Increased caudate dopamine D2 receptor availability as a genetic marker for schizophrenia.
Schizophrenia has a heritability of about 80%, but the detailed molecular genetic basis of the disorder has remained elusive. Relative hyperfunction of the subcortical dopamine system has been previously suggested to be one of the key pathophysiologic mechanisms in schizophrenic psychosis. To examine the contributions of genetic vulnerability for schizophrenia to the dopamine system in the human brain. Population-based twin cohort study. Finland. Six monozygotic and 5 dizygotic unaffected co-twins of patients with schizophrenia were ascertained, along with 4 monozygotic and 3 dizygotic healthy control twins with no family history of psychotic disorders. Striatal dopamine D(2) receptor availability estimated with positron emission tomographic imaging and carbon 11 ((11)C)-labeled raclopride, and performance on neuropsychological tests sensitive to frontal lobe functioning and to schizophrenia vulnerability. Unaffected monozygotic co-twins had increased caudate D(2) density compared with unaffected dizygotic co-twins and healthy control twins. Higher D(2) receptor binding in caudate was associated with a poor performance on cognitive tasks related to schizophrenia vulnerability in the whole sample. The caudate dopamine D(2) receptor up-regulation is related to genetic risk for schizophrenia. Higher dopamine D(2) receptor density in caudate is also associated with poorer performance on cognitive tasks involving corticostriatal pathways. This finding suggests that caudate dopamine dysregulation is also a trait phenomenon related to psychosis vulnerability. This pattern of results provides a theoretical rationale for early pharmacologic intervention approaches using dopamine D(2) receptor blocking drugs.


Effects of fenbendazole on routine immune response parameters of BALB/c mice.
Fenbendazole (FBZ) is an anthelmintic drug widely used to treat and prevent pinworm outbreaks in laboratory rodents. