μFE models can represent microdamaged regions of healthy and metastatically involved whole vertebrae identified through histology and contrast enhanced μCT imaging.
Micro-damage formation within the skeleton is an important stimulant for bone remodeling, however abnormal build-up of micro-damage can lead to skeletal fragility. In this study, µCT imaging based micro finite element (μFE) models were used to evaluate tissue level damage criteria in whole healthy and metastatically-involved vertebrae. T13-L2 spinal segments were excised from osteolytic (n=3) and healthy (n=3) female athymic rnu/rnu rats. Osteolytic metastasis was generated by intercardiac injection of HeLa cancer cells. Micro-mechanical axial loading was applied to the spinal motion segments under μCT imaging. Vertebral samples underwent BaSO4 staining and sequential calcein/fuchsin staining to identify load induced micro-damage. μCT imaging was used generate specimen specific μFE models of the healthy and osteolytic whole rat vertebrae. Model boundary conditions were generated through deformable image registration of loaded and unloaded scans. Elevated stresses and strains were detected in regions of micro-damage identified through histological and BaSO4 staining within healthy and osteolytic vertebral models, as compared to undamaged regions. Additionally, damaged regions of metastatic vertebrae experienced significantly higher local stresses and strains than those in the damaged regions of healthy specimens. Areas identified by BaSO4 staining, however, yielded lower levels of stress and strain in damaged and undamaged regions of healthy and metastatic vertebrae as compared to fuschin staining. The multimodal (experimental, image-based and computational) techniques used in this study demonstrated the ability of local stresses and strains computed through µFE analysis to identify trabecular micro-damage, that can be applied to biomechanical analyses of healthy and diseased whole bones.


Gastrointestinal pharmacology: irritable bowel syndrome.
Over the past 30 years, the main treatment of irritable bowel syndrome has aimed to normalize gastrointestinal transit using either laxatives or antidiarrheal agents, with or without the concurrent use of spasmolytics. The recent introduction of serotonin-related drugs has stimulated investigations into the pathophysiology of irritable bowel syndrome, including an evaluation of visceral sensitivity. At the same time, more information has been acquired on the status of the local immune system as a possible cause for sensitization of nerve terminals. Such investigations have stimulated the emergence of new concepts and original candidate drugs for the treatment of this functional disorder. Particular attention is devoted to the correction of visceral hyperalgesia.


Urethroplasty, by perineal approach, for bulbar and membranous urethral strictures in children and adolescents.
To evaluate the safety and efficacy of urethroplasty for bulbar and membranous urethral strictures using the perineal approach in children and adolescents. Urethroplasty by the perineal approach is considered the best treatment for bulbar and membranous urethral strictures in adults. It is not as clear whether this also holds true in children, because the published data addressing this question are scant. We retrospectively reviewed our urethroplasty database to identify patients who had undergone urethroplasty using the perineal approach surgery at age 1 to 13 years (children) and 14 to 18 years (adolescents). A total of 14 patients who had undergone urethroplasty by the perineal approach were identified, including 5 children (mean age 10.8 years) and 9 adolescents (mean age 16.7 years). Of the 14 patients, 7 had membranous and 7 bulbar urethral strictures. The membranous strictures were all secondary to pelvic fractures. The bulbar strictures were "idiopathic" in 57%, traumatic in 29%, and secondary to hypospadias in 14%. All bulbar strictures had been previously treated for 2.5 years, on average, by repeated dilation or urethrotomy that failed. Anastomotic urethroplasty was used in 79% of the patients and tissue transfer techniques in the remainder. The mean follow-up was 30 months (range 12 to 54). Surgery was primarily successful in 93% of the patients, and subsequently successful in 100%. The mean maximal urinary flow increased from 2.65 mL/s preoperatively to 27.65 mL/s postoperatively. No significant complications occurred, and success was similar in both groups. In pediatric patients, as in adults, bulbar and membranous strictures can be treated successfully with urethroplasty using the perineal approach. These patients should probably not be treated "conservatively" with urethral dilation or endoscopic incision. Longer follow-up is needed to confirm that these good results are maintained as these patients cross into adulthood, especially for those who underwent repair before puberty.


Clinicopathological features and survival of rare primary pulmonary lymphoepithelial carcinoma: A cohort from a single center.
Primary pulmonary lymphoepithelial carcinoma (PLEC) is a rare subtype of nonsmall cell lung cancer. This study aimed to investigate the clinicopathological and prognostic characteristics of resected primary PLEC. In this retrospective study, 95 consecutive patients with primary PLEC, who received radical surgical resection treatment, were examined from October 2009 to January 2022. The clinicopathological features and their association with survival outcomes were analyzed. Primary PLEC predominated in relatively younger patients and nonsmokers, who lacked driver mutations and were always positive for immunohistochemical markers of the squamous cell lineage. Further, 21.1% of patients had abnormally elevated preoperative serum marker fragments of cytokeratin 19 (Cyfra21-1). The median follow-up time was 43.5 months. The 1-, 3-, and 5-year recurrence-free survival (RFS) rates were 96.5%, 81.8%, and 64.3%, respectively. The median RFS time was not reached. Cox univariate survival analysis showed that patients with positive lymph nodes had significantly worse RFS than those with negative ones (p = 0.017). The patients with open surgery experienced significantly worse RFS than those with video-assisted thoracoscopic surgery (p = 0.038). The multivariate survival analysis confirmed that only lymph node involvement (hazard ratio: 2.769; 95% confidence interval: 1.171-6.548, p = 0.020) was an independent prognostic factor. Primary PLEC is a rare type of lung cancer with a favorable outcome, more common in young and nonsmoking Asian populations. Driver gene mutations are rare. Regional lymph node metastasis is an independent prognostic factor for RFS after radical surgical resection.


A DGCR8-Independent Stable MicroRNA Expression Strategy Reveals Important Functions of miR-290 and miR-183-182 Families in Mouse Embryonic Stem Cells.
Dgcr8 knockout cells provide a great means to understand the function of microRNAs (miRNAs) in vitro and in vivo. Current strategies to study miRNA function in Dgcr8 knockout cells depend on transient transfection of chemically synthesized miRNA mimics, which is costly and not suitable for long-term study and genetic selection of miRNA function. Here, we developed a cost-effective DGCR8-independent stable miRNA expression (DISME) strategy based on a short hairpin RNA vector that can be precisely processed by DICER. Using DISME, we found that miR-294 promoted the formation of meso-endoderm lineages during embryonic stem cell differentiation. Furthermore, DISME allowed for a pooled screen of miRNA function and identified an miR-183-182 cluster of miRNAs promoting self-renewal and pluripotency in mouse embryonic stem cells. Altogether, our study demonstrates that DISME is a robust and cost-effective strategy that allows for long-term study and genetic selection of miRNA function in a Dgcr8 knockout background.


Ultrasonography and computed tomography of extracapsular invasion in cervical lymph nodes of squamous cell carcinoma in the oral cavity.
This study evaluates the efficacy of imaging for diagnosing extracapsular invasion by metastatic cervical lymph nodes. The subjects were 47 patients with squamous cell carcinoma of the oral cavity examined by both ultrasonography and computed tomography. All subjects had undergone neck dissection, and histopathologic examination confirmed the presence of one or more metastatic cervical lymph nodes in each of them. Histopathologic examination confirmed extracapsular invasion in 25 of 48 metastatic nodes; in the remaining 23 nodes, the tumor was confined within the capsule. Average short-axis diameters of the lymph nodes of these groups differed significantly (17.4 ± 10.6 and 12.7 ± 4.4 mm, respectively; P = 0.046). The diagnostic sensitivity, specificity, and accuracy of ultrasonography for detecting macroscopic extracapsular invasion were 80%, 74%, and 77%, respectively. The corresponding values for computed tomography were 85%, 83%, and 84%. Diagnostic images are effective for the preoperative diagnosis of extracapsular invasion of metastatic cervical nodes of carcinoma of the oral cavity.


Blood supply of the flexor pollicis longus tendon.
The blood supply of the flexor pollicis longus tendon was investigated by arterial injections of india ink--latex-barium-solution in 15 fresh, adult cadaver upper extremities. In the digital area vascularity was by way of two vincula (V1 and V2). V1, located just proximal to the metacarpophalangeal joint and the A1 pulley, originated either from both digital arteries or from the princeps pollicis artery alone. V2, located at the interphalangeal joint level under the A2 pulley, originated from both digital arteries. In the predigital area branches of the median nerve artery supplied the area from the base of the thumb to the musculotendinous junction; in addition, the microvascular network of the mesotendon contributed to this area. Intratendinous vascularization was more abundant in the predigital area.


A Rare Case Report of a Tail-Gut Cyst from a Gynecological Point of View.
A tail-gut cyst can be often a misleading clinical entity. In half of the patients there is no presenting symptom. On the other half, the patients most commonly present with a variety of symptoms such as rectal pain, constipation, lower back pain, dysuria or dyspareunia. The recommended treatment of choice for the tail-gut cyst is complete surgical excision without rupture of the cyst. We present the case of a 29-year-old female with history of dyspareunia over a 5-month period, who discovered an "ovarian" cyst during an annual scheduled ultrasound appointment. However, the intraoperative findings were surprising. The bottomline is always to keep in mind the Pandora's Box of the retrorectal space.


Serological diagnosis of subcutaneous dirofilariosis.
We present serological evidence that Dirofilaria immitis was the causative agent of subcutaneous nodules in two patients. ELISA and enzyme-linked immunoblot analysis showed that the two human sera had a reactivity pattern similar to that of D. immitis rabbit immune serum. The reaction pattern of the human sera was very different to that of Dirofilaria repens rabbit immune serum. In the light of our findings, the traditional view that subcutaneous filarial nodules are due to D. repens has to be revised.


Evidence Review for Centers for Disease Control and Prevention Guidance Development on Laboratory Testing to Detect Treponema pallidum Infection (Syphilis).
The articles in this supplement address key questions on syphilis diagnostics, provide reference tables of test performances, and discuss optimal specimens and knowledge gaps. Laboratory-developed genetic direct detection tests could be most useful at the point of care and add to the currently available serologic methods of nontreponemal and treponemal tests.


Aryl-indolyl maleimides as inhibitors of CaMKIIdelta. Part 1: SAR of the aryl region.
A family of aryl-substituted maleimides was prepared and studied for their activity against calmodulin dependant kinase. Inhibitory activities against the enzyme ranged from 34nM to >20microM and were dependant upon both the nature of the aryl group and the hydrogen bond donating potential of the maleimide ring. Key interactions with the kinase ATP site and hinge region were found to be consistent with homology modeling predictions.


Results of hatching and rearing broiler chickens in different incubation systems.
Hatchery efficiency is based on hatchability and the number of salable chicks. The hatchery sector has been seeking new alternatives to optimize production rates, including the use of different systems (multistage [MS] or single-stage [SS] machines) to improve incubation conditions. The present study aimed to compare results for hatchability, chick quality, and broiler performance of chicks from 2 incubator systems-MS and SS. The experimental design for hatchability, hatch window, egg weight loss, and chick performance variables was completely randomized with 2 treatments (MS and SS). Performance variables were analyzed as a 2 x 2 factorial arrangement (incubator type x chick sex). Egg weight loss between incubation and transfer was higher for eggs incubated in MS (P < 0.05). Hatchability was higher for eggs incubated in SS (P < 0.05), and chicks in SS had a longer hatch window (P < 0.05). Embryo diagnosis revealed higher final mortality for embryos incubated in MS (P < 0.05), as well as higher percentages of alive and dead pipped and cracked eggs (P < 0.05). Physical quality was better for chicks from SS (P < 0.05). There was no interaction between the studied factors for performance results (P > 0.05). Incubator type did not affect broiler performance for any of the studied ages (P > 0.05), whereas male broilers had better performance than females (P < 0.05). The SS incubation system proved better than the MS system at meeting embryo requirements during embryo development, with better hatching rates and chick quality, although performance variables were not influenced by incubation type.


Influence of 2-Methoxyestradiol and Sex on Hypoxia-Induced Pulmonary Hypertension and Hypoxia-Inducible Factor-1-α.
Background Women are at greater risk of developing pulmonary arterial hypertension, with estrogen and its downstream metabolites playing a potential role in the pathogenesis of the disease. Hypoxia-inducible factor-1-α (HIF 1α) is a pro-proliferative mediator and may be involved in the development of human pulmonary arterial hypertension . The estrogen metabolite 2-methoxyestradiol (2 ME 2) has antiproliferative properties and is also an inhibitor of HIF 1α. Here, we examine sex differences in HIF 1α signaling in the rat and human pulmonary circulation and determine if 2 ME 2 can inhibit HIF 1α in vivo and in vitro. Methods and Results HIF 1α signaling was assessed in male and female distal human pulmonary artery smooth muscle cells ( hPASMC s), and the effects of 2 ME 2 were also studied in female hPASMC s. The in vivo effects of 2 ME 2 in the chronic hypoxic rat (male and female) model of pulmonary hypertension were also determined. Basal HIF 1α protein expression was higher in female hPASMC s compared with male. Both factor-inhibiting HIF and prolyl hydroxylase-2 (hydroxylates HIF leading to proteosomal degradation) protein levels were significantly lower in female hPASMC s when compared with males. In vivo, 2 ME 2 ablated hypoxia-induced pulmonary hypertension in male and female rats while decreasing protein expression of HIF 1α. 2 ME 2 reduced proliferation in hPASMC s and reduced basal protein expression of HIF 1α. Furthermore, 2 ME 2 caused apoptosis and significant disruption to the microtubule network. Conclusions Higher basal HIF 1α in female hPASMC s may increase susceptibility to developing pulmonary arterial hypertension . These data also demonstrate that the antiproliferative and therapeutic effects of 2 ME 2 in pulmonary hypertension may involve inhibition of HIF 1α and/or microtubular disruption in PASMC s.


Clinical Outcomes Related to Background Diuretic Use and New Diuretic Initiation in Patients With HFrEF.
Up to 20% of patients in heart failure with reduced ejection fraction (HFrEF) trials are not taking diuretic agents at baseline, but little is known about them. The aim of this study was to examine outcomes in patients with HFrEF not taking diuretic medications and after diuretic medications are started. Patient characteristics and outcomes were compared between patients taking or not taking diuretic drugs at baseline in the ATMOSPHERE (Aliskiren Trial of Minimizing Outcomes for Patients With Heart Failure) and PARADIGM-HF (Prospective Comparison of ARNI With ACEI to Determine Impact on Global Mortality and Morbidity in Heart Failure Trial) trials combined. Patients starting diuretic medications were also compared with those remaining off diuretic drugs during follow-up. Symptoms (Kansas City Cardiomyopathy Questionnaire Clinical Summary Score [KCCQ-CSS]), hospitalization for worsening heart failure (HF), mortality, and kidney function (estimated glomerular filtration rate slope) were examined. At baseline, the 3,079 of 15,415 patients (20%) not taking diuretic medications had a less severe HF profile, less neurohumoral activation, and better kidney function. They were less likely to experience the primary outcome (hospitalization for HF or cardiovascular death) than patients taking diuretic agents (adjusted HR: 0.77; 95% CI: 0.74-0.80; P < 0.001) and death of any cause. Commencement of a diuretic drug was associated with higher subsequent risk for death (adjusted HR: 2.05; 95% CI: 1.99-2.11; P < 0.001) and greater decreases in KCCQ-CSS and estimated glomerular filtration rate. The 5 strongest predictors of initiation of diuretic medications were higher N-terminal pro-B-type natriuretic peptide, higher body mass index, older age, history of diabetes, and worse KCCQ-CSS. In PARADIGM-HF, fewer patients who were treated with sacubitril/valsartan commenced diuretic agents (OR: 0.72; 95% CI: 0.58-0.88; P = 0.002). Patients with HFrEF not taking diuretic medications and those who remained off them had better outcomes than patients treated with diuretic agents or who commenced them.


The peritoneal leukocytes of the germ-free mouse.
The cytology of the lavage-recoverable peritoneal cell population of germ-free mice is similar to that of conventional controls. The microbial status has no effect on the total counts, differential counts or 3H-thymidine labelling index of peritoneal leukocytes.


Complex Transformer Network for Single-Angle Plane-Wave Imaging.
Plane-wave imaging (PWI) is a high-frame-rate imaging technique that sacrifices image quality. Deep learning can potentially enhance plane-wave image quality, but processing complex in-phase and quadrature (IQ) data and suppressing incoherent signals pose challenges. To address these challenges, we present a complex transformer network (CTN) that integrates complex convolution and complex self-attention (CSA) modules. The CTN operates in a four-step process: delaying complex IQ data from a 0° single-angle plane wave for each pixel as CTN input data; extracting reconstruction features with a complex convolution layer; suppressing irrelevant features derived from incoherent signals with two CSA modules; and forming output images with another complex convolution layer. The training labels are generated by minimum variance (MV). Simulation, phantom and in vivo experiments revealed that CTN produced comparable- or even higher-quality images than MV, but with much shorter computation time. Evaluation metrics included contrast ratio, contrast-to-noise ratio, generalized contrast-to-noise ratio and lateral and axial full width at half-maximum and were -11.59 dB, 1.16, 0.68, 278 μm and 329 μm for simulation, respectively, and 9.87 dB, 0.96, 0.62, 357 μm and 305 μm for the phantom experiment, respectively. In vivo experiments further indicated that CTN could significantly improve details that were previously vague or even invisible in DAS and MV images. And after being accelerated by GPU, the CTN runtime (76.03 ms) was comparable to that of delay-and-sum (DAS, 61.24 ms). The proposed CTN significantly improved the image contrast, resolution and some unclear details by the MV beamformer, making it an efficient tool for high-frame-rate imaging.


Deaths as a result of work-related injury in Australia, 1982-1984.
A comprehensive study of deaths of work-related injuries which occurred throughout Australia in the years 1982-1984 was undertaken by means of coroners' records. Of 16,246 coroner-certified deaths that were attributed to trauma or to acute poisoning (excluding deaths of suicide or medical misadventure), the coroners' files were located for 15,462 (95.2%) deaths. From the files, a total of 1738 fatalities was judged to be work-related during the three-year period; of these, 1544 fatalities occurred in persons who were employed in the civilian labour force, which gave an average annual fatality incidence of 8.06 fatalities per 100,000 persons in the labour force. The death rate was much higher in men (12.05 fatalities per 100,000 men) than it was in women (1.34 fatalities per 100,000 women), increased with age, and was highest for the traditionally-dangerous occupations (such as mining, transport and rural occupations). The distribution of work fatalities by the main cause of death, and the nature of the injury event is described.


Conditional power for assessing population interventions.
To calculate conditional power in comparative two-period studies with previously observed baseline data. Isolate the variability attributable to the yet-to-observed data and modify the standard power formulae. For illustration, we examine rates of opioid overdose before and after a reformulation of one opioid product. The null hypothesis posited no impact of the reformulation, alternative hypotheses posited possible impacts, and ancillary hypotheses posited different secular pre-post changes directly observable in comparators. Conditional power varied with the size of the comparator population and with the assumed pre-post change for the comparator. Pre-post designs can be initiated after the baseline period is over. Power calculations that are conditioned on observed baseline data account differently for variability in the baseline and follow-up periods.


Mild photothermal/radiation therapy potentiates ferroptosis effect for ablation of breast cancer via MRI/PA imaging guided all-in-one strategy.
Nanotheranostics advances anticancer management by providing therapeutic and diagnostic functions, that combine programmed cell death (PCD) initiation and imaging-guided treatment, thus increasing the efficacy of tumor ablation and efficiently fighting against cancer. However, mild photothermal/radiation therapy with imaging-guided precise mediating PCD in solid tumors, involving processes related to apoptosis and ferroptosis, enhanced the effect of breast cancer inhibition is not fully understood. Herein, targeted peptide conjugated gold nano cages, iRGD-PEG/AuNCs@FePt NPs ternary metallic nanoparticles (Au@FePt NPs) were designed to achieve photoacoustic imaging (PAI)/Magnetic resonance imaging (MRI) guided synergistic therapy. Tumor-targeting Au@FePt forms reactive oxygen species (ROS), initiated by X-ray-induced dynamic therapy (XDT) in collaboration with photothermal therapy (PTT), inducing ferroptosis-augmented apoptosis to realize effective antitumor therapeutics. The relatively high photothermal conversion ability of Au@FePt increases the temperature in the tumor region and hastens Fenton-like processes to achieve enhanced synergistic therapy. Especially, RNA sequencing found Au@FePt inducting the apoptosis pathway in the transcriptome profile. Au@FePt combined XDT/PTT therapy activate apoptosis and ferroptosis related proteins in tumors to achieve breast cancer ablation in vitro and in vivo. PAI/MRI images demonstrated Au@FePt has real-time guidance for monitoring synergistic anti-cancer therapy effect. Therefore, we have provided a multifunctional nanotheranostics modality for tumor inhibition and cancer management with high efficacy and limited side effects.


Diversification of the barley and wheat blt101/wpi6 promoters by the Xumet element without affecting stress responsiveness.
BLT101-family plasma membrane proteins are found in a wide range of organisms from bacteria to nematodes and are involved in the regulation of cellular cation concentration under stress conditions. A comparison of the promoter regions of barley blt101 and its wheat ortholog, wpi6, revealed highly conserved nucleotide sequences between both genes and a unique insertion of a Xumet element in the blt101 promoter. The Xumet insertion occurred between a putative abscisic acid-responsive element (ABRE) and the dehydration-responsive element/c-repeat (DRE/CRT) within the blt101 promoter. However, blt101 and wpi6 were induced similarly in response to ABA, drought and low temperature, suggesting that the insertion does not affect promoter functions. The Xumet insertion in the blt101/wpi6 promoter region was detected in five barley cultivars, but absent in two wheat cultivars tested, suggesting that the insertion is barley-specific. Genomic Southern blot analysis revealed a large number of Xumet sequences interspersed in the barley genome, whereas only one or very few copies are present in the wheat genome. The data suggested that an expansion in copy number of Xumet elements occurred in the barley genome through evolution.


Structural basis for amino acid exchange by a human heteromeric amino acid transporter.
Heteromeric amino acid transporters (HATs) comprise a group of membrane proteins that belong to the solute carrier (SLC) superfamily. They are formed by two different protein components: a light chain subunit from an SLC7 family member and a heavy chain subunit from the SLC3 family. The light chain constitutes the transport subunit whereas the heavy chain mediates trafficking to the plasma membrane and maturation of the functional complex. Mutation, malfunction, and dysregulation of HATs are associated with a wide range of pathologies or represent the direct cause of inherited and acquired disorders. Here we report the cryogenic electron microscopy structure of the neutral and basic amino acid transport complex (b[0,+]AT1-rBAT) which reveals a heterotetrameric protein assembly composed of two heavy and light chain subunits, respectively. The previously uncharacterized interaction between two HAT units is mediated via dimerization of the heavy chain subunits and does not include participation of the light chain subunits. The b(0,+)AT1 transporter adopts a LeuT fold and is captured in an inward-facing conformation. We identify an amino-acid-binding pocket that is formed by transmembrane helices 1, 6, and 10 and conserved among SLC7 transporters.


Light distributions in artery tissue: Monte Carlo simulations for finite-diameter laser beams.
Finite-width light distributions in arterial tissue during Argon laser irradiation (476 nm) are simulated using the Monte Carlo method. Edge effects caused by radial diffusion of the light extend +/- 1.5 mm inward from the perimeter of a uniform incident beam. For beam diameters exceeding 3 mm the light distribution along the central axis can be described by the one-dimensional solution for an infinitely wide beam. The overlapping edge effects for beam diameters smaller than 3 mm reduce the penetration of the irradiance in the tissue. The beam profile influences the light distribution significantly. The fluence rates near the surface for a Gaussian beam are two times higher on the central axis and decrease faster radially than for a flat profile. The diverging light from a fiber penetrates tissue in a manner similar to collimated light.


Regulation of neurite outgrowth in N1E-115 cells through PDZ-mediated recruitment of diacylglycerol kinase zeta.
Syntrophins are scaffold proteins that regulate the subcellular localization of diacylglycerol kinase zeta (DGK-zeta), an enzyme that phosphorylates the lipid second-messenger diacylglycerol to yield phosphatidic acid. DGK-zeta and syntrophins are abundantly expressed in neurons of the developing and adult brain, but their function is unclear. Here, we show that they are present in cell bodies, neurites, and growth cones of cultured cortical neurons and differentiated N1E-115 neuroblastoma cells. Overexpression of DGK-zeta in N1E-115 cells induced neurite formation in the presence of serum, which normally prevents neurite outgrowth. This effect was independent of DGK-zeta kinase activity but dependent on a functional C-terminal PDZ-binding motif, which specifically interacts with syntrophin PDZ domains. DGK-zeta mutants with a blocked C terminus acted as dominant-negative inhibitors of outgrowth from serum-deprived N1E-115 cells and cortical neurons. Several lines of evidence suggest DGK-zeta promotes neurite outgrowth through association with the GTPase Rac1. DGK-zeta colocalized with Rac1 in neuronal processes and DGK-zeta-induced outgrowth was inhibited by dominant-negative Rac1. Moreover, DGK-zeta directly interacts with Rac1 through a binding site located within its C1 domains. Together with syntrophin, these proteins form a tertiary complex in N1E-115 cells. A DGK-zeta mutant that mimics phosphorylation of the MARCKS domain was unable to bind an activated Rac1 mutant (Rac1(V12)) and phorbol myristate acetate-induced protein kinase C activation inhibited the interaction of DGK-zeta with Rac1(V12), suggesting protein kinase C-mediated phosphorylation of the MARCKS domain negatively regulates DGK-zeta binding to active Rac1. Collectively, these findings suggest DGK-zeta, syntrophin, and Rac1 form a regulated signaling complex that controls polarized outgrowth in neuronal cells.


Identification and remediation of pediatric fluency and voice disorders.
Early identification of pediatric disfluency and voice disorders is advisable because these disorders may progress to lifelong communicative impairments if left untreated. Especially with disfluency or stuttering, it is critical that an informed differential diagnosis be made to determine whether a speech pattern represents normal disfluency or actual stuttering. Voice disorders can be overlooked as laryngitis, when in fact the problem may be organic in origin. This article describes characteristics of both disorders, etiologic factors, and checklists to assess children for referral to an otolaryngologist and/or speech-language pathologist. Medical and therapeutic treatment recommendations also are discussed.


Increased expression of annexin I and thioredoxin detected by two-dimensional gel electrophoresis of drug resistant human stomach cancer cells.
The therapy of advanced cancer using chemotherapy alone or in combination with radiation or hyperthermia yields an overall response rate of about 20-50%. This success is often marred by the development of resistance to cytostatic drugs. Our aim was to study the global analysis of protein expression in the development of chemoresistance in vitro. We therefore used a cell culture model derived from the gastric carcinoma cell line EPG 85-257P. A classical multidrug-resistant subline EPG85-257RDB selected to daunorubicin and an atypical multidrug-resistant cell variant EPG85-257RNOV selected to mitoxantrone, were analysed using two-dimensional electrophoresis in immobilized pH-gradients (pH 4.0-8.0) in the first dimension and linear polyacrylamide gels (12%) in the second dimension. After staining with coomassie brilliant blue, image analysis was performed using the PDQuest system. Spots of interest were isolated using preparative two-dimensional electrophoresis and subjected to microsequencing. A total of 241 spots from the EPG85-257RDB-standard and 289 spots from the EPG85-257RNOV-standard could be matched to the EPG85-257P-standard. Microsequencing after enzymatic hydrolysis in gel, mass spectrometric data and sequencing of the peptides after their fractionation using microbore HPLC identified that two proteins annexin I and thioredoxin were overexpressed in chemoresistant cell lines. Annexin I was present in both the classical and the atypical multidrug-resistant cells. Thioredoxin was found to be overexpressed only in the atypical multidrug-resistant cell line.


Application of enzyme-linked-immuno-electro-diffusion-assay (ELIEDA) in diagnosis of bilharziasis.
The sensitivity of the technique of immunoelectrodiffusion has been enhanced by combining it with an enzyme method. The precipitated immuno-complexes are treated with antibodies conjugated with an enzyme specific for each class of immuno-globulins. This association realizes the ELIEDA and enable the class of immunoglobulins involved in the reactions to be determined. The technique allows the follow up and the qualitative evaluation of the bilharzial antibody in natural and experimental affections.


Association of HADHA with human RNA silencing machinery.
In RNA silencing, small RNAs produced by dicer mediate target repression guided by RNA induced silencing complex (RISC). To effectively mediate this response, one or more proteins are employed at each stage. In the present study, we investigated HADHA, a new component in the human RNA silencing machinery. Immunoprecipitation analysis revealed that, HADHA associates with dicer complex and immunohistochemical studies confirmed its co localization with Dicer in the cytoplasm. Further, over expression of HADHA resulted in higher abundance levels of mature miRNA against a reduction in respective precursor levels and vice versa in HADHA knocked down cells. These findings suggest an auxiliary role for HADHA in miRNA biogenesis and help in better understanding of molecular mechanisms underlying RNAi pathway in mammals.


Comparison of different hypervariable regions of 16S rRNA for taxonomic profiling of vaginal microbiota using next-generation sequencing.
The exploration of vaginal microbiota by using next-generation sequencing (NGS) of 16S ribosomal RNA (rRNA) gene is widely used. Up to now, different hypervariable regions have been selected to study vaginal microbiota by NGS and there is no standard method for analysis. The study aimed to characterize vaginal microbiota from clinical samples using NGS targeting the 16S rRNA gene and to determine the performance of individual and concatenated hypervariable region sequences to generate the taxonomic profiles of the vaginal microbiota. Fifty-one vaginal DNA samples were subjected to 16S rRNA gene NGS based on the Ion Torrent PGM platform with the use of two primer sets spanning seven hypervariable regions of the 16S rRNA gene. Our analysis revealed that the predominant bacterial genera were Lactobacillus, Gardnerella and Atopobium, which accounted for 78%, 14% and 2%, respectively, of sequences from all vaginal bacterial genera. At the species level, Lactobacillus iners, Gardnerella vaginalis and Atopobium vaginae accounted for 72%, 10% and 6%, respectively, of the bacterial cells present. Analyses using the V3 region generally indicated the highest bacterial diversity followed by the V6-V7 and V4 regions, while the V9 region gave the lowest bacterial resolution. NGS based on the 16S rRNA gene can give comprehensive estimates of the diversity of vaginal bacterial communities. Selection of sequences from appropriate hypervariable regions is necessary to provide reliable information on bacterial community diversity.


Malignant thymoma metastatic to the pelvis: a rare case and considerations for management.
Thymoma is the most common tumor of the anterior mediastinum. Due to its relative rarity, its inconsistent clinical course and its frequent proximity to vital thoracic structures, management of this generally indolent tumor varies considerably. We present an unusual case of recurrent thymoma metastatic to the pelvis and review treatment experience employing surgical, radiotherapeutic and medical modalities. The present case is that of a 46-year-old woman with recurrent thymoma metastatic to a distal pelvic lymph node. Resection of the pelvic recurrence followed many years of local and systemic treatment for her thoracic primary tumor. Her case is unique for its involvement of pelvic anatomy and her clinical course marked by treatment-related congestive heart failure. While the indolent clinical course of thyomoma frequently necessitates re-treatment and multi-modality therapy in patients suffering recurrences, treatment selection must take into account potential long-term morbidity and attendant quality of life. When anatomically and technically feasible, resection of recurrent disease should be considered in attempts to avoid potential cumulative and long-term toxicity resultant from radiotherapy and chemotherapy.


Small-Scale Recombinant Adeno-Associated Virus Purification.
Recombinant adeno-associated virus (rAAV) vectors have become increasingly popular in research and clinical trials due to their efficient gene transfer and long-term expression in tissues including brain. In addition, rAAV has demonstrated an impressive safety profile in gene therapy trials. The emergence of rAAV serotypes with different cell tropisms and distribution properties has allowed scientists to tailor serotypes to specific experimental needs. AAV does not have a cytopathic effect; therefore, purification methods require extraction of the viral vector from the cell. This involves gradient ultracentrifugation of the cellular extract sometimes followed by chromatography. This chapter describes a small-scale production method for rAAV purification from ten to twenty 15 cm plates of human embryonic kidney-derived 293B cells (HEK 293) cells that can yield approximately 300 μl of a 5 × 10(12) to 1 × 10(13) genome copies/ml viral preparation final concentration.


Comparing data sources in estimating disability-adjusted life years (DALYs) for ischemic heart disease and chronic obstructive pulmonary disease in a cross-sectional setting in Finland.
The disability-adjusted life years (DALYs) summarize the burden of years of life lost (YLL) due to premature mortality and years lost due to disability (YLD). Our aim was to estimate the burden of ischemic heart disease (IHD) and chronic obstructive pulmonary disease (COPD) in Finland in 2012, and to examine, how much the YLD are affected by the use of different data sources. The YLL were calculated using mortality data for the Finnish 25-74-year-old population in 2012. The YLD were calculated using data from the FINRISK 2012 survey (3041 males, 3383 females aged 25-74 years) and then directly adjusted to the corresponding population. Different administrative registers on 1) hospital inpatient episodes and specialist outpatient visits, 2) entitlement to specially reimbursed medicines, and 3) purchases of prescribed medicines were used for estimation of the YLD in addition to self-reported data. The DALYs were calculated without age-weighting. The YLL for IHD were 37.5 for males and 9.1 for females per 1000 population among 25-74-year-old people in Finland in 2012. The YLD for IHD varied markedly depending on which data sources were used. All data sources combined, the YLD per 1000 were 5.3 for males and 2.5 for females resulting in estimated 42.8 and 11.6 DALYs per 1000 due to IHD among males and females, respectively. For COPD, the YLL were 4.7 for males and 2.0 for females per 1000. Also for COPD, the YLD varied markedly depending on data sources used. The YLD per 1000 based on all data sources combined were 2.0 for males and 1.6 for females. As a result, estimated 6.7 and 3.6 DALYs per 1000 were due to COPD among males and females, respectively. Especially for COPD, all mild disease cases could probably not be identified from the included registers. Thereby, including survey data improved the coverage of the data. The YLD of IHD and COPD varied markedly between the data sources used in the calculations. However, as YLL constituted a major part of DALYs for these diseases, the variation in YLD did not lead to substantial variation in DALYs.


Initial series of robotic radical nephrectomy with vena caval tumor thrombectomy.
Laparoscopy has become a standard modality for most renal tumors but not as yet for renal cell carcinoma (RCC) involving the inferior vena cava (IVC). Robotic technology may facilitate such complex procedures. We report the first series of robotic nephrectomy with IVC tumor thrombectomy including the first cases requiring cross-clamping of the IVC in a minimally invasive fashion. Five patients underwent robotic nephrectomy with IVC tumor thrombectomy including one patient having two renal veins, each with an IVC thrombus, for a total of six IVC thrombi. The IVC was opened in all patients, and tumor thrombi were delivered intact, followed by sutured closure. The mean patient age was 64 yr (53-70 yr) with a mean body mass index of 36.6 kg/m(2) (22-43 kg/m(2)). Thrombi protruded 1 cm, 2 cm, 4 cm, and 5 cm into the IVC in five patients and 3 cm and 2 cm in the patient with two thrombi. The mean estimated blood loss was 170 ml (50-400 ml). Mean operative time was 327 min (240-411 min). Mean length of stay was 1.2 d. There were no complications, transfusions, or readmissions. This early series represents a limited experience by a single surgeon with a new procedure and may not be reproducible in larger numbers or by all surgeons. Further experience is necessary to validate this application.


Neurodegenerative disease.
This report from the Society of Neuroscience meeting in Los Angeles is limited to a small cohort of presentations which reflect this reporter's research interests, namely mechanisms of cell death in neurodegenerative disease, and in particular Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), and Parkinson's disease (PD).


Renal tubular differentiation in mouse and mouse metanephric culture. I. Ultrastructural studies.
We studied the morphologic features of renal tubular differentiation in transfilter metanephric culture. Differentiation of portions of the S-shaped loop into proximal convoluted tubule was detected shortly after 72 h of culture by the appearance of microvilli, coated membrane invaginations, and an apical vacuolar-microtubular network. These features developed synchronously, and the microvilli became progressively more numerous and more compact to form a brush border. Computer-assisted morphometric analysis showed only minor differences between proximal tubular cells from 18-day embryos and tubular cells from 7-day cultures of blastema taken from 11-day embryos. Segments of tubule corresponding to distal convoluted tubules were lined with relatively simple cells that contained few differentiating characteristics. Morphometric modeling of the tubular cells indicated a simple shape consistent with an inherent transport function.


[Normal cells and dykaryoses in vaginal smears using a high- resolution system for quantitative analysis].
In pap-stained cervical smears the cells were identified, destained and afterwards stained according to the Feulgen procedure. The measurements of the nuclei were performed on a scanning microscope photometer and 4 features were extracted for cell classification: total DNA-amount, nuclear area, degree of condensation of chromatin and granularity of chromatin. The comparison and separation of the cell groups was performed at the Computing Center of the Univ. of Ulm by 2 programs, a discriminant analysis and a specially adapted histogram. The distinction of normal cells and dyscaryoses was possible to a high degree by the features total DNA-amount and nuclear area; the 2 parameters describing the chromatin structure proved to be useless in these cases. Nuclear structure parameters were helpful in distinguishing certain types of normal cells.


Hospital Radiopharmacy in Argentina during the COVID-19 pandemic. Criteria and rationale for the organization of work.
This publication presents criteria and bases for the work organization in the safe practice of Hospital Radiopharmacy, in order to minimize the risk of viral transmission during the COVID-19 pandemic, in a reference facility of the National Energy Commission Atomic of Argentina, while continuing to perform essential services for the health system. For this purpose, documents from the National Energy Commission Atomic, IAEA, WHO and other scientific publications were consulted as reference. These recommendations are under constant review and are permanently updated. Within this framework, the present model of work organization for this essential activity is proposed, including general and specific recommendations and its epidemiological and immunological basis.


The role of neuropeptides in the sacral autonomic reflex pathways of the cat.
Immunohistochemical and pharmacological studies were conducted to examine the origin and function of peptidergic nerves in the sacral autonomic system of the cat. Leucine-enkephalin (L-Enk) immunoreactivity was identified in nerve terminals in peripheral ganglia on the surface of the urinary bladder and in the parasympathetic nucleus in the sacral spinal cord. In colchicine-treated animals L-Enk was also detected in sacral preganglionic neurons (sPGN) identified by retrograde transport of a fluorescent dye. L-Enk terminals in bladder ganglia are believed to arise from sPGN since the terminals were eliminated by transection of the sacral ventral roots. Pharmacological studies indicated that exogenous as well as endogenously released enkephalins have an inhibitory action at both ganglionic and spinal sites in the sacral outflow to the urinary bladder. Peptides were also associated with afferents nerves in the sacral autonomic system. The distribution of substance P, VIP and cholecystokinin in the sacral dorsal horn paralleled the distribution of visceral afferent projections as demonstrated with HRP techniques. Dye labeling combined with immunohistochemistry revealed that some dorsal root ganglion cells projecting to the pelvic viscera contain substance P or VIP.


Mouse thymocyte apoptosis and cell loss in response to exercise and antioxidant administration.
Various physical and psychological stressors can cause thymocyte apoptosis and cell loss in rodents. Although glucocorticoids (GC) are commonly implicated, oxidative stress may also play a role. The purpose of this study was to examine the effect of an acute bout of strenuous treadmill running, and the antioxidant N-acetyl-L-cysteine (NAC) on thymocyte loss and apoptosis. Eighty-eight female C57BL/6 mice were given NAC (1 g/kg, i.p.) or saline (SAL) 30 min before 90 min of treadmill exercise at a 2 degrees slope (EX; 30 min at 22 m/min; 30 min at 25 m/min; and 30 min at 28 m/min) and sacrificed immediately (Imm) or 24 h following EX. Control mice (NonEX) were exposed to treadmill noise and vibration without running. Thymocytes were isolated and analyzed for phosphatidylserine (PS) externalization (Annexin V), loss of membrane integrity, mitochondria membrane depolarization, intracellular hydrogen peroxide (H(2)O(2)) production, and intracellular glutathione (GSH) as well as protein levels of caspase 3, Bcl-2, and cytosolic cytochrome c. Blood was analyzed for corticosterone (CORT) concentrations by radioimmunoassay. Exercise stress caused a significant increase in plasma CORT concentrations in EX + SAL + Imm and EX + NAC + Imm groups compared to NonEX mice. Relative to NonEX mice, thymocytes isolated from EX + SAL + Imm mice showed signs of an early apoptotic profile as indicated by decreased GSH stores and increased mitochondrial membrane depolarization. These effects were followed by a 50% reduction in thymocyte numbers 24 h post-exercise (EX + SAL + 24 h). Alterations in GSH levels, mitochondrial membrane depolarization, and thymocyte loss were not observed in mice receiving NAC. These results suggest that exercise-induced thymocyte apoptosis and cell loss may be mediated via an oxidative stress pathway.


Hypoplastic Left Heart Syndrome Across the Lifespan: Clinical Considerations for Care of the Fetus, Child, and Adult.
Hypoplastic left heart syndrome (HLHS) is the most common anatomic lesion in children born with single-ventricle physiology and is characterised by the presence of a dominant right ventricle and a hypoplastic left ventricle along with small left-sided heart structures. Diagnostic subgroups of HLHS reflect the extent of inflow and outflow obstruction at the aortic and mitral valves, specifically stenosis or atresia. If left unpalliated, HLHS is a uniformly fatal lesion in infancy. Following introduction of the Norwood operation, early survival has steadily improved over the past 4 decades, mirroring advances in operative and perioperative management as well as reflecting refinements in patient surveillance and interstage clinical care. Notably, survival after staged palliation has increased from 0% to a 5-year survival of 60%-65% for children in some centres. Despite the prevalence of HLHS in childhood with relatively favourable surgical outcomes in contemporary series, this cohort is only now reaching early adult life and longer-term outcomes have yet to be elucidated. In this article we focus on contemporary clinical management strategies for patients with HLHS across the lifespan, from fetal to adult life. Nomenclature and diagnostic considerations are discussed and current literature pertaining to putative genetic etiologies is reviewed. The spectrum of fetal and paediatric interventional strategies, both percutaneous and surgical, is described. Clinical, patient-reported, and neurodevelopmental outcomes of HLHS are delineated. Finally, note is made of current areas of clinical uncertainty, and suggested directions for future research are highlighted.


Neurologic deterioration in patients with acute ischemic stroke or transient ischemic attack.
To improve epidemiologic knowledge of neurologic deterioration (ND) in patients with acute ischemic stroke (AIS). In this prospective observational study, we captured ND prospectively in 29,446 patients with AIS admitted to 15 hospitals in Korea within 7 days of stroke onset. ND was defined as an increase in NIH Stroke Scale (NIHSS) score ≥2 (total), or ≥1 (motor or consciousness), or any new neurologic symptoms. Change in incidence rate after stroke onset, causes, factors associated with ND, modified Rankin Scale (mRS) score at 3 months and 1 year, and a composite of stroke, myocardial infarction, and all-cause death at 1 year were assessed. ND occurred in 4,299 (14.6%) patients. The highest rate, 6.95 per 1,000 person-hours incidence, was within the first 6 hours, which decreased to 2.09 within 24-48 hours, and 0.66 within 72-96 hours after stroke onset. Old age, female sex, diabetes, early arrival, large artery atherosclerosis as a stroke subtype, high NIHSS scores, glucose level, systolic blood pressure, leukocytosis at admission, recanalization therapy, TIA without a relevant lesion, and steno-occlusion of relevant arteries were associated with ND. The causes were stroke progression (71.8%) followed by recurrence (8.5%). Adjusted relative risks (95% CI) for poor outcome (mRS 3-6) at 3 months and 1 year were 1.75 (1.70-1.80) and 1.70 (1.65-1.75), respectively. The adjusted hazard ratio (95% CI) for the composite event was 1.59 (1.45-1.74). ND should be taken into consideration as a factor that may influence the outcome in acute ischemic stroke.


[Leptospira infections in cats].
Leptospirosis is a widespread zoonosis found in over 150 mammalian species. Subclinically infected rodents and farm animals serve as reservoir hosts. Cats are exposed to leptospires regularly, particularly through hunting infected rodents. However, clinical leptospirosis is rare in cats. Nevertheless, infected cats can shed the pathogen through urine and thus, cats could play an important role in the epidemiology of leptospires.


Medical dataset classification for Kurdish short text over social media.
The Facebook application is used as a resource for collecting the comments of this dataset, The dataset consists of 6756 comments to create a Medical Kurdish Dataset (MKD). The samples are comments of users, which are gathered from different posts of pages (Medical, News, Economy, Education, and Sport). Six steps as a preprocessing technique are performed on the raw dataset to clean and remove noise in the comments by replacing characters. The comments (short text) are labeled for positive class (medical comment) and negative class (non-medical comment) as text classification. The percentage ratio of the negative class is 55% while the positive class is 45%.


[Value of the discover mental defects for the pathogenesis of psychosocial disorders by normal pupils--a study about patients of a neuropsychiatric station for children (author's transl)].
200 normal pupils were tested, to discover mental defects by HAWIK IQ less than or equal to 84. All tested pupils were treated stationary for psychosocial disorders. 37 mental defects were demonstrable. The mental defect gets the third rank in the quantity of pathological causes for psychosocial disorders. The atmosphere reactive disorders head the list followed by partial psychological disorders of performance. The early recognition of these defects is of necessity in medical, psychological and pedagogic view.


Contact allergy due to colophony (VII). Sensitizing studies with oxidation products of abietic and related acids.
9 oxidation products of abietic acid, dehydroabietic acid, and levopimaric acid were prepared synthetically to determine their sensitizing potential in guinea pigs. It was found that compounds with epoxy and peroxo groups in rings A and B had a notable sensitizing potential. The same result was found with 7-oxode-hydroabietic acid identified earlier in rosin (3) and a polar fraction obtained from commercial abietic acid, suggesting the presence of still unidentified oxidation products. Hydroxylation of rings A or B, or conversion to the methyl esters, considerably decreases the sensitizing potential. A model is presented underlining the importance of hydrophobic and polar domains, in addition to chemically reactive groupings, in the allergen. Insertion into the lipid bilayer may play an important rôle in contact sensitivity.


Effects on serotonin of (-)nicotine and dimethylphenylpiperazinium in the dorsal raphe and nucleus accumbens of freely behaving rats.
The aim of this study was to investigate the neurochemical mechanism underlying the effect of nicotine and dimethylphenylpiperazinium (DMPP) on 5-hydroxytryptamine (5-HT) release in the dorsal raphe nucleus and nucleus accumbens of freely behaving rats. For comparison, lobeline, cytisine and RJR-2403 were also investigated. It was found that all drugs, when infused locally, evoked an increase of 5-HT in the dorsal raphe nucleus. However, the magnitudes of the 5-HT increase were comparatively different between the drugs in the ranking of their potency: DMPP>RJR 2403>>nicotine>lobeline>cytisine. Both methyllycaconitine, a nicotinic acetylcholine receptor (nAChR) antagonist and methyllycaconitine, a selective alpha7-containing nAChR antagonist blocked the effects of nicotine and DMPP, suggesting that alpha7 subunit mediated the increases in 5-HT. However, DMPP was reported to increase 5-HT using non-nAChR mechanism [Lendvai B, Sershen H, Lajtha A, Santha E, Baranyi M, Vizi ES (1996) Differential mechanisms involved in the effect of nicotinic agonists DMPP and lobeline to release [3H]5-HT from rat hippocampal slices. Neuropharmacology 35:1769-1777]. To test if 5-HT carriers were involved, a selective 5-HT reuptake inhibitor citalopram (1 microM) was infused into the dorsal raphe nucleus before administration of nicotine or DMPP. As a result, citalopram significantly blocked the effect of DMPP, whereas it had no influence on nicotine. Finally, the 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) was used to test whether the increases in 5-HT were depolarization-dependent. Administration of 8-OH-DPAT (0.1 mg/kg, s.c.) produced significant decreases in 5-HT in the animals treated with nicotine. In contrast, the effect of DMPP was not altered by 8-OH-DPAT, suggesting that the increases in 5-HT were independent of cell membrane depolarization. In conclusion, there are different mechanisms involved in nicotine- and DMPP-evoked increases in 5-HT. This is consistent with prior work suggesting DMPP may primarily act on 5-HT carriers.


An autopsy case of hemilaterally dominant and systematic/extensive border zone infarction: sequela of preceding atherosclerotic obstruction of one common carotid artery followed by repeated episodes of systemic hypotension.
A 68-year-old man was admitted to St Marianna University Hospital on account of loss of consciousness with left hemiplegia. During the hospital recovery course with a rehabilitation procedure, the patient's blood pressure was very unstable, fluctuating between high (210/110 mmHg) and low (110/70 mmHg) values accompanied by a fainting sensation. A second stroke of left hemiplegia took place 1 month later. Afterwards, his condition worsened to tetraplegia with dysarthria. Three months later, lung cancer with multiple metastasis including his left neck was found and he died from adynamic ileus 6 months after the onset of the present illness. Autopsy revealed nearly complete atheromatous obstruction and more than 50% stenosis, respectively, of his right common and internal/external carotid arteries. His intracranial arterial trunks and main branches were all patent with localized atherosclerosis of only moderate degree. The pathology of the brain existed predominantly in the right hemisphere in the border zone area between the anterior and middle cerebral arteries systematically with numerous disseminated foci of complete or incomplete necrosis, white matter and gray matter being involved independently. Involvement of centrum semiovale white matter is more extensive and intensive than that of gray matter. Of the gray matter, cerebral cortex as well as striatum, periventricular (the third ventricle) gray and cerebellar cortex was involved. The specific characteristic topography and distribution of the lesions together with their histopathology are described in detail with illustration. It is concluded that this case represents an outstanding example of hemodynamic cerebral circulatory insufficiency doubly caused by hemilateral carotid artery stenosis and repeated episodes of systemic hypotension.


How do policy levers shape the quality of a national health system?
Poor quality of care may have a detrimental effect on access and take-up and can become a serious barrier to the universality of health services. This consideration is of particular interest in view of the fact that health systems in many countries must address a growing public-sector deficit and respond to increasing pressures due to COVID-19 and aging population, among other factors. In line with a rapidly emerging literature, we focus on patient satisfaction as a proxy for quality of health care. Drawing on rich longitudinal and cross-sectional data for Spain and multilevel estimation techniques, we show that in addition to individual level differences, policy levers (such as public health spending and the patient-doctor ratio, in particular) exert a considerable influence on the quality of a health care system. Our results suggest that policymakers seeking to enhance the quality of care should be cautious when compromising the level of health resources, and in particular, health personnel, as a response to economic downturns in a sector that traditionally had insufficient human resources in many countries, which have become even more evident in the light of the current health crisis. Additionally, we provide evidence that the increasing reliance on the private health sector may be indicative of inefficiencies in the public system and/or the existence of features of private insurance which are deemed important by patients.


Modelling the effects of water-point closure and fencing removal: a GIS approach.
Artificial water-points in the form of troughs or ground tanks are used to augment natural water supplies within rangelands in many parts of the world. Access to such water-points leads to the development of a distinct ecological sub-system, the piosphere, where trampling and grazing impact modify the vegetation. This study aims to consolidate existing information in a GIS based model to investigate grazing patterns within the landscape. The model focuses on the closure of water-points and removal of fences on Nanya Station, New South Wales, Australia. We found that the manipulation of water-points and fences in one management intervention may change grazing activity in a way different to that which would be experienced if each had been modified separately. Such effects are further modified by the spatial distribution of the water-points and the underlying vegetation.


Increased concentration of the complement split product C5a in acute pyelonephritis during pregnancy.
Pregnant women with acute pyelonephritis develop acute respiratory distress syndrome (ARDS) more frequently than non-pregnant women. The reasons for this remain unknown. The complement system is a complex set of self-assembling proteins that have been implicated in the pathophysiology of ARDS and sepsis. The purpose of this study was to determine if activation of the complement system occurs in pregnant women with acute pyelonephritis. A cross-sectional study was conducted to determine the plasma concentrations of C3a, C4a and C5a (i.e., complement split products) in pregnant patients with acute pyelonephritis (n=38) and normal pregnant women (n=38). The complement split products C3a, C4a and C5a were measured using ELISA. Data were analyzed using non-parametric statistics. 1) The median plasma concentration of C5a in pregnant patients with acute pyelonephritis was significantly higher than that in normal pregnant women (p<0.001); 2) there was no statistical difference in the median plasma concentration of C3a and C4a between the two groups (p>0.05); and 3) concentrations of C3a, C4a and C5a were not different among patients with acute pyelonephritis with and without bacteremia. 1) Pyelonephritis in pregnant women is associated with an increased plasma concentration of C5a, but not C3a and C4a; and 2) an excess of C5a can predispose pregnant women to develop ARDS and multi-organ failure in pyelonephritis. This finding may have clinical implications since blocking C5a improves ARDS in experimental sepsis.


Salt-induced absorbance changes of P-515 in broken chloroplasts.
Absorbance changes, caused by adding KCl to a suspension of broken chloroplasts in the presence of a low concentration of MgCl2, have been measured in the wavelength region 460-540 nm. The magnitude of the KCl-induced absorbance changes is shown to be proportional to the logarithm of the KCL concentration gradient initially induced across the thylakoid membrane. The difference spectrum of these absorbance changes is shown to be identical with the spectrum of the light-induced absorbance changes, which has been attributed to an electrochromic shift of p-515. This is interpreted as evidence that under these conditions salt-induced absorbance changes of P-515 occur in response to a membrane diffusion potential. The results indicate that the electrogenic potential across the thylakoid membrane, generated by a single turnover light flash, is in the range between 15 and 35 mV.


The Oxa1 protein forms a homooligomeric complex and is an essential part of the mitochondrial export translocase in Neurospora crassa.
The Oxa1 protein is a ubiquitous constituent of the inner membrane of mitochondria. Oxa1 was identified in yeast as a crucial component of the protein export machinery known as the OXA translocase, which facilitates the integration of proteins from the mitochondrial matrix into the inner membrane. We have identified the Neurospora crassa Oxa1 protein which shows a sequence identity of 22% to the yeast homologue. Despite the low level of identity, the function of the homologues is conserved as the N. crassa gene fully complemented a yeast null mutant. Genetic analysis revealed that Oxa1 is essential for viability in N. crassa. Cells propagated under conditions that severely reduce Oxa1 levels grew extremely slowly and were deficient in subunits of complex I and complex IV. Isolation of the Oxa1 complex from N. crassa mitochondria revealed a 170-180-kDa complex that contained exclusively Oxa1. Since the Oxa1 monomer has a molecular weight of 43,000, our data suggest that the OXA translocase consists of a homooligomer most likely containing four Oxa1 subunits.


Denosumab did not improve computerized tomography erosion scores when added to intensive urate-lowering therapy in gout: Results from a pilot randomized controlled trial.
Disordered osteoclast activity has been implicated in the pathogenesis of gouty bone erosion. We sought to determine if the addition of denosumab (a monoclonal antibody targeting the receptor activator of nuclear factor kappa-B ligand - RANKL) to intensive urate-lowering therapy (ULT) improves gouty bone erosion. Open-label, parallel-group pilot randomized controlled trial in which 20 participants with gout with at least one confirmed conventional radiographic foot bone erosion were assigned in a 1:1 allocation to receive denosumab (60 mg subcutaneous every 6 months) added to intensive ULT (serum urate ≤5 mg/dL or 300 µmol/L at the time of randomization and continued for the duration of the study), or intensive ULT alone. The primary outcome was the change in the bilateral foot and ankle computed tomography (CT) bone erosion score from baseline to 12 months, assessed by an experienced musculoskeletal radiologist blinded to study assignment. Secondary outcomes included change in serum C-terminal telopeptide (CTX), and patient reported outcomes of pain and function. Although serum CTX declined markedly in the denosumab/ULT group compared with the ULT alone group, there was no interval change in CT erosion score in either the denosumab/ULT or ULT alone group after one year of follow-up. Other secondary outcomes did not differ between groups. There were two severe adverse events: One patient developed atrial fibrillation (on denosumab/ULT) and another atrial flutter (on ULT alone). In this pilot study, denosumab did not offer additional benefit to intensive urate lowering therapy for gouty bone erosion.


[The Bridgelok system for additional retention of bonded prostheses].
The partial protesis with metal carving and cemented with composite resin is used frequently as a reversible method in order to substitute teeth, due to the light preparation necessary for the original teeth. Recently, a new system (Bridgelok) with pins has been proposed as safer for this kind of protesis. This paper studies if this new system originates a higher resistance compared to other system without pins.


Role of eukaryotic initiation factor 5 in the formation of 80 S initiation complexes.
The function of eukaryotic initiation factor 5 (eIF-5) from rabbit reticulocyte lysate has been studied by sucrose gradient preparation of 40 S and 80 S initiation complexes. eIF-5 is required for transfer of initiator tRNA from 40 S preinitiation complexes to puromycin-reactive 80 S complexes. The transfer is dependent upon GTP hydrolysis and is associated with release of eIF-2 and eIF-3 from the 40 S subunit. The GTP-dependent loss of eIF-2 and eIF-3 is catalyzed by eIF-5 in the absence of 60 S subunits or when subunit joining is prevented by edeine, but not when GTP is replaced by GuoPP(NH)P. Unstable 40 S subunit . Met-tRNAf complexes generated by eIF-5 can form puromycin-reactive 80 S complexes when 60 S subunits are added in the absence of added GTP. In addition, kinetic evidence is presented that indicates GTP hydrolysis occurs prior to 80 S complex formation.


Teriparatide Improves Trabecular Osteoporosis but Simultaneously Promotes Ankylosis of the Spine in the Twy Mouse Model for Diffuse Idiopathic Skeletal Hyperostosis.
Diffuse idiopathic skeletal hyperostosis (DISH) is a common skeletal disorder in the elderly, which can develop into periosteal hyperostosis and paradoxically into immobilization-associated trabecular osteoporosis. The bone anabolic agent, teriparatide (TPD), seems to be a rational treatment for the immobilization-associated osteoporosis. However, it can lead to development of hyperostosis lesions in DISH patients. Here, we demonstrate TPD effectively treats trabecular osteoporosis while simultaneously promoting ankylosis of the spine in DISH model tiptoe-walking Yoshimura (twy) mice, compared with the ICR mice. Eighteen male twy mice were divided into three groups, and ICR mice were used as a normal control. Subcutaneous injections of TPD or phosphate-buffered saline (PBS) were performed according to three dosing regimens; 40 µg/kg once daily (TPD × 1 group), 40 µg/kg three times daily (TPD × 3 group), and PBS (control; Ctl group). Treatment was commenced at the age of 7 weeks and continued for 5 weeks. Micro-computed tomography (µCT) and histological analysis were performed. Longitudinal µCT study revealed that trabecular bone volume in both the vertebral body and distal femur decreased with time in the Ctl group, but increased dramatically in the TPD × 3 group. The twy mice developed ankylosis of the spine, the progression of which was accelerated with TPD therapy. We also confirmed that TPD therapy promoted ossification of spinal ligaments. Histomorphometrical study revealed that TPD treatment increased bone formation at the vertebrae enthesis region and in the trabecular bone. TPD therapy effectively treats trabecular osteoporosis, but potentially promotes ankylosis of the spine in patients with DISH.


Case report: The management of advanced oral cancer in a Jehovah's Witness using the Ultracision Harmonic Scalpel.
We present the first case of a head and neck oncological procedure accomplished in a Jehovah's Witness using the Ultracision Harmonic Scalpel (Ethicon, Cincinnati, OH). Jehovah's Witnesses present a serious challenge to the head and neck cancer surgeon due to their refusal to accept transfusion of any blood products. However, our experience reinforces the view that surgical management of head and neck cancer is possible in these patients. We show the Harmonic Scalpel, an ultrasonic tissue dissector, to be a useful surgical tool in obviating the need for blood transfusion. Preoperative optimisation, intra-operative surgical and anaesthetic techniques are also fully discussed.


Ionic permeation and conduction properties of neuronal KCNQ2/KCNQ3 potassium channels.
Heteromeric KCNQ2/3 potassium channels are thought to underlie the M-current, a subthreshold potassium current involved in the regulation of neuronal excitability. KCNQ channel subunits are structurally unique, but it is unknown whether these structural differences result in unique conduction properties. Heterologously expressed KCNQ2/3 channels showed a permeation sequence of while showing a conduction sequence of A differential contribution of component subunits to the properties of heteromeric KCNQ2/3 channels was demonstrated by studying homomeric KCNQ2 and KCNQ3 channels, which displayed contrasting ionic selectivities. KCNQ2/3 channels did not exhibit an anomalous mole-fraction effect in mixtures of K(+) and Rb(+). However, extreme voltage-dependence of block by external Cs(+) was indicative of multi-ion pore behavior. Block of KCNQ2/3 channels by external Ba(2+) ions was voltage-independent, demonstrating unusual ionic occupation of the outer pore. Selectivity properties and block of KCNQ2 were altered by mutation of outer pore residues in a manner consistent with the presence of multiple ion-binding sites. KCNQ2/3 channel deactivation kinetics were slowed exclusively by Rb(+), whereas activation of KCNQ2/3 channels was altered by a variety of external permeant ions. These data indicate that KCNQ2/3 channels are multi-ion pores which exhibit distinctive mechanisms of ion conduction and gating.


Calculating cancer latency using data from a nested case-control study of prostatic cancer.
Assessing latency for neoplastic diseases is crucial for determining the causal effects of a complex mix of carcinogenic exposures. An initial assessment of cancer risks in a U.S. tire and rubber plant revealed a significant SMR of 140 for prostatic cancer. Using an industry-based, case-control death certificate study of prostatic malignancies, we found matched odds ratios of about 3 (p less than 0.025) for Batch Preparation, the work area with the greatest exposure to carbon black, solvents, and heavy metal oxides. To assess latency, we used the matched case-control series to calculate annual estimates of the odds ratio by determining the proportion of cases and controls employed for greater than 1 month in Batch Preparation during each year under study. This approach produced a plot with great fluctuations. To reduce variability in the resulting curve, a method was developed that measured the "etiologic fraction", which is its highest point represents an estimate of the peak of the latency distribution. For Batch Preparation the modal point was 29 years before death with the greatest risk occurring from employment in the mid-1940's. The latency method allows risk assessment for time and year of greatest exposure difference, thus suggesting appropriate prevention strategies. Applications of this method for other types of studies and exposures are discussed.


A framework for the description of charge order, pseudo and superconducting gap, critical temperature and pairing interaction in cuprate superconductors.
A unified phenomenological description framework is proposed for the evaluation of some of the most important observables of the cuprate superconductors: the pseudogap (PG) ΔPG, the local superconducting amplitudes ΔSC(r i ), the critical temperature T c and charge ordering (CO) parameters. Recent detailed measurements of CO structures and CO wavelengths λ CO are faithfully reproduced by solutions of a Cahn-Hilliard differential equation with a free energy potential V GL that produces alternating small charge modulations. The charge oscillations induce atomic fluctuations that mediate the SC pair interaction proportional to the V GL amplitude. The local SC amplitude and phase θ i are connected by Josephson coupling E J(r ij ) and the SC long-range order transition occurs when [Formula: see text]. The calculated results of the wavelength λ CO, ΔPG, [Formula: see text] and T c calculations are in good agreement with a variety of experiments.


A Critical Role for the Paraventricular Nucleus of the Hypothalamus in the Regulation of the Volume Reflex in Normal and Various Cardiovascular Disease States.
This review focuses on studies implicating forebrain neural pathways and neuromodulator systems, particularly, the nitric oxide system within the paraventricular nucleus of the hypothalamus in regulating neurohumoral drive, autonomic pathways, and fluid balance. Accumulating evidence from animals with experimental models of hypertension and heart failure as well as humans with hypertension suggests that alterations in central neural pathways, particularly, within the PVN neuromodulated by neuronal nitric oxide, are involved in regulating sympathetic outflow particularly to the kidney resulting in alterations in fluid balance commonly observed in hypertension and heart failure states. The characteristics of the hypertensive and heart failure states include alterations in neuronal nitric oxide within the PVN to cause an increase in renal sympathetic nerve activity to result in sodium and fluid retention in these diseases. A comprehensive understanding of these mechanisms will enhance our ability to treat hypertensive and heart failure conditions and their cardiovascular complications more efficiently.


Investigating Social Media to Evaluate Emergency Medicine Physicians' Emotional Well-being During COVID-19.
Emergency medicine (EM) physicians experience tremendous emotional health strain, which has been exacerbated during COVID-19, and many have taken to social media to express themselves. To analyze social media content from academic EM physicians and resident physicians to investigate changes in content and language as indicators of their emotional well-being. This cross-sectional study used machine learning and natural language processing of Twitter posts from self-described academic EM physicians and resident physicians between March 2018 and March 2022. Participants included academic EM physicians and resident physicians with publicly accessible posts (at least 300 total words across the posts) from the US counties with the top 10 COVID-19 case burdens. Data analysis was performed from June to September 2022. Being an EM physician or resident physician who posted on Twitter. Social media content themes during the prepandemic period, during the pandemic, and across the phases of the pandemic were analyzed. Psychological constructs evaluated included anxiety, anger, depression, and loneliness. Positive and negative language sentiment within posts was measured. This study identified 471 physicians with a total of 198 867 posts (mean [SD], 11 403 [18 998] words across posts; median [IQR], 3445 [1100-11 591] words across posts). The top 5 prepandemic themes included free open-access medical education (Cohen d, 0.44; 95% CI, 0.38-0.50), residency education (Cohen d, 0.43; 95% CI, 0.37-0.49), gun violence (Cohen d, 0.37; 95% CI, 0.32-0.44), quality improvement in health care (Cohen d, 0.33; 95% CI, 0.27-0.39), and professional resident associations (Cohen d, 0.33; 95% CI, 0.27-0.39). During the pandemic, themes were significantly related to healthy behaviors during COVID-19 (Cohen d, 0.83; 95% CI, 0.77-0.90), pandemic response (Cohen d, 0.71; 95% CI, 0.65-0.77), vaccines and vaccination (Cohen d, 0.60; 95% CI, 0.53-0.66), unstable housing and homelessness (Cohen d, 0.40; 95% CI, 0.34-0.47), and emotional support for others (Cohen d, 0.40; 95% CI, 0.34-0.46). Across the phases of the pandemic, thematic content within social media posts changed significantly. Compared with the prepandemic period, there was significantly less positive, and concordantly more negative, language used during COVID-19. Estimates of loneliness, anxiety, anger, and depression also increased significantly during COVID-19. In this cross-sectional study, key thematic shifts and increases in language related to anxiety, anger, depression, and loneliness were identified in the content posted on social media by academic EM physicians and resident physicians during the pandemic. Social media may provide a real-time and evolving landscape to evaluate thematic content and linguistics related to emotions and sentiment for health care workers.


The use of pedometers in stroke survivors: are they feasible and how well do they detect steps?
To determine (1) the feasibility of pedometers for stroke patients and (2) the level of agreement between pedometers and actual step count. Observational agreement study. Six stroke units. Independently mobile stroke patients (N=50) ready for hospital discharge. Patients were asked to apply 3 pedometers: 1 around the neck and 1 above each hip. Patients performed a short walk lasting 20 seconds, then a 6-minute walk test 6MWT. Video recordings determined the criterion standard step count. Agreement between the step count recorded by pedometers and the step count recorded by viewing the criterion standard video recordings of the 2 walks. Five patients (10%) needed assistance to put on the pedometers, and 5 (10%) could not read the step count. Thirty-nine (78%) would use pedometers again. Below a gait speed of about 0.5 m/s, pedometers did not generally detect steps. Agreement analyses showed that even above 0.5 m/s, pedometers undercounted steps for both the short walk and 6MWT; for example, the mean difference between the video recorder and pedometer around the neck was 5.93 steps during the short walk and 32.4 steps during the 6MWT. Pedometers are feasible but generally do not detect steps at gait speeds below about 0.5 m/s, and they undercount steps at gait speeds above 0.5 m/s.


Postoperative neuroleptic malignant syndrome. A case report.
A case of postoperative neuroleptic malignant syndrome is presented. A healthy 23-year-old male underwent a shoulder repair under uneventful fentanyl, halothane, nitrous oxide and oxygen anaesthesia. He received droperidol 5 mg IV and metoclopramide 10 mg IV intraoperatively to prevent postoperative nausea. Postoperatively, the patient developed autonomic instability, fever and generalized muscle rigidity. His level of consciousness was depressed. These findings were consistent with the diagnosis of neuroleptic malignant syndrome. The supportive treatment of the patient included active cooling measures, muscle relaxation and mechanical ventilation. The ability of anti-dopaminergic agents, including metoclopramide and droperidol, to precipitate the neuroleptic malignant syndrome is discussed. Treatment of the neuroleptic malignant syndrome is briefly discussed.


Effects of increasing calcium propionate in a finishing diet on dry matter intake and glucose metabolism in steers.
The objective of this study was to determine whether increasing propionate alters dry matter intake (DMI), glucose clearance rate, blood metabolites, insulin concentrations, and hepatic gene expression in steers fed a finishing diet. Holstein steers (n = 15; BW = 243 ± 3.6 kg) were individually fed a finishing diet ad libitum. Steers were allocated by body weight (BW) to receive: no Ca propionate (Control), 100 g/d Ca propionate (Low), or 300 g/d Ca propionate (High) in the diet. Orts were collected and weighed daily to determine DMI. Blood samples were collected on days 0, 7, and 21, and BW recorded on days 0, 14, and 28. An intravenous glucose tolerance test (IVGTT) was conducted on days 14 and 28 of the trial. Liver biopsies were collected on day 33 for gene expression analysis. Blood samples were analyzed for whole blood glucose and lactate, plasma non-esterified fatty acids (NEFAs), and insulin concentrations. Data were analyzed using a mixed model with treatment, day and their interaction included, with day and minute as a repeated measure. The control treatment had greater (P < 0.01) DMI than low and high steers. Body weight was increased in control steers on days 14 and 28 compared with the steers receiving the High treatment (P = 0.03 for the interaction). Blood glucose concentrations tended (P = 0.09) to be higher on day 21 than days 0 and 7 but was not affected by treatment (P = 0.58). Plasma NEFA concentrations were lower (P = 0.05) for control steers than other treatments, and greater (P = 0.002) on day 0 than days 7 and 21. Blood lactate concentrations were greater (P = 0.05) on day 7, than days 0 and 21, but not affected by treatment (P = 0.13). High steers had greater plasma insulin concentrations in response to the IVGTT than steers on the other treatments (P = 0.001). There was no treatment (P ≥ 0.16) or day effect (P ≥ 0.36) on glucose peak, plateau, or clearance rate. High steers had greater expression of solute carrier family 16 member 1 (SLC16A1; P = 0.05) and tended to have greater hepatic expression of solute carrier family 2 member 2 (SLC2A2; P = 0.07). These data indicate that increased propionate may decrease DMI and insulin sensitivity.


(Butane-1,2,3,4-tetraol-κ(3) O (1),O (2),O (3))(ethanol-κO)tris-(nitrato-κ(2) O,O')erbium(III).
In the title Er(III)-erythritol complex, [Er(NO3)3(C2H5OH)(C4H10O4)], the Er(III) cation is chelated by one erythritol mol-ecule, three nitrate anions and an ethanol mol-ecule, completing an irregular ErO10 coordination geometry. The Er-O bond lengths are in the range 2.348 (3)-2.583 (3) Å. In the crystal, extensive O-H⋯O hydrogen bonding links the mol-ecules into a three-dimensional supra-molecular structure.


Identification and characterization of the murine TRPM4 channel.
The transient receptor potential (TRP) channels form a superfamily with six transmembrane structures, which is common in other types of voltage-dependent channels. The TRP-melastatin (TRPM) subfamily includes the putative tumor-suppressor melastatin, which was originally found as a down-regulated protein in melanoma tumor cell lines. Here, we report a novel TRP-related protein that is a murine orthologue of human TRPM4. The function of the novel murine TRPM4 was studied in HEK-293 cells using a fluorescent calcium indicator, fura-2. The removal and re-introduction of extracellular calcium triggered changes in the intracellular calcium only in cells expressing TRPM4a, which suggests that this novel channel plays a role in the calcium entry process. We also isolated a splice variant of TRPM4 that was proven to be non-functional. Both TRPM4 variants integrated into the plasma membrane. Furthermore, FRET analysis revealed that TRPM4a and TRPM4b localized close together, suggesting a multimerization of the two molecules.


Cytomegalovirus as an immunomodulator across the lifespan.
Human cytomegalovirus (HCMV) is a nearly ubiquitous β-herpesvirus that establishes latent infection in the majority of the world's population. HCMV infection profoundly influences the host immune system and, perhaps more than any other human pathogen, has been shown to create a lasting imprint on human T and NK cell compartments. HCMV-seropositivity has been associated with both beneficial effects, such as increased vaccine responsiveness or heterologous protection against infections, and deleterious effects, such as pathological neurodevelopmental sequelae from congenital infection in utero and cumulative damage from chronic lifelong latency into old age. The significance of many of these associations is unclear, as studies into the causal mechanisms linking HCMV and these disease outcomes are lacking; however, HCMV-mediated changes to the immune system may play a key role. This review examines how HCMV impacts the host immune system in an age-dependent manner with important implications for human immunophenotypes and long-term disease risk.


Cardiovascular reactivity and smoking in coronary prone (type A) and non-coronary prone (type B) subjects.
Twenty-nine Type A subjects, (greater than 75 percentile on the Jenkins Activity Survey) and 28 Type B's (less than 25 percentile of Jenkins) were asked to smoke 1 cigarette (1.0 mg nicotine, 16 mg tar, 14 mg CO) using a standardized smoking procedure. The results showed that both Type A and B subjects experienced significant smoking induced changes in heart rate, skin temperature and pulse transit time. An analysis of covariance also revealed that the Type A subjects exhibited greater heart rate reactivity due to smoking than their type B counter parts. The results are discussed in terms of the possible role of cardiovascular reactivity in the mediation of the subjective and behavioral effects of smoking in the type A individual.


Utilization of elongation factor Tu gene (tuf) sequencing and species-specific PCR (SS-PCR) for the molecular identification of Acetobacter species complex.
The aim of this study was to use tuf gene as a molecular target for species discrimination in the Acetobacter genus, as well as to develop species-specific PCR method for direct species identification of Acetobacter aceti. The results showed that most Acetobacter species could be clearly distinguished, and the average sequence similarity for the tuf gene (89.5%) among type strains was significantly lower than that of the 16S rRNA gene sequence (98.0%). A pair of species-specific primers were designed and used to specifically identify A. aceti, but none of the other Acetobacter strains. Our data indicate that the phylogenetic relationships of most strains in the Acetobacter genus can be resolved using tuf gene sequencing, and the novel species-specific primer pair could be used to rapidly and accurately identify the species of A. aceti by the PCR based assay.


[The effect of local corticosteroid therapy on ophthalmic tonus and ocular hydro- and hemodynamics in experimental inflammatory pathology].
The laboratory samples of 0.1% dexamethasone and 1% hydrocortisone eye drops which had been designed at the State Research Center of Drugs were investigated for their effects on intraocular pressure in the presence of photokeratitis caused by UV irradiation. Producing pronounced antiinflammatory effects, the daily triple instillations of corticosteroids for 9 days tended to gradually increase the ophthalmotone induced both by the imbalance of production and outflow of intraocular fluid and by ocular hemodynamic disorders associated with the enhanced blood filling of the choroidal vessels and with the higher intensity of pulse oscillations.


Measurement of ketamine and xylazine in rat brain by liquid-liquid extraction and gas chromatography-mass spectrometry.
In human and veterinary medicine, the injectable drugs ketamine and xylazine are mainly used in combination to induce, and then maintain general anaesthesia; they also provide pain and stress relief. Some side-effects have been reported on the auditory brainstem response, a method is therefore required to determine their concentrations in the brain. This paper presents a method to determine nanogramme quantities of ketamine and xylazine in rat brain using liquid-liquid extraction and gas chromatography-mass spectrometry in selective ion monitoring mode. The technique requires only 0.5 g of sample, and uses xylazine d6 as an internal standard. The method was linear between 0.86 and 34.4 μg/g of brain. Limits of quantification were 378 and 87 ng (approximately 0.76 and 0.17 μg/g of brain) for ketamine and xylazine, respectively. The reliability of the method in terms of accuracy, within-day and between-day precision was also demonstrated. For xylazine, bias and intra-day precision were good (<3.0%), as was between-day precision (<10.5%); the equivalent values for ketamine were 7%, 11.1% and 20.9%, respectively. Stability of the analytes in the matrix at -80 °C was assessed over five months; both compounds were found to be stable for at least 1 month, even at very low concentrations. The procedure was successfully applied to determine (for the first time) the in vivo brain levels of both drugs in animals following systemic administration. The procedure will be useful in future studies of the side-effects of these drugs, and their interactions with other compounds.


Targeting Cryptosporidium parvum capture.
Polymer microarrays offer a high-throughput approach to the screening and assessment of a large number of polymeric materials. Here, we report the first study of protozoan-polymer interactions using a microarray approach. Specifically, from screening hundreds of synthetic polymers, we identified materials that either trap the waterborne protozoan parasite, Cryptosporidium parvum, or prevent its adhesion, both of which have major practical applications. Comparison of array results revealed differences in the adhesion characteristics of viable and non-viable C. parvum oocysts. Material properties, including polymer composition, wettability and surface chemistry, allowed correlation of binding and identification of structure function relationships. Understanding C. parvum binding interactions could assist in improved water treatment processes and the identified polymers could find applications in sensor and filter materials.


Comparative study of the effectiveness of oral fluconazole and intravaginal clotrimazole in the treatment of vaginal candidiasis.
A study was carried out to compare 3 treatment regimens for vaginal candidiasis. A total of 150 women with clinical and mycological evidence of vaginal candidiasis were randomized to receive 50 mg of oral fluconazole daily for 6 days (50 women), a single oral 150 mg dose of fluconazole (50 women), or 100 mg of intravaginal clotrimazole daily for 6 days (50 women). They were assessed at 5-15 days (short-term assessment) and again at 30-60 days (long-term assessment) after the completion of treatment. Candida species were completely eradicated from the vagina in 88% or 80% in the 6-day oral fluconzaole group, 76% or 70% in the single oral fluconazole group, and 72% or 60% in the intravaginal clotrimazole group at short-term or long-term assessment, respectively. The rates of clinical effectiveness were 92% or 88% in the 6-day oral fluconzaole group, 80% or 76% in the single oral fluconazole group, and 72% or 58% in the intravaginal clotrimazole group at the short-term or long-term assessment, respectively. Treatment-related side effects were not found in any group. This study suggests that the treatment of vaginal candidiasis with oral fluconazole is effective and that a single oral fluconazole dose might be one choice in the treatment of vaginal candidiasis.


Diagnostic value of serum procalcitonin and C-reactive protein for infections after allogeneic hematopoietic stem cell transplantation versus nontransplant setting.
Procalcitonin (PCT) has been increasingly used as a biomarker of infection. The purpose of this study was to evaluate its diagnostic value after hematopoietic stem cell transplantation (HSCT), where non-infectious febrile complications such as graft-versus-host disease frequently develop. We retrospectively analyzed 144 febrile episodes (infections: 82, and noninfections: 62) in adult patients with hematological disorders, including 57 and 87 episodes in HSCT and non-HSCT patients, respectively. Of 57 febrile episodes in HSCT patients, 46 (86%) and 25 (44%) revealed positivity for C-reactive protein (CRP) and PCT, respectively. Among 87 febrile episodes in non-HSCT patients, 81 (93%) and 22 (25%) events showed positive results of CRP and PCT. Both of these biomarkers were associated with infectious episodes in univariate analysis. Multivariate analysis showed that a high cut-off level (>9.5 mg/dL) of CRP was a better indicator for infections than PCT in HSCT patients, while PCT positivity was more diagnostic for infections than any cutoff CRP level in non-HSCT patients. It may be necessary to interpret the results of these biomarkers with different orders of priority in transplant versus nontransplant patients.


Allocation and stoichiometric regulation of phosphorus in a freshwater zooplankton under limited conditions: Implication for nutrient cycling.
Because zooplankton is potentially limited by phosphorus (P) in freshwater, they may modify their body P distributions in different biochemical and anatomic components depending on the environmental P levels. In the present study, we quantified the distribution and regulation of P in a freshwater zooplankton Daphnia magna under P-limited conditions by using 33P as a radiotracer. We demonstrated that the P allocation patterns in D. magna were independent of the ontogenic development. Carapace accounted for 35-54% of total body P, followed by small molecules and nucleic acids (11-30%), whereas phospholipids represented only a minor P pool. The proportion of body P allocated into carapace decreased from 51.8% in +P adults to 16.5% in the -P adults, and a lower proportion of body P was also allocated to eggs in the -P adults than in the +P adults (3.8 vs. 16.5%). Meanwhile, no difference in allocation pattern was detected in the juveniles under +P and -P conditions, demonstrating an interaction between effects of P condition and ontogeny. Furthermore, the P turnover rates of nucleic acids and phospholipids in the -P juveniles were only half of those found in the +P individuals, suggesting a reduced metabolic rate under P-deficient conditions. However, the P turnover rate of small molecules, nucleic acids and phospholipids did not vary with the P condition in adults. It appeared that the adults could maintain their basic P metabolism by down-regulating the P allocation to carapace and eggs. Our results provide an insight into the tolerance of zooplankton to P-deficiency and bear implications on involvement of Daphnia in regulation of P cycling and availability in the epilimnion.


Results of Expanded-Criteria Donor Kidneys: A Single-Center Experience in Hungary.
To ease organ shortage, many transplant centers accept kidneys from expanded-criteria donors (ECDs). Our aim was to analyze the results of ECD grafts in our center. Data on cadaveric donors were retrospectively analyzed between January 2011 and September 2014. Definition of ECD was: (1) donor age ≥60 years, (2) donor age 50 to 59 years, and (3) the presence of 2 among the following criteria: hypertension, serum creatinine >1.5 mg/dL, or death from cerebrovascular accident. Standard-criteria donors (SCDs) were those who did not meet the criteria for an ECD. During the observation period, 215 cadaveric donors were reported within our region, and 14 kidneys were offered to our center from Eurotransplant. Ninety-one (40%) among the reported donors were ECDs and 123 (54%) were SCDs. The rates of delayed graft function (DGF) and acute rejection (ARE) were not influenced by transplantation of an ECD graft. The cumulative patient and graft survival rates for ECDs were comparable with those of patients who received an optimal graft. ECD grafts can be transplanted safely, without the increased risk of DGF, ARE, and inferior patient and graft survival, in the case of careful patient allocation, and with the use of induction therapy.


Sequential changes in cell-mediated immune responses to herpes simplex virus following primary herpetic infection in man.
During primary herpes simlex virus infections, sensitization to the virus was demonstrated at the time of presentation by lymphocyte blastogenesis. Recovery from infection coincided with development of the macrophage migration inhibition response, and it is suggested that macrophage activation by products of stimulated lymphocytes contributes to control of the infection.