged and the LWR increased in the 9 h group as compared with those in the 3 h group. In the thrombolysis plus Naomaitong group, the pathological changes were improved, the CIS diminished and the LWR reduced. Injuries of the lung and stomach can be caused by cerebral ischemia, and the impairment was exacerbated following the prolongation of the ischemia. Thrombolysis therapy can cause brain and stomach hemorrhage. Thrombolysis therapy used early can perform protection against the injuries. Naomaitong, used together with thrombolysis, can reduce the mortality, the brain and stomach hemorrhage ratios, and perform protective effects on the injuries of cerebral ischemia.


Multivariate statistical analysis to assess the surface water quality of a snow and glacier-fed river: A case from Alaknanda River basin.
The water quality of Himalayan rivers has declined due to human activities, untreated effluent discharge, and poor sewage and drainage systems. The current study aimed to assess the water quality of these rivers using multivariate statistical analysis throughout four seasons. The analyses of 44 surface water samples taken during the monsoon, winter, spring, and summer seasons are well within the ranges acceptable for drinking and domestic use after the sedimentation. The suspended soils and turbidity are highly correlated and affect the water quality index (WQI). The WQI of headwater streams is good during low water flow seasons and poor during high water flow seasons. This is due to the number of melting glaciers and suspended solids/turbidity. Principal component analysis shows that in all the seasons, human activities such as road-cutting projects across the river and natural causes such as intense rainfall and melting of moraine-filled glaciers both impact the WQI. The findings of this study provide important information for future research and policy decisions aimed at improving the water quality of the Himalayan rivers.


Commercial Transactions in Children: The Case of Ghana.
The current article represents an examination of commercial transactions involving the sale of children in contemporary Ghana. It presents the results of a criminological analysis of 20 cases of commercial transactions in children in Ghana. It describes the sociodemographic characteristics of offenders and victims, victim-offender relationships, offender motivations, public reactions to the phenomenon, as well as the criminal justice system's responses to the crime. The data were extracted from Ghanaian print and electronic presses. The data show that more boys than girls were sold and that the ages of the victims ranged from 1 month to 19 years, although younger, prepubescent children were more likely to be sold than adolescents and younger adults. The results further show that the relationship between the offender and the child victim was a primary one, with parent-child relationships being dominant, followed by uncle-nephew. Pecuniary reasons were the primary motive for the crime, with offenders invariably expressing the need for money to satisfy pressing financial needs or personal enrichment. The data show that offenders were subject to prompt arrest, prosecution, and incarceration. A summary is provided for each of the 20 cases analyzed in the study.


GXXXG and AXXXA: common alpha-helical interaction motifs in proteins, particularly in extremophiles.
The GXXXG motif is a frequently occurring sequence of residues that is known to favor helix-helix interactions in membrane proteins. Here we show that the GXXXG motif is also prevalent in soluble proteins whose structures have been determined. Some 152 proteins from a non-redundant PDB set contain at least one alpha-helix with the GXXXG motif, 41 +/- 9% more than expected if glycine residues were uniformly distributed in those alpha-helices. More than 50% of the GXXXG-containing alpha-helices participate in helix-helix interactions. In fact, 26 of those helix-helix interactions are structurally similar to the helix-helix interaction of the glycophorin A dimer, where two transmembrane helices associate to form a dimer stabilized by the GXXXG motif. As for the glycophorin A structure, we find backbone-to-backbone atomic contacts of the C alpha-H...O type in each of these 26 helix-helix interactions that display the stereochemical hallmarks of hydrogen bond formation. These glycophorin A-like helix-helix interactions are enriched in the general set of helix-helix interactions containing the GXXXG motif, suggesting that the inferred C alpha-H...O hydrogen bonds stabilize the helix-helix interactions. In addition to the GXXXG motif, some 808 proteins from the non-redundant PDB set contain at least one alpha-helix with the AXXXA motif (30 +/- 3% greater than expected). Both the GXXXG and AXXXA motifs occur frequently in predicted alpha-helices from 24 fully sequenced genomes. Occurrence of the AXXXA motif is enhanced to a greater extent in thermophiles than in mesophiles, suggesting that helical interaction based on the AXXXA motif may be a common mechanism of thermostability in protein structures. We conclude that the GXXXG sequence motif stabilizes helix-helix interactions in proteins, and that the AXXXA sequence motif also stabilizes the folded state of proteins.


More americans living longer with cardiovascular disease will increase costs while lowering quality of life.
In the past several decades, some risk factors for cardiovascular disease have improved, while others have worsened. For example, smoking rates have dropped and treatment rates for cardiovascular disease have increased-factors that have made the disease less fatal. At the same time, Americans' average body mass index and incidence of diabetes have increased as the population continues to live longer-factors that have made cardiovascular disease more prevalent. To assess the aggregate impact of these opposing trends, we used the nine National Health and Nutrition Examination Survey waves from 1973 to 2010 to forecast total cardiovascular disease risk and prevalence from 2015 to 2030. We found that continued improvements in cardiovascular disease treatment and declining smoking rates will not outweigh the influence of increasing population age and obesity on cardiovascular disease risk. Given an aging population, an obesity epidemic, and declining mortality from the disease, the United States should expect to see a sharp rise in the health care costs, disability, and reductions in quality of life associated with increased prevalence of cardiovascular disease. Policies that target the treatment of high blood pressure and cholesterol and the reduction of obesity will be necessary to curb the imminent spike in cardiovascular disease prevalence.


Comparison and validation of point spread models for imaging in natural waters.
It is known that scattering by particulates within natural waters is the main cause of the blur in underwater images. Underwater images can be better restored or enhanced with knowledge of the point spread function (PSF) of the water. This will extend the performance range as well as the information retrieval from underwater electro-optical systems, which is critical in many civilian and military applications, including target and especially mine detection, search and rescue, and diver visibility. A better understanding of the physical process involved also helps to predict system performance and simulate it accurately on demand. The presented effort first reviews several PSF models, including the introduction of a semi-analytical PSF given optical properties of the medium, including scattering albedo, mean scattering angles and the optical range. The models under comparison include the empirical model of Duntley, a modified PSF model by Dolin et al, as well as the numerical integration of analytical forms from Wells, as a benchmark of theoretical results. For experimental results, in addition to that of Duntley, we validate the above models with measured point spread functions by applying field measured scattering properties with Monte Carlo simulations. Results from these comparisons suggest it is sufficient but necessary to have the three parameters listed above to model PSFs. The simplified approach introduced also provides adequate accuracy and flexibility for imaging applications, as shown by examples of restored underwater images.


Regulation of phosphorylation of the c-erbB-2/HER2 gene product by a monoclonal antibody and serum growth factor(s) in human mammary carcinoma cells.
Monoclonal antibody (MAb) 4D5 was used to analyze the phosphorylation of p185HER2, the gene product of c-erbB-2/HER2, in SK-BR-3 cells. Culture in the continuous presence of 4D5 reduced the in vivo steady-state levels of p185HER2 phosphorylation by 80% in a dose-dependent manner, suggesting that MAb 4D5 may have interfered with the activation of phosphorylation of p185HER2. The observed MAb-mediated reduction of p185HER2 phosphorylation could not be completely accounted for by down-regulation. When cultures were grown under serum-free conditions, the steady-state levels of p185HER2 phosphorylation were reduced by 56%, and addition of 4D5 further inhibited phosphorylation to 20% of steady-state levels. With continuous exposure to increasing concentrations of newborn calf serum in these cultures, there was a linear increase in tyrosine-specific phosphorylation of p185HER2, reaching a 5.4-fold increase with 10% newborn calf serum. Phosphorylation of p185HER2 in the presence of newborn calf serum was not attributable to stimulation of the epidermal growth factor receptor by epidermal growth factor or by transforming growth factor-alpha. Extension of these observations to two other mammary carcinoma cell lines. MDA-MB-453 and BT-474, also demonstrated a significant capacity of serum to induce p185HER2 phosphorylation. The demonstration of antibody-mediated partial inhibition of phosphorylation under serum-free conditions suggests that mammary carcinoma cells may also produce and secrete a factor or factors which may activate p185HER2. Our observation that growth-inhibitory MAb 4D5 is able to reduce the phosphorylation of p185HER2 by newborn calf serum and by a cellular-derived factor(s) suggests the existence of a growth factor(s) which uses phosphorylation of p185HER2 as a signal transduction pathway to regulate cell proliferation.


