Red Blood Cells Homeostatically Bind Mitochondrial DNA through TLR9 to Maintain Quiescence and to Prevent Lung Injury.
Potentially hazardous CpG-containing cell-free mitochondrial DNA (cf-mtDNA) is routinely released into the circulation and is associated with morbidity and mortality in critically ill patients. How the body avoids inappropriate innate immune activation by cf-mtDNA remains unknown. Because red blood cells (RBCs) modulate innate immune responses by scavenging chemokines, we hypothesized that RBCs may attenuate CpG-induced lung inflammation through direct scavenging of CpG-containing DNA. To determine the mechanisms of CpG-DNA binding to RBCs and the effects of RBC-mediated DNA scavenging on lung inflammation. mtDNA on murine RBCs was measured under basal conditions and after systemic inflammation. mtDNA content on human RBCs from healthy control subjects and trauma patients was measured. Toll-like receptor 9 (TLR9) expression on RBCs and TLR9-dependent binding of CpG-DNA to RBCs were determined. A murine model of RBC transfusion after CpG-DNA-induced lung injury was used to investigate the role of RBC-mediated DNA scavenging in mitigating lung injury in vivo. Under basal conditions, RBCs bind CpG-DNA. The plasma-to-RBC mtDNA ratio is low in naive mice and in healthy volunteers but increases after systemic inflammation, demonstrating that the majority of cf-mtDNA is RBC-bound under homeostatic conditions and that the unbound fraction increases during inflammation. RBCs express TLR9 and bind CpG-DNA through TLR9. Loss of TLR9-dependent RBC-mediated CpG-DNA scavenging increased lung injury in vivo. RBCs homeostatically bind mtDNA, and RBC-mediated DNA scavenging is essential in mitigating lung injury after CpG-DNA. Our data suggest a role for RBCs in regulating lung inflammation during disease states where cf-mtDNA is elevated, such as sepsis and trauma.


Five new species of the genus Philydrodes Bernhauer, 1929 from China (Coleoptera: Staphylinidae: Omaliinae: Anthophagini).
Five new species of the subgenus Minyphilydrodes of the genus Philydrodes Bernhauer, 1929 (Coleoptera: Staphylinidae: Omaliinae: Anthophagini) from China are described and illustrated: Ph. tridentatus sp.n. (Hubei, Gansu, Shaanxi and Sichuan), Ph. tibetanus sp.n. (Yunnan), Ph. reticulatus sp.n. (Shaanxi), Ph. wrasei sp.n. (Sichuan) and Ph. michaeli sp.n. (Sichuan). The morphological features of Philydrodes, differences between Japanese and Chinese species, the relations of the genus with other taxa of the tribe Anthophagini Thomson, 1859 are briefly discussed. A key to the species of the genus from China and a distribution map are provided. The genus Philydrodes is recorded from continental Asia and China for the first time.


Postcardiac injury syndrome. An immunologic pleural fluid analysis.
The postcardiac injury syndrome (PCIS) is characterized by inflammation of the pericardium, pleura, and pulmonary parenchyma following a variety of cardiac injuries. Although it has been clinically recognized for decades, confirmation of the syndrome has been problematic owing to lack of a sufficiently diagnostic test. Previously, we have reported pleural fluid characteristics which help to exclude other diagnoses that may mimic the syndrome. We describe the first immunologic assessment, including antimyocardial antibody testing, of pleural fluid from a patient with PCIS which supports a local immunologic mechanism in the pathogenesis of the syndrome. These results support the important role of pleural fluid analysis in the diagnosis of PCIS.


Angiogenic inhibition mediated by a DNAzyme that targets vascular endothelial growth factor receptor 2.
The vascular endothelial growth factor receptor (VEGFR) is an important angiogenic target for cancer gene therapy. In this study, we designed an mRNA-cleaving oligodeoxynucleotide that targets the VEGF receptor 2 (VEGFR2) transcript (VEGFR2 DNAzyme). This DNAzyme was found to digest efficiently mRNA substrates of VEGFR2 in a concentration- and time-dependent manner. We also showed that the DNAzyme induces apoptosis and markedly inhibits endothelial cell growth compared with a disabled DNAzyme and untreated controls. In contrast, the DNAzyme did not inhibit the growth of MDA-MB-435 cells in vitro. The DNAzyme in complex with a nonviral carrier also significantly inhibited tumor growth in vivo. After the fourth injection, there was nearly a 75% reduction of tumor size in the DNAzyme-treated group compared with the saline-injected control group (P = 0.024). Marked cell death in the peripheral regions of the tumor accompanied by a reduction in blood vessel density is consistent with the antiangiogenic mechanism of the DNAzyme. This study indicates that DNAzymes, targeting angiogenic growth factors of tumors, show promise as antitumor agents.


Real-Time Monitoring and Quantitative Evaluation of Resin In-Filtrant Repairing Enamel White Spot Lesions Based on Optical Coherence Tomography.
The aim of the present study was to explore the feasibility of real-time monitoring and quantitative guiding the repair of enamel white spot lesions (WSLs) with resin infiltration by optical coherence tomography (OCT). Seven New Zealand rabbits were treated with 37% phosphoric acid etchant for 15 min to establish the model of enamel demineralization chalk spots of upper incisors, which were repaired by Icon resin infiltrant. OCT, stereo microscope (SM) imaging, scanning electron microscope (SEM) imaging and hematoxylin eosin (HE) staining were used to image each operation step. The changes of WSLs of enamel before and in the process of restoration with resin infiltrant showed specific performance in OCT images, which were consistent with the corresponding results of stereomicroscope and SEM. OCT can non-invasively and accurately image the whole process of repairing enamel demineralization layer with resin infiltration real-time, which can effectively guide the clinical use of resin infiltrant to repair enamel WSLs and be used as an imaging tool to evaluate the process and effect of restoration with resin infiltrant at the same time.


Zebrafish Mesp family genes, mesp-a and mesp-b are segmentally expressed in the presomitic mesoderm, and Mesp-b confers the anterior identity to the developing somites.
Segmentation of a vertebrate embryo begins with the subdivision of the paraxial mesoderm into somites through a not-well-understood process. Recent studies provided evidence that the Notch-Delta and the FGFR (fibroblast growth factor receptor) signalling pathways are required for segmentation. In addition, the Mesp family of bHLH transcription factors have been implicated in establishing a segmental prepattern in the presomitic mesoderm. In this study, we have characterized zebrafish mesp-a and mesp-b genes that are closely related to Mesp family genes in other vertebrates. During gastrulation, only mesp-a is expressed in the paraxial mesoderm at the blastoderm margin. During the segmentation period, both genes are segmentally expressed in one to three stripes in the anterior parts of somite primordia. In fused somites (fss) embryos, in which all early somite boundary formation is blocked, initial mesp-a expression at the gastrula stage remains intact, but the expression of mesp-a and mesp-b is not detected during the segmentation period. This suggests that these genes are downstream targets of fss at the segmentation stage. Comparison with her1 expression (Müller, M., von Weizsäcker, E. and Campos-Ortega, J. A. (1996) Development 122, 2071-2078) suggests that, like her1, mesp genes are not expressed in primordia of the first several somites. Furthermore, we found that zebrafish her1 expression oscillates in the presomitic mesoderm. The her1 stripe, which first appears in the tailbud region, moves in a caudal to rostral direction, and it finally overlaps the most rostral mesp stripe. Thus, in the trunk region, both her1 and mesp transcripts are detected in every somite primordium posterior to the forming somites. Ectopic expression of Mesp-b in embryos causes a loss of the posterior identity within the somite primordium, leading to a segmentation defect. These embryos show a reduction in expression of the posterior genes, myoD and notch5, with uniform expression of the anterior genes, FGFR1, papc and notch6. These observations suggest that zebrafish mesp genes are involved in anteroposterior specification within the presumptive somites, by regulating the essential signalling pathways mediated by Notch-Delta and FGFR.


Pyruvate orthophosphate dikinase mRNA organ specificity in wheat and maize.
Polyadenylated RNA was isolated from leaves and seeds of a C(3) plant (Triticum aestivum L. cv Cheyenne, CI 8885) and from a C(4) plant (Zea mays L. cv Golden bantam). Each polyadenylated RNA preparation was translated in vitro with micrococcal nuclease-treated reticulocyte lysate. When the in vitro translation products were probed with antibodies to pyruvate orthophosphate dikinase (PPDK) (EC 126.96.36.199), two sizes of polypeptide were identified. A 110 kilodalton polypeptide was found in the in vitro translation products of mRNA isolated exclusively from leaves of both wheat and maize. A 94 kilodalton polypeptide, similar to the PPDK polypeptide which can be extracted after in vivo synthesis in maize and wheat leaves and seeds, was found in the in vitro translation products obtained from wheat seeds and maize kernels.These results indicate that the mRNAs for PPDK polypeptides are organ-specific in both a C(4) and a C(3) plant. Hague et al. (1983 Nucleic Acids Res 11: 4853-4865) proposed that the larger size polypeptide of the in vitro translation polypeptide from maize leaf RNA contains a ;transit sequence' which permits entry into the chloroplasts of a polypeptide synthesized in vivo in maize leaf cell cytoplasm. It appears that in wheat leaves also the transit of synthesized PPDK polypeptide through an intracellular membrane may be required, while such a transit sequence seems not to be required within cells of wheat and maize seeds.


Cecropins, antibacterial peptides from insects and mammals, are potently fungicidal against Candida albicans.
Natural products are the major source of lead compounds for drugs against human pathogens. Among the first natural peptides from animals for which a potent antibacterial activity has been recognized were the cecropins. The 30- to 40-residue alpha-helical peptides display their activity by permeabilizing the membranes of bacteria. Although originally isolated from insect hemolymph, a structural and functional correlate was also found in a mammal. Here, we report on the finding that cecropin A and B from the silk moth Cecropia as well as the porcine cecropin P1 are capable of inhibiting the growth of and to kill yeast-phase Candida albicans. The peptides were tested in radial diffusion and microbroth dilution assays. They displayed potent activity against a clinical isolate as well as against defined culture strains of the pathogenic yeast but are of exceedingly low cytotoxicity towards the human cell line Jurkat. The candidacidal properties of the intensely studied molecules known to be highly active against Gram-positive and Gram-negative bacteria may renew the interest in these natural broad-spectrum peptide antibiotics and their limited cytotoxicity to human cells may be exploited for the development of topical therapeutics against pathogens resistant to classical antibiotics.


[INFLUENCE OF ALCOHOLISM AND ALCOHOLIC ADDICTION ON INCREASING THE RATE OF CRIME IN SOCIETY (MEDICAL AND LEGAL PREVENTIVE MEASURES)].
The purpose of the study is to determine universal medical and legal measures for individual and nationwide prevention of fortification of the population for practical application by developing and adopting international normative legal act - the Convention on Combating Fortification in Society. To achieve this purpose, the authors have used general scientific and special methods of cognition. Thus, the statistical method was used to analyze the data of official statistics for 2014-2019 regarding the number of crimes committed while intoxication; the method of comparison was used in the analysis of data on studying the development of adolescents' behavior due to alcohol consumption and the effect of special programs on the prevention of alcoholism; the method of questioning was used when interviewing students of higher educational institutions regarding the prevalence of alcohol consumption among young people. The authors of the article have offered to develop the Convention on Combating Fortification in Society, which should include binding international standards for the prevention and treatment of alcoholism in the Member States of this Convention. It has been indicated that the normative and legal acts of the future Member States of the Convention should contain: a definition of the concepts of "fortification" and "alcoholism"; medical, socio-pedagogical, criminal and other measures to counteract alcoholism; provisions for the treatment of alcohol-dependent persons; measures of universal access to information resources in the field of fortification, etc. It has been offered to amend the national regulatory acts of the Member States of the suggested Convention.


Holonomic Quantum Control with Continuous Variable Systems.
Universal computation of a quantum system consisting of superpositions of well-separated coherent states of multiple harmonic oscillators can be achieved by three families of adiabatic holonomic gates. The first gate consists of moving a coherent state around a closed path in phase space, resulting in a relative Berry phase between that state and the other states. The second gate consists of "colliding" two coherent states of the same oscillator, resulting in coherent population transfer between them. The third gate is an effective controlled-phase gate on coherent states of two different oscillators. Such gates should be realizable via reservoir engineering of systems that support tunable nonlinearities, such as trapped ions and circuit QED.


Age dependence and prognostic impact of neuron specific enolase (NSE) in children with neuroblastoma.
Serum neuron specific enolase (NSE) was determined in 159 patients with neuroblastoma at diagnosis and in 183 children of various age groups. We found an age dependence of reference intervals for NSE and defined the 95th percentiles as upper normal limits. The specificity was 91.3% and the sensitivity 73.0%. The incidence of abnormal NSE levels increased with stage. The NSE serum levels were not influenced by histologic differentiation. Neuron specific enolase proved to be a reliable tumor-marker for monitoring the disease. Moreover, abnormal NSE values at diagnosis were of prognostic significance for patients with localized neuroblastoma (stages I-III) and for children with metastatic disease (stage IV), but not for infants with stage IV S. In comparison to catecholamine metabolite determination neuron specific enolase appeared to be a slightly less specific, equally sensitive tumor marker but with prognostic information for children with neuroblastoma.


Levothyroxine Supplementation in Euthyroid Pregnant Women With Positive Autoantibodies: A Systematic Review and Meta-Analysis.
To explore the role of levothyroxine (LT4) supplementation in affecting the outcome of pregnant euthyroid women with thyroperoxidase (TPO) antibodies. MEDLINE, EMBASE, Google Scholar, and the Web of Science databases were searched. The primary outcome was pre-term birth (PTB), defined as live birth before 37 weeks of gestation; secondary outcomes were gestational hypertension, pre-eclampsia (PE), placental abruption, miscarriage, intra-uterine death (IUD), and admission to neonatal intensive care unit (NICU). All these outcomes were explored in euthyroid women with TPO antibodies receiving compared to those not receiving LT4 supplementation in pregnancy. Random-effect meta-analyses were used to analyze the data and results reported as pooled odds ratios (OR) with their 95% confidence intervals (CI). The risk of PTB was lower in women with TPO antibodies receiving compared to those not receiving LT4 supplementation (OR of 0.60 (95% CI 0.4-0.9). However, this association came mainly from observational studies (OR: 0.29, 95% CI 0.1-0), while RCTs did not show any beneficial effect of LT4 supplementation in affecting such outcomes. Conversely, there was no difference in the risk of gestational hypertension, preeclampsia, placental abruption, miscarriage, and admission to NICU between the two groups. LT4 supplementation in TPO euthyroid women is not associated with a reduced risk of PTB in TPO-positive women with normal thyroid function.


Detection of hypoxic fractions in murine tumors by comet assay: comparison with other techniques.
The alkaline comet assay was used to detect the hypoxic fractions of murine tumors. A total of four tumor types were tested using needle aspiration biopsies taken immediately after a radiation dose of 15 Gy. Initial studies confirmed that the normalized tail moment, a parameter reflecting single-strand DNA breaks induced by the radiation, was linearly related to radiation dose. Further, it was shown that for a mixed population (1:1) of cells irradiated under air-breathing or hypoxic conditions, the histogram of normal tail moment values obtained from analyzing 400 cells in the population had a double peak which, when fitted with two Gaussian distributions, gave a good estimate of the proportion of the two subpopulations. For the four tumor types, the means of the calculated hypoxic fractions from four or five individual tumors were 0.15 +/- 0.04 for B16F1, 0.08 +/- 0.04 for KHT-LP1, 0.17 +/- 0.04 for RIF-1 and 0.14 +/- 0.01 for SCCVII. Analysis of variance showed that the hypoxic fraction in KHT-LP1 tumors is significantly lower than those of the other three tumors (P = 0.026) but that there is no significant difference in hypoxic fraction between B16F1, RIF-1 and SCCVII tumors (P = 0.574). Results from multiple samples taken from each of five RIF-1 tumors showed that the intertumor heterogeneity of hypoxic fractions was greater than that within the same tumor. The mean hypoxic fraction obtained using the comet assay for the four tumor types was compared with the hypoxic fraction determined by the clonogenic assay, or median pO2 values, or [3H]misonidazole binding in the same tumor types.(ABSTRACT TRUNCATED AT 250 WORDS)


Comparing the EQ-5D and the SF-6D descriptive systems to assess their ceiling effects in the US general population.
The EuroQol (EQ-5D) and SF-6D (derived from the SF-12) were compared to assess any ceiling effect in the EQ-5D and the SF-6D descriptive systems. In addition, the Physical Component Summary (PCS-12), the Mental Component Summary (MCS-12) and the EuroQol Visual Analog Scale (EQ-VAS) were compared on their discriminative ability to detect differences among individuals with different morbidities and sociodemographic characteristics. Data from the 2000 Medical Expenditure Panel Survey were used for the analysis. A total of 11,248 individuals that were 18 years or older and had data on all the study variables were included in the analysis. A total of 5104 individuals (47%) reported no limitations on all of the EQ-5D dimensions and only 683 (5.8%) were classified in full health based on the SF-6D descriptive system. Approximately 49% of the respondents that reported no limitations on the EQ-5D reported feeling "tense or downhearted and low,""a little," (level 2) or "some" (level 3) of the time on SF-6D. PCS-12 scores and EQ-VAS scores among individuals reporting no limitations on the EQ-5D descriptive system were significantly lower for respondents reporting coronary heart disease, angina, diabetes, myocardial infarction, high blood pressure or joint pain compared with respondents that reported no medical condition. Effect sizes for medical conditions using the PCS-12 were larger than the effect sizes using the EQ-VAS. Unlike the EQ-5D descriptive system, the SF-6D descriptive system derived from the SF-12 does not seem to have a ceiling effect. Nevertheless, the SF-6D does not discriminate between individuals with different morbidities who report full health on the EQ-5D, as does the PCS-12 and the EQ-VAS.


Observations on amphotericin B treatment of kala-azar given in a rural set up in Bihar, India.
In a kala-azar endemic area in rural Bihar with a large number of patients unresponsive to sodium antimony gluconate and pentamidine, treatment with amphotericin B was tried in a rural set-up with the objective to cure these patients, and to assess whether such a centre could be run successfully in a rural set-up. After thorough clinical examination and biochemical investigations, parasitologically confirmed patients who had haemoglobin above 5 g/dl, electrolyte imbalance if any, corrected, and ECG changes suggestive of myocardial damage stabilised after 10 days of bed rest were treated. Amphotericin B deoxycholate was infused at a dose of 1 mg/kg body weight, daily for 20 days. The adverse events were closely monitored. All 7 (100%) untreated patients of kala-azar, 258 (97%) of the 266 antimony and pentamidine resistant patients, and 31 (86%) of the 36 patients who had relapsed after a low dose regimen of amphotericin B, were cured with 20 infusions of amphotericin B. Eight (3%) patients of the antimony and pentamidine resistant group and 5 (14%) patients who had relapsed after low dose amphotericin B regimen required 25 infusions of amphotericin B to achieve parasitological cure. With some precautions and proper management, all patients of kala-azar could be cured with amphotericin B in a rural set-up. A significant (P < 0.05) percentage of patients of group C relapsing after a low dose amphotericin B regime requiring 25 infusions for cure suggests that an adequate dose regime of amphotericin B should be given during the first course of treatment to prevent emergence of drug resistance.


HDAC5 catalytic activity suppresses cardiomyocyte oxidative stress and NRF2 target gene expression.
Histone deacetylase 5 (HDAC5) and HDAC9 are class IIa HDACs that function as signal-responsive repressors of the epigenetic program for pathological cardiomyocyte hypertrophy. The conserved deacetylase domains of HDAC5 and HDAC9 are not required for inhibition of cardiac hypertrophy. Thus, the biological function of class IIa HDAC catalytic activity in the heart remains unknown. Here we demonstrate that catalytic activity of HDAC5, but not HDAC9, suppresses mitochondrial reactive oxygen species generation and subsequent induction of NF-E2-related factor 2 (NRF2)-dependent antioxidant gene expression in cardiomyocytes. Treatment of cardiomyocytes with TMP195 or TMP269, which are selective class IIa HDAC inhibitors, or shRNA-mediated knockdown of HDAC5 but not HDAC9 leads to stimulation of NRF2-mediated transcription in a reactive oxygen species-dependent manner. Conversely, ectopic expression of catalytically active HDAC5 decreases cardiomyocyte oxidative stress and represses NRF2 activation. These findings establish a role of the catalytic domain of HDAC5 in the control of cardiomyocyte redox homeostasis and define TMP195 and TMP269 as a novel class of NRF2 activators that function by suppressing the enzymatic activity of an epigenetic regulator.


Quality of Life in Gastroesophageal Reflux Disease Three Months After Laparoscopic Nissen's Fundoplication.
Gastroesophageal reflux disease (GERD) affects various elements of life including sleep, daily and social functioning, and physical and emotional activities. This study aims to determine the impact of laparoscopic Nissen's fundoplication (LNF) on health-related quality of life. This prospective study was conducted in a tertiary care hospital, Pakistan, from Jan 2019 to Feb 2020. Forty-seven participants completed the study. All patients completed the Gastroesophageal Reflux Disease Health-Related Quality of Life (GERD-HRQoL) questionnaire both pre-operatively and three months after LNF. There was significant difference in pre- and post-operative median Health-Related Quality of Life score (p value: 0.0073). There was improvement in items related to heartburn in HRQoL questionnaire, while questions related to swallowing and bloating either showed no change or worsening. LNF has a significant impact on health-related quality of life. It is important for the physician to consider the impact of GERD in daily life. Management goals for GERD should also include improvement in quality of life of the patient.


[The looks of Gendai Kamada].
We realized the looks of Gendai Kamada. At first, we found in a figure, in "Seishu Hanaoka and His Surgery" by Syuzo Kure, that the portrait described as that of Gendai is his father's. And we discovered the illustrations that illustrate the looks of Gendai in "Gekakihaizufu", which was a textbook of clinical anesthesia and surgery, printed in 1840. Using these illustrations, we realized the looks of Gendai Kamada.


Evaluation of two mobile telemedicine systems in the emergency room.
Two different prototype mobile telemedicine systems were constructed for use in the emergency room. They could transmit physiological signals as well as video pictures and sound. One device, the mobile emergency bed (MEB), was powered by battery and had a wireless connection to the local-area network (LAN). For the other, the mobile emergency server (MES), a patient monitor, video-camera and microphone were connected by a radio-frequency link to a server. A functional evaluation and a clinical evaluation (by 12 emergency doctors in six emergency centres) were performed on both prototypes. The bandwidth and the video quality of the MEB were better than those of the MES, because of the digital transmission of the wireless LAN. The MES was better for directing patient treatment and teleconsultation; the MEB was better for static patients in the emergency centre. In general, the MES was more suitable for practical emergency telemedicine work.


Competitive polymerase chain reaction quantitation of c-erbA beta 1, c-erbA alpha 1, and c-erbA alpha 2 messenger ribonucleic acid levels in normal, heterozygous, and homozygous fibroblasts of kindred S with thyroid hormone resistance.
Mutations in the T3-binding domain of the thyroid hormone receptor gene c-erbA beta result in dominant negative proteins and thyroid hormone resistance syndromes. Variable clinical manifestations of resistance to thyroid hormones have been reported, including short stature and neuropsychological abnormalities. The molecular bases for heterogeneity of phenotype among and within kindreds have not been fully elucidated. Recent investigations have considered differential expression of mutant and wild-type beta 1-receptor alleles and the regulation thereof as a mechanism to explain differential sensitivity to thyroid hormones. We used reverse transcription-competitive polymerase chain reaction (PCR) to measure c-erbA beta 1, c-erbA alpha 1, and c-erbA alpha 2 mRNAs in skin fibroblasts cultured from normal subjects, heterozygotes, and a severely affected homozygous mutant of kindred S. The homozygous mutant of kindred S had severe growth and mental retardation. After reverse transcription with primers specific for each of the c-erbA mRNAs, first strand cDNAs were amplified by PCR using subtype-specific amplimers. Primer design allowed simultaneous detection of wild-type and mutant messages in heterozygous fibroblasts and showed an approximately 1:1 ratio of these mRNAs in three patients. Inclusion of competitive standard cDNAs of known concentration in the PCR reactions allowed quantitation of the absolute levels of the beta 1-, alpha 1-, and alpha 2 mRNAs by comparison of products on ethidium bromide-stained agarose gels. These studies showed no effect of the presence of the mutant beta 1-allele, as fibroblast RNA from normal subjects, heterozygotes, and the homozygote gave values of 56-184, 2.8-12, and 23-40 attomol/5 micrograms total RNA for beta 1-, alpha 1-, and alpha 2 mRNAs, respectively. We conclude that these sensitive methods allow the detection of molecular species present at levels as low as 10 molecules/cell, and that this potent dominant negative receptor does not disrupt c-erbA expression at the level of mRNA. The neuropsychological sequelae of the kindred S mutation are not due to relative overexpression of the mutant allele.


Silent brain infarcts and early cognitive outcomes after transcatheter aortic valve implantation: a systematic review and meta-analysis.
Silent brain infarcts (SBIs) are frequently identified after transcatheter aortic valve implantation (TAVI), when patients are screened with diffusion-weighted magnetic resonance imaging (DW-MRI). Outside the cardiac literature, SBIs have been correlated with progressive cognitive dysfunction; however, their prognostic utility after TAVI remains uncertain. This study's main goals were to explore (i) the incidence of and potential risk factors for SBI after TAVI; and (ii) the effect of SBI on early post-procedural cognitive dysfunction (PCD). A systematic literature review was performed to identify all publications reporting SBI incidence, as detected by DW-MRI after TAVI. Silent brain infarct incidence, baseline characteristics, and the incidence of early PCD were evaluated via meta-analysis and meta-regression models. We identified 39 relevant studies encapsulating 2408 patients. Out of 2171 patients who underwent post-procedural DW-MRI, 1601 were found to have at least one new SBI (pooled effect size 0.76, 95% CI: 0.72-0.81). The incidence of reported stroke with focal neurological deficits was 3%. Meta-regression noted that diabetes, chronic renal disease, 3-Tesla MRI, and pre-dilation were associated with increased SBI risk. The prevalence of early PCD increased during follow-up, from 16% at 10.0 ± 6.3 days to 26% at 6.1 ± 1.7 months and meta-regression suggested an association between the mean number of new SBI and incidence of PCD. The use of cerebral embolic protection devices (CEPDs) appeared to decrease the volume of SBI, but not their overall incidence. Silent brain infarcts are common after TAVI; and diabetes, kidney disease, and pre-dilation increase overall SBI risk. While higher numbers of new SBIs appear to adversely affect early neurocognitive outcomes, long-term follow-up studies remain necessary as TAVI expands to low-risk patient populations. The use of CEPD did not result in a significant decrease in the occurrence of SBI.


A molecular dynamics study on the transport of a charged biomolecule in a polymeric adsorbent medium and its adsorption onto a charged ligand.
The transport of a charged adsorbate biomolecule in a porous polymeric adsorbent medium and its adsorption onto the covalently immobilized ligands have been modeled and investigated using molecular dynamics modeling and simulations as the third part of a novel fundamental methodology developed for studying ion-exchange chromatography based bioseparations. To overcome computational challenges, a novel simulation approach is devised where appropriate atomistic and coarse grain models are employed simultaneously and the transport of the adsorbate is characterized through a number of locations representative of the progress of the transport process. The adsorbate biomolecule for the system studied in this work changes shape, orientation, and lateral position in order to proceed toward the site where adsorption occurs and exhibits decreased mass transport coefficients as it approaches closer to the immobilized ligand. Furthermore, because the ligands are surrounded by counterions carrying the same type of charge as the adsorbate biomolecule, it takes the biomolecule repeated attempts to approach toward a ligand in order to displace the counterions in the proximity of the ligand and to finally become adsorbed. The formed adsorbate-ligand complex interacts with the counterions and polymeric molecules and is found to evolve slowly and continuously from one-site (monovalent) interaction to multisite (multivalent) interactions. Such a transition of the nature of adsorption reduces the overall adsorption capacity of the ligands in the adsorbent medium and results in a type of surface exclusion effect. Also, the adsorption of the biomolecule also presents certain volume exclusion effects by not only directly reducing the pore volume and the availability of the ligands in the adjacent regions, but also causing the polymeric molecules to change to more compact structures that could further shield certain ligands from being accessible to subsequent adsorbate molecules. These findings have significant practical implications to the design and construction of polymeric porous adsorbent media for effective bioseparations and to the synthesis and operation of processes employed in the separation of biomolecules. The modeling and analysis methods presented in this work could also be suitable for the study of biocatalysis where an enzyme is immobilized on the surface of the pores of a porous medium.


Prestimulus EEG influence on late ERP components.
Ten healthy volunteers were submitted to an auditory oddball event related potentials (ERP) paradigm. Single trial 500 ms poststimulus ERPs (Pz, Cz, Fz--linked earlobes) along with the correspondent 1000 ms prestimulus EEG (O1-Cz) were stored. EEG epochs were submitted to spectral analysis and a slow wave index (SWI = delta + theta/total) was computed. Three selective ERP averages corresponding to low, medium and high SWI were computed. N2 latency was longer and P3a amplitude was lower in high SWI averages as compared to low SWI averages.


Influence of Polymer Relaxation Time on the Electrospinning Process: Numerical Investigation.
"Electrospinnability", or the ease with which a solution can be used to obtain bead-free uniform fibers, depends on a large number of parameters, including solution properties, process parameters and ambient conditions. In this study, the effect of the polymer relaxation time on electrospinning of dilute polymer solutions is investigated numerically. It is shown that elastic stresses (ES) increase exponentially with the Deborah number ( D e ). For each polymer concentration there exists a critical D e below which the ES are insufficient to overcome capillary stresses (CS) and lead to the formation of beaded fibers. However, above the critical D e , there is a higher probability of the ES overcoming the CS and leading to the formation of uniform fibers. This analysis suggests the possibility of improved electrospinnability even with dilute polymer solutions, provided the relaxation time is sufficiently large. It is also found that changes in the drag coefficient due to change in the polymer conformation and self-concentration of polymer molecules would become significant for the electrospinning of polymer solutions with a longer relaxation time and high conductivity.


Role of nitric oxide in the regulation of glucose kinetics in response to endotoxin in dogs.
The purpose of the present in vivo study was to determine the role of nitric oxide (NO) in the regulation of glucose metabolism in response to endotoxin by blocking NO synthesis with N(G)-monomethyl-L-arginine (L-NMMA). In five dogs, the appearance and disappearance rates of glucose (by infusion of [6,6-(2)H(2)]glucose), plasma glucose concentration, and plasma hormone concentrations were measured on five different occasions: saline infusion, endotoxin alone (E coli, 1.0 microg/kg i.v.), and endotoxin administration plus three different doses of primed, continuous infusion of L-NMMA. Endotoxin increased rate of appearance of glucose from 13.7 +/- 1.6 to 23.6 +/- 3.3 micromol x kg(-1) x min(-1) (P < 0.05), rate of disappearance of glucose from 13.9 +/- 1.1 to 24.8 +/- 3.1 micromol x kg(-1) x min(-1) (P < 0.001), plasma lactate from 0.5 +/- 0.1 to 1.7 +/- 0.1 mmol/l (P < 0.01), and counterregulatory hormone concentrations. L-NMMA did not affect the rise in rate of appearance and disappearance of glucose, plasma lactate, or the counterregulatory hormone response to endoxin. Plasma glucose levels were not affected by endotoxin with or without L-NMMA. In conclusion, in vivo inhibition of NO synthesis by high doses of L-NMMA does not affect glucose metabolism in response to endotoxin, indicating that NO is not a major mediator of glucose metabolism during endotoxemia in dogs.


Leptin, visfatin, insulin resistance, and body composition change in chronic obstructive pulmonary disease.
The aim of the study was to compare endocrine parameters such as leptin, visfatin, insulin resistance, exercise capacity and body composition change, the pulmonary functions test (PFT) and arterial blood gases (ABG) parameters of chronic obstructive pulmonary disease (COPD) patients and in healthy controls. Fifty-five patients with COPD and without malnutrition and 25 healthy controls were included in our study. The serum leptin, visfatin, tumor necrosis factor alpha (TNF-alpha) and insulin resistance, body fat-free mass (FFM) and fat mass (FM) were measured in the groups. Additionally, body mass index (BMI) was calculated and the 6-minute walk test (6MWT), PFT and ABG analyses were performed in all of the cases. No difference in BMI between the COPD group and controls was determined. Serum leptin and visfatin levels, FFM and 6MWT distance were significantly lower in the patients with COPD (p < 0.001, p = 0.001, p = 0.032, p < 0.001, respectively). A correlation was found between serum leptin levels and BMI (r = 0.333, p = 0.027), and with FM (r = 0.365, p = 0.029). Serum visfatin level was correlated with the percentage of forced expiratory volume in the first second in the patients with COPD (r = 0.371, p = 0.013). HOMA-IR (Homeostasis model assessment of insulin resistance) and serum TNF-alpha levels in the patients with COPD were found to be significantly higher than controls (p = 0.001, p < 0.001). These results may be earlier signs for further diseases that can emerge in the advanced stages in patients with COPD. Evaluating the patients not only with the pulmonary function and also systemically, contributes to minimizing the mortality and morbidity.


Localization of alpha-actinin-4 during infections by actin remodeling bacteria.
Bacterial pathogens cause disease by subverting the structure and function of their target host cells. Several foodborne agents such as Listeria monocytogenes (L. monocytogenes), Shigella flexneri (S. flexneri), Salmonella enterica serovar Typhimurium (S. Typhimurium) and enteropathogenic Escherichia coli (EPEC) manipulate the host actin cytoskeleton to cause diarrheal (and systemic) infections. During infections, these invasive and adherent pathogens hijack the actin filaments of their host cells and rearrange them into discrete actin-rich structures that promote bacterial adhesion (via pedestals), invasion (via membrane ruffles and endocytic cups), intracellular motility (via comet/rocket tails) and/or intercellular dissemination (via membrane protrusions and invaginations). We have previously shown that actin-rich structures generated by L. monocytogenes contain the host actin cross-linker α-actinin-4. Here we set out to examine α-actinin-4 during other key steps of the L. monocytogenes infectious cycle as well as characterize the subcellular distribution of α-actinin-4 during infections with other model actin-hijacking bacterial pathogens (S. flexneri, S. Typhimurium and EPEC). Although α-actinin-4 is absent at sites of initial L. monocytogenes invasion, we show that it is a new component of the membrane invaginations formed during secondary infections of neighboring host cells. Importantly, we reveal that α-actinin-4 also localizes to the major actin-rich structures generated during cell culture infections with S. flexneri (comet/rocket tails and membrane protrusions), S. Typhimurium (membrane ruffles) and EPEC (pedestals). Taken together, these findings suggest that α-actinin-4 is a host factor that is exploited by an assortment of actin-hijacking bacterial pathogens.


A novel role for protein tyrosine phosphatase 1B as a positive regulator of neuroinflammation.
Protein tyrosine phosphatase 1B (PTP1B) is a member of the non-transmembrane phosphotyrosine phosphatase family. Recently, PTP1B has been proposed to be a novel target of anti-cancer and anti-diabetic drugs. However, the role of PTP1B in the central nervous system is not clearly understood. Therefore, in this study, we sought to define PTP1B's role in brain inflammation. PTP1B messenger RNA (mRNA) and protein expression levels were examined in mouse brain and microglial cells after LPS treatment using RT-PCR and western blotting. Pharmacological inhibitors of PTP1B, NF-κB, and Src kinase were used to analyze these signal transduction pathways in microglia. A Griess reaction protocol was used to determine nitric oxide (NO) concentrations in primary microglia cultures and microglial cell lines. Proinflammatory cytokine production was measured by RT-PCR. Western blotting was used to assess Src phosphorylation levels. Immunostaining for Iba-1 was used to determine microglial activation in the mouse brain. PTP1B expression levels were significantly increased in the brain 24 h after LPS injection, suggesting a functional role for PTP1B in brain inflammation. Microglial cells overexpressing PTP1B exhibited an enhanced production of NO and gene expression levels of TNF-α, iNOS, and IL-6 following LPS exposure, suggesting that PTP1B potentiates the microglial proinflammatory response. To confirm the role of PTP1B in neuroinflammation, we employed a highly potent and selective inhibitor of PTP1B (PTP1Bi). In LPS- or TNF-α-stimulated microglial cells, in vitro blockade of PTP1B activity using PTP1Bi markedly attenuated NO production. PTP1Bi also suppressed the expression levels of iNOS, COX-2, TNF-α, and IL-1β. PTP1B activated Src by dephosphorylating the Src protein at a negative regulatory site. PTP1B-mediated Src activation led to an enhanced proinflammatory response in the microglial cells. An intracerebroventricular injection of PTP1Bi significantly attenuated microglial activation in the hippocampus and cortex of LPS-injected mice compared to vehicle-injected mice. The gene expression levels of proinflammatory cytokines were also significantly suppressed in the brain by a PTP1Bi injection. Together, these data suggest that PTP1Bi has an anti-inflammatory effect in a mouse model of neuroinflammation. This study demonstrates that PTP1B is an important positive regulator of neuroinflammation and is a promising therapeutic target for neuroinflammatory and neurodegenerative diseases.


Determining witnessed status for out-of-hospital cardiac arrest.
Witnessed status is considered a core variable in reporting cardiac arrest data and can be ascertained from either the emergency dispatch recording or the pre-hospital record. The purpose of this study is to compare and assess the quality and consistency of these information sources. This retrospective analysis included 1896 cases of out-of-hospital cardiac arrest occurring between September 1, 2012 and December 31, 2014. We found that there was minimal (kappa=0.30, 95% CI 0.27-0.33) to moderate (kappa=0.64, 95% CI 0.59-0.69) agreement between the pre-hospital record and the emergency dispatch recording when these sources of information are used to determine witnessed status. Witnessed status could not be determined from the emergency dispatch recording in 36.2% (n=684) of eligible cases. Survival was similar regardless of the method used to determine witnessed status. Using a combination of the pre-hospital record and the emergency dispatch recording yielded the highest number of witnessed cases. The determination of witnessed status in out-of-hospital cardiac arrest may be challenging, as evidenced by the discrepancies in witnessed status when comparing different sources of information. The large number of cases where the witnessed status could not be determined from the emergency dispatch recording precludes its use as the sole source of information. It is reasonable to use the patient care record alone, however it should be recognized that there is misclassification of witnessed status regardless of the method used and this may affect the strength of association between witnessed status and survival.


Polio Endgame: Lessons Learned From the Immunization Systems Management Group.
The Immunization Systems Management Group (IMG) was established to coordinate and oversee objective 2 of the Polio Eradication and Endgame Strategic Plan 2013-2018, namely, (1) introduction of ≥1 dose of inactivated poliovirus vaccine in all 126 countries using oral poliovirus vaccine (OPV) only as of 2012, (2) full withdrawal of OPV, starting with the withdrawal of its type 2 component, and (3) using polio assets to strengthen immunization systems in 10 priority countries. The IMG's inclusive, transparent, and partnership-focused approach proved an effective means of leveraging the comparative and complementary strengths of each IMG member agency. This article outlines 10 key factors behind the IMG's success, providing a potential set of guiding principles for the establishment and implementation of other interagency collaborations and initiatives beyond the polio sphere.


Evaluation of the in-hospital hemovigilance by introduction of the information technology-based system.
Hemovigilance is an important aspect of transfusion medicine. However, the frequency of the adverse reactions often varies using different reporters. Recently, we have employed a new information technology (IT)-based in-hospital hemovigilance system. Here, we evaluated changes in practice after implementation of an IT-based reporting system. We compared the rate of frequency and details of blood transfusion-related adverse reactions 3 years before and after introduction of the IT-based reporting system. Contents and severity of the adverse reactions were reported in a paper-based reporting system, but input by selecting items in an IT-based reporting system. The details of adverse reactions are immediately sent to the blood transfusion unit online. After we introduced the IT-based reporting system, the reported rate of transfusion-related adverse reactions increased approximately 10-fold from 0.20% to 2.18% (p < 0.001), and frequencies of urticaria, pruritus, rash, fever (p < 0.001), hypertension (p = 0.001), tachycardia (p = 0.003), and nausea and vomiting (p = 0.010) increased significantly. Although there was no error report in the paper-based reporting, incorrect reports were observed in 90 cases (0.52%) in the IT-based reporting (p < 0.001). The advantages of IT-based reporting were: 1) a significant increase in the frequency of adverse reaction reporting and 2) a significant decrease in underreporting, although the true frequency has yet to be clarified. The disadvantage of the IT-based reporting was an increased incidence of incorrect inputs, all of which was unnoticed by the reporters. Our results showed several important points in need of monitoring after introduction of an IT-based reporting system.


Human liver sulphotransferase and UDP-glucuronosyltransferase: structure-activity relationship for phenolic substrates.
1. Human liver sulphotransferase and UDP-glucuronosyltransferase were studied with phenol, methyl-, ethyl-, propyl-, butyl-, phenyl-, nitro-, amino-phenols and hydroxybenzoic acids as substrates. 2. The Michaelis-Menten constants (Km) and the maximum velocities of reaction (Vmax) of sulphotransferase and UDP-glucuronosyltransferase for each substrate were measured. 3. The Km values for sulphotransferase varied over 5000-fold whereas they varied over 25-fold for UDP-glucuronosyltransferase. 4. Sulphotransferase and UDP-glucuronosyltransferase have different structure-activity relationships with phenolic substrates.


Down-regulation of cyclin E expression by caffeine promotes cancer cell entry into the S-phase of the cell cycle.
Expression of cyclin E is believed to be a critical factor promoting cell entry into the S-phase and cell proliferation. Indeed, normal proliferating cells and most tumor cell lines are characterized by the existence of a minimal cyclin E threshold level in the G1-phase, and only those cells expressing cyclin E over this threshold enter into the S-phase of the cell cycle. However, through studying clinical tumor tissue specimens, we recently observed that some cancer cells can enter into the S-phase with minimal levels of cyclin E expression. In an effort to establish an in vitro cell model system for studying the mechanisms underlying this phenomenon, we treated MOLT-4 lymphocyte leukemia cells with 50 mM caffeine and found that the levels of cyclin E expression were decreased markedly in these cells following 2 to 4-h exposure to caffeine. Quite unexpectedly, we observed that the percentage of the cells progressing through the S-phase increased despite the reduced levels of cyclin E, as analyzed for the cellular DNA contents, expression of nuclear-bound PCNA, immunolabelling with Ki-67 antibody and incorporation of BrdU. In fact, these cells entered into the S-phase with a level of cyclin E well below the threshold level for untreated cells, thus suggesting that lower levels of cyclin E expression are associated with cell proliferation under certain circumstances. We speculate that caffeine may enhance MOLT-4 cell entrance into the S-phase through activation of Cdc25, which in turn activates cyclin-dependent protein kinases (CDKs) including CDK2 and drives the cell cycle progression; while degradation of cyclin E by the ubiquitin/proteasome pathway may account for the decreased levels of cyclin E in these cells. Our findings from both the MOLT-4 cell line and patients' cancer tissues may help decipher the mystery of the deregulation of cell cycle progression and carcinogenesis in some malignant tumors.


Anoxygenic photo- and chemo-synthesis of phototrophic sulfur bacteria from an alpine meromictic lake.
Meromictic lakes are interesting ecosystems to study anaerobic microorganisms due their permanent stratification allowing the formation of a stable anoxic environment. The crenogenic meromictic Lake Cadagno harbors an important community of anoxygenic phototrophic sulfur bacteria responsible for almost half of its total productivity. Besides their ability to fix CO2 through photosynthesis, these microorganisms also showed high rates of dark carbon fixation via chemosyntesis. Here, we grew in pure cultures three populations of anoxygenic phototrophic sulfur bacteria previously isolated from the lake, accounting for 72.8% of the total microbial community and exibiting different phenotypes: (1) the motile, large-celled purple sulfur bacterium (PSB) Chromatium okenii, (2) the small-celled PSB Thiodictyon syntrophicum and (3) the green sulfur bacterium (GSB) Chlorobium phaeobacteroides. We measured their ability to fix CO2 through photo- and chemo-synthesis, both in situ in the lake and in laboratory under different incubation conditions. We also evaluated the efficiency and velocity of H2S photo-oxidation, an important reaction in the anoxygenic photosynthesis process. Our results confirm that phototrophic sulfur bacteria strongly fix CO2 in the presence of light and that oxygen increases chemosynthesis at night, in laboratory conditions. Moreover, substancial differences were displayed between the three selected populations in terms of activity and abundance.


Trichuris muris: antigen recognition and transfer of immunity in mice by IgA monoclonal antibodies.
Mesenteric node lymphocytes from mice that had been infected with the nematode Trichuris muris, and then boosted with adult worm excretory-secretory antigens were fused with myeloma cells to produce a panel of 9 monoclonal antibodies (MoAbs). Five of the MoAbs were of the IgA isotype. The antigen recognition profiles of these MoAbs were studied using SDS-PAGE and immunoblotting; three major profile patterns were identified. Five MoAbs recognized a major band in the MW range 43-48 kD; all recognized a range of antigens. Three MoAbs were used to localize antigens in the bodies of adult worms. Granules within the anterior stichocytes were recognized strongly, as was material within the eggs and pseudocoelom. Two MoAbs stained the cuticle. Although the phosphorylcholine (PC) determinant was widely distributed within worm tissues none of the MoAbs tested recognized PC. Passive transfer of immunity was achieved using two of the IgA monoclonals; no immunity was transferred by the IgM and IgG MoAbs used. The limited recognition profiles of these IgA MoAbs, and the ability to stain stichocyte granules, suggest that their protective activity results from an interaction with ES antigens.


Antagonism of hypotensive and bradycardic effects of centrally administered guanfacine by metiamide in anesthetized rabbits.
In anesthetized rabbits, intracerebroventricular (ICV) administration of the alpha-adrenoceptor agonist, guanfacine, like clonidine, elicited dose-related falls in arterial blood pressure and heart rate. ICV pretreatment with the histamine H2-receptor antagonist, metiamide, significantly reduced the hypotensive and bradycardic responses caused by ICV injection of either guanfacine or clonidine. The present results seem to indicate that the antagonistic effect of metiamide against either guanfacine or clonidine is probably due to its alpha-adrenoceptor blocking action rather than its histamine H2-receptor blocking action, suggesting that these interactions might be of central origin.


Inequalities in cervical cancer screening in Eastern Europe: perspectives from Bulgaria and Romania.
The incidence of cervical cancer in Eastern Europe has been on the rise, in contrast to the reduction in incidence in most countries of Western Europe. The objectives of the paper are to delineate the inequalities in cervical cancer screening in Romania and Bulgaria and identify explanations for these inequalities. Representative samples of women - 1,099 in Bulgaria and 1,053 in Romania, were interviewed through a structured questionnaire. We found multiple dimensions of inequalities in cervical cancer prevalence and prevention, including disparities in comparison to other countries, disparities due to socioeconomic status, education, residency and ethnicity, as well as differential barriers faced by women in access to screening and in relationships with providers. We identified mediators of the effects of socio-economic status on screening history. The study concludes that the effect of SES on screening is mediated mainly by the structural barriers in accessing the healthcare system, as well as women's perceptions of the multiple costs of the smear. These conclusions are relevant to the development of national screening programs and health promotion in the two countries.


One cornea for two patients: case report.
Case reporting the use of one donor cornea for two transplantation procedures: deep anterior lamellar keratoplasty (DALK) in a case of an imminent corneal perforation caused by herpetic stromal necrosis, and Descemet stripping with endothelial keratoplasty (DSEK) in an eye with pseudophakic bullous keratopathy (PBK). Descemet's membrane (DM), denuded by stromal necrosis, served as the starting point for dissection plane and creation of the recipient bed for DALK. The next steps were excision of the diseased stroma along the edge of trephination, and transplantation of a 400-450 microm thick, manually dissected lamellar graft. The remaining posterior layers of the donor cornea, 100-150 microm thick, were used as a graft in the DSEK procedure for PBK. The integrity of the globe was saved, and best-corrected visual acuity (BCVA) of 20/40 was reached after DALK in the eye with an imminent corneal perforation. A subnormal central corneal thickness (CCT) of 430 microm did not interfere with corneal shape (43.50 x 45.50 D) and function. The graft remained attached and clear after DSEK in the eye with PBK, with BCVA of 20/30 and a CCT of 653 microm. One donor cornea can be used for two lamellar keratoplasties, DALK and DSEK. Although the described obstacles may prevent this approach from becoming widely used, it may prove useful when one is confronted with a need for an urgent anterior lamellar keratoplasty, a long list of cases for DSEK, and a shortage of donor corneas.


A new skin suture technique for multiple Z-plasty.
A new skin suture technique was developed for multiple Z-plasty. In this technique, the wound of multiple Z-plasty is closed by several continuous locking sutures. With use of this technique, the operative time is reduced, and suture removal is facilitated to alleviate pain and discomfort on the part of the patient.


Potency testing of inactivated tissue culture vaccine against Japanese encephalitis using antigen capture ELISA.
The potency of vaccines against Japanese encephalitis (JE) is usually determined by a mouse challenge test. In the present study, an antigen capture enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies (MoAbs) was used to screen inactivated tissue culture JE vaccine lots. This test is simple, quick and reliable besides being very sensitive and specific.


RLS in middle aged women and attention deficit/hyperactivity disorder in their offspring.
Previous studies have suggested that attention deficit/hyperactivity disorder (ADHD) and restless legs syndrome (RLS) could share some common genetic backgrounds, but the effect of these genetic components could be modest. To test this hypothesis, we conducted a large-scaled cross-sectional study to examine whether women with a child with ADHD had a higher risk of having RLS than women of unaffected children. We included 65,554 women free of diabetes, arthritis, and pregnancy in the current analyses. Information on RLS was assessed using a set of standardized questions. Participants were considered to have RLS if they met four RLS diagnostic criteria recommended by the International RLS Study Group and had restless legs ≥5 times/month. Information on ADHD in offspring was collected via questionnaire. We observed a significant association between presence of ADHD in the offspring and risk of having RLS; the multivariate-adjusted OR for RLS was 1.27 (95% CI: 1.15, 1.41; P<0.0001), after adjusting for age, body mass index, number of deliveries during life time and other covariates. We found that mothers of children with ADHD had an increased risk of having RLS. Further studies are warranted to explore biological mechanisms underling this association.


[Effect of boanmycin on apoptosis and cell cycle of human esophageal cancer(Eca-109) cells].
The anticancer mechanism of boanmycin was thought to inhibit the synthesis of protein in human esophageal cancer cell line. But it is not clear that there are other mechanisms of boanmycin to produce its anticancer effect on human esophageal cancer. This study was designed to examine the effect of boanmycin on apoptosis and cell cycle of human esophageal cancer cells (Eca-109). Microscopic observation, flow cytometry, and agarose gel electrophoresis were used to determine the induction of apoptosis and the changes of cell cycle. The human esophageal cancer xenografts in nude mice treated by boanmycin showed morphological changes of different apoptotic phases. In vitro, boanmycin (50-125 micrograms/ml, 30-36 hours) elicited typical apoptotic changes in Eca-109 cells and typical DNA ladder of apoptotic cells appeared. The Eca-109 cells treated by boanmycin for 42 hours were blocked at G2/M phase. Boanmycin could induce apoptosis of Eca-109 cells and block the cell cycle at G2/M phase.


The Yersinia pestis V antigen is a regulatory protein necessary for Ca2(+)-dependent growth and maximal expression of low-Ca2+ response virulence genes.
The low-Ca2+ response is a multicomponent virulence regulon of the human-pathogenic yersiniae in which 12 known virulence genes are coordinately regulated in response to environmental cues of temperature, Ca2+, and nucleotides such as ATP. Yersinial growth also is regulated, with full growth yield being permitted at 37 degrees C only if Ca2+ or a nucleotide is present. In this study, we constructed and characterized a mutant Yersinia pestis specifically defective in the gene encoding the V antigen, one of the virulence genes of the low-Ca2+ response. An in-frame internal deletion-insertion mutation was made by removing bases 51 through 645 of lcrV and inserting 61 new bases. The altered lcrV was introduced into the low-Ca2+ response plasmid in Y. pestis by allelic exchange, and the resulting mutant was characterized for its two-dimensional protein profiles, growth, expression of an operon fusion to another low-Ca2+ response virulence operon, and virulence in mice. The mutant had lost its Ca2+ and nucleotide requirement for growth, showed diminished expression of Ca2(+)-and nucleotide-regulated virulence genes, and was avirulent in mice. The mutation could be complemented with respect to the growth property by supplying native V antigen operon sequences in trans in high copy number (on pBR322). Partial complementation of the growth defect and almost complete complementation of the virulence defect were seen with a lower-copy-number complementing replicon (a pACYC184 derivative). The data are consistent with the interpretation that V antigen is bifunctional, with a role in regulating growth and expression of low-Ca2+ response virulence genes in addition to its putative role as a secreted virulence protein.


Approach to Common Visual Complaints Presenting to the Outpatient Neurologist.
Visual complaints are commonly encountered by the practicing neurologist. We review assessment of vision loss, diplopia, and positive visual phenomena, all of which require a thoughtful evaluation to localize disease and refine management. While many causative entities are unlikely to cause poor visual outcomes, including dry eyes, migraine, and congenital strabismus, others may threaten vision, life, or both, such as posterior communicating artery aneurysms, pituitary apoplexy, or temporal arteritis. A systematic approach to vision loss and diplopia is reviewed along with focused differential diagnoses.


The nhTMEM16 Scramblase Is Also a Nonselective Ion Channel.
The TMEM16 family comprises Ca2+-activated Cl- channels and phospholipid scramblases. The crystal structure of a fungal homolog, nhTMEM16, revealed an important architectural feature of this protein family in the form of a bilayer-spanning hydrophilic groove that is directly exposed to the membrane. This groove likely provides a pathway for lipid translocation. As mutations that alter ion channel activity of the TMEM16 proteins localize around the groove, it was suggested that the ion and lipid pathways coincide such that the ion pore is partly lined by phospholipids. However, this proposal was not supported by the observation that nhTMEM16 does not mediate ion transport. Here we show that nhTMEM16 mediates both ion and lipid transport and that its properties closely resemble those of a previously characterized fungal homolog, afTMEM16. We show that the reported lack of ion transport activity of nhTMEM16 is due to the lipid composition of the reconstitution membranes and to the presence of a GFP tag. Thus, nhTMEM16, like afTMEM16 and the mammalian TMEM16F, mediates simultaneous lipid scrambling and nonspecific ion transport. This supports the hypothesis that these two processes are tightly correlated and likely to be a general functional feature of the TMEM16 scramblases and therefore of general importance in understanding their biological roles.


Consistent latencies of vestibular evoked myogenic potentials.
This study investigated the association between neck length and vestibular evoked myogenic potential (VEMP) latencies in healthy children, adolescents, and adults to elucidate when VEMP latencies reach consistent levels. Findings of VEMP tests in 14 healthy children, seven healthy adolescents, and 14 healthy adults were analyzed for correlations with neck length, which was measured as the distance of a line dropping vertically from the mastoid tip to the horizontal plane passing through the clavicle. All healthy children, adolescents, and adults exhibited present VEMP responses. Children, adolescents, and adults significantly differed in p13 latency, n23 latency, and p13-n23 interval. According to receiver operating characteristic curve analysis, the optimal cutoff values of p13 and n23 latencies between children and adults were 12.6 and 19.8 msec, respectively. Because the odds ratio of p13 latency was less than that of n23 latency, n23 latency was used to discriminate VEMP latencies between children and adults. Accordingly, a cutoff value of 15.3 cm for neck length was proposed as a criterion for predicting VEMP latency within the adult range. Consequently, a positive correlation between neck length and VEMP latency was observed when neck length was <15.3 cm, while above which level one need not account for neck length in evaluating VEMP latency. The intra-subject variability of norms can be enhanced if the normative data for VEMP characteristic parameters take structural variance into account. This study suggests that the adult range of VEMP latencies can be anticipated if neck length is >15.3 cm.


Substrate-induced assembly of fibronectin into networks: influence of surface chemistry and effect on osteoblast adhesion.
The influence of surface chemistry-substrates with controlled surface density of -OH groups-on fibronectin (FN) conformation and distribution is directly observed by atomic force microscopy (AFM). FN fibrillogenesis, which is known to be a process triggered by interaction with integrins, is shown in our case to be induced by the substrate (in absence of cells), which is able to enhance FN-FN interactions leading to the formation of a protein network on the material surface. This phenomenon depends both on surface chemistry and protein concentration. The level of the FN fibrillogenesis was quantified by calculating the fractal dimension of the adsorbed protein from image analysis of the AFM results. The total amount of adsorbed FN is obtained by making use of a methodology that employs Western blotting combined with image analysis of the corresponding protein bands, with the lowest sensitivity threshold equal to 15 ng of adsorbed protein. Further, FN adsorption is correlated to human osteoblast adhesion through morphology and actin cytoskeleton formation. Actin polymerization is in need of the formation of the protein network on the substrate's surface. Cell morphology is more rounded (as quantified by calculating the circularity of the cells by image analysis) when the degree of FN fibrillogenesis on the substrate is lower.


The effect of class II gene transfection on the tumourigenicity of the H-2K-negative mouse leukaemia cell line K36.16.
There is much evidence to suggest that potentially immunogenic tumour cells can escape cytolytic immune destruction by loss of class I antigen expression. Many tumours are allele-specific class I negative and, in murine systems, reconstitution of class I expression by gene transfection leads to an increase in tumour immunogenicity. In many systems where mice have rejected class I transfected tumour cells they are also immune to a subsequent challenge with the untransfected parent tumour. In this study we have examined the effect of stable class II antigen expression (induced by gene transfection) on a class I loss mutant (H-2Kk negative) murine cell line, K36.16. We show that H-2Ek expression is more effective at increasing tumour immunogenicity than the reconstitution of H-2Kk expression in these cells. This suggests that the induction of class II antigen expression on tumour cells may provide an effective way of enhancing tumour-specific immune responses in vivo.


Sequence of a turkey BAC clone identifies MHC Class III orthologs and supports ancient origins of immunological gene clusters.
Two genetically unlinked gene clusters currently define the turkey major histocompatibility complex (MHC). Previous studies identified turkey bacterial artificial chromosome (BAC) clones hypothesized as orthologs of the MHC-B and MHC-Y regions of the chicken. Physical mapping assigned these clones to the same microchromosome (MGA18) and sequencing of the MHC-B BAC found near synteny with a portion of the chicken B-locus. This study examines the sequence of the second MHC BAC clone that was hypothesized, based on subclone sequences, to be orthologous to the MHC-Y. Sequencing of this clone identified a class I locus and orthologs of additional genes found in the mammalian class III region. Approximately 50% of the BAC insert is comprised of sequence corresponding to the centromeric repeat, MGASat2. This turkey MHC BAC sequence is unique from sequences assigned to the MHC-Y in the chicken. Based on sequence comparisons, the class I gene appears to be a nonfunctional pseudogene. The class III genes (BAT1, BAT3, STK19, and a G4-like locus) represent the second class III gene cluster identified in the galliform genome. This cluster appears to be of ancient origin and provides insight into the evolution of the avian MHC.


Relationship of plasma N-terminal pro-brain natriuretic peptide concentrations to heart failure classification and cause of respiratory distress in dogs using a 2nd generation ELISA assay.
Cardiac biomarkers provide objective data that augments clinical assessment of heart disease (HD). Determine the utility of plasma N-terminal pro-brain natriuretic peptide concentration [NT-proBNP] measured by a 2nd generation canine ELISA assay to discriminate cardiac from noncardiac respiratory distress and evaluate HD severity. Client-owned dogs (n = 291). Multicenter, cross-sectional, prospective investigation. Medical history, physical examination, echocardiography, and thoracic radiography classified 113 asymptomatic dogs (group 1, n = 39 without HD; group 2, n = 74 with HD), and 178 with respiratory distress (group 3, n = 104 respiratory disease, either with or without concurrent HD; group 4, n = 74 with congestive heart failure [CHF]). HD severity was graded using International Small Animal Cardiac Health Council (ISACHC) and ACVIM Consensus (ACVIM-HD) schemes without knowledge of [NT-proBNP] results. Receiver-operating characteristic curve analysis assessed the capacity of [NT-proBNP] to discriminate between dogs with cardiac and noncardiac respiratory distress. Multivariate general linear models containing key clinical variables tested associations between [NT-proBNP] and HD severity. Plasma [NT-proBNP] (median; IQR) was higher in CHF dogs (5,110; 2,769-8,466 pmol/L) compared to those with noncardiac respiratory distress (1,287; 672-2,704 pmol/L; P < .0001). A cut-off >2,447 pmol/L discriminated CHF from noncardiac respiratory distress (81.1% sensitivity; 73.1% specificity; area under curve, 0.84). A multivariate model comprising left atrial to aortic ratio, heart rate, left ventricular diameter, end-systole, and ACVIM-HD scheme most accurately associated average plasma [NT-proBNP] with HD severity. Plasma [NT-proBNP] was useful for discriminating CHF from noncardiac respiratory distress. Average plasma [NT-BNP] increased significantly as a function of HD severity using the ACVIM-HD classification scheme.


Ultrastructural localization of basic lysine-rich proteins during the nucleologenesis in preimplantation mouse embryos.
The paper describes nucleologenesis in preimplantation mouse embryos studied by electron microscopy after staining with ethanolic phosphotungstic acid (E-PTA), a substance which binds to basic lysine-rich proteins and makes it thus possible to follow their localization and distribution. A nucleolar precursor body (NPB) homogeneously stained with E-PTA first appears in pronuclei of a fertilized oocyte. In 2-cell stage embryos, the staining intensity of the central part of NPB is reduced, whereas the outer zone retains the PTA positivity. In the course of further development, reticulation takes place in the peripheral zone and all structures of a functional nucleolus are gradually formed. Tiny spherical bodies, called "remnant NPBs", can still be observed in reticular nucleoli of cells at the morula stage. The similarities in reaction and morphological appearance of the NPB and the corresponding structures in oocytes of the mouse and rat antral follicle are discussed.


Diabetes modulates capacitative calcium entry and expression of transient receptor potential canonical channels in human saphenous vein.
Diabetes is associated with a perturbation of signaling pathways in vascular tissue, which causes vasomotor dysfunction such as hypertension. We have previously demonstrated that vessels from diabetic patients were more contractile than those from non-diabetic. However, in human vessels, the receptor-stimulated contraction is mainly due to enzymatic, rather than calcium signaling pathway. In this study, we hypothesized that the differential contractile response between diabetic and non-diabetic human vessels could be due to the receptor signaling to sarcoplasmic reticulum and the regulation of capacitative calcium entry. In saphenous vein samples (n=20) collected from diabetic patients undergoing bypass surgery, the contraction initiated by the addition of the sarco-endoplasmatic reticulum calcium ATPase blocker, cyclopiazonic acid, was significantly higher than that in the vessels from non-diabetic patients (n=26) (84.0+/-14.9% vs 44.2+/-9.2%), and this contraction was inhibited by SKF-96365, an inhibitor of store-operated calcium channels. Pre-incubation with indomethacin reduced the cyclopiazonic acid-induced contraction in the non-diabetic veins, but had no effect on the diabetic ones. The gene expression of transient receptor potential canonical channels (TRPC)4 was upregulated by 22% in the diabetic vessels compared with the non-diabetic ones. However, the protein expression of TRPC1 and TRPC6 was downregulated in the diabetic group by 50%. We concluded that diabetes would modulate the capacitative calcium entry likely through the store-operated calcium channel specifically via the regulation of TRPC.


FISH analysis in cell touch preparations and cytological specimens from formalin-fixed fetal autopsies.
Postmortem studies on still-borns and miscarriages are important to define the sex and eventually the morphologic anomalies correlated to chromosomal aberrations. When the conditions for carrying out a cytogenetic study do not exist, these chromosomal alterations can be investigated by nucleic acid fluorescent in situ hybridization (FISH), which can be performed on interphase nuclei, usually on formalin-fixed paraffin embedded tissues or on fresh cytological specimens. The objective of the present study is to prove whether this technique can be successfully applied to formalin-fixed cell touch preparations and cytological specimens obtained from foetal autopsies. The study was carried out 12 abortions some of which were spontaneous and some of which were therapeutic. The materials were formalin-fixed. Cell touch preparations and cytological specimens were obtained. The FISH was performed using X/Y probes (Vysis) and the Aneuvysion Kit (05J38-030, Vysis), the probes being for chromosomes 13/21 and X/Y/18. To verify the reliability of the technique, the same reactions were also performed on fresh analogous materials. The slides were evaluable, and the probes hybridized to interphase nuclei showed distinct signals. All the samples were adequate for FISH analysis without any notable difference in the results. Moreover, it is technically possible to perform this analysis not only on fresh but particularly on formalin-fixed cytological specimens. On the other hand, the use of this type of cytological samples, as compared to formalin-fixed and paraffin embedded tissue sections, has the advantage of presenting intact, noncut nuclei with preserved cytomorphology, avoiding the problems of overlapping nuclei and making the identification of the real chromosomal arrangement easier.


[IPEX syndrome and human Treg cells].
CD4(+)CD25(+) T cells which have also been described as regulatory T cells (Treg), have immune inhibitory functions in the immune system. This population inhibits excessive immune responses, such as those present in patients with autoimmune disease, allergy and inflammation, and plays an important role in maintenance of immunological homeostasis. It has been demonstrated that the FOXP3 gene is a master gene for a transcriptional factor of Tregs. This finding has led to the elucidation of the Treg functions during development, differentiation and immune suppression. Either a deficiency or dysfunction of Tregs results in IPEX (immune dysregulation, polyendocrinopathy, enteropathy, and X-linked) syndrome. The clinical features of IPEX syndrome include chronic dermatitis, enteropathy characterized by severe and refractory diarrhea, and autoimmune endocrinopathy, such as early-onset insulin-dependent diabetes mellitus, thyroiditis, or both. This syndrome is also associated with various symptoms such as anemia, thrombocytopenia and nephritis which may be caused by an autoimmune response. We herein describe the clinical and molecular characteristics of patients with IPEX syndrome and also elucidate the function of human Treg cells.


Morphological, biological and molecular characterization of three strains of Trypanosoma cruzi Chagas, 1909 (Kinetoplastida, Trypanosomatidae) isolated from Triatoma sordida (Stal) 1859 (Hemiptera, Reduviidae) and a domestic cat.
A study was conducted of the biological, morphological and molecular characters of 3 strains of Trypanosoma cruzi (SI(5), SI(8) and SIGR(3)) isolated from specimens of Triatoma sordida collected in Santo Inácio and a domestic cat. In order to carry out the study, the following parameters were evaluated: pre-patent period, parasitaemia curves, morphology of the parasites, mortality rates, histopathological lesions and molecular typing. The strains presented variable pre-patent periods, low parasitaemia and no animal mortality. The morphological study of trypomastigotes showed a predominance of intermediate-width and short-length forms, as well as low nuclear index. Epimastigotes presented a low nuclear index, intermediate-width forms in strains SI(5) and SI(8), and large-width forms in SIGR(3). A shorter length could be noted in strains SI(8) and SIGR3, whereas SI(5) displayed an intermediate length. The histopathological study did not detect amastigote nests in tissues. The amplification of the divergent domain of 24Sα rRNA, HSP60 and GPI genes of strains SI(5), SI(8) and SIGR(3) classified the 3 strains into Group II. Biological parameters made it possible to classify the strains isolated in Santo Inácio (BA) into Biodeme III, Zymodeme 1 and Group II of T. cruzi.


The influence of a single acetic acid and lactic acid injection on rat normal thyroid tissue.
Until now, the use of acetic and lactic acid for the treatment of thyroid nodular lesions, as well as histopathologic changes in the thyroid tissue after their intrathyroidal injections has not been described. The aim of this study is to evaluate macro- and microscopic changes in rat normal thyroid tissue induced by single intrathyroidal injection of acetic acid (AA) and lactic acid (LA) in different concentrations, as well as the influence of intrathyroidal acid injection on function of the recurrent laryngeal nerves. Male Wistar rats were divided into two groups of 50 (Group A and B). Rats of each group were divided into the subgroups of five. In each subgroup of Group A and B, AA and LA water solutions of different concentrations (0%, 2.5%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 80%), respectively, were injected into the right thyroid lobe. From Group A and B, 7 and 8 rats died, respectively (after injections of 50-80% AA and LA solutions). After 4 weeks post AA or LA injection the thyroid along with the surrounding tissues was excised and assessed macro- and microscopically in all rats that survived. Before the injection and just before the second operation, the vocal cord function was evaluated. Our results show that both, acetic and lactic acid of different concentrations damage rat normal thyroid tissue after a single intrathyroidal injection. The minimal concentrations of acetic and lactic acid that cause lethal changes in rat normal thyroid tissue are 20% and 10%, respectively. Acetic and lactic acid in the concentrations of 40% and 50%, respectively, and higher can cause injury of the tissues adjacent to the thyroid.


A novel pathway for sensory-mediated arousal involves splicing of an intron in the period clock gene.
D. melanogaster is an excellent animal model to study how the circadian (≅24-h) timing system and sleep regulate daily wake-sleep cycles. Splicing of a temperature-sensitive 3'-terminal intron (termed dmpi8) from the circadian clock gene period (per) regulates the distribution of daily activity in Drosophila. The role of dmpi8 splicing on daily behavior was further evaluated by analyzing sleep. Transgenic flies of the same genetic background but expressing either a wild-type recombinant per gene or one where the efficiency of dmpi8 splicing was increased were exposed to different temperatures in daily light-dark cycles and sleep parameters measured. In addition, transgenic flies were briefly exposed to a variety of sensory-mediated stimuli to measure arousal responses. Surprisingly, we show that the effect of dmpi8 splicing on daytime activity levels does not involve a circadian role for per but is linked to adjustments in sensory-dependent arousal and sleep behavior. Genetically altered flies with high dmpi8 splicing efficiency remain aroused longer following short treatments with light and non-photic cues such as mechanical stimulation. We propose that the thermal regulation of dmpi8 splicing acts as a temperature-calibrated rheostat in a novel arousal mechanism, so that on warm days the inefficient splicing of the dmpi8 intron triggers an increase in quiescence by decreasing sensory-mediated arousal, thus ensuring flies minimize being active during the hot midday sun despite the presence of light in the environment, which is usually a strong arousal cue for diurnal animals.


Late-onset muscle weakness in partial phosphofructokinase deficiency: a unique myopathy with vacuoles, abnormal mitochondria, and absence of the common exon 5/intron 5 junction point mutation.
Three patients (ages 51, 59, and 79) from two generations of an Ashkenazi Jewish family had partial (33% activity) phosphofructokinase (PFK) deficiency that presented with fixed muscle weakness after the age of 50 years. MR spectroscopy revealed accumulation of phosphomonoesters during exercise. Muscle biopsy showed a vacuolar myopathy with increased autophagic activity and several ragged-red and cytochrome c oxidase-negative fibers. The older patient, age 79 at biopsy, had several necrotic fibers. Electron microscopy revealed subsarcolemmal and intermyofibrillar glycogen accumulation and proliferation of mitochondria with paracrystalline inclusions, probably related to reduced availability of energy due to impaired glycolysis. The common point mutation of exon 5/intron 5 junction seen in Jewish Ashkenazi patients with PFK deficiency was excluded. We conclude that late-onset fixed muscle weakness occurs in partial PFK deficiency and it may represent the end result of continuing episodes of muscle fiber destruction. Partial enzyme deficiency in two successive generations suggests a unique molecular mechanism.


Pulmonary-renal syndrome in systemic sclerosis.
Renal failure, pulmonary hypertension, and interstitial lung disease are major causes of morbidity and mortality in systemic sclerosis (SSc). However, the concomitant occurrence of pulmonary hemorrhage associated with acute renal failure in SSc has been rarely described. The present study is the first analysis of pulmonary-renal syndrome in SSc. We present a 44-year-old woman with SSc who died of a fulminant course of acute renal failure associated with diffuse alveolar hemorrhage. We termed this uncommon and fatal complication of SSc scleroderma-pulmonary-renal syndrome (SPRS). A search of the English-written literature yielded reports of 10 additional similar cases. These patients, together with our present case, form the basis of the present analysis. The average age of the patients with SPRS was 46 years. The majority of the patients (80%) were women, and most had diffuse SSc. SPRS occurred an average of 6.4 years after disease onset and was associated with prior fibrosing alveolitis and/or D-penicillamine treatment. Interestingly, normotensive renal failure seems to characterize the scleroderma patients, because 9 of 11 (82%) had normal blood pressure. SPRS bears a poor prognosis: all of the 11 patients (100%) died within 12 months of admission. However, only 60% of the 5 patients for whom we have treatment data received corticosteroids. Pulmonary-renal syndrome is a rare but fatal complication of SSc. Because the treatment data are scarce and the prognosis is poor, aggressive treatment with pulse corticosteroids, cyclophosphamide, and possibly plasmapheresis is suggested.


Lowering Contact Resistances of Two-Dimensional Semiconductors by Memristive Forming.
Two-dimensional semiconductors have great potential for beyond-silicon electronics. However, because of the lack of controllable doping methods, Fermi level pinning, and van der Waals (vdW) gaps at the metal-semiconductor interfaces, these devices exhibit high electrical contact resistances, restricting their practical applications. Here, we report a general contact-resistance-lowering strategy by constructing vertical metal-semiconductor-metal memristor structures at the contact regions and setting them into a nonvolatile low-resistance state through a memristive forming process. Through this, we reduce the contact resistances of MoS2 field-effect transistors (FETs) by at least one order of magnitude and improve the on-state current densities of MoTe2 FETs by about two orders of magnitude. We also demonstrate that this strategy is applicable to other two-dimensional semiconductors, including MoSe2, WS2, and WSe2, and a variety of contact metals, including Au, Cu, Ni, and Pd. The good stability and universality indicate the great potential for technological applications.


Immune correlates of the Thai RV144 HIV vaccine regimen in South Africa.
One of the most successful HIV vaccines to date, the RV144 vaccine tested in Thailand, demonstrated correlates of protection including cross-clade V1V2 immunoglobulin G (IgG) breadth, Env-specific CD4+ T cell polyfunctionality, and antibody-dependent cellular cytotoxicity (ADCC) in vaccinees with low IgA binding. The HIV Vaccine Trials Network (HVTN) 097 trial evaluated this vaccine regimen in South Africa, where clade C HIV-1 predominates. We compared cellular and humoral responses at peak and durability immunogenicity time points in HVTN 097 and RV144 vaccinee samples, and evaluated vaccine-matched and cross-clade immune responses. At peak immunogenicity, HVTN 097 vaccinees exhibited significantly higher cellular and humoral immune responses than RV144 vaccinees. CD4+ T cell responses were more frequent in HVTN 097 irrespective of age and sex, and CD4+ T cell Env-specific functionality scores were higher in HVTN 097. Env-specific CD40L+ CD4+ T cells were more common in HVTN 097, with individuals having this pattern of expression demonstrating higher median antibody responses to HIV-1 Env. IgG and IgG3 binding antibody rates and response magnitude to gp120 vaccine- and V1V2 vaccine-matched antigens were higher or comparable in HVTN 097 than in RV144 ADCC, and ADCP functional antibody responses were elicited in HVTN 097. Env-specific IgG and CD4+ Env responses declined significantly over time in both trials. Overall, cross-clade immune responses associated with protection were better than expected in South Africa, suggesting wider applicability of this regimen.


Computer errors in electrocardiography.
Computer interpretation of electrocardiograms is far from adequate. This paper highlights a particular type of computer error that is merely symbolic of the large number of errors the computer can make. In addition to computer errors in electrocardiography, it is likely that computer interpretation has diminished the ability of physicians to interpret electrocardiograms.


Presentation, Diagnostic Testing and Initial Treatment of Vitreoretinal Lymphoma.
Vitreoretinal lymphoma is a malignancy with high mortality. Incidence is rare, and there is a lack of medical evidence to direct management. This work describes presentation, diagnostic testing, and first treatment approaches in a recently diagnosed and treated patient cohort. Clinical registry-based observational study. Forty-eight women and 32 men (age range, 32-91 years; median age, 64 years) diagnosed with vitreoretinal lymphoma. An international network of ophthalmologists reported clinical features and management of patients presenting with vitreoretinal lymphoma between January 1, 2020 and December 31, 2022 via an electronic platform. Visual acuity at presentation (logarithm of the minimum angle of resolution [logMAR]); basis for diagnosis; first treatment. Vitreoretinal lymphoma was bilateral at presentation in 65% of patients (n = 52) and an initial site of lymphoma in 78% (n = 62). Of 127 eyes with lymphoma at presentation, vitreous was involved in 89% (n = 113) and was the only involved eye tissue in 40% (n = 51), and retina was involved in 46% (n = 59) and was the only involved eye tissue in 9% (n = 11). Median logMAR visual acuity of the worse-seeing eye was 0.50. The lymphoma was diagnosed from ocular specimens in 80% of patients (64/80), usually vitreous (57/64 patients [89%]), and on other clinical information in 20% of patients (16/80). Cellular studies were performed on ocular specimens from 59 of 64 patients (92%), most often cytology. Tumor gene analysis was used in 21 of 64 patients (33%), and cytokine assays were used in 13 of 64 patients (20%). For 76 patients (95%), treatment was initiated within 6 months of diagnosis and included ocular (38/76 [48%]), extraocular (17/76 [21%]), and ocular plus extraocular (21/76 [26%]) approaches. Intravitreal methotrexate was the most common ocular treatment (83/87 eyes [95%]). Using data collected from 80 patients diagnosed with vitreoretinal lymphoma since 2020, we show that visual impairment is common, and that management often involves diagnosis by cellular tests and treatment with intravitreal chemotherapy. Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.


An outbreak of orthopoxvirus-associated disease in horses in southern Brazil.
An outbreak of severe cutaneous disease associated with an orthopoxvirus in horses in southern Brazil is described. Fourteen Crioulo mares and foals from a husbandry farm developed papules, and vesicles progressing to proliferative and exudative lesions on the muzzle, external nares, and external and internal lips. The vesicles eroded, and the proliferative lesions eventually bled and progressed to moist crusts and scars. The clinical signs lasted approximately 6-12 days, after which the animals progressively recovered. Direct electron microscopy of skin biopsies revealed brick-shaped, 250-300-nm virus particles with orthopoxvirus morphology. Histological examination of the lesions revealed vacuolar degeneration of the cells of the stratum spinosum and the presence of large intracytoplasmic, eosinophilic inclusion bodies. Polymerase chain reaction amplification of poxvirus A-type inclusion body gene confirmed the presence of orthopoxvirus DNA in horse tissues. Inoculation of tissue homogenates into the chorioallantoic membrane of chicken embryonated eggs and intraperitoneally in mice resulted in pock-like lesions with the microscopic appearance of poxvirus-induced histopathology. Taken together, these results demonstrate the association of an orthopoxvirus with the outbreak of cutaneous disease in horses. The origin of the agent causing the outbreak is uncertain because no similar condition has been reported in Brazil.


Effect of corticotropin-releasing factor on the pituitary-ovary axis in human luteal phase.
In order to test the possible negative effect of corticotropin-releasing factor (CRF) on the pituitary-gonadal axis, two gonadotropin-releasing hormone (GnRH) tests, on two consecutive days, were performed in five normally cycling women in the midluteal phase of the menstrual cycle. The first GnRH test was performed after 2 h during which time basal blood samples were collected every 10 min. On the second day, the GnRH test was repeated after a 2-h CRF infusion which was continued throughout the GnRH test. Our results demonstrate that while CRF increases cortisol levels it affects neither the basal nor the GnRH-stimulated gonadotropin levels. Similarly, both basal and luteinizing hormone (LH)-stimulated progesterone concentrations were not modified by the CRF infusion thereby also excluding a possible direct effect of the peptide on corpus luteum steroidogenesis.


The chi-square test of independence.
The Chi-square statistic is a non-parametric (distribution free) tool designed to analyze group differences when the dependent variable is measured at a nominal level. Like all non-parametric statistics, the Chi-square is robust with respect to the distribution of the data. Specifically, it does not require equality of variances among the study groups or homoscedasticity in the data. It permits evaluation of both dichotomous independent variables, and of multiple group studies. Unlike many other non-parametric and some parametric statistics, the calculations needed to compute the Chi-square provide considerable information about how each of the groups performed in the study. This richness of detail allows the researcher to understand the results and thus to derive more detailed information from this statistic than from many others. The Chi-square is a significance statistic, and should be followed with a strength statistic. The Cramer's V is the most common strength test used to test the data when a significant Chi-square result has been obtained. Advantages of the Chi-square include its robustness with respect to distribution of the data, its ease of computation, the detailed information that can be derived from the test, its use in studies for which parametric assumptions cannot be met, and its flexibility in handling data from both two group and multiple group studies. Limitations include its sample size requirements, difficulty of interpretation when there are large numbers of categories (20 or more) in the independent or dependent variables, and tendency of the Cramer's V to produce relative low correlation measures, even for highly significant results.


Cavitary lung cancer lined with normal bronchial epithelium and cancer cells.
Reports of cavitary lung cancer are not uncommon, and the cavity generally contains either dilated bronchi or cancer cells. Recently, we encountered a surgical case of cavitary lung cancer whose cavity tended to enlarge during long-term follow-up, and was found to be lined with normal bronchial epithelium and adenocarcinoma cells.


Mutational tuning of galectin-3 specificity and biological function.
Galectins are defined by a conserved β-galactoside binding site that has been linked to many of their important functions in e.g. cell adhesion, signaling, and intracellular trafficking. Weak adjacent sites may enhance or decrease affinity for natural β-galactoside-containing glycoconjugates, but little is known about the biological role of this modulation of affinity (fine specificity). We have now produced 10 mutants of human galectin-3, with changes in these adjacent sites that have altered carbohydrate-binding fine specificity but that retain the basic β-galactoside binding activity as shown by glycan-array binding and a solution-based fluorescence anisotropy assay. Each mutant was also tested in two biological assays to provide a correlation between fine specificity and function. Galectin-3 R186S, which has selectively lost affinity for LacNAc, a disaccharide moiety commonly found on glycoprotein glycans, has lost the ability to activate neutrophil leukocytes and intracellular targeting into vesicles. K176L has increased affinity for β-galactosides substituted with GlcNAcβ1-3, as found in poly-N-acetyllactosaminoglycans, and increased potency to activate neutrophil leukocytes even though it has lost other aspects of galectin-3 fine specificity. G182A has altered carbohydrate-binding fine specificity and altered intracellular targeting into vesicles, a possible link to the intracellular galectin-3-mediated anti-apoptotic effect known to be lost by this mutant. Finally, the mutants have helped to define the differences in fine specificity shown by Xenopus, mouse, and human galectin-3 and, as such, the evidence for adaptive change during evolution.


Molecular voids formed from effective attraction in submonolayer DNA deposited on Au(111).
The development of DNA-based biosensors requires a deep understanding of how DNA molecules adsorb and organize on solid state surfaces as well as the electronic properties of individual and aggregates of DNA molecules. Using scanning tunneling microscopy (STM) and atomic force microscopy (AFM), we have successfully characterized an attractive force driven molecular void formation for DNA chemically adsorbed on Au(111) as a function of strand length and deposition conditions. Here we report the observation of these void structures formed on the Au(111) surface by adsorption of both 45 and 90 base pair long, thiolated double-stranded DNA. We found that the average void diameter decreases when increasing the number of base pairs exposed to the surface. The critical determinant in the molecular void formation is the total charge delivered to the surface via the adsorption of the DNA strands and the related counterions, which can ultimately be quantified by the number of base pairs in each adsorbed DNA molecule. Complementary measurements involving STM and AFM suggest that an intact Au(111) surface area is preserved inside the void and is surrounded by a submonolayer of DNA molecules adsorbed on the surface. The discussion of the possible mechanisms for the void formation implies an effective attraction between the DNA molecules.