Productivity of Three Pea (Pisum sativum L.) Varieties as Influenced by Nutrient Supply and Meteorological Conditions in Boreal Environmental Zone.
In order to grow crops that reduce the negative impact on the environment, as well as meet the nutritional needs of the increasing human population, it is necessary to include new and more sustainable production strategies into current agricultural systems. The aim of our study was to evaluate the optimal nutritional conditions of semi-leafless pea productivity and ascertain the influence of meteorological factors on the productivity of these plants under boreal environmental conditions. The test involved three semi-leafless pea varieties, one of which was a new variety, and eight N fertilization treatments were used: (1) without fertilizers (N0), (2) without N fertilizers (N0), (3) N15, (4) N30, (5) N45, (6) N15+15, (7) N60, and (8) N60. Plots of the second-seventh treatment received a base application of P40K80; the eighth treatment received P80K160. Fertilizer efficiency depended on the meteorological conditions. Based on their productivity, the pea varieties were arranged in the following descending order: Ieva DS ˃ Respect ˃ Simona. Compared with unfertilized peas, NPK fertilizers enhanced the seed yield by 10.6-12.9% on average. Splitting the N30 rate and applying N60, under a background of P40K80, was not efficient. The optimal rate of N15-45P40K80 fertilizers for peas was determined. Meteorological factors significantly influenced seed yield by 75.2%, 44.1%, and 79.9% for varieties Ieva DS, Simona, and Respect, respectively.


Factors contributing to amplitude variability of the steady-state visual evoked response.
Ten steady-state visual evoked responses (VER's) were recorded from each of 47 normal, adult subjects. For each subject, the mean and SD for the 10 VER amplitudes were calculated and used to determine amplitude variability. Although some subjects produced extremely reliable VER's, data from the majority showed a considerable degree of variability. A number of factors including trend (the tendency of the VER amplitudes to increase or decrease with repeated measurements), noise, attention, binocularity, accommodation, eye movements, artifacts, and electrode placement were evaluated to determine their relative contributions to this variability. Noise and trend factors produced a large proportion of the variability (75%) whereas the other factors were found to be relatively insignificant.


[Research progress on mechanism of acupuncture and moxibustion in treatment of irritable bowel syndrome].
The current clinical evidence and underlying mechanisms of acupuncture and moxibustion in the treatment of irritable bowel syndrome (IBS) were summarized, so as to better optimize clinical treatment. The relevant articles of acupuncture and moxibustion in the treatment of IBS in recent years were retrieved and summarized. We found that the clinical efficacy of acupuncture and moxibustion in the treatment of IBS was relatively reliable. However, the mutual relationships among various mechanisms of action such as abnormal gastrointestinal motility, high visceral sensitivity, intestinal microenvironment disorders, and abnormal intestinal-brain interactions need to be further explored. The authors believe that in-depth explorations of the bidirectional regulation of "gut-brain axis", the law of changes in the abundance and diversity of intestinal flora, and the establishment of a more ideal animal model of TCM syndrome differentiation are useful ideas for subsequent research.


The 40-kDa component of the phagocyte NADPH oxidase (p40phox) is phosphorylated during activation in differentiated HL60 cells.
The superoxide-generating NADPH oxidase complex of phagocytic cells is a multicomponent system containing a membrane-bound flavocytochrome b and a small G protein Rac as well as cytosolic factors p67phox, p47phox and p40phox which translocate to the membrane upon activation. Known mechanisms underlying the translocation of these proteins include polyphosphorylation of p47phox and specific Src homology 3/polyproline motif interactions. In this study, through two-dimensionnal electrophoresis and immunoprecipitation experiments, we show using dimethylsulfoxide-differentiated HL60 promyelocytes that p40phox is in a basal phosphorylated state in resting cells and undergoes further phosphorylation on multiple sites upon stimulation of the NADPH oxidase by either phorbol myristate acetate or by the formyl peptide fMet-Leu-Phe-Lys. Moreover, the extent of phosphorylation is strongly correlated with the level of superoxide production. Typically, in cells transiently activated by fMet-Leu-Phe-Lys, onset of superoxide production coincides with the appearance of new phosphorylated species of p40phox and, at the end of the respiratory burst, dephosphorylation of p40phox is observed. In vitro assays show that the kinase(s) involved in the phosphorylation of p40phox differ from those which participate in the phosphorylation of p47phox. This suggests that, in the cell, the phosphorylation of p40phox and of p47phox are under the control of two different kinase pathways.


Differentiation of peripheral nerve sheath tumors in patients with neurofibromatosis type 1 using diffusion-weighted magnetic resonance imaging.
We sought to determine the value of diffusion-weighted (DW) magnetic resonance imaging (MRI) for characterization of benign and malignant peripheral nerve sheath tumors (PNSTs) in patients with neurofibromatosis type 1 (NF1). Twenty-six patients with NF1 and suspicion of malignant transformation of PNSTs were prospectively enrolled and underwent DW MRI at 3T. For a set of benign (n = 55) and malignant (n = 12) PNSTs, functional MRI parameters were derived from both biexponential intravoxel incoherent motion (diffusion coefficient D and perfusion fraction f) and monoexponential data analysis (apparent diffusion coefficients [ADCs]). A panel of morphological MRI features was evaluated using T1- and T2-weighted imaging. Mann-Whitney U-test, Fisher's exact test, and receiver operating characteristic (ROC) analyses were applied to assess the diagnostic accuracy of quantitative and qualitative MRI. Cohen's kappa was used to determine interrater reliability. Malignant PNSTs demonstrated significantly lower diffusivity (P < 0.0001) compared with benign PNSTs. The perfusion fraction f was significantly higher in malignant PNSTs (P < 0.001). In ROC analysis, functional MRI parameters showed high diagnostic accuracy for differentiation of PNSTs (eg, ADCmean, 92% sensitivity with 98% specificity, AUC 0.98; Dmean, 92% sensitivity with 98% specificity, AUC 0.98). By contrast, morphological imaging features had only limited sensitivity (18-94%) and specificity (18-82%) for identification of malignancy. Interrater reliability was higher for monoexponential data analysis. DW imaging shows better diagnostic performance than morphological features and allows accurate differentiation of benign and malignant peripheral nerve sheath tumors in NF1.


Haemodynamic effects of pindolol in essential hypertension with special reference to resistance vessels in the forearm during maximal vasodilatation.
Eleven patients, mean age 45 years, with previously untreated hypertension (WHO stage I) were treated over 16 months with pindolol in dosages of 5-30 mg/day. The effects on blood pressure and peripheral vascular resistance (PR) were studied before and after 2 and 16 months of treatment. Recordings were performed at rest, and after maximal vasodilatation obtained by ischaemic work to exhaustion with a hand-ergometer. Blood pressure was significantly reduced after both 2 and 16 months of treatment (P < 0.001). During treatment small reductions in heart rate were seen at rest but not after maximal vasodilatation (P < 0.02 and NS, respectively). At rest there was a great variability and no significant changes in PR. Peripheral vascular resistance after maximal vasodilatation was significantly reduced after both 2 and 16 months of treatment (P < 0.005 and P < 0.01, respectively). We conclude that long-term treatment with pindolol may give a regression of structural vascular changes.


Plasma Metabolite Profiles of Red Meat, Poultry, and Fish Consumption, and Their Associations with Colorectal Cancer Risk.
Red and processed meat consumption has been consistently associated with increased risk of colorectal cancer (CRC), but the association for fish intake is unclear. Evidence using objective dietary assessment approaches to evaluate these associations is sparse. We aim to investigate the plasma metabolite profiles related to red meat, poultry, and fish consumption and examine their associations with CRC risk. We measured plasma metabolites among 5269 participants from the Nurses' Health Study (NHS), NHSII, and Health Professionals Follow-Up study (HPFS). We calculated partial Spearman correlations between each metabolite and self-reported intake of seven red meat, poultry, and fish groups. Metabolite profile scores correlated to self-reported dietary intakes were developed using elastic net regression. Associations between self-reported intakes, metabolite profile scores, and subsequent CRC risk were further evaluated using conditional logistic regression among 559 matched (1:1) case-control pairs in NHS/HPFS and replicated among 266 pairs in Women's Health Study. Plasma metabolites, especially highly unsaturated lipids, were differentially associated with red meat and fish groups. Metabolite profile scores for each food group were significantly correlated with the corresponding self-reported dietary intake. A higher dietary intake of processed red meat was associated with a higher risk of CRC (pooled OR per 1 SD, 1.15; 95% CI: 1.03, 1.29). In contrast, higher metabolite profile scores for all fish groups, not dietary intakes, were consistently associated with a lower CRC risk: the pooled OR per 1 SD was 0.86 (95% CI: 0.78, 0.96) for total fish, 0.86 (95% CI: 0.77, 0.96) for dark meat fish, and 0.87 (95% CI: 0.78, 0.97) for canned tuna fish. No significant associations were found for other food groups. Red meat and fish intake exhibited systematically different plasma metabolite profiles. Plasma metabolite profile of fish intake was inversely associated with CRC risk.


Dimensions of quality of life and self-monitoring therapy with oral anticoagulants--still research or everyday practice?
In correlation with increased life expectancy of patients, quality of life (QOL) has become a factor of increasing interest by the patient himself and also of importance in health-care planning and recruitment of financial resources. In this context, self-monitoring of long-term anticoagulant treatment might be a strategy that could mean a step forward in health-related as well as general life satisfaction for patients participating in self-monitoring programs. Also, the new strategy of increased home-control of anticoagulant treatment illustrates the complexity of multiple factors that can lead to changes in the subjective feeling and objective aspects of QOL. Our intention in a pilot study was to probe the feasibility of QOL research and relevant factors of influence by retrospectively evaluating data from two groups of outpatients seen in a large treatment center. The high frequency (n = 8 in sample 2) of disturbed sleep as a simple screening indicator stresses the probable importance of undetected depression, which might require treatment and could confound research as to QOL. Instruments to measure QOL in oral anticoagulation self-monitoring should therefore be adapted to the heterogeneous structure of factors in the target population, and include psychological parameters, especially in regard to health-related locus of control and mood.


Determinants of the inhibition of a Taiwan habu venom metalloproteinase by its endogenous inhibitors revealed by X-ray crystallography and synthetic inhibitor analogues.
Venoms from crotalid and viperid snakes contain several peptide inhibitors which regulate the proteolytic activities of their snake-venom metalloproteinases (SVMPs) in a reversible manner under physiological conditions. In this report, we describe the high-resolution crystal structures of a SVMP, TM-3, from Taiwan habu (Trimeresurus mucrosquamatus) cocrystallized with the endogenous inhibitors pyroGlu-Asn-Trp (pENW), pyroGlu-Gln-Trp (pEQW) or pyroGlu-Lys-Trp (pEKW). The binding of inhibitors causes some of the residues around the inhibitor-binding environment of TM-3 to slightly move away from the active-site center, and displaces two metal-coordinated water molecules by the C-terminal carboxylic group of the inhibitors. This binding adopts a retro-manner principally stabilized by four possible hydrogen bonds. The Trp indole ring of the inhibitors is stacked against the imidazole of His143 in the S-1 site of the proteinase. Results from the study of synthetic inhibitor analogues showed the primary specificity of Trp residue of the inhibitors at the P-1 site, corroborating the stacking effect observed in our structures. Furthermore, we have made a detailed comparison of our structures with the binding modes of other inhibitors including batimastat, a hydroxamate inhibitor, and a barbiturate derivative. It suggests a close correlation between the inhibitory activity of an inhibitor and its ability to fill the S-1 pocket of the proteinase. Our work may provide insights into the rational design of small molecules that bind to this class of zinc-metalloproteinases.


Histamine dihydrochloride and interleukin-2 in the treatment of acute myeloid leukemia.
Despite successful chemotherapeutic remission induction, most patients with acute myeloid leukemia still are destined to die from recurrent and refractory disease. Strategies to prolong remission and improve survival include high-dose chemotherapy, autologous or allogeneic stem cell transplantation, and other immunotherapeutic approaches. The focus on interleukin-2 (IL-2) has arisen from in vitro demonstration of enhancement of tumor-specific cytotoxic T-lymphocyte and natural killer-cell activity after cytokine activation. The local secretion of inhibitory substances and oxidative stress by monocyte/macrophage populations may inhibit the cytotoxic capacity of these effector cells. Recently, agents capable of modulating cytotoxic effector function have been identified, including histamine dihydrochloride, which has been shown to act as an inhibitor of cellular-mediated free-radical production. In the presence of histamine, cytotoxic effector function may be restored. The addition of histamine to IL-2-based cytokine therapies has resulted in enhanced in vitro cytotoxic T-cell and natural killer-cell function and has allowed for the development of lower, and thus, less toxic, IL-2 regimens. Preliminary studies of this combination in patients with acute myeloid leukemia in remission suggest improved survival with tolerable toxicity. The strategy for implementation of IL-2-based immunotherapy in patients with acute myeloid leukemia is reviewed.


Metabolic Engineering of the Native Monoterpene Pathway in Spearmint for Production of Heterologous Monoterpenes Reveals Complex Metabolism and Pathway Interactions.
Spearmint produces and stores large amounts of monoterpenes, mainly limonene and carvone, in glandular trichomes and is the major natural source of these compounds. Towards producing heterologous monoterpenes in spearmint, we first reduced the flux into the native limonene pathway by knocking down the expression of limonene synthase (MsLS) by RNAi method. The MsLS RNAi lines exhibited a huge reduction in the synthesis of limonene and carvone. Detailed GC-MS and LC-MS analysis revealed that MsLS RNAi plants also showed an increase in sesquiterpene, phytosterols, fatty acids, flavonoids, and phenolic metabolites, suggesting an interaction between the MEP, MVA shikimate and fatty acid pathways in spearmint. Three different heterologous monoterpene synthases namely, linalool synthase and myrcene synthase from Picea abies and geraniol synthase from Cananga odorata were cloned and introduced independently into the MsLS RNAi mutant background. The expression of these heterologous terpene synthases resulted mainly in production of monoterpene derivatives. Of all the introduced monoterpenes geraniol showed the maximum number of derivatives. Our results provide new insights into MEP pathway interactions and regulation and reveals the existence of mechanisms for complex metabolism of monoterpenes in spearmint.


PEDF Is Associated with the Termination of Chondrocyte Phenotype and Catabolism of Cartilage Tissue.
Objective. To investigate the expression and target genes of pigment epithelium-derived factor (PEDF) in cartilage and chondrocytes, respectively. Methods. We analyzed the expression pattern of PEDF in different human cartilaginous tissues including articular cartilage, osteophytic cartilage, and fetal epiphyseal and growth plate cartilage, by immunohistochemistry and quantitative real-time (qRT) PCR. Transcriptome analysis after stimulation of human articular chondrocytes with rhPEDF was performed by RNA sequencing (RNA-Seq) and confirmed by qRT-PCR. Results. Immunohistochemically, PEDF could be detected in transient cartilaginous tissue that is prone to undergo endochondral ossification, including epiphyseal cartilage, growth plate cartilage, and osteophytic cartilage. In contrast, PEDF was hardly detected in healthy articular cartilage and in the superficial zone of epiphyses, regions that are characterized by a permanent stable chondrocyte phenotype. RNA-Seq analysis and qRT-PCR demonstrated that rhPEDF significantly induced the expression of a number of matrix-degrading factors including SAA1, MMP1, MMP3, and MMP13. Simultaneously, a number of cartilage-specific genes including COL2A1, COL9A2, COMP, and LECT were among the most significantly downregulated genes. Conclusions. PEDF represents a marker for transient cartilage during all neonatal and postnatal developmental stages and promotes the termination of cartilage tissue by upregulation of matrix-degrading factors and downregulation of cartilage-specific genes. These data provide the basis for novel strategies to stabilize the phenotype of articular cartilage and prevent its degradation.


Neurofilament light chain as a disease severity biomarker in ATTRv: data from a single-centre experience.
Hereditary transthyretin amyloidosis (ATTRv) is a treatable multisystem disorder with prevalent peripheral nervous system impairment. Besides neurophysiological measures, there are few markers to monitor disease progression. Neurofilament light chain (NfL) has recently been considered a sensitive biomarker for neuroaxonal damage in this setting. To evaluate NfL levels in a cohort of ATTRv patients and pre-symptomatic carriers and correlate the serum concentrations with other markers of disease severity. We analysed NfL serum from 17 ATTRv patients or carriers and 26 controls. An exhaustive clinical and instrumental evaluation was performed in all patients. NfL levels were significantly higher in ATTRv cases when compared with controls. A significant correlation was found between NfL values and NIS scale, Sudoscan values from feet, interventricular septum thickness, and Quality of Life-Diabetic Neuropathy (Norfolk QoL-DN) questionnaire. We confirm that NfL is a reliable and promising biomarker to evaluate the ATTRv severity and monitor its progression.


Evidence of hydrogen embrittlement of tungsten carbide.
Tungsten carbide vessels containing materials at high temperature and high pressure are used in many laboratories. We note that any oils at medium to high temperature which can break down and liberate hydrogen cause rapid failure of the pressure vessel, whereas perfluorated kerosenes used as lubricants inside a pressure vessel give sharply increased life of the vessel.


Dynamics of genomic change during evolutionary rescue in the seed beetle Callosobruchus maculatus.
Rapid adaptation can prevent extinction when populations are exposed to extremely marginal or stressful environments. Factors that affect the likelihood of evolutionary rescue from extinction have been identified, but much less is known about the evolutionary dynamics (e.g., rates and patterns of allele frequency change) and genomic basis of successful rescue, particularly in multicellular organisms. We conducted an evolve-and-resequence experiment to investigate the dynamics of evolutionary rescue at the genetic level in the cowpea seed beetle, Callosobruchus maculatus, when it is experimentally shifted to a stressful host plant, lentil. Low survival (~1%) at the onset of the experiment caused population decline. But adaptive evolution quickly rescued the population, with survival rates climbing to 69% by the F5 generation and 90% by the F10 generation. Population genomic data showed that rescue likely was caused by rapid evolutionary change at multiple loci, with many alleles fixing or nearly fixing within five generations of selection on lentil. Selection on these loci was only moderately consistent in time, but parallel evolutionary changes were evident in sublines formed after the lentil line had passed through a bottleneck. By comparing estimates of selection and genomic change on lentil across five independent C. maculatus lines (the new lentil-adapted line, three long-established lines and one case of failed evolutionary rescue), we found that adaptation on lentil occurred via somewhat idiosyncratic evolutionary changes. Overall, our results suggest that evolutionary rescue in this system can be caused by very strong selection on multiple loci driving rapid and pronounced genomic change.


Reappraisal of thromboangiitis obliterans--a pathological contribution.
Thromboangiitis obliterans (TAO), the Winiwarter-Buerger disease, is a vasoocclusive disease of unknown etiology which typically affects medium-sized extremital vessels of young male smokers. While the diagnosis of TAO is largely based on patients' presentation and clinical criteria, pathological substrates have been poorly defined and repeatedly disputed. Comparing the histology and immunohistochemistry of TAO especially with those of arteriosclerosis obliterans (ASO) and thromboembolism in two larger studies, we recently identified several features with significant meaning for the differential diagnosis. The unique tissue appearance of TAO is in favor of a general disorder of minute vessels indicating periarteritis rather than pure endarteritis, whereas an inflammatory reaction directed to the internal elastic lamina corresponds with the severity of disease. Since TAO or its syndromic equivalent is probably more common than currently diagnosed, the presented pathological criteria may help to identify latent cases, the overlap with ASO, and untypical organ involvement of TAO.


Modeling of wave fields generated by ultrasonic transducers using a quasi-Monte Carlo method.
The sound fields generated by ultrasonic transducers are modeled using the quasi-Monte Carlo (QMC) method, which is found to overcome the conflict between accuracy and efficiency that occurs in existing wave field calculation methods. The RI equation, which is frequently used as a model equation in ultrasonic field calculation, is used here as an exact method and for comparison purposes. In the QMC method, the judgment sampling method and Halton sequence are used for pseudo-random sampling from the sound source, and then the sound field distributions are found by solving the integral solution using the sample mean. Numerical examples and results are presented when modeling unfocused, focused, and steered and focused beam fields. The accuracy and efficiency of the QMC method are discussed by comparing the results obtained using different modeling methods. The results show that the proposed method has a high level of efficiency due to the nature of the QMC algorithm and a high level of accuracy because no approximation is required. In addition, wave fields can be modeled with the QMC method as long as sound sources can be effectively pseudo-randomly sampled, allowing the proposed method to be applied to various types of transducers.