The characterisation of hyalocytes: the origin, phenotype, and turnover.
To determine the characterisation of hyalocytes: the origin, phenotype, and turnover in the rodent. To characterise the ultrastructure and distribution of hyalocytes, transmission and scanning electron microscopy was performed in rat eyes. Immunophenotypical analysis was performed by either anti-ED1 or ED2 antibodies. To examine the origin of the hyalocytes, the chimeric mice were created and were used to transplant the bone marrow (BM) cells from enhanced green fluorescent protein (EGFP) transgenic mice. The turnover of hyalocytes was examined at 0, 4, 6, 7, and 12 months after BM transplantation. Hyalocytes were distributed especially in the vitreous cortex and had an irregular shape with a spherical granule. Immunophenotypical studies demonstrated that most of the hyalocytes in rat eyes expressed ED2 but not ED1. In the chimeric mice, the hyalocytes were GFP negative right after BM transplantation. Interestingly, more than 60% of hyalocytes were replaced within 4 months and approximately 90% within 7 months after BM transplantation. The rodent hyalocytes were shown to express tissue macrophage marker, were derived from BM, and totally replaced within 7 months. These data provide the characterisation of hyalocytes in physiological conditions, especially their origin, distribution, and turnover, and may contribute to the better understanding of the pathogenesis of vitreoretinal disease.


sRNAbase: a Web resource for small RNAs in Enterobacteriaceae.
Bacterial small RNAs (sRNAs) have gained considerable attention due to their multivalent roles in the survival and pathogenesis of bacteria and mostly identified through bio-computational methods. A manually curated web-resource, sRNAbase has been constructed to give comprehensive and exhaustive information on non-coding small RNAs excluding tRNAs and rRNAs in Enterobacteriaceae family. The sRNA entries curated in sRNAbase contain experimentally verified small RNAs available in the literature and their partial/non-homologs reported within the related genomes from our earlier studies. The sRNAbase aims to facilitate the scientific community by providing information on the physical genomic location of the non-coding small RNAs, its alias names, sequences, strand orientation, gene identification numbers of the conserved genes that sandwiches the particular sRNA with possible functional role and a link to the PubMed literatures. Currently, sRNAbase holding information on 1986 entries belongs to 80 sRNA families spread over 45 Enterobacteriaceae genomes. The sRNAbase is accessible on the web at http://bicmku.in:8081/srnabase/.


Susceptibility allele-specific loss of miR-1324-mediated silencing of the INO80B chromatin-assembly complex gene in pre-eclampsia.
In humans, the elucidation of the genetics underlying multifactorial diseases such as pre-eclampsia remains complex. Given the current day availability of genome-wide linkage- and expression data pools, we applied pathway-guided genome-wide meta-analysis guided by the premise that the functional network underlying these multifactorial syndromes is under selective genetic pressure. This approach drastically reduced the genomic region of interest, i.e. 2p13 linked with pre-eclampsia in Icelandic families, from 8 679 641 bp (region with linkage) to 45 264 bp (coding exons of prioritized genes) (0.83%). Mutation screening of the candidate genes (n = 13) rapidly reduced the minimal critical region and showed the INO80B gene, encoding a novel winged helix domain (pfam14465) and part of the chromatin-remodeling complex, to be linked to pre-eclampsia. The functional defect in placental cells involved a susceptibility allele-dependent loss-of-gene silencing due to increased INO80B RNA stability as a consequence of differential binding of miR-1324 to the susceptibility allele of rs34174194. This risk allele is located at position 1 in an absolutely conserved 7-mer (UUGUCUG) in the 3-UTR of INO80B immediately downstream of a variant Pumillio Recognition Element (UGUANAAG). These data support that pre-eclampsia genes affect a conserved fundamental mechanism that evolved as a consequence of hemochorial placentation. Functionally, this involves founder-dependent, placentally expressed paralogous genes that regulate an essential trophoblast differentiation pathway but act at different entry points.


Virus-induced diabetes mellitus in mice and the thymus-dependent immune system.
The present study concerns the effect of the experimental diabetogenic encephalomyocarditis (EMC) virus on normal and athymic nude mice of BALB/c origin. The effect of simultaneous immunosuppressive pharmacological treatment with a derivative of cyclophosphamide in a relatively low dose (3 mg/mouse) was also studied. After inoculation with EMC virus, 36% of the normal mice, but none of the nude mice, developed diabetes mellitus and 93% of the normal mice, but none of the nude mice, developed paresis of one or more leg(s). When lower doses of EMC virus were given, few or none of the normal mice developed diabetes or paresis. After treatment with a cyclophosphamide-derivative, the number of paralysed mice increased. EMC virus in abundant amounts could be isolated from the pancreas and heart of all virus-inoculated mice, including the non-diabetic nude mice. Antibodies against EMC virus were found in all groups of virus-inoculated mice, although only in small amounts in nude and immunosuppressed normal mice. Histological examination revealed no significant differences between the islets of Langerhans of the experimental mice, diabetic as well as non-diabetic, and the control mice with respect to lymphocytic infiltration. It is concluded that the thymus-dependent immune system seems to be of decisive importance for the development of diabetes in this virus model.


Strand breaks arising from the repair of the 5-bromodeoxyuridine-substituted template and methyl methanesulphonate-induced lesions can explain the formation of sister chromatid exchanges.
5-Bromodeoxyuridine (BrdU)-induced sister chromatid exchanges (SCEs) are mainly determined during replication on a BrdU-substituted template. The BrdU, once incorporated, is rapidly excised as uracil (U), and the gap is repaired with the incorporation of BrdU from the medium, which leads to further repair. During the second S period in BrdU medium, this process continues as the strand acts as template. Experiments suggest that 3-aminobenzamide (3AB) delays the ligation of the gaps formed after U excision, resulting in enhanced SCE levels during the second cycle of BrdU incorporation. When normal templates of G1 cells are treated before BrdU introduction with methyl methanesulphonate (MMS), 3AB in the first cycle doubles the MMS-induced SCEs but has no effect on them during the second cycle. When the BrdU-substituted template is treated with MMS in G1 of the second cycle, 3AB again doubles the SCEs due to MMS and also enhances the SCEs resulting from delays in ligation of the gaps following U excision in the BrdU-substituted template. The repair processes of MMS lesions that are sensitive to 3AB and lead to SCEs take place rapidly, while the repair process of late repairing lesions that lead to SCEs appear to be insensitive to 3AB. A model for SCE induction is proposed involving a single-strand break or gap as the initial requirement for SCE initiation at the replicating fork. Subsequent events represent natural stages in the repair process of a lesion, ensuring replication without loss of genetic information.(ABSTRACT TRUNCATED AT 250 WORDS)


Transitional care: Concept analysis using Rodgers' evolutionary approach.
The process of moving patients from a hospital to a home or another care setting, also called transitional care, can lead to unwanted complications and negative outcomes. The term "transitional care" involves a wide range of conditions and services to ensure the continuity of care and prevention of unwanted consequences in vulnerable individuals, who are affected by any changes in care settings or caregivers. The purpose of this concept analysis is to clarify the concept of transitional care by considering its application in different studies and its changes over time. A concept analysis. Literature from 2008 to 2018 were sought using Medline, PubMed, Google Scholar and Cochrane databases with terms "transitional care", "transitional care" OR "care transitions", "transitional care" AND "nursing", "transitional care" AND "discharge planning". Rodgers' evolutionary concept analysis method was used to clarify the antecedents, attributes and consequences. The 46 eligible articles were fully studied and findings were categorized into antecedents, attributes and consequences. Transitional care antecedents were classified into three categories: patient/family/caregivers related factors, factors related to hospital system (inpatient care), and social factors. The defining attributes include patient-oriented transitional care, transitional care with a hospital-based approach and transitional care with a community-based approach. Consequences categorized into patient-related consequences, family/caregiver-related consequences, and hospital-related consequences. The results of the analysis of the transitional care concept have shown that this concept is not limited to care provided by nurses at discharge, and that its proper implementation requires considering many factors including the status of patients and their families, different members of the healthcare team, and environmental and social conditions and facilities. A proper understanding of transitional care not only specifies the role of care providers, but also creates a basis for designing an evidence based care program.


Diethyldithiocarbamate enhances production of nitric oxide and TNF-alpha by lipopolysaccharide-stimulated rat Kupffer cells.
Previous studies have shown that large doses of diethyldithiocarbamate (DDC) cause liver injury in rats and the pathogenesis of this injury involves, in part, release of superoxide anion by Kupffer cells. The purpose of this study was to evaluate if DDC was able to stimulate other potentially toxic mediators such as nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) using isolated rat Kupffer cells. DDC alone did not stimulate the release of NO and TNF-alpha by Kupffer cells. Interestingly, when Kupffer cells were stimulated by lipopolysaccharide (LPS), DDC (0-30 microM) enhanced the production of both NO and TNF-alpha in a concentration-dependent manner. Therefore, we further studied how DDC modulated the response of Kupffer cells to LPS. Immunocytochemical studies revealed that DDC increased the amount of inducible NO synthase and TNF-alpha protein in Kupffer cells after their exposure to LPS. The enhanced effects of DDC on the release of NO and TNF-alpha from Kupffer cells was inhibited by N-acetyl-L-cysteine (an inhibitor of transcription factor NF-kappaB activation). By using a specific antibody for NF-kappaBp65, it was found that DDC enhanced the LPS-activated nuclear translocation of NF-kappaB. There was no evidence of intracellular oxidative stress following either LPS alone or DDC + LPS exposure. The stimulatory effect of DDC on both NO and TNF-alpha release was inhibited by H-7 (an inhibitor of protein kinase C) but not H-8 (an inhibitor of cAMP-dependent protein kinase). These findings demonstrate that DDC enhances the production of NO and TNF-alpha by LPS-stimulated Kupffer cells and suggest that protein kinase C plays a critical role in mediating these effects of DDC.


Practice patterns and complications of hysterectomy for invasive cervical cancer after the Laparoscopic Approach to Cervical Cancer trial.
After the publication of the Laparoscopic Approach to Cervical Cancer trial, the standard surgical approach for early-stage cervical cancer is open radical hysterectomy. Only limited data were available regarding whether the change to open abdominal hysterectomy observed after the Laparoscopic Approach to Cervical Cancer trial led to an increase in postoperative complication rates as a consequence of the decrease in the use of the minimally invasive approach. This study aimed to analyze whether there was a correlation between the publication of the Laparoscopic Approach to Cervical Cancer trial and an increase in the 30-day complications associated with surgical treatment of invasive cervical cancer. Data from the American College of Surgeons National Surgical Quality Improvement Program were used to compare the results in the pre-Laparoscopic Approach to Cervical Cancer period (January 2016 to December 2017) vs the results in the post-Laparoscopic Approach to Cervical Cancer period (January 2019 to December 2020). The rates of each surgical approach (open abdominal or minimally invasive) hysterectomy for invasive cervical cancer during the 2 periods were assessed. Subsequently, 30-day major complication, minor complication, unplanned hospital readmission, and intra- or postoperative transfusion rates before and after the publication of the Laparoscopic Approach to Cervical Cancer trial were compared. Overall, 3024 patients undergoing either open abdominal hysterectomy or minimally invasive hysterectomy for invasive cervical cancer were included in the study. Of the patients, 1515 (50.1%) were treated in the pre-Laparoscopic Approach to Cervical Cancer period, and 1509 (49.9%) were treated in the post-Laparoscopic Approach to Cervical Cancer period. The rate of minimally invasive approaches decreased significantly from 75.6% (1145/1515) in the pre-Laparoscopic Approach to Cervical Cancer period to 41.1% (620/1509) in the post-Laparoscopic Approach to Cervical Cancer period, whereas the rate of open abdominal approach increased from 24.4% (370/1515) in the pre-Laparoscopic Approach to Cervical Cancer period to 58.9% (889/1509) in the post-Laparoscopic Approach to Cervical Cancer period (P<.001). The overall 30-day major complications remained stable between the pre-Laparoscopic Approach to Cervical Cancer period (85/1515 [5.6%]) and the post-Laparoscopic Approach to Cervical Cancer period (74/1509 [4.9%]) (adjusted odds ratio, 0.85; 95% confidence interval, 0.61-1.17). The overall 30-day minor complications were similar in the pre-Laparoscopic Approach to Cervical Cancer period (103/1515 [6.8%]) vs the post-Laparoscopic Approach to Cervical Cancer period (120/1509 [8.0%]) (adjusted odds ratio, 1.17; 95% confidence interval, 0.89-1.55). The unplanned hospital readmission rate remained stable during the pre-Laparoscopic Approach to Cervical Cancer period (7.9% per 30 person-days) and during the post-Laparoscopic Approach to Cervical Cancer period (6.3% per 30 person-days) (adjusted hazard ratio, 0.78; 95% confidence interval, 0.58-1.04)]. The intra- and postoperative transfusion rates increased significantly from 3.8% (58/1515) in the pre-Laparoscopic Approach to Cervical Cancer period to 6.7% (101/1509) in the post-Laparoscopic Approach to Cervical Cancer period (adjusted odds ratio, 1.79; 95% confidence interval, 1.27-2.53). This study observed a significant shift in the surgical approach for invasive cervical cancer after the publication of the Laparoscopic Approach to Cervical Cancer trial, with a reduction in the minimally invasive abdominal approach and an increase in the open abdominal approach. The change in surgical approach was not associated with an increase in the rate of 30-day major or minor complications and unplanned hospital readmission, although it was associated with an increase in the transfusion rate.


TLR activation pathways in HIV-1-exposed seronegative individuals.
TLRs trigger innate immunity that recognizes conserved motifs of invading pathogens, resulting in cellular activation and release of inflammatory factors. The influence of TLR activation on resistance to HIV-1 infection has not been investigated in HIV-1 exposed seronegative (ESN) individuals. PBMCs isolated from heterosexually ESN individuals were stimulated with agonists specific for TLR3 (poly I:C), TLR4 (LPS), TLR7 (imiquimod), and TLR7/8 (ssRNA40). We evaluated expression of factors involved in TLR signaling cascades, production of downstream effector immune mediators, and TLR-expression in CD4+ and CD14+ cells. Results were compared with those obtained in healthy controls (HCs). ESN individuals showed: 1) comparable percentages of CD14+/TLR4+ and CD4+/TLR8+ CD14+/TLR8+ cells; 2) higher responsiveness to poly I:C, LPS, imiquimod, and ssRNA40 stimulation, associated with significantly increased production of IL-1beta, IL-6, TNF-alpha, and CCL3; 3) augmented expression of mRNA specific for other targets (CCL2, CSF3, CSF2, IL-1alpha, IL-8, IL-10, IL-12, cyclooxygenase 2) demonstrated by broader TLRs pathway expression analyses; and 4) increased MyD88/MyD88s(short) ratio, mainly following TLR7/8 stimulation. We also compared TLR-agonist-stimulated cytokine/chemokine production in CD14+ PBMCs and observed decreased IFN-beta production in ESN individuals compared with HCs upon TLR7/8-agonist stimulation. These data suggest that TLR stimulation in ESN individuals results in a more robust release of immunologic factors that can influence the induction of stronger adaptive antiviral immune responses and might represent a virus-exposure-induced innate immune protective phenotype against HIV-1.


Brain morphometry of preschool age children affected by autism spectrum disorder: Correlation with clinical findings.
The aim of our study was to use a combined imaging and clinical approach to identify possible patterns of clinical and imaging findings in a cohort of preschool age autism spectrum disorder (ASD) patients. In order to identify imaging patterns that could be related to specific clinical features, a selected group of ASD patients (age range 3-6 years) without dysmorphic features, epilepsy or other major neurological signs, malformations or other lesions at MRI was subjected to brain volumetric analysis using semiautomatic brain segmentation. An age-matched group of typically developing children was subjected to the same analysis. Our results were consistent with previous literature: Total gray matter volume, total cortical gray matter volume and amygdalar volumes were significantly greater in the ASD group than the control group. When we divided the study group into subgroups on the basis of clinical findings such as high- or low-functioning, or verbal and nonverbal, the only significant difference between verbal and nonverbal subjects was in cerebellar hemispheric size. In conclusions, our results confirm that newer brain MRI techniques using semiautomatic brain segmentation can provide information useful for defining the differences between ASD patients and controls, particularly if they form part of an integrated approach between MRI and cognitive-behavioral and genetic data. Clin. Anat. 32:143-150, 2019. © 2018 Wiley Periodicals, Inc. HIGHLIGHTS: Combined imaging and clinical approach in autism spectrum disorders Semiautomatic brain segmentation in a selected preschool age ASD group Reduced total cerebellar white matter volume in non-verbal ASD patients.


A vestibular prosthesis with highly-isolated parallel multichannel stimulation.
This paper presents an implantable vestibular stimulation system capable of providing high flexibility independent parallel stimulation to the semicircular canals in the inner ear for restoring three-dimensional sensation of head movements. To minimize channel interaction during parallel stimulation, the system is implemented with a power isolation method for crosstalk reduction. Experimental results demonstrate that, with this method, electrodes for different stimulation channels located in close proximity ( mm) can deliver current pulses simultaneously with minimum inter-channel crosstalk. The design features a memory-based scheme that manages stimulation to the three canals in parallel. A vestibular evoked potential (VEP) recording unit is included for closed-loop adaptive stimulation control. The main components of the prototype vestibular prosthesis are three ASICs, all implemented in a 0.6- μm high-voltage CMOS technology. The measured performance was verified using vestibular electrodes in vitro.


Penetration of chlorhexidine into human skin.
This study evaluated a model of skin permeation to determine the depth of delivery of chlorhexidine into full-thickness excised human skin following topical application of 2% (wt/vol) aqueous chlorhexidine digluconate. Skin permeation studies were performed on full-thickness human skin using Franz diffusion cells with exposure to chlorhexidine for 2 min, 30 min, and 24 h. The concentration of chlorhexidine extracted from skin sections was determined to a depth of 1,500 microm following serial sectioning of the skin using a microtome and analysis by high-performance liquid chromatography. Poor penetration of chlorhexidine into skin following 2-min and 30-min exposures to chlorhexidine was observed (0.157 +/- 0.047 and 0.077 +/- 0.015 microg/mg tissue within the top 100 microm), and levels of chlorhexidine were minimal at deeper skin depths (less than 0.002 microg/mg tissue below 300 microm). After 24 h of exposure, there was more chlorhexidine within the upper 100-microm sections (7.88 +/- 1.37 microg/mg tissue); however, the levels remained low (less than 1 microg/mg tissue) at depths below 300 microm. There was no detectable penetration through the full-thickness skin. The model presented in this study can be used to assess the permeation of antiseptic agents through various layers of skin in vitro. Aqueous chlorhexidine demonstrated poor permeation into the deeper layers of the skin, which may restrict the efficacy of skin antisepsis with this agent. This study lays the foundation for further research in adopting alternative strategies for enhanced skin antisepsis in clinical practice.


Obesity and symptoms of depression contribute independently to the poor asthma control of obesity.
Obesity is a major risk factor for poorly controlled asthma, but the reasons for poor asthma control in this patient population are unclear. Symptoms of depression have been associated with poor asthma control, and increase with higher body mass index (BMI). The purpose of this study was to assess whether depressive symptoms underlie poor asthma control in obesity. We determined the relationship between BMI, psychological morbidity and asthma control at baseline in a well-characterized patient population participating in a clinical trial conducted by the American Lung Association-Asthma Clinical Research Centers. Obese asthmatic participants had increased symptoms of depression (Center for Epidemiologic Studies Depression Scale score in lean 10.1 ± 8.1, overweight 10.0 ± 8.1, obese 12.4 ± 9.9; p = 0.03), worse asthma control (Juniper Asthma Control Questionnaire score in lean 1.43 ± 0.68, overweight 1.52 ± 0.71, obese 1.76 ± 0.75; p < 0.0001), and worse asthma quality of life (scores in lean 5.21 ± 1.08, overweight 5.08 ± 1.05, obese 4.64 ± 1.09; p < 0.0001). Asthmatics with obesity and those with symptoms of depression both had a higher risk of having poorly controlled asthma (adjusted odds ratio of 1.83 CI 1.23-3.52 for obesity, and 2.08 CI 1.23-3.52 for depression), but there was no interaction between the two. Obesity and symptoms of depression are independently associated with poor asthma control. As depression is increased in obese asthmatics it may be an important co-morbidity contributing to poor asthma control in this population, but factors other than depression also contribute to poor asthma control in obesity.


[Planned sedation with dexmedetomidine hydrochloride after pediatric cardiac surgery; an institutional experience].
Dexmedetomidine hydrochloride (DEX) is a newly developed alpha-2 adrenergic agonist sedative and has been shown to be effective in post-surgical patients, providing not only unique sedation but also stabilization of hemodynamic and respiratory function. We investigated the hemodynamic and respiratory effects and efficacy of DEX in 84 consecutive patients (age <6 months: 18, 6-12 months: 13, 1-3 years: 29, 4-9 years: 18, >10 years: 5, male:female = 44:40) who were sedated by DEX in combination with a small dose of midazolam and morphine. DEX was commenced at an initial dose of 0.7 microg/kg/hr during surgery, approximately 1 hour prior to transfer to the intensive care unit (ICU). DEX infusion was maintained at a rate of 0.2-0.7 microg/kg/hr after ICU admission throughout weaning from mechanical ventilation and extubation. The dose of the sedatives was optimized by scoring on Ramsay's sedative scale. There were no undesirable hemodynamic changes throughout the DEX infusion. Respiration was maintained and all patients were extubated uneventfully. Optimal level of sedation was achieved in all patients. There were no adverse events related to DEX administration. Moreover, junctional ectopic tachycardia (JET) and severe pulmonary hypertension (PH) leading to clinical deterioration, which are the major causes of postoperative morbidity in pediatric cardiac surgery, occurred at a low incidence in this series. Our DEX protocol provided 1) satisfactory postoperative sedation without compromising hemodynamics and respiration, and 2) prevention and amelioration of postoperative morbidity caused by sympathomimetic stimulation, in pediatric cardiac surgery.


Differential effects of myosin-antibody complexes on contractile rings and circumferential belts in epitheloid cells.
The role of myosin filaments during assembly and activity of microfilament rings was analyzed by microinjecting epitheloid cells (PtK2 and LLC-PK1 kidney cell lines) with specific anti-myosins. Six monoclonal antibodies directed against the light meromyosin (LMM) region of the myosin molecule were characterized with respect to epitope location, and their effects on actin-activated MgATPase as well as on assembly, structural integrity and stability of myosin filaments. All of these antibodies recognized LLC-PK1 myosin, but only three reacted with PtK2 myosin. The remaining three served as matching controls in experiments with this cell line. When injected in amounts sufficient to yield an excess of antibody over myosin, the reactive antibodies significantly delayed formation and constriction of the contractile ring in mitotic cells. These rings contained less myosin, but not less actin, than the controls. This indicates that the recruitment and alignment of actin in the cleavage furrow can occur independently of other components of the contractile ring. After completion of cytokinesis, the majority of the injected cells was unable to assemble a normal circumferential belt. This resulted in defective epitheloid sheets. Approximately one third of these cells showed grossly distorted cell shapes and an increase in locomotory activity. All these changes were fully reversible with time, suggesting that the effects of the antibodies were overcome by protein synthesis. The differential sensitivity seen between contractile rings and peripheral belts is discussed with respect to differences in their architecture, stability and proposed function.


Transrectal osteopathic manipulation treatment for chronic coccydynia: feasibility, acceptability and patient-oriented outcomes in a quality improvement project.
Pain of the coccyx, coccydynia, is a common condition with a substantial impact on the quality of life. Although most cases resolve with conservative care, 10 % become chronic and are more debilitating. Treatment for chronic coccydynia is limited; surgery is not definitive. Osteopathic manipulative treatment (OMT) is the application of manually guided forces to areas of somatic dysfunction to improve physiologic function and support homeostasis including for coccydynia, but its use as a transrectal procedure for coccydynia in a primary care clinic setting is not well documented. We aimed to conduct a quality improvement (QI) study to explore the feasibility, acceptability, and clinical effects of transrectal OMT for chronic coccydynia in a primary care setting. This QI project prospectively treated and assessed 16 patients with chronic coccydynia in a primary care outpatient clinic. The intervention was transrectal OMT as typically practiced in our clinic, and included myofascial release and balanced ligamentous tension in combination with active patient movement of the head and neck. The outcome measures included: acceptance, as assessed by the response rate (yes/no) to utilize OMT for coccydynia; acceptability, as assessed by satisfaction with treatment; and coccygeal pain, as assessed by self-report on a 0-10 numerical rating scale (NRS) for coccydynia while lying down, seated, standing, and walking. Sixteen consecutive patients with coccydynia were offered and accepted OMT; six patients also received other procedural care. Ten patients (two males, eight females) received only OMT intervention for their coccydynia and were included in the per-protocol analysis. Posttreatment scores immediately after one procedure (acute model) and in follow-up were significantly improved compared with pretreatment scores. Follow-up pain scores provided by five of the 10 patients demonstrated significant improvement. The study supports transrectal OMT as a feasible and acceptable treatment option for coccydynia. Patients were satisfied with the procedure and reported improvement. There were no side effects or adverse events. These data suggest that the use of transrectal OMT for chronic coccydynia is feasible and acceptable; self-reported improvement suggests utility in this clinic setting. Further evaluation in controlled studies is warranted.


Effects of Tagetes patula on Active and Inactive Stages of Root-Knot Nematodes.
Although marigold (Tagetes patula) is known to produce allelopathic compounds toxic to plant-parasitic nematodes, suppression of Meloidogyne incognita can be inconsistent. Two greenhouse experiments were conducted to test whether marigold is more effective in suppressing Meloidogyne spp. when it is active rather than dormant. Soils infested with Meloidogyne spp. were collected and conditioned in the greenhouse either by 1) keeping the soil dry (DRY), 2) irrigating with water (IRR), or 3) drenching with cucumber (Cucumis sativus) leachate (CL) for 5 wk. These soils were then either planted with cucumber, marigold or remained bare for 10 wk. Suppression of nematode by marigold was then assayed using cucumber. DRY conditioning resulted in the highest number of inactive nematodes, whereas CL and IRR had higher numbers of active nematodes than DRY. At the end of the cucumber bioassay, marigold suppressed the numbers of Meloidogyne females in cucumber roots if the soil was conditioned in IRR or CL, but not in DRY. However, in separate laboratory assays, marigold root leachate slightly reduced M. incognita J2 activity but did not reduce egg hatch (P > 0.05). These finding suggest that marigold can only suppress Meloidogyne spp. when marigold is actively growing. This further suggests that marigold will more efficiently suppress Meloidogyne spp. if planted when these nematodes are in active stage.


[Lumbar phlebography. Contribution to the diagnosis of lumbosciatica of disk origin].
Lumbar phlebography: its diagnostic value in lumbosciatica due to disc disease. The authors' purpose was to define the diagnostic value of lumbar phlebography in the investigation of intervertebral disc lesions and to compare these results with those obtained from clinical examination and plain X-rays. They conducted a retrospective study of 200 cases of patients hospitalized for sciatica who were investigated by lumbar phlebography using selective catheterization, without any prior neuro-radiological examination. 104 of these cases were operated on. The lumbar phlebography was interpretable in 99 p. cent of cases. Severe thrombo-embolic episodes occurred in two cases. In the 104 patients who went to operation, the surgical procedure confirmed the presence of a herniated disc (suspected on phlebography) in 96 p. cent of cases. Topographical agreement between the phlebographic data and the operative findings was observed in 81 p. cent of cases. Phlebography proved to be more reliable at L4-L5 than at L5-S1. In the patients in whom clinical examination and plain X-ray findings provided coherent information as to the site of the herniated disc, phlebography was considered to be of no use. In those patients for whom this examination did not provide coherent information, phlebography proved to be valuable in the topographical diagnosis of the herniated disc.


A Recombinant Fab Antibody Against Kwakhurin as a Tool for Sensitive Indirect Competitive ELISA.
Pueraria candollei var. mirifica (P. candollei), known as White Kwao Krua in Thai, has long been used in traditional Thai medicine for the symptoms of menopause due to the potent estrogenic activity exhibited by the isoflavonoids and chromenes it contains. Recently, health hazards caused by P. candollei-derived products have arisen in Japan, and demands for analytical methods to standardize the P. candollei have been increasing. Previously, we have focused on quantifying the unique P. candollei-derived isoflavonoid kwakhurin (Kwa) and developed an indirect competitive enzyme- linked immunosorbent assay (icELISA) using a monoclonal antibody (MAb) against Kwa. However, MAb preparation requires the use of costly culture medium and sophisticated techniques. In this study, we produced a recombinant antigen-binding fragment (Fab) against Kwa, as an alternative to MAb, for use in icELISA for quantitative analysis of Kwa. The VHCH1 and VL-CL proteins were individually expressed in Escherichia coli BL21 (DE3) strain and were then refolded to form active anti-Kwa Fab. Characterization of anti-Kwa Fab revealed that it possessed high specificity (cross-reactivities with other Kwa-related compounds, <0.03%) and high sensitivity (limit of detection, 8.16 ng/mL). Additionally, validation analyses indicated that icELISA using anti-Kwa Fab is highly precise, accurate, and sufficiently reliable for use in quantitative analysis of Kwa. Consequently, an icELISA incorporating anti-Kwa Fab was developed for the analysis of P. candollei-derived products, to assure consumer safety.


Stabilization of a straight longitudinal dune under bimodal wind with large directional variation.
It has been observed that the direction in which a sand dune extends its crest line depends on seasonal variation of wind direction; when the variation is small, the crest line develops more or less perpendicularly to the mean wind direction to form a transverse dune with some undulation. In the case of bimodal wind with a large relative angle, however, the dune extends its crest along the mean wind direction and evolves into an almost straight longitudinal dune. Motivated by these observations, we investigate the dynamical stability of isolated dunes using the crest line model, where the dune dynamics is represented by its crest line motion. First, we extend the previous linear stability analysis under the unidirectional wind to the case with nonzero slant angle between the wind direction and the normal direction of the crest line, and show that the stability diagram does not depend on the slant angle. Second, we examine how the linear stability is affected by the seasonal changes of wind direction in the case of bimodal wind with equal strength and duration. For the transverse dune, we find that the stability is virtually the same with that for the unidirectional wind as long as the dune evolution during a season is small. On the other hand, in the case of the longitudinal dune, the dispersions of the growth rates for the perturbation are drastically different from those of the unidirectional wind, and we find that the largest growth rate is always located at k=0. This is because the growth of the perturbation with k≠0 is canceled by the alternating wind from opposite sides of the crest line even though it grows during each duration period of the bimodal wind. For a realistic parameter set, the system is in the wavy unstable regime of the stability diagram for the unidirectional wind, thus the straight transverse dune is unstable to develop undulation and eventually evolves into a string of barchans when the seasonal variation of wind direction is small, but the straight longitudinal dune is stabilized under the large variation of bimodal wind direction. We also perform numerical simulations on the crest line model, and find that the results are consistent with our linear analysis and the previous reports that show that the longitudinal dunes tend to have a straight ridge elongating over time.


A simplified table using validated diagnostic criteria is effective to improve characterization of colorectal polyps: the CONECCT teaching program.
Introduction and study aims Accurate real-time endoscopic characterization of colorectal polyps is key to choosing the most appropriate treatment. Mastering the currently available classifications is challenging. We used validated criteria for these classifications to create a single table, named CONECCT, and evaluated the impact of a teaching program based on this tool. Methods A prospective multicenter study involving GI fellows and attending physicians was conducted. During the first session, each trainee completed a pretest consisting in histological prediction and choice of treatment of 20 colorectal polyps still frames. This was followed by a 30-minute course on the CONECCT table, before taking a post-test using the same still frames reshuffled. During a second session at 3 - 6 months, a last test (T3 M) was performed, including these same still frames and 20 new ones. Results A total 419 participants followed the teaching program between April 2017 and April 2018. The mean proportion of correctly predicted/treated lesions improved significantly from pretest to post-test and to T3 M, from 51.0 % to 74.0 % and to 66.6 % respectively ( P < 0.001). Between pretest and post-test, 343 (86.6 %) trainees improved, and 153 (75.4 %) at T3 M. Significant improvement occurred for each subtype of polyp for fellows and attending physicians. Between the two sessions, trainees continued to progress in the histology prediction and treatment choice of polyps CONECCT IIA. Over-treatment decreased significantly from 30.1 % to 15.5 % at post-test and to 18.5 % at T3 M ( P < 0.001). Conclusion The CONECCT teaching program is effective to improve the histology prediction and the treatment choice by gastroenterologists, for each subtype of colorectal polyp.


Lung cancer chemoprevention with retinol palmitate. Preliminary data from a randomized trial on stage Ia non small-cell lung cancer.
On the basis of epidemiologic and experimental evidence of an anticancer activity of vitamin A, a randomized clinical trial was activated in Milan with the aim of evaluating if retinol palmitate administration (per os, 300,000 I.U. daily) after complete resection of stage Ia non small-cell lung cancer could reduce the occurrence of cancer relapses (within 3 years) and/or the occurrence of new primary tumors (beyond 3 years). By September 1987, 181 patients had entered the trial: 87 in the treatment arm and 94 in the control arm. After a median follow-up of 14 months, the interim analysis was focused on the evaluation of toxicity, compliance, and early recurrences. Although the large majority of patients were affected by skin and mucous membrane desquamation and dryness during treatment, these symptoms were generally mild and well tolerated, and never induced the patient to stop the treatment. Other side effects like headache, hair loss, itching, or dyspepsia were detected at a much lower frequency. Only in 3 patients the treatment was interrupted, because of signs or symptoms potentially related to vitamin A administration. At the time of the analysis, a total of 42 (23%) patients had relapsed; 16 (18%) in the treated arm, and 26 (28%) in the control arm. The largest difference between treated patients and controls was observed for bone metastases (2 vs. 7) and brain metastases (3 vs. 6), and for squamous histology (6 vs. 11). Only 2 cases of new primary cancer were detected, both in the control arm. These results are promising both in terms of tolerance and efficacy of treatment, but given the short median follow-up they must be very cautiously interpreted. A longer follow-up is necessary to establish whether a significant proportion of early recurrences could be prevented, or only delayed, by vitamin A administration.


Validating the AMRFinder Tool and Resistance Gene Database by Using Antimicrobial Resistance Genotype-Phenotype Correlations in a Collection of Isolates.
Antimicrobial resistance (AMR) is a major public health problem that requires publicly available tools for rapid analysis. To identify AMR genes in whole-genome sequences, the National Center for Biotechnology Information (NCBI) has produced AMRFinder, a tool that identifies AMR genes using a high-quality curated AMR gene reference database. The Bacterial Antimicrobial Resistance Reference Gene Database consists of up-to-date gene nomenclature, a set of hidden Markov models (HMMs), and a curated protein family hierarchy. Currently, it contains 4,579 antimicrobial resistance proteins and more than 560 HMMs. Here, we describe AMRFinder and its associated database. To assess the predictive ability of AMRFinder, we measured the consistency between predicted AMR genotypes from AMRFinder and resistance phenotypes of 6,242 isolates from the National Antimicrobial Resistance Monitoring System (NARMS). This included 5,425 Salmonella enterica, 770 Campylobacter spp., and 47 Escherichia coli isolates phenotypically tested against various antimicrobial agents. Of 87,679 susceptibility tests performed, 98.4% were consistent with predictions. To assess the accuracy of AMRFinder, we compared its gene symbol output with that of a 2017 version of ResFinder, another publicly available resistance gene detection system. Most gene calls were identical, but there were 1,229 gene symbol differences (8.8%) between them, with differences due to both algorithmic differences and database composition. AMRFinder missed 16 loci that ResFinder found, while ResFinder missed 216 loci that AMRFinder identified. Based on these results, AMRFinder appears to be a highly accurate AMR gene detection system.


Comparative immune responses to candidate arthritogenic bacteria do not confirm a dominant role for Klebsiella pneumonia in the pathogenesis of familial ankylosing spondylitis.
Using humoral immune responses, Klebsiella pneumoniae has been implicated as a candidate microbial trigger in ankylosing spondylitis (AS) by several investigators but refuted by others. The objective of this case-control study was to compare the cellular (T-cell proliferation) and humoral (IgG and IgA, by ELISA) immune responses of affected individuals in multiplex AS families with those of unaffected family members and normal healthy controls in order to find out whether affected individuals exhibit a predominant immune response to K. pneumoniae. Twenty-five families with two or more individuals affected with AS and 34 normal healthy controls matched with the affected family members for age, sex and ethnicity were enrolled in the study. All affected (n = 57) and unaffected (n = 37) family members had a detailed clinical evaluation. Peripheral blood was drawn to determine T-lymphocyte proliferation and the IgG and IgA (by ELISA analysis) immune responses to K. pneumoniae, Salmonella typhimurium, Yersinia enterocolitica and Chlamydia trachomatis. Immune responses to each of the four candidate organisms were compared in affected and unaffected individuals. Each individual was classified by the predominant antigenic immune response that they showed when comparison was made among the same concentrations of the four candidate microbial antigens. This stratification was then used (i) to compare immune responses in affected and unaffected family members and (ii) to compare clinical characteristics of affected family members. There was no difference in mean stimulation indices or antibody responses between affected and unaffected family members for each of the candidate organisms. In terms of predominant cellular immune responses to these organisms, there was no difference between affected and unaffected family members with respect to K. pneumoniae, C. trachomatis or Y. enterocolitica. However, a higher percentage of affected family members (25.9%) exhibited a predominant response to S. typhimurium compared with unaffected family members (5.9%, P < 0.02). In assessing antibody titres, K. pneumoniae was the predominant amongst these four organisms, but there was no difference between affected family members, unaffected family members and normal healthy controls. There was no relationship between immune responses and clinical characteristics. Our analysis of affected and unaffected family members in familial AS demonstrated no significant differences with respect to cellular or humoral immune responses to K. pneumoniae and three control microbes. In addition, K. pneumoniae did not exhibit a predominant immune response in affected individuals. Thus we find no supportive evidence to implicate a causal role for K. pneumoniae in familial AS.


Hospitals face tough issues in years following mergers.
Despite fears in recent years that the federal government and courts would make hospital mergers impossible, mergers are continuing to take place at a regular pace. But what happens in the aftermath of a hospital merger--both to the hospitals involved and to their communities? Four case studies explore the difficult issues that hospitals face. In addition, we take a look at the Roanoke and Rockford cases, and what the hospitals involved are doing today.


A mutant ataxin-3 fragment results from processing at a site N-terminal to amino acid 190 in brain of Machado-Joseph disease-like transgenic mice.
Machado-Joseph disease also called spinocerebellar ataxia type 3 (MJD/SCA3) is a hereditary and neurodegenerative movement disorder caused by ataxin-3 with a polyglutamine expansion (mutant ataxin-3). Neuronal loss in MJD/SCA3 is associated with a mutant ataxin-3 toxic fragment. Defining mutant ataxin-3 proteolytic site(s) could facilitate the identification of the corresponding enzyme(s). Previously, we reported a mutant ataxin-3 mjd1a fragment in the brain of transgenic mice (Q71) that contained epitopes C-terminal to amino acid 220. In this study, we generated and characterized neuroblastoma cells and transgenic mice expressing mutant ataxin-3 mjd1a lacking amino acids 190-220 (deltaQ71). Less deltaQ71 than Q71 fragments were detected in the cell but not mouse model. The transgenic mice developed an MJD/SCA3-like phenotype and their brain homogenates had a fragment containing epitopes C-terminal to amino acid 220. Our results support the toxic fragment hypothesis and narrow the mutant ataxin-3 cleavage site to the N-terminus of amino acid 190.


Extended-spectrum beta-lactamases in Klebsiella pneumoniae bloodstream isolates from seven countries: dominance and widespread prevalence of SHV- and CTX-M-type beta-lactamases.
A huge variety of extended-spectrum beta-lactamases (ESBLs) have been detected during the last 20 years. The majority of these have been of the TEM or SHV lineage. We have assessed ESBLs occurring among a collection of 455 bloodstream isolates of Klebsiella pneumoniae, collected from 12 hospitals in seven countries. Multiple beta-lactamases were produced by isolates with phenotypic evidence of ESBL production (mean of 2.7 beta-lactamases per isolate; range, 1 to 5). SHV-type ESBLs were the most common ESBL, occurring in 67.1% (49 of 73) of isolates with phenotypic evidence of ESBL production. In contrast, TEM-type ESBLs (TEM-10 type, -12 type, -26 type, and -63 type) were found in just 16.4% (12 of 73) of isolates. The finding of TEM-10 type and TEM-12 type represents the first detection of a TEM-type ESBL in South America. PER (for Pseudomonas extended resistance)-type beta-lactamases were detected in five of the nine isolates from Turkey and were found with SHV-2-type and SHV-5-type ESBLs in two of the isolates. CTX-M-type ESBLs (bla(CTX-M-2) type and bla(CTX-M-3) type) were found in 23.3% (17 of 73) of isolates and were found in all study countries except for the United States. We also detected CTX-M-type ESBLs in four countries where they have previously not been described-Australia, Belgium, Turkey, and South Africa. The widespread emergence and proliferation of CTX-M-type ESBLs is particularly noteworthy and may have important implications for clinical microbiology laboratories and for physicians treating patients with serious K. pneumoniae infections.


Influence of the waist diameters on transmission characteristics and strain sensitivity of microtapered long-period fiber gratings.
Transmission characteristics of microtapered long-period fiber gratings (MTLPGs) and their strain and temperature sensitivities with variations in the waist diameters are investigated theoretically and experimentally. Transmission characteristics of MTLPGs strongly depend on the waist diameter of the tapered optical fiber (TOF) because of the modification of the effective index difference between the core and the cladding modes. Based on the photoelastic effect, the resonant wavelengths of MTLPGs with variations in strain shift to shorter wavelengths. The strain sensitivity of the MTLPG with a waist diameter of 25 μm is improved by a factor of 20 compared with that of a 125 μm long-period fiber grating. The temperature sensitivities of MTLPGs are also enhanced by reducing the waist diameter of the TOF.


An Improved Antagonist Radiotracer for the κ-Opioid Receptor: Synthesis and Characterization of (11)C-LY2459989.
The κ-opioid receptors (KORs) are implicated in several neuropsychiatric diseases and addictive disorders. PET with radioligands provides a means to image the KOR in vivo and investigate its function in health and disease. The purpose of this study was to develop the selective KOR antagonist (11)C-LY2459989 as a PET radioligand and characterize its imaging performance in nonhuman primates. LY2459989 was synthesized and assayed for in vitro binding to opioid receptors. Ex vivo studies in rodents were conducted to assess its potential as a tracer candidate. (11)C-LY2459989 was synthesized by reaction of its iodophenyl precursor with (11)C-cyanide, followed by partial hydrolysis of the resulting (11)C-cyanophenyl intermediate. Imaging experiments with (11)C-LY2459989 were performed in rhesus monkeys with arterial input function measurement. Imaging data were analyzed with kinetic models to derive in vivo binding parameters. LY2459989 is a full antagonist with high binding affinity and selectivity for KOR (0.18, 7.68, and 91.3 nM, respectively, for κ, μ, and δ receptors). Ex vivo studies in rats indicated LY2459989 as an appropriate tracer candidate with high specific binding signals and confirmed its KOR binding selectivity in vivo. (11)C-LY2459989 was synthesized in high radiochemical purity and good specific activity. In rhesus monkeys, (11)C-LY2459989 displayed a fast rate of peripheral metabolism. Similarly, (11)C-LY2459989 displayed fast uptake kinetics in the brain and an uptake pattern consistent with the distribution of KOR in primates. Pretreatment with naloxone (1 mg/kg, intravenously) resulted in a uniform distribution of radioactivity in the brain. Further, specific binding of (11)C-LY2459989 was dose-dependently reduced by the selective KOR antagonist LY2456302 and the unlabeled LY2459989. Regional binding potential values derived from the multilinear analysis-1 (MA1) method, as a measure of in vivo specific binding signal, were 2.18, 1.39, 1.08, 1.04, 1.03, 0.59, 0.51, and 0.50, respectively, for the globus pallidus, cingulate cortex, insula, caudate, putamen, frontal cortex, temporal cortex, and thalamus. The novel PET radioligand (11)C-LY2459989 displayed favorable pharmacokinetic properties, a specific and KOR-selective binding profile, and high specific binding signals in vivo, thus making it a promising PET imaging agent for KOR.


Sewage bacteriophage inactivation by cationic porphyrins: influence of light parameters.
Photodynamic therapy has been used to inactivate microorganisms through the use of targeted photosensitizers. Although the photoinactivation of microorganisms has already been studied under different conditions, a systematic evaluation of irradiation characteristics is still limited. The goal of this study was to test how the light dose, fluence rate and irradiation source affect the viral photoinactivation of a T4-like sewage bacteriophage. The experiments were carried out using white PAR light delivered by fluorescent PAR lamps (40 W m(-2)), sun light (600 W m(-2)) and an halogen lamp (40-1690 W m(-2)). Phage suspensions and two cationic photosensitizers (Tetra-Py(+)-Me, Tri-Py(+)-Me-PF) at concentrations of 0.5, 1.0 and 5.0 microM were used. The results showed that the efficacy of the bacteriophage photoinactivation is correlated not only with the sensitizer and its concentration but also with the light source, energy dose and fluence rate applied. Both photosensitizers at 5.0 microM were able to inactivate the T4-like phage to the limit of detection for each light source and fluence rate. However, depending of the light parameters, different irradiation times are required. The efficiency of photoinactivation is dependent on the spectral emission distribution of the light sources used. Considering the same light source and a fixed light dose applied at different fluence rates, phage inactivation was significantly higher when low fluence rates were used. In this way, the light source, fluence rate and total light dose play an important role in the effectiveness of the antimicrobial photodynamic therapy and should always be considered when establishing an optimal antimicrobial protocol.


Parent relationship quality buffers against the effect of peer stressors on depressive symptoms from middle childhood to adolescence.
During the transition to adolescence, several developmental trends converge to increase the importance of peer relationships, the likelihood of peer-related stressors, and the experience of depressive symptoms. Simultaneously, there are significant changes in parent-child relationships. The current study sought to evaluate whether positive relationship quality with parents continued to serve a protective effect by buffering the relationship between stressful life events, especially peer stress, and increases in depressive symptoms throughout the transition to adolescence. Participants in a large (N = 692) 2-site accelerated longitudinal study were recruited in 3rd, 6th, and 9th grade and followed every 3 months for 1 year. At baseline, parents and youth reported on parent-child relationship quality, and every 3 months thereafter reported on their levels of stressors and depressive symptoms. Parent relationship quality moderated the relationship of person-level fluctuations in peer stressors, such that there was a stronger association between peer stressors and increases in depressive symptoms in youth with lower levels of positive parental relationship quality. This effect was specific to peer stressors. These results suggest that low levels of parent relationship quality leave youth particularly vulnerable to the depressogenic effects of peer stressors from childhood through adolescence.


Current veterinary regulations and way ahead.
This article aims to perform a comparative systematic review of regulations in veterinary medicine between the years 2016 to 2023. It explores the complex web of veterinary medicine regulations in various agencies and the nations, including USA (United States of America), EU (European Union), UK (United Kingdom), Japan, Australia, and India. Current article provides the comparative study on the veterinary regulations of different countries, including acts, directives, and drug approval processes. Such as, the specific legislation is needed to address zoonotic diseases. The strategic and regulated stockpiling of the veterinary drugs especially in chronic veterinary disease outbreak. It is essential to develop the dedicated Veterinary Pharmacopoeia for the regulated standardization of the raw materials as well as the formulations. Veterinary medical device is a field which is highly unregulated. There is a need to have regulations for the same. It is important to have dedicated veterinary pharmacovigilance centers which help in improving quality of medications to the livestock farms. After comparing the regulations of different countries. We observed that there is the absence of the zoonotic diseases and pharma stockpiling in every country. There is also an absence of the dedicated veterinary pharmacopoeia in every country. USA and Australia have the veterinary medical device regulation which is not there in other countries. Around the globe only Australia has the dedicated pharmacovigilance center. Including these recommendations into regulatory framework enhances the quality and safety of veterinary medicine. The current article adds a valuable resource for policymakers, veterinarians, and stakeholders in the field of animal health care.


Transcriptome sequencing promotes insights on the molecular mechanism of SKP-SC-EVs mitigating denervation-induced muscle atrophy.
Complex pathophysiological changes accompany denervation-induced skeletal muscle atrophy, but no effective treatment strategies exist. Our previous study indicated that extracellular vesicles derived from skin-derived precursors-derived Schwann cells (SKP-SC-EVs) can effectively mitigate denervation-induced muscle atrophy. However, the specific molecular mechanism remains unclear. In this study, we used bioinformatics methods to scrutinize the impact of SKP-SC-EVs on gene expression in denervation-induced skeletal muscle atrophy. We found that SKP-SC-EVs altered the expression of 358 genes in denervated skeletal muscles. The differentially expressed genes were predominantly participated in biological processes, including cell cycle, inflammation, immunity, and adhesion, and signaling pathways, such as FoxO and PI3K.Using the Molecular Complex Detection (MCODE) plugin, we identified the two clusters with the highest score: cluster 1 comprised 37 genes, and Cluster 2 consisted of 24 genes. Then, fifty hub genes were identified using CytoHubba. The intersection of Hub genes and genes obtained by MCODE showed that all 23 genes related to the cell cycle in Cluster 1 were hub genes, and 5 genes in Cluster 2 were hub genes and associated with inflammation. Overall, the differentially expressed genes in denervated skeletal muscle following SKP-SC-EVs treatment are primarily linked to the cell cycle and inflammation. Consequently, promoting proliferation and inhibiting inflammation may be the critical process in which SKP-SC-EVs delay denervation-induced muscle atrophy. Our findings contribute to a better understanding of the molecular mechanism of SKP-SC-EVs delaying denervation-induced muscle atrophy, offering a promising new avenue for muscle atrophy treatment.


Prostaglandin involvement in hypersensitivity of ischemic hearts to arrhythmogenic influence of ouabain.
Arrhythmias were produced by ouabain (2.7 X 10(-7) M) in isolated perfused guinea pig hearts. In control hearts the average time to onset of arrhythmias following exposure to ouabain was approximately 35 min. Ligation of the left anterior descending coronary artery significantly shortened the time to arrhythmias to about 15 min. This effect of ligation was attenuated significantly by pretreating the hearts with acetylsalicylic acid (ASA) and other nonsteroidal anti-inflammatory drugs. In contrast, the protective influence of ASA was reversed by either prostaglandin (PG) E2, PGF2 alpha, or PGI2 (2.8 X 10(-9) M). When PGE2 or PGI2 were present in the absence of ASA, the time to arrhythmias following ouabain administration was significantly shortened. PGF2 alpha had no effect in these experiments. The sensitivity was enhanced significantly by treatment with arachidonic acid. This effect was reduced by treatment with ASA or indomethacin. Prostaglandin production during the experiments was estimated by monitoring release of 6-keto-PGF1 alpha, the hydrolysis product of prostacyclin. Abbreviation of time to arrhythmias by ligation was greatest in hearts that released large amounts of 6-keto-PGF1 alpha and least in those that released low amounts. These results suggest that coronary ligation potentiates the arrhythmogenic effects of ouabain by a mechanism probably mediated by prostaglandin(s).


4-Nitrochalcone as a potential drug in non-clinical breast cancer studies.
Breast cancer is a high-magnitude public health problem, continually challenging physicians and scientists worldwide in the field of drug therapy. 4-nitrochalcone (4NC) is a phenolic compound that has promising antitumor activity in vitro, but its application in breast cancer treatment is still poorly explored. This study aimed to evaluate the action of 4NC in vitro and in vivo breast cancer models. The cytotoxic potential of 4NC was tested towards MCF-7 and MDA-MD-231 breast cancer cells, with a lower impact in the non-tumor lineage HB4a. For in vivo studies, solid Ehrlich carcinoma (SEC) was used, a syngeneic mouse model with non-nuclear estrogen and progesterone positivity, characterized by immunohistochemistry. Daily oral administration of 4NC (25 mg kg-1) for 21 days led to a consistent reduction in tumor growth compared to the vehicle group. No signs of toxicity evaluated by hematological, biochemical, histological, and oxidative stress parameters were observed in mice, and the DL50 was >2000 mg kg-1. The effectors Raptor and S6K1 showed decreased activation, with a consequent reduction in protein synthesis; concomitantly, there was an increase in LC3-II levels, but the protective autophagic response was not completed, with the maintenance of p62 levels and cell death. These results open new possibilities for the use of 4NC as a tumor cell metabolism modulating agent.


Detecting Risk of Low Health Literacy in Disadvantaged Populations Using Area-based Measures.
Socio-economic status (SES) and low health literacy (LHL) are closely correlated. Both are directly associated with clinical and behavioral risk factors and healthcare outcomes. Learning healthcare systems are introducing small-area measures to address the challenges associated with maintaining patient-reported measures of SES and LHL. This study's purpose was to measure the association between two available census block measures associated with SES and LHL. Understanding the relationship can guide the identification of a multi-purpose area based measure for delivery system use. A retrospective observational design was deployed using all US Census block groups in Utah. The principal dependent variable was a nationally-standardized health literacy score (HLS). The primary explanatory variable was a state-standardized area deprivation index (ADI). Statistical methods included linear regression and tests of association. Receiver operating characteristic (ROC) analysis was used to develop LHL criteria using ADI. A significant negative association between the HLS and the ADI score remained after adjusting for area-level risk factors (β: -0.21 (95% CI: -0.22, -0.19) p < .001). Eighteen block groups (<1%) were identified as having LHL using HLS. A combination of three or more ADI components correlated with LHL predicted 78% of HLS LHL block groups and 35 additional block groups not identified using HLS (c-statistic: 0.64; 95% CI: 0.62, 0.66). HLS and ADI use differing measurement criteria but are closely correlated. A state-based ADI detected additional neighborhoods with risk of LHL compared to use of a national HLS. An ADI represents a multi-purpose area measure of social determinants useful for learning health systems tailoring care.


Acute Convulsive Seizures: When is It Too Early to Treat?
Acute convulsive seizures are overwhelming events that require immediate attention from clinicians and practitioners, especially when witnessed in a hospital setting. Adequate management of inpatient-witnessed seizures lies in understanding the time-related pathophysiologic stages of the seizure. The anatomical location of the seizure origin is as important as seizure stage but might not be easy to determine in the acute setting, especially if the seizure is nonfocal. Although investigating and treating the cause of a seizure has high priority, understanding the interplay between the pathophysiologic responses and the various bodily responses are crucial in treatment. This interplay has interesting dynamics that change within minutes. Knowing such dynamics allows clinicians and practitioners to choose their best treatment options in the best time interval when a seizure occurs in an acute care facility. Our commentary provides an overview of such dynamics and how they can change the misconceptions that many clinicians and practitioners have in dealing with an acute convulsive seizure.


Studying of drug solubility in water and alcohols using drug-ammonium ionic liquid-compounds.
Synthesis of three mefenamic acid (MEF) derivatives - ionic liquid compounds composed of MEF in an anionic form and ammonium cation (choline, MEF1), or {di(2-hydroxyethyl)dimethyl ammonium (MEF2)}, or {tri(2-hydroxyethyl)methyl ammonium compound (MEF3)} is presented. The basic thermal properties of pure compounds i.e. fusion temperatures, and the enthalpy of fusion of these compounds have been measured with differential scanning microcalorimetry technique (DSC). Molar volumes have been calculated with the Barton group contribution method. The solubilities of MEF1, MEF2 and MEF3 using the dynamic method were measured at constant pH in a range of temperature from (290 to 370) K in three solvents: water, ethanol and 1-octanol. The experimental solubility data have been correlated by means of three commonly known GE equations: the Wilson, NRTL and UNIQUAC with the assumption that the systems studied here present simple eutectic behaviour. The activity coefficients of pharmaceuticals at saturated solutions in each binary mixture were calculated from the experimental data. The formation of MEF-ionic liquid compounds greatly increases the solubility in water in comparison with pure MEF or complexes with 2-hydroxypropyl-β-cyclodextrin. The development of these compounds formulations will assist in medication taking into account oral solid or gel medicines.