The Effect of Heat Transfer and Polymer Concentration on Non-Newtonian Fluid from Pore-Scale Simulation of Rock X-ray Micro-CT.
Most of the pore-scale imaging and simulations of non-Newtonian fluid are based on the simplifying geometry of network modeling and overlook the fluid rheology and heat transfer. In the present paper, we developed a non-isothermal and non-Newtonian numerical model of the flow properties at pore-scale by simulation of the 3D micro-CT images using a Finite Volume Method (FVM). The numerical model is based on the resolution of the momentum and energy conservation equations. Owing to an adaptive mesh generation technique and appropriate boundary conditions, rock permeability and mobility are accurately computed. A temperature and concentration-dependent power-law viscosity model in line with the experimental measurement of the fluid rheology is adopted. The model is first applied at isothermal condition to 2 benchmark samples, namely Fontainebleau sandstone and Grosmont carbonate, and is found to be in good agreement with the Lattice Boltzmann method (LBM). Finally, at non-isothermal conditions, an effective mobility is introduced that enables to perform a numerical sensitivity study to fluid rheology, heat transfer, and operating conditions. While the mobility seems to evolve linearly with polymer concentration in agreement with a derived theoretical model, the effect of the temperature seems negligible by comparison. However, a sharp contrast is found between carbonate and sandstone under the effect of a constant temperature gradient. Besides concerning the flow index and consistency factor, a master curve is derived when normalizing the mobility for both the carbonate and the sandstone.


FIGO Statement: Vaccination in pregnancy.
Pregnant women and their fetuses are among the vulnerable populations that can be severely affected by communicable diseases. As such, some vaccines such as the influenza and the Tdap (tetanus toxoid, reduced diphtheria toxoid, and acellular pertussis) vaccines are strongly recommended in each pregnancy, with generally safe profiles. Other vaccines can be offered based on risk factors, and only when the benefits of receiving them outweigh the risks. Development of vaccines against group B streptococcus infection and respiratory syncytial virus infection are of great importance. In this paper, the recommendations for administration of each vaccine during pregnancy are discussed. The FIGO Committee for Safe Motherhood and Newborn Health Committee endorses the recommendations to vaccinate all pregnant women against influenza during the influenza season at any time during the pregnancy and against Tdap preferably between the 27th and 36th weeks of pregnancy in each pregnancy.


A probable case of multiple myeloma from Bronze Age China.
Paleopathological evidence of cancer from past populations is rare, especially outside of Europe and North Africa. This study expands upon the current temporal and spatial distribution of cancer by presenting a probable case of multiple myeloma from Bronze Age China. The human skeletal remains of an adult male from the Qijia culture horizon (1750-1400 BCE) of the Bronze Age cemetery of Mogou (), located in Gansu Province, Northwest China. The human skeletal remains were assessed macroscopically and radiographically using plain x-rays. Multiple ovoid-shaped osteolytic lesions with sharply demarcated margins were observed. The axial skeletal had the greatest involvement, specifically the vertebrae, ribs, and sternum. Radiographic imaging revealed more extensive destruction of cancellous than cortical bone, indicating that the marrow was the focal point of the disease. Based on the nature, distribution, and radiographic appearance of the lesions, the most likely diagnosis is multiple myeloma. This is one of the only cases of cancer identified in archaeological human skeletal remains from East Asia and is the first published case of a hematopoietic malignancy from mainland China. The analysis and publication of examples of neoplasia from areas that expand upon the current known temporal and spatial distribution is necessary in order to better reconstruct the history and evolution of cancer. Poor skeletal preservation prevented the full extent of osteolytic lesions to be observed. By placing case studies such as this into a temporal and spatial framework, it is possible for future research to begin to interrogate possible underlying causes of cancer in ancient populations within the context of changing environmental conditions and subsistence strategies.


Steroidal Glycosides from the Aerial Parts of Avena sativa L. and Their Cytotoxic Activity.
Twelve steroidal glycosides (1-12) were isolated from the aerial parts of Avena sativa L. (Poaceae). Among the isolated compounds, 1 was directly isolated from the plant for the first time, and 2-6 were new steroidal glycosides. The structures of 1-6 were determined by analysis of their spectroscopic data, chemical transformations, and chromatographic and spectroscopic analyses of the hydrolyzed products. Compounds 5 and 6 were novel steroidal glycosides with a B-ring contracted skeleton (B-nor steroid). Compounds 1, 9, 11, and 12 were cytotoxic to HL-60 human promyelocytic leukemia cells, MIA PaCa-2 human pancreatic carcinoma cells, and A549 human lung adenocarcinoma cells with IC50 values ranging from 0.79 to 13.5 μM. HL-60 cells treated with 1 exhibited apoptotic characteristics, namely, condensed nuclear chromatin, accumulation of sub-G1 cells, and activation of caspase-3. Additionally, the loss of the mitochondrial membrane potential and the release of cytochrome c into the cytoplasm in 1-treated HL-60 cells suggested that 1 induced apoptosis through a mitochondrial-dependent apoptotic pathway.


LINC00974 sponges miR-33a to facilitate cell proliferation, invasion, and EMT of ovarian cancer through HMGB2 upregulation.
The function and mechanism of long intergenic non-protein coding RNA 974 (LINC00974) are rarely reported in ovarian cancer (OC). The study aimed to investigate how LINC00974 affects the progression of OC. The expression levels of LINC00974, microRNA-33a (miR-33a), and high mobility group box 2 (HMGB2) mRNA were detected by qRT-PCR. The LINC00974/miR-33a/HMGB2 axis was confirmed by dual-luciferase reporter, RNA-binding protein immunoprecipitation (RIP), and biotinylated RNA pull-down assays. A series of in vitro experiments were employed to assess the effects of LINC00974/miR-33a/HMGB2 axis on the proliferation, invasion and epithelial mesenchymal transition (EMT) of OC cells. Results showed that LINC00974 and HMGB2 mRNA expression were upregulated in OC cells, while miR-33a expression was downregulated. HMGB2 was a direct target gene of miR-33a. LINC00974 act as a competing endogenous RNA (ceRNA) to directly bind with miR-33a, thereby upregulated HMGB2 expression. Notably, silencing of LINC00974 suppressed cell proliferation, invasion and EMT of OC cells, whereas miR-33a knockdown partially reversed the phenotypes of LINC00974 on OC cells. Overall, our study demonstrated that LINC00974 sponges miR-33a to promote cell proliferation, invasion, and EMT of OC through HMGB2 upregulation. LINC00974/miR-33a/HMGB2 axis may be an important signaling pathway in the progression of OC.


Intralesional treatment of cutaneous leishmaniasis with sodium stibogluconate antimony.
Cutaneous leishmaniasis represents a difficult disease to manage in endemic areas. Systemic treatment is hampered by both expense and compliance. Side effects may play a major role in this aspect as well. The effectiveness of intralesional treatment of leishmaniasis was investigated. Seven hundred and ten patients were treated with injections of sodium stibogluconate intralesionally. The clinical diagnosis was confirmed by demonstrating the parasite in the smears obtained from the lesion. Fine insulin needle was used to infiltrate the lesion with sodium stibogluconate (0.5 to 1.0 mL). Generally eight injections were sufficient, but some of the complicated lesions needed up to 24 injections. Sixty-two percent of patients were men. The majority of the study population (64%) were children below 15 years of age. The results showed that 72% of lesions healed completely, 23.9% showed some improvement, while 4.1% showed some deterioration. Lesions of the lips, cheeks, chin, and neck healed faster than lesions in other parts of the body. Side effects were mild and limited to pain at the site of the injection and hyperpigmentation in those who were treated by folk medicine. Intralesional treatment is as effective as the standard systemic antimonials. It offers a less expensive alternative and a low side effects profile. Our findings confirmed the findings of earlier workers. It is recommended for treatment of cutaneous leishmaniasis in endemic areas.


Effect of cyanide on the plasmalemma potential of mnium.
By centrifuging Mnium cuspidatum leaf cells, the cytoplasm can be distinguished from the vacuole and a microelectrode tip can be located unambiguously in the cytoplasm. The site of the electrogenic pump is clearly demonstrated to reside in the plasmalemma as shown by depolarization of the cell electropotential induced by CN(-).


Increased expression of p34cdc2 and its kinase activity in human gastric and colonic carcinomas.
We examined the expression of p34cdc2 and its kinase activity in human gastric and colonic carcinoma cell lines and carcinoma tissues and studied its relation with a tumor-suppressor gene product, p53. All the gastric and colonic cancer cell lines expressed p34cdc2 and showed its kinase activity at various levels. When the cells were arrested in mitotic metaphase by the use of nocodazole, p34cdc2 kinase activity was induced and p53 was apparently phosphorylated. Of 12 gastric carcinoma cases, 11 (91.7%) showed higher p34cdc2 kinase activity in tumor tissues than in corresponding non-neoplastic mucosa. The protein kinase activities in the individual cases were well correlated with the levels of p34cdc2 protein expression. A good correlation was also found between the expression of p34cdc2 and proliferating cell nuclear antigen (PCNA). Almost all the colonic carcinomas showed higher cdc2 kinase activity and increased p34 expression when compared with non-neoplastic mucosa. Interestingly, most of the gastric and colonic carcinomas having high cdc2 kinase activity expressed high levels of p53. These findings suggest that the increased p34cdc2 kinase activity might cause the development and proliferation of gastric and colonic carcinomas, partly through abnormal p53 accumulation.


Anti-Xa versus time-guided anticoagulation strategies in extracorporeal membrane oxygenation: a systematic review and meta-analysis.
The purpose was to compare time-based vs anti-Xa-based anticoagulation strategies in patients on ECMO. We conducted a systematic review and meta-analysis using multiple electronic databases and included studies from inception to July 19, 2019. The proportion of bleeding, thrombosis, and mortality were evaluated.Twenty-six studies (2,086 patients) were included. Bleeding occurred in 34.2% (95%CI 25.1;43.9) of the patients with anti-Xa-based versus 41.6% (95%CI 24.9;59.4) of the patients with time-based anticoagulation strategies. Thrombosis occurred in 32.6% (95%CI 19.1;47.7) of the patients with anti-Xa-based versus 38.4% (95%CI 22.2;56.1) of the patients with time-based anticoagulation strategies. And mortality rate was 35.4% (95%CI 28.9;42.1) of the patients with anti-Xa-based versus 42.9% (95%CI 36.9;48.9) of the patients with time-based anticoagulation strategies. Among the seven studies providing results from both anticoagulation strategies, significantly fewer bleeding events occurred in the anti-Xa-based anticoagulation strategy (adjusted OR 0.49 (95%CI 0.32;0.74), p < 0.001) and a significantly lower mortality rate (adjusted OR 0.61 (95%CI 0.40;0.95), p = 0.03). There was no significant difference in thrombotic events (adjusted OR 0.91 (95%CI 0.56;1.49), p = 0.71). In these seven observational studies, only a small fraction of the patients were adults, and data were insufficient to analyze the effect of the type of ECMO.In this meta-analysis of observational studies of patients on ECMO, an anti-Xa-based anticoagulation strategy, when compared to a time-based strategy, was associated with fewer bleeding events and mortality rate, without an increase in thrombotic events.


Functional epitope analysis of the human CD4 molecule: antibodies that inhibit human immunodeficiency virus type 1 gene expression bind to the immunoglobulin CDR3-like region of CD4.
We recently demonstrated that monoclonal antibody (MAb) 13B8-2, specific for the immunoglobulin (Ig) complementary determining region 3 (CDR3)-like region of the CD4 molecule, inhibits viral transcription in human immunodeficiency virus (HIV)-infected CEM cells and HIV type 1 (HIV-1) promoter activity. Here, we have studied the capacity of several MAb specific for the D1 domain of CD4, including anti-CDR2-like (Leu-3a and ST4) and anti-CDR3-like (13B8-2 and ST40) MAb, and for the D2 domain of CD4 (BL4) to inhibit both provirus transcription in HIV-1LAI-infected CEM cells and transcription of the chloramphenicol acetyltransferase (CAT) gene under control of the HIV-1 long terminal repeat in transiently transfected CEM cells. We found that HIV-1 promoter activity and provirus transcription are inhibited only by MAb that bind to the CDR3-like region in domain 1 of CD4. Moreover, we demonstrated that the Fab fragment of an anti-CDR3-like region-specific anti-CD4 MAb is a powerful inhibitor of HIV-1 promoter activity. These results have implications for understanding the role of the CDR3-like region in CD4 T-cell signaling, which controls provirus transcription.


A PVP-extract fungal protein of Omphalia lapideacens and its antitumor activity on human gastric tumors and normal cells.
Omphalia lapidescens is an important medicinal fungus as well as traditional Chinese medicine used for disease treatment. It is mainly used as a vermifuge for anthelmintic therapy, but it has not been hitherto reported to possess antitumor activity. In this study, a purified bioactive protein in O. lapidescens (pPeOp) was obtained using polyvinylpyrrolidone (PVP) followed by gel filtration chromatography. To evaluate the in vitro antitumor activity of pPeOp in human gastric tumor cells (MC-4 and SGC-7901) and normal cells (MC-1), MTT assay and FCM assay were used and the morphological changes, cell viability, cell death rate and cell apoptosis rate of MC-4, SGC-7901 and MC-1 cells were estimated. The results showed that pPeOp could significantly reduce the cell viability of MC-4 and SGC-7901 cells in a concentration-dependent manner, with IC50 values of 236.05 and 156.28 µg/ml, respectively. The morphological observation also indicated a similar result. In FCM assays, a significant increase of cell death rate and cell apoptosis rate of the tumor cells were observed, indicating probable necrosis-inducing effects and/or apoptosis-inducing effects of pPeOp. Importantly, there was no significant effect of pPeOp on MC-1 cells in each assay, showing that pPeOp has no adverse effects on the normal cells. In conclusion, pPeOp is a newly discovered bioactive protein in O. lapidescens and this is the first report on antitumor activity of such a fungal protein. This may provide a meaningful basis for developing a new protein drug for treatment against cancer, especially gastric cancer.


Vaccinia virus WR53.5/F14.5 protein is a new component of intracellular mature virus and is important for calcium-independent cell adhesion and vaccinia virus virulence in mice.
The vaccinia virus WR53.5L/F14.5L gene encodes a small conserved protein that was not detected previously. However, additional proteomic analyses of different vaccinia virus isolates and strains revealed that the WR53.5 protein was incorporated into intracellular mature virus (IMV). The WR53.5 protein contains a putative N-terminal transmembrane region and a short C-terminal region. Protease digestion removed the C terminus of WR53.5 protein from IMV particles, suggesting a similar topology to that of the IMV type II transmembrane protein. We generated a recombinant vaccinia virus, vi53.5L, that expressed WR53.5 protein under isopropyl-beta-d-thiogalactopyranoside (IPTG) regulation and found that the vaccinia virus life cycle proceeded normally with or without IPTG, suggesting that WR53.5 protein is not essential for vaccinia virus growth in cell cultures. Interestingly, the C-terminal region of WR53.5 protein was exposed on the cell surface of infected cells and mediated calcium-independent cell adhesion. Finally, viruses with inactivated WR53.5L gene expression exhibited reduced virulence in mice when animals were inoculated intranasally, demonstrating that WR53.5 protein was required for virus virulence in vivo. In summary, we identified a new vaccinia IMV envelope protein, WR53.5, that mediates cell adhesion and is important for virus virulence in vivo.


The kinetics of the visible growth of a primary melanoma reflects the tumor aggressiveness and is an independent prognostic marker: a prospective study.
Primary melanoma (MM) could be a good model to test an intuitive concept: a cancer that is growing fast in its early phase is likely to have a high aggressiveness. Since MMs are visible tumors, many patients can provide information to indirectly assess the kinetics of their lesion. A prospective study was designed to assess if the kinetics of the visible growth of a primary MM, as described by the patient, could be a noninvasive prognostic marker. The ratio of MM thickness to delay between MM appearance and MM removal was used as a surrogate value for the kinetics of the MM growth. To assess the delay between MM appearance and removal, 362 patients with self-detected invasive MM fulfilled a detailed questionnaire, which provided 2 types of estimations of this delay and thus 2 melanoma kinetics indexes (MKI and MKI*). After a median follow-up of 4 years, univariate and multivariate analyses assessed whether relapse-free survival was linked to MKI or MKI*. MKI was significantly predictive of relapse-free survival (HR = 1.84 [1.51-2.25]) and relapse at 1 year (RR = 2.93 [1.84-4.69]), independently from Breslow thickness. MKI was retained in multivariate prognostic models, just after thickness and before other usual markers. MKI* was also a significant independent risk marker, although less predictive. In this model, the initial growth kinetics of a cancer reflects its aggressiveness and a high index predicts a short-term relapse. The "subjective" data obtained from patients about their MM history, although usually neglected, can thus provide a better prognostic marker than many "objective" tests.


The role of the bladder tumour antigen test in the management of gross haematuria.
The bladder tumour antigen (BTA) test has been found to be more sensitive than urine cytology for the detection of recurrent bladder cancer. We chose to evaluate the role of the bladder tumour antigen test in the management of gross haematuria on initial presentation. A prospective analysis over a 3-month period of consecutive cases of gross haematuria was conducted. Routine investigations including intravenous urogram, flexible cystoscopy, urine cytology testing and BTA testing were performed. Results were correlated with the final diagnosis, and also with histology and tumour stage in the cases of malignant outcome. The prevalence of malignancy in our series was 25.5%. Sensitivity of the BTA test was 67% and specificity 66%. This was compared to cytology which yielded a sensitivity of 66% and specificity of 100%. There were 12 cases of falsely positive results on BTA testing, giving a positive predictive value of 40%. Four cases of falsely negative results were all from superficial tumours. In view of its poor sensitivity and positive predictive value, the BTA test has a limited role in the initial management of gross haematuria.


Evaluation of clinical diagnostic criteria for psychogenic movement disorders.
Previous studies showed that there are certain features suggestive of a psychogenic disorder, and Fahn and Williams proposed criteria for psychogenic movement disorders. Data on the sensitivity and specificity of these criteria are lacking. We expanded on the Fahn and Williams criteria to create a new set of diagnostic criteria. We retrospectively reviewed 79 patients in a movement disorders specialty clinic. We applied the proposed diagnostic criteria to both cases and controls and analyzed sensitivity and specificity. The diagnostic criteria correctly identified "clinically probable" (or better) psychogenic movement disorders with a sensitivity of 83% and specificity of 100%. For "clinically possible" or greater, sensitivity was 97% and specificity was 96%. In addition, a significantly higher proportion of patients with psychogenic disease were female and reported either a family history of neurological disease or other exposure to neurological disease (P = 0.001 and 0.01, respectively). The diagnostic criteria are simple to apply and have high sensitivity and specificity for psychogenic movement disorders. Also, we report a correlation between previous exposure to a disease model and psychogenic disease.


The ab initio ground-state potential energy function of beryllium monohydride, BeH.
The accurate ground-state potential energy function of beryllium monohydride, BeH, has been determined from large-scale ab initio calculations using the multi-reference averaged coupled-pair functional (MR-ACPF) method in conjunction with the correlation-consistent core-valence basis sets up to septuple-zeta quality. The effects of electron correlation beyond the MR-ACPF level of approximation were taken into account. The scalar relativistic and adiabatic (the diagonal correction) effects, as well as some of the nonadiabatic effects, were also discussed. The vibration-rotation energy levels of three isotopologues, BeH, BeD, and BeT, were predicted to sub-cm(-1) accuracy.


Stem cell generation and choice of fate: role of cytokines and cellular microenvironment.
Hematopoietic stem cells (HSC) have provided a model for the isolation, enrichment and transplantation of stem cells. Gene targeting studies in mice have shown that expression of the thrombopoietin receptor (TpoR) is linked to the accumulation of HSCs capable to generate long-term blood repopulation when injected into irradiated mice. The powerful increase in vivo in HSC numbers by retrovirally transduced HOX4B, a homeotic gene, along with the role of the TpoR, suggested that stem cell fate, renewal, differentiation and number can be controlled. The discovery of the precise region of the mouse embryo where HSCs originate and the isolation of supporting stromal cell lines open the possibility of identifying the precise signals required for HSC choice of fate. The completion of human genome sequencing coupled with advances in gene expression profiling using DNA microarrays will enable the identification of key genes deciding the fate of stem cells. Downstream from HSCs, multipotent hematopoietic progenitor cells appear to co-express a multiplicity of genes characteristic of different blood lineages. Genomic approaches will permit the identification of the select group of genes consolidated by the commitment of these multipotent progenitors towards one or the other of the blood lineages. Studies with neural stem cells pointed to the unexpected plastic nature of these cells. Isolation of stem cells from multiple tissues may suggest that, providing the appropriate environment/ signal, tissues could be regenerated in the laboratory and used for transplantation. A spectacular example of influence of the environment on cell fate was revealed decades ago by using mouse embryonic stem cells (ES). Injected into blastocysts, ES cells contribute to the formation of all adult tissues. Injected into adult mice, ES cells become cancer cells. After multiple passages as ascites, when injected back into the blastocyst environment, ES- derived cancer cells behaved again as ES cells. More recently, the successful cloning of mammals and reprogramming of transferred nuclei by factors in the cytoplasm of oocytes turned back the clock by showing that differentiated nuclei can be "re-booted" to generate again the stem cells for different tissues.


Upregulated expression of ADAM12 is associated with progression of oral squamous cell carcinoma.
ADAMs are a disintegrin and metalloproteinase family of membrane-associated metalloproteinases characterized by their multidomain structure, and have been reported to be associated with various malignant tumors. The aim of this study was to identify crucial members of the ADAM family in oral squamous cell carcinoma (OSCC), and to reveal their biological function and clinical significance. To clarify whether ADAM family genes are involved in OSCC, changes in the expression profile were investigated by real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) analysis and immunohistochemical analysis. Functional analysis was performed by comparing cellular proliferation of siADAM-transfected cell lines and parental cell lines. Real-time qRT-PCR analysis identified significantly upregulated expression of ADAM12 in OSCC-derived cell lines. This was validated in OSCC samples using real-time qRT-PCR and immuno-histochemical staining. ADAM12 expression was correlated with TNM classification; significantly greater expression of ADAM12 was observed in tumors with higher T classification and more advanced stages. Moreover, siADAM12-transfected cells showed both a suppressed proliferation rate and increased transforming growth factor (TGF)-β3 expression. Our data indicate that ADAM12 is overexpressed in OSCC and might accelerate cellular proliferation. Its function may be associated with TGF-β signaling. This study suggests that controlling the expression or activity of ADAM12 could be a useful strategy in the development of an effective cure for OSCC.


[Radical surgery of locally advanced prostatic cancer].
Retrospective study in 51 patient with locally advanced prostate cancer (34 pT3 and 17 N+) selected from 88 radical prostatectomies performed between February 1985 and December 1993. An evaluation is made of the clinical/pathological stage correlation and the effectiveness of the different diagnostic methods (61% of understaging). The possibility to rescue locally spread tumours is analyzed, 7/51 (13.72%) patients receiving pre-operative neoadjuvant hormonal therapy. Estimated survival at 8 years was 82.6% of all patients treated with radical surgery and, at the end of the study, the situation of 30/46 (65.2%) patients assessed by digital rectal examination, quality of life index, PSA follow-up, abdominal ultrasound and/or CT, bone scan and chest X-ray, was an estimated 8-year disease-free evolution. It is observed that radical surgery and the post-surgical hormonal approach permit good survival in locally advanced prostate cancer and that the use of a neoadjuvant hormonal approach permit good survival in locally advanced prostate cancer and that the use of a neoadjuvant hormonal approach allows in many cases the rescue of patients for radical surgery.


'They all supported me but I felt like I suddenly didn't belong anymore': an exploration of perceived disadvantages to online support seeking.
In recent years, there has been a significant rise in the number of online support communities dedicated to issues surrounding infertility. The aim of this study was to focus on investigating the perceived disadvantages of online infertility support communities from the perspective of those who access and participate in them. A total of 295 participants completed an online questionnaire about their use of online support communities. Responses were analysed using inductive content analysis. Since the majority of respondents were women, the results of the study largely reflect the experiences of the female population. Over half of the sample (57.9%) reported experiencing disadvantages to online support. Content analysis revealed that the most commonly cited disadvantages were reading about negative experiences (10.9%), reading about other peoples pregnancies (8.8%), inaccurate information (7.8%) and its addictive (5.8%). These results suggest that there are many perceived disadvantages to online infertility support communities. While some of these disadvantages reflect fears commonly cited in the literature, there are also unique disadvantages associated with the experience of infertility and its treatment. The study highlights a number of important areas in which health professionals and community moderators could intervene to better support and improve the online experiences of patients experiencing infertility.


Reliable and reproducible murine models for commonly used abdominal plastic surgical flaps.
Animal models have been used for many years in surgical research to develop different surgical techniques, improve understanding of anatomy and physiology and hone surgical skills. The benefit of such models has been particularly important in developing relatively young specialties like plastic surgery and many plastic surgical techniques are designed and studied in animals long before they are used in humans. We describe techniques for raising several reliable and reproducible abdominal flaps in rodents, including transverse rectus abdominis myocutaneous flaps in rats and mice, superficial inferior epigastric artery flaps in rats and perforator flaps in rats. The intention of this paper is to act as a point of reference for any microvascular or plastic surgeon who is planning to perform abdominal plastic surgical flap research or further microvascular skills.


Ivabradine as a heart rate-lowering agent in a patient with end-stage renal failure after heart transplantation.
A 56 year-old woman with a transplanted heart, with arterial hypertension and chronic pulmonary obstructive disease, was hospitalised because of palpitations, dyspnea, chest pain and oedema. After cyclosporine treatment she was diagnosed with renal failure, which was treated by hemodialysis. Heart rate (HR) at admission was 100, mean HR in 24-hour Holter monitoring was 106 bpm. Ivabradine was added to the treatment. The dose of 2.5 mg bid was doubled after three days. Mean HR in control Holter monitoring was 81. Ivabradine was well tolerated in this patient. The clinical benefits were observed soon after application and maintained during the follow-up.


Contrast ultrasound perfusion imaging of lower extremities in peripheral arterial disease: a novel diagnostic method.
The purpose of this study was to establish contrast-enhanced ultrasound perfusion imaging (CUPI) of the lower extremities as a novel non-invasive diagnostic tool for patients with peripheral arterial disease (PAD). Ultrasound contrast agent (SonoVue) was injected into a peripheral vein of 16 control subjects and 16 PAD patients and its appearance in the calf muscle was detected by low-energy harmonic ultrasound. Analysis of the wash-in curves revealed that PAD patients had a significantly longer time to peak intensity (TTP), i.e. duration of maximum contrast perfusion [37 s (19-79 s) in control subjects vs. 56 s (32-104 s) in PAD patients at rest, age-adjusted P=0.002]. Exercise stress test of the calf muscle resulted in a decrease of the TTP, maintaining the significant difference in TTP between the groups [19 s (8-37 s) in control subjects vs. 32 s (18-48 s) in PAD patients after exercise, age-adjusted P=0.004]. Neither ankle-brachial index and TTP nor age and TTP showed a significant correlation. CUPI reflects the regional blood circulation of the calf muscle. In this pilot study, PAD patients show a significantly longer TTP than control subjects. The clinical relevance of CUPI is topic of ongoing studies.


Identification of the major protein components in breast secretions from women with benign and malignant breast diseases.
The protein composition of breast secretions from 99 premenopausal women with benign or malignant breast diseases and from 70 control women without breast pathologies has been studied by using polyacrylamide gel electrophoresis. These fluids have been classified into two types according to their major polypeptide components. Type I fluids are defined by three major distinctive bands at Mr 44,000, 24,000, and 17,000, while those designated Type II present distinctive bands at Mr 80,000, 15,000, and 14,000. Amino acid sequencing and immunoblotting analysis demonstrated that proteins in Type I secretions correspond to Zn-alpha 2-glycoprotein, apolipoprotein D, and gross cystic disease fluid protein-15, while those from Type II fluids have been identified as lactoferrin, lysozyme, and alpha-lactalbumin. Most women (93%) without breast pathology and most patients (88%) with benign diseases had secretions with a Type I polypeptide pattern. By contrast, a large percentage (57%) of secretions from women with breast carcinoma presented a Type II protein pattern. Further studies with a large number of women will be useful for corroborating the potential clinical interest of breast fluid protein analysis.


Distinct Mechanisms of IgM Antibody-Mediated Acquired von Willebrand Syndrome and Successful Treatment with Recombinant von Willebrand Factor in One Patient.
Acquired von Willebrand Syndrome (AVWS) is a rare coagulation disorder which can be associated with IgM paraproteinaemia. Recently, recombinant von Willebrand factor (rVWF) has become available for the treatment of bleedings in patients with inherited von Willebrand disease, but experience in patients with AVWS is limited. We report on 2 patients with AVWS with underlying IgM paraproteinaemia with distinct underlying pathomechanisms. In 1 patient, the paraprotein built unspecific complexes with von Willebrand factor (VWF). In the other patient, we were able to detect an IgM antibody against VWF. Bleeding in this patient was successfully treated with rVWF. To our knowledge, this is the first report about the successful use of rVWF in a patient with AVWS with the detection of a VWF-specific antibody.


Studies on penem and carbapenem. I. Syntheses and oral absorption of ester-type prodrugs of sodium (5R,6S)-2-(2-fluoroethylthio)-6-[(1R)-1-hydroxyethyl]penem-3-c arboxylat e.
Acyloxyalkyl esters (2a-d), alkyloxycarbonyloxyalkyl esters (2e-g) and (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl ester (2h) of (5R,6S)-2-(2-fluoroethylthio)-6-[(1R)-1-hydroxyethyl]penem-3- carboxylic acid (1) were synthesized. Enhanced oral absorption was observed in mice reflecting increased lipophilicity, compared with the parent 1 itself. Among them, the ester 2h showed a prolonged plasma level and a large area under the blood concentration-time curve (AUC) in rats. These ester-type prodrugs of penem 1 in phosphate buffer (pH 6.86) were much more stable than those of cephalosporins which easily degraded via isomerization to delta 2 cephalosporins.


Guillain-Barré as an autoimmune disease.
Guillain-Barré syndrome (GBS) entails a demyelinating process of the peripheral nervous system. The etiopathogenesis of the syndrome is still a matter of debate although considerable progress has been accomplished in the recent years. Abundant evidence has been put forward so as to support the role of the immune system in initiating and perpetuating the ongoing damage culminating in the emergence of the clinically overt syndrome. As such, data on the involvement of the humoral immune pathways add to the information already presented with regard to cell-mediated mechanisms participating in disruption of peripheral nerve. The following review will focus on the current knowledge of these complex mechanisms and the relative significance of each in the pathogenesis of GBS.


Discovery of an Antarctic Ascidian-Associated Uncultivated Verrucomicrobia with Antimelanoma Palmerolide Biosynthetic Potential.
The Antarctic marine ecosystem harbors a wealth of biological and chemical innovation that has risen in concert over millennia since the isolation of the continent and formation of the Antarctic circumpolar current. Scientific inquiry into the novelty of marine natural products produced by Antarctic benthic invertebrates led to the discovery of a bioactive macrolide, palmerolide A, that has specific activity against melanoma and holds considerable promise as an anticancer therapeutic. While this compound was isolated from the Antarctic ascidian Synoicum adareanum, its biosynthesis has since been hypothesized to be microbially mediated, given structural similarities to microbially produced hybrid nonribosomal peptide-polyketide macrolides. Here, we describe a metagenome-enabled investigation aimed at identifying the biosynthetic gene cluster (BGC) and palmerolide A-producing organism. A 74-kbp candidate BGC encoding the multimodular enzymatic machinery (hybrid type I-trans-AT polyketide synthase-nonribosomal peptide synthetase and tailoring functional domains) was identified and found to harbor key features predicted as necessary for palmerolide A biosynthesis. Surveys of ascidian microbiome samples targeting the candidate BGC revealed a high correlation between palmerolide gene targets and a single 16S rRNA gene variant (R = 0.83 to 0.99). Through repeated rounds of metagenome sequencing followed by binning contigs into metagenome-assembled genomes, we were able to retrieve a nearly complete genome (10 contigs) of the BGC-producing organism, a novel verrucomicrobium within the Opitutaceae family that we propose here as "Candidatus Synoicihabitans palmerolidicus." The refined genome assembly harbors five highly similar BGC copies, along with structural and functional features that shed light on the host-associated nature of this unique bacterium. IMPORTANCE Palmerolide A has potential as a chemotherapeutic agent to target melanoma. We interrogated the microbiome of the Antarctic ascidian, Synoicum adareanum, using a cultivation-independent high-throughput sequencing and bioinformatic strategy. The metagenome-encoded biosynthetic machinery predicted to produce palmerolide A was found to be associated with the genome of a member of the S. adareanum core microbiome. Phylogenomic analysis suggests the organism represents a new deeply branching genus, "Candidatus Synoicihabitans palmerolidicus," in the Opitutaceae family of the Verrucomicrobia phylum. The Ca. Synoicihabitans palmerolidicus 4.29-Mb genome encodes a repertoire of carbohydrate-utilizing and transport pathways, a chemotaxis system, flagellar biosynthetic capacity, and other regulatory elements enabling its ascidian-associated lifestyle. The palmerolide producer's genome also contains five distinct copies of the large palmerolide biosynthetic gene cluster that may provide structural complexity of palmerolide variants.


Mechanism, measurement, and prevention of oxidative stress in male reproductive physiology.
Numerous factors influence male fertility. Among these factors is oxidative stress (OS), which has elicited an enormous interest in researchers in recent period. Reactive oxygen species (ROS) are continuously produced by various metabolic and physiologic processes. OS occurs when the delicate balance between the production of ROS and the inherent antioxidant capacity of the organism is distorted. Spermatozoa are particularly sensitive to ROS as their plasma membrane contains polyunsaturated fatty acids (PUFA), which oxidizes easily. They also lack cytoplasm to generate a robust preventive and repair mechanism against ROS. The transition metal ions that are found in the body have a catalytic effect in the generation of ROS. Lifestyle behaviours such as smoking and alcohol use and environmental pollution further enhance the generation of ROS and thus, cause destructive effects on various cellular organelles like mitochondria, sperm DNA etc. This article analyzes the detrimental effects of OS on male fertility, measurement of OS and effective ways to decrease or eliminate them completely. We have also provided information on oxidative stress in other systems of the body, which may be applied to future research in the field of reproductive biology.


Urgent COVID-19 Vaccination of Healthcare Workers via a Quality Improvement Initiative.
The COVID-19 pandemic has posed a significant threat to US healthcare workers' mental and physical health. The US Food and Drug Administration approved the first mRNA COVID-19 vaccine for Emergency Use Authorization on December 11, 2020. High-risk healthcare workers were determined to be Phase 1a. Goal: Complete the two-dose vaccine series in all interested phase 1a staff immediately after the COVID-19 vaccine was available and distributed to our institution, December 14, 2020. A multidisciplinary team involving key stakeholders performed process mapping to develop four key drivers for vaccination success: rapid vaccine procurement, proper storage and handling, well-defined vaccine administration and follow-up plan, and system preparation. We tested interventions using plan-do-study-act cycles. We included employees and providers with direct patient care responsibilities, age 18 years or older, employed at the children's health system, or the affiliated academic medical center. We examined the total number of dose 1 and dose 2 vaccines administered for our primary outcome, and the balancing measure included the percent of wasted vaccines. Three thousand nine-hundred twenty-one healthcare personnel completed the survey, and 73% reported intent to receive the COVID-19 vaccine immediately or at a later time. After 57 clinic days, we vaccinated 83% (n = 5,231) of healthcare personnel at our institution, and 99% completed the two-dose series. Due to surplus vaccines, we vaccinated an additional 1,258 community members with 99% completion of the two-dose series. Vaccine waste was minimal 0.1%. We describe the development and implementation of a successful COVID-19 employee and community vaccination program.


[Therapeutic difficulties in a 3 year-old boy with Kawasaki disease].
A case of a three year-old by with delayed diagnosis of disease is presented. Treatment with immunoglobulins and methylpreduisolone was affective, however, no regression in coronary artery lesions was observed. The diagnosis and treatment of Kawasaki disease are discussed.


Coupling of an aldehyde or ketone to pyridine mediated by a tungsten imido complex.
The reactivity of W(NPh)(o-(Me3SiN)2C6H4)(py)2 and W(NPh)(o-(Me3SiN)2C6H4)(pic)2 (py=pyridine; pic=4-picoline) with unsaturated substrates has been investigated. Treatment of W(NPh)(o-(Me3SiN)2C6H4)(py)2 with diphenylacetylene or 2,3-dimethyl-1,3-butadiene generates W(NPh)(o-(Me3SiN)2C6H4)(eta2-PhCCPh) and W(NPh)(o-(Me3SiN)2C6H4)(eta4-CH2=C(Me)C(Me)=CH2), respectively, while the addition of ethylene to W(NPh)(o-(Me3SiN)2C6H4)(py)2 generates the known metallacycle W(NPh)(o-(Me3SiN)2C6H4)(CH2CH2CH2CH2). The addition of 2 equiv of acetone to W(NPh)(o-(Me3SiN)2C6H4)(pic)2 provides the azaoxymetallacycle W(NPh)(o-(Me3SiN)2C6H4)(OCH(Me)2)(OC(Me)2-o-C5H3N-p-Me), the result of acetone insertion into the ortho C-H bond of picoline. Similarily, the addition of 2 equiv of RC(O)H [R=Ph, tBu] to W(NPh)(o-(Me3SiN)2C6H4)(py)2 generates W(NPh)(o-(Me3SiN)2C6H4)(OCH2R)(OCHR-o-C5H4N) [R=Ph, tBu,]. In contrast, reaction between W(NPh)(o-(Me3SiN)2C6H4)(py)2 and 2-pyridine carboxaldehyde yields the diolate W(NPh)(o-(Me3SiN)2C6H4)(OCH(C5H4N)CH(C5H4N)O). The synthesis of W(NPh)(o-(Me3SiN)2C6H4)(PMe3)(py)(eta2-OC(H)C6H4-p-Me), formed by the addition of p-tolualdehyde to a mixture of W(NPh)(o-(Me3SiN)2C6H4)(py)2 and PMe3, suggests that an eta2-aldehyde intermediate is involved in the formation of the azaoxymetallacycle, while the isolation of W(NPh)(o-(Me3SiN)2C6H4)(Cl)(OC(Me)(CMe3)-o-C5H4N), formed by the reaction of pinacolone with W(NPh)(o-(Me3SiN)2C6H4)(py)2, in the presence of adventitious CH2Cl2, suggests that the reaction proceeds via the hydride W(NPh)(o-(Me3SiN)2C6H4)(H)(OC(Me)(CMe3)-o-C5H4N).


Predictors of Mortality in Neonates with Seizures; a Prospective Cohort Study.
The aim of this study was to analyze prognostic indicators for mortality in neonates with seizures in a level III Neonatal Intensive Care Unit (NICU). A cohort of 100 neonates with clinically manifested seizures hospitalized in the NICU during 4 years period was prospectively monitored for the first year of life. The cohort consisted of 33 preterm and 67 full-term babies with 60 male and 40 female infants. The mortality rate in the first year of life of infants with seizures in the neonatal period was 23%. The most common cause of seizures was birth asphyxia for full-term infants and intra-periventricular hemorrhage for preterm infants. Death was more common in pre-term than term infants (p <0,005). Simple regression demonstrated statistically significant associations between death in the first year of life and a cluster of highly associated variables: resuscitation (p<0, 01), mechanical ventilation (p<0,01) and asphyxia (p<0,05). This cluster of variables significantly correlates with: gestational age (p<0, 05), birth weight (p<0, 05) and intracranial hemorrhage (p<0, 05). In this cohort of neonates with seizures asphyxia requiring neonatal resuscitation was the primary risk factor for death.


[Detection of occult malignant melanoma by 18F-FDG PET-CT and gamma probe].
Malignant melanoma has a high relapse rate, and PET is very useful for its detection, above all when its size is small or it is in locations that are difficult to resect. The radioguided surgery being implanted in many sites, new expectations are opening in these situations--the intraoperative location of the lesions by 18F-FDG and detection probe. This is a case report of a 44 years old woman diagnosed of malignant melanoma in her left leg 12 years ago. At present, she has a tumor in her left thigh, which was no longer palpable after a fine needle puncture was performed. This was reported as a mesenchymal tumor. Due to the possibility of recurrent melanoma, need for excision was suggested. In order to locate it, a PET-CT study with 18F-FDG was performed and the lesion was marked on the skin with the help of metallic points. The lesion was located in surgery with a gamma probe, showing relapse of melanoma. The importance of the case is found in the fact that it is the first step to extend the indications of radioguided surgery with the use of PET, a technique that offers a ride range of possibilities.


Hospitalization for anorexia nervosa in Italy.
This study provides: a) rates of hospitalization for anorexia nervosa (AN) in Italy in 2004-2005 period; b) an estimate of the incidence of moderate to severe hospitalized AN in the same period among persons aged 10-19 years. 9863 hospital discharges of patients aged 10-59 years with a diagnosis of AN (ICD9CM code 307.1) were extrapolated from the Italian Hospital Discharges Database. Patients aged 10-19 years, first admitted in 2004-2005, never hospitalized for AN in 2001-2003, were assumed to be a reasonable proxy of incident cases. Crude rate of AN associated hospitalization was 24.2 per 100,000 person-years among women and 1.6 per 100,000 person-years among men. Estimated incidence rate of AN was 22.8 per 100,000 among women compared with 2.0 per 100,000 among men in the 10-19 years age group. This study provides, for the first time, nationwide incidence estimates of AN in Italy.


The Role of Acetaldehyde in the Increased Acceptance of Ethanol after Prenatal Ethanol Exposure.
Recent studies show that acetaldehyde, the first metabolite in the oxidation of ethanol, can be responsible for both, the appetitive and the aversive effects produced by ethanol intoxication. More specifically, it has been hypothesized that acetaldehyde produced in the periphery by the liver is responsible for the aversive effects of ethanol, while the appetitive effects relate to the acetaldehyde produced centrally through the catalase system. On the other hand, from studies in our and other laboratories, it is known that ethanol exposure during the last gestational days (GD) consistently enhances the postnatal acceptance of ethanol when measured during early ontogeny in the rat. This increased liking of ethanol is a conditioned appetitive response acquired by the fetus by the association of ethanol's flavor and an appetitive reinforcer. Although this reinforcer has not yet been fully identified, one possibility points to acetaldehyde produced centrally in the fetus as a likely candidate. This hypothesis is supported by data showing that very early in the rat's ontogeny brain catalases are functional, while the liver's enzymatic system is still immature. In this study, rat dams were administered on GD 17-20 with water or ethanol, together with an acetaldehyde-sequestering agent (D-penicillamine). The offspring's responses to ethanol was then assessed at different postnatal stages with procedures adequate for each developmental stage: on day 1, using the "odor crawling locomotion test" to measure ethanol's odor attractiveness; on day 5, in an operant conditioning procedure with ethanol as the reinforcer; and on day 14 in an ethanol intake test. Results show that the absence of acetaldehyde during prenatal ethanol exposure impeded the observation of the increased acceptance of ethanol at any age. This seems to confirm the crucial role of acetaldehyde as a reinforcer in the appetitive learning occurring during prenatal ethanol exposure.


A global clustering algorithm to identify long intergenic non-coding RNA--with applications in mouse macrophages.
Identification of diffuse signals from the chromatin immunoprecipitation and high-throughput massively parallel sequencing (ChIP-Seq) technology poses significant computational challenges, and there are few methods currently available. We present a novel global clustering approach to enrich diffuse CHIP-Seq signals of RNA polymerase II and histone 3 lysine 4 trimethylation (H3K4Me3) and apply it to identify putative long intergenic non-coding RNAs (lincRNAs) in macrophage cells. Our global clustering method compares favorably to the local clustering method SICER that was also designed to identify diffuse CHIP-Seq signals. The validity of the algorithm is confirmed at several levels. First, 8 out of a total of 11 selected putative lincRNA regions in primary macrophages respond to lipopolysaccharides (LPS) treatment as predicted by our computational method. Second, the genes nearest to lincRNAs are enriched with biological functions related to metabolic processes under resting conditions but with developmental and immune-related functions under LPS treatment. Third, the putative lincRNAs have conserved promoters, modestly conserved exons, and expected secondary structures by prediction. Last, they are enriched with motifs of transcription factors such as PU.1 and AP.1, previously shown to be important lineage determining factors in macrophages, and 83% of them overlap with distal enhancers markers. In summary, GCLS based on RNA polymerase II and H3K4Me3 CHIP-Seq method can effectively detect putative lincRNAs that exhibit expected characteristics, as exemplified by macrophages in the study.


Single-dose versus multiple-dose ciprofloxacin plus metronidazole prophylaxis in transrectal ultrasound-guided biopsy of the prostate: a randomized controlled trial.
To investigate and compare the infectious and non-infectious complications of single-dose versus multiple-dose antibiotic therapy for trans-rectal ultrasound (TRUS)-guided biopsy of the prostate. Patients were enrolled in a prospective randomized study that was designed to investigate the effects of single-dose versus multiple-dose antimicrobial prophylaxis regimen mainly on asymptomatic bacteriuria, urinary tract infection (UTI) without fever, fever and urinary septicemia. The single-dose group received one ciprofloxacin 500 mg tablet and two metronidazole 250 mg tablets at 2 hours before the biopsy, while the multiple-doses group received those every 12 hours from 3 days before the biopsy. One-hundred and sixty patients were evaluated in two groups and bacteriuria in urinalysis was encountered in 12 patients (15%) in the single-dose group and four patients (5%) in the multiple-dose group, with a significant difference (P=0.035). UTI without fever occurred in six patients (7.5%) in the single-dose group and one patient (1.25%) in the multiple-dose group, with no significant difference (borderline P=0.053). After biopsy, three patients (3.75%) returned with fever due to UTI and bacteremia in the single-dose group and none in the multiple-dose group, but with no significant difference (P=0.08). Regarding non-infectious complications, there were no significant differences between the two groups. Using prophylactic antibiotics for prostate biopsy in multiple doses, and at least 3 days before the procedure significantly reduces the rate of bacteriuria compared with a single-dose regimen.


Effect of macrophage colony-stimulating factor on in vitro osteoclast generation and bone resorption.
To investigate the role of recombinant human macrophage colony-stimulating factor (rhM-CSF) on in vitro bone resorption, two bone explants, each at a different developmental stage, were adopted, namely 1) radii and 2) metatarsals of 17-day-old embryonic mice. At this stage of gestation, bone resorption in the radii is exclusively dependent upon fusion of osteoclast precursors and activation of mature osteoclasts, whereas in metatarsals it is dependent upon the generation of new osteoclasts. rhM-CSF showed no effect in radii, but stimulated 45Ca release in metatarsals, when they were either intact or periosteum stripped in coculture with embryonal liver as a source of hemopoietic progenitors. The rhM-CSF-induced increase in 45Ca release was paralleled by a higher number of osteoclasts. The stimulating effect was found to be in a concentration range between 250-500 U/ml M-CSF. The action of rhM-CSF was blocked by irradiation, indicating that it is dependent upon cell proliferation. These results, thus, show that M-CSF stimulates bone resorption only when it is dependent on generation of new osteoclasts. M-CSF does not appear to have any effect on the activity of mature osteoclasts. The mechanism of action might be direct on osteoclast precursors or indirect on accessory cells influencing osteoclast generation.


Human bullous pemphigoid antigen (BPAG1). Amino acid sequences deduced from cloned cDNAs predict biologically important peptide segments and protein domains.
Bullous pemphigoid antigens are defined as the autoantigens in a blistering skin disease, bullous pemphigoid. One of them, a 230-kDa protein (BPAG1), is associated with hemidesmosomes, attachment complexes at the basal keratinocyte-lamina lucida interface within the dermal-epidermal basement membrane zone. The precise functions and cellular compartmentalization of BPAG1 are unknown. In this study, a human keratinocyte lambda gt11 cDNA library was screened for clones corresponding to BPAG1. The composite of overlapping cDNAs delineated 8,930 base pairs of nucleotide sequences that contained an open reading frame encoding 2,649 amino acids. Analysis of the deduced amino acid sequences predicted a putative signal peptide of 43 amino acids and the presence of a membrane-associated sequence of 17 amino acids. Several potential sites for N-glycosylation, as well as for protein kinase C or cAMP- and cGMP-dependent protein kinase-mediated phosphorylation were identified. Three peptide segments were predicted to be highly antigenic, potentially serving as epitopes for the formation of autoantibodies. Eight repeat segments of 38 residues each with a high degree of homology with sequences in desmoplakin I, a component of desmosomal cytoplasmic plaques, were detected in the carboxyl-terminal end of the molecule. In addition, the presence of three subdomains characterized by heptad repeats predicted an alpha-helical coiled coil dimer structure in the central portion of the protein. These data suggest that BPAG1 may be a membrane-associated protein that plays a role in the attachment of basal keratinocytes to the underlying basement membrane.


Pathophysiology of altered consciousness during seizures: Subtraction SPECT study.
The mechanisms underlying altered consciousness during seizures are poorly understood. Previous clinicopathologic studies suggest a role for the thalamus and upper brainstem in consciousness mechanisms. To examine blood flow changes associated with altered consciousness during seizures. Seventy-one patients with epilepsy who underwent video-EEG monitoring and ictal/interictal SPECT were studied. Patients were divided into three groups depending on their conscious state during seizures: 1) complete impairment of consciousness (CI), 2) no impairment of consciousness (NI), or 3) uncertain impairment of consciousness (UI). The distribution of blood flow changes during these seizures was assessed by subtraction (ictal - interictal) SPECT co-registered to MRI. Conscious state was assessed in relation to secondary ictal hyperperfusion in subcortical regions (i.e., thalamus and upper brainstem). Impairment of consciousness showed a strong association with secondary hyperperfusion in the thalamic/upper brainstem region (p = 0.01), occurring in 92% (45/49) of CI, 69% (9/13) of UI, and 11% (1/9) of NI. These findings are consistent with a role for the thalamus and upper brainstem in consciousness mechanisms. The authors suggest that the spread of epileptic discharges or a trans-synaptic activation (diaschisis) of these structures is an important mechanism in the alteration of consciousness during seizures. Variance in the results may be due to differences in timing of radioisotope injection, sensitivity of the subtraction SPECT technique, and the ability to clinically assess the conscious state.


X-ray photoelectron spectroscopy characterization of gold nanoparticles functionalized with amine-terminated alkanethiols.
Gold nanoparticles (AuNPs) functionalized with a short chain amine-terminated alkanethiol (HS-(CH(2))(2)NH(2) or C2 NH(2)-thiol) are prepared via a direct synthesis method and then ligand-exchanged with a long chain amine-terminated alkanethiol (HS-(CH(2))(11)NH(2) or C11 NH(2)-thiol). Transmission electron microscopy analysis showed the AuNPs were relatively spherical with a median diameter of 24.2 ± 4.3 nm. X-ray photoelectron spectroscopy was used to determine surface chemistry of the functionalized and purified AuNPs. The ligand-exchange process was monitored within the time range from 30 min to 61 days. By the fourth day of exchange all the C2 NH(2)-thiol molecules had been replaced by C11 NH(2)-thiol molecules. C11 NH(2)-thiol molecules continued to be incorporated into the C11 NH(2) self-assembled monolayer between days 4 and 14 of ligand-exchange. As the length of the exchange time increased, the functionalized AuNPs became more stable against aggregation. The samples were purified by a centrifugation and resuspension method. The C2 NH(2) covered AuNPs aggregated immediately when purification was attempted. The C11 NH(2) covered AuNPs could be purified with minimal or no aggregation. Small amounts of unbound thiol (∼15%) and oxidized sulfur (∼20%) species were detected on the ligand-exchanged AuNPs. Some of the unbound thiol and all of the oxidized sulfur could be removed by treating the functionalized AuNPs with HCl.


Utility of diffusion-weighted and contrast-enhanced magnetic resonance imaging in diagnosing and differentiating between high- and low-grade uterine endometrial stromal sarcoma.
Endometrial stromal sarcoma (ESS) is a rare uterine malignancy that features different prognoses for its high- and low-grade subtypes. We investigated the diagnostic accuracy of magnetic resonance (MR) imaging in diagnosing and differentiating between high- and low-grade ESS. We retrospectively reviewed the preoperative pelvic MR images of consecutive patients who received histologically confirmed diagnoses of high-grade ESS (n = 11) and low-grade ESS (n = 9) and T2-hyperintense leiomyoma (n = 16). Two radiologists independently evaluated imaging features in T1-, T2-, and diffusion-weighted and contrast-enhanced MR images. Statistical analysis included Mann-Whitney tests and Fisher's exact test, with sensitivity, specificity and diagnostic accuracy of imaging features. High-grade ESS was associated with significantly more extensive necrosis and hemorrhage and distinct feather-like enhancement compared with low-grade ESS (P < .05 for all). The feather-like enhancement pattern yielded a diagnostic accuracy of 95%, sensitivity of 91%, and specificity of 100% in differentiating high-grade from low-grade ESS. This imaging characteristic was significantly superior to the necrosis (80%, P = .033) or hemorrhage (75%, P = .007). Both high- and low-grade ESS demonstrated T2 hypointense bands, marginal nodules, intratumoral nodules, and worm-like intra-myometrial nodules, and their tumor apparent diffusion coefficient (ADC) values were significantly lower than those of T2-hyperintense leiomyomas (P < .001). Diffusion-weighted MR imaging is useful in diagnosing ESS against T2-hyperintense leiomyomas, and contrast enhancement aids in further differentiating between high- and low-grade ESS.


Comparative genomics using microarrays reveals divergence and loss of virulence-associated genes in host-specific strains of the insect pathogen Metarhizium anisopliae.
Many strains of Metarhizium anisopliae have broad host ranges, but others are specialists and adapted to particular hosts. Patterns of gene duplication, divergence, and deletion in three generalist and three specialist strains were investigated by heterologous hybridization of genomic DNA to genes from the generalist strain Ma2575. As expected, major life processes are highly conserved, presumably due to purifying selection. However, up to 7% of Ma2575 genes were highly divergent or absent in specialist strains. Many of these sequences are conserved in other fungal species, suggesting that there has been rapid evolution and loss in specialist Metarhizium genomes. Some poorly hybridizing genes in specialists were functionally coordinated, indicative of reductive evolution. These included several involved in toxin biosynthesis and sugar metabolism in root exudates, suggesting that specialists are losing genes required to live in alternative hosts or as saprophytes. Several components of mobile genetic elements were also highly divergent or lost in specialists. Exceptionally, the genome of the specialist cricket pathogen Ma443 contained extra insertion elements that might play a role in generating evolutionary novelty. This study throws light on the abundance of orphans in genomes, as 15% of orphan sequences were found to be rapidly evolving in the Ma2575 lineage.


Effects of oral monosodium (L)-glutamate on insulin secretion and glucose tolerance in healthy volunteers.
To investigate the effects of glutamate on insulin secretion and glucose tolerance in humans. Monosodium (L)-glutamate (10 g) was given orally in a double-blind placebo-controlled cross-over study to 18 healthy volunteers, aged 19-28 years, with an oral (75 g) glucose load. The 75 min insulin response (AUC(0,75 min)), up to tmax of glutamate kinetics, was significantly correlated with the AUC(0,75 min) of glutamate concentrations (r=0.485, P=0.049). Glucose tolerance was not affected. Oral (L)-glutamate enhances glucose-induced insulin secretion in healthy volunteers in a concentration-dependent manner.


Japan's Academic Barriers to Gender Equality as Seen in a Comparison of Public and Private Medical Schools: A Cross-Sectional Study.
Background: Gender inequalities persist in Japanese academic medicine. Some public medical schools have introduced various types of career support for women physicians, whereas few private schools have. Few studies describe the representation of women at different academic ranks and adequacy of career support in public and private medical schools in Japan. Study Design: Cross-sectional descriptive study. Methods: We used publicly available data from the 2018 National Survey on Career Support for Japanese Women Physicians published by the Association of Japanese Medical Colleges in March 2019, which was answered by departments regarding supporting women physicians. Participants represented 51 public and 29 private medical schools in Japan. The proportion of women at academic ranks and career support availability in private and public medical schools were determined using chi-squared test or Fisher's exact test. Results: The proportion of women in senior ranks was significantly higher in private (28.2%) than in public medical schools (25.4%) (p < 0.001). Excluding associate professors, the proportion of professors, lecturers, and assistant professors was significantly higher in private medical schools (3.8% vs. 5.8%, p = 0.002; 12.2% vs. 16.0%, p < 0.001; 20.5% vs. 29.9%, p < 0.001). More public medical schools provided position support and support for other job aspects (43.1% vs. 20.7%, p = 0.043; 70.6% vs. 20.7%, p < 0.001). Conclusions: Public medical schools have lower proportions of women in the academic hierarchy but provide more career support than do private medical schools. Further study is needed to reveal the possible causes of this pattern.


Sunitinib alone or in combination with chemotherapy for the treatment of advanced breast cancer: A systematic review and meta-analysis.
Sunitinib is an orally delivered tyrosine kinase inhibitor that exhibits antiangiogenic effects. FDA has approved sunitinib for the treatment of metastatic renal cell carcinoma. However, its efficacy for the treatment of advanced breast cancer (ABC) remains controversial. Therefore, we performed this systematic review and meta-analysis to synthesize evidence from published randomized controlled trials (RCTs) about the efficacy of sunitinib alone and in combination with chemotherapy for the treatment of ABC. We followed PRISMA statement guidelines during the preparation of this systematic review and meta-analysis. A computer literature search of PubMed, SCOPUS, web of knowledge, and Cochrane Central Register of Controlled Trials (CENTRAL) has been conducted using relevant keywords. Studies were screened for eligibility and data were extracted to an online data extraction form. Progression free survival (PFS) and overall survival (OS) were pooled as Hazard ratio (HR) in a meta-analysis model using generic inverse variance method. Objective response rate (ORR) and complications were pooled as relative risk (RR) in a random effect model meta-analysis using Mantel-Haenzel method. Six RCTs, with a total sample size of 2273 patients, met our eligibility criteria and were included in this meta-analysis. Sunitinib monotherapy was not superior to chemotherapy in terms of PFS (HR = 1.00, 95% CI [0.86 to 1.16], P = 0.99), OS (HR = 1.07; 95% CI [0.87 to 1.32], P = 0.5), or ORR (RR = 0.70, 95% CI [0.74 to 1.03], P = 0.07). Sunitinib in combination with chemotherapy did not show superiority to chemotherapy in terms of PFS (HR = 0.99, 95% CI [0.86 to 1.14], P = 0.89) and OS (HR = 1.04, 95% CI [0.85 to 1.28], P = 0.69). However, the ORR favored sunitinib in combination with chemotherapy group (RR = 1.15, 95% CI [1.01 to 1.31]) with a statistically significant P value (P = 0.03). Current evidence shows that sunitinib, either alone or in combination with chemotherapy, has no clinical benefit for patients with advanced breast cancer. However, previous studies did not considered patient stratification and outcome assessment based on molecular markers. In terms of safety, toxicity was common with sunitinib treatment.


Evaluating the validity and reliability of a modified schedule of sexist events: implications for public health research on women's HIV risk behaviors.
This study evaluated the validity and reliability of a modified version of the Schedule of Sexist Events-Lifetime (SSE, Klonoff & Landrine, 1995) (SSE-LM) and its applicability to women's HIV risk behaviors. Participants included a multi-ethnic baseline sample of women (N = 754) between the ages of 18 and 39 years (M = 22.43) who attended family planning clinics in the San Francisco Bay area. Confirmatory factor analysis demonstrated that a higher-order factor model provided a satisfactory and a more parsimonious account for correlations among the measure's four sexism factors than a single latent factor model. Scale reliability for the SSE-LM was excellent (Cronbach' s alpha = .89). Structural equation modeling results demonstrated that the SSE-LM is associated with women's sexual risk behaviors with secondary partners, but not primary partners.


[Diagnostic utility of anti-cyclic citrullinated peptide antibody in early rheumatoid arthritis].
Current therapeutic strategies against rheumatoid arthritis (RA) employ increasingly aggressive regimens from an early stage of the disease ; thus, serological markers more specific than IgM-rheumatoid factor (IgM-RF) are desirable. Anti-cyclic citrullinated peptide (anti-CCP) antibody has been reported as a useful and highly specificity marker for the diagnosis of RA. To clarify the diagnostic utility of anti-CCP antibody in early RA, we measured serum concentrations of anti-CCP antibody, IgM-RF, anti-agalactosyl IgG antibody and matrix metalloproteinase (MMP)-3 in 184 polyarthritis patients who showed onset symptoms within the previous 2 years. The diagnostic sensitivity of anti-CCP antibody in early RA was 60.0%, equivalent to IgM-RF (66.3%) and anti-agalactosyl IgG antibody (66.0%). Specificity, positive predictive values and diagnostic accuracy of anti-CCP antibody were the best among the four tested makers. In 38 patients who initially did not meet the ACR criteria for RA, but were diagnosed with RA during the course, the diagnostic sensitivity of anti-CCP antibody was 55.3%. On the other hand, the disease activity score (DAS) 28 of anti-CCP antibody positive and the negative patients was 5.16 and 5.34, respectively. Our data indicated that determination of anti-CCP antibody was useful for early diagnosis of RA.


Multiple imputation technique applied to appropriateness ratings in cataract surgery.
Missing data such as appropriateness ratings in clinical research are a common problem and this often yields a biased result. This paper aims to introduce the multiple imputation method to handle missing data in clinical research and to suggest that the multiple imputation technique can give more accurate estimates than those of a complete-case analysis. The idea of multiple imputation is that each missing value is replaced with more than one plausible value. The appropriateness method was developed as a pragmatic solution to problem of trying to assess "appropriate" surgical and medical procedures for patients. Cataract surgery was selected as one of four procedures that were evaluated as a part of the Clinical Appropriateness Initiative. We created mild to high missing rates of 10%, 30% and 50% and compared the performance of logistic regression in cataract surgery. We treated the coefficients in the original data as true parameters and compared them with the other results. In the mild missing rate (10%), the deviation from the true coefficients was quite small and ignorable. After removing the missing data, the complete-case analysis did not reveal any serious bias. However, as the missing rate increased, the bias was not ignorable and it distorted the result. This simulation study suggests that a multiple imputation technique can give more accurate estimates than those of a complete-case analysis, especially for moderate to high missing rates (30 - 50%). In addition, the multiple imputation technique yields better accuracy than a single imputation technique. Therefore, multiple imputation is useful and efficient for a situation in clinical research where there is large amounts of missing data.


Tubulointerstitial damage and progression of renal failure.
The present work reviews the mechanisms and close association between glomerular and tubular damage and its relationship to renal functional impairment. In addition, we present an overview of the pathways involved in the progression of tubulointerstitial fibrosis and a brief summary of the treatments used to retard the progression to end-stage renal failure.


Reducing post-lumbar puncture headaches with small bore atraumatic needles.
Lumbar puncture for testing of Alzheimer's disease pathophysiology for diagnostic confirmation is likely to become more common in the coming years. Minimizing adverse effects from this testing will be essential for clinical practice. Small bore, atraumatic needles reduce the occurrence of post-lumbar puncture headache (PLPH). Our goal was to extend this recommendation specifically to a well-characterized aging population. We assessed PLPH in the Alzheimer's Disease Neuroimaging Initiative cohort and found that PLPH occurrence was reduced only when using a 24 gauge atraumatic needle. We recommend that lumbar punctures for clinical and research purposes in Alzheimer's disease be conducted with 24 gauge atraumatic needles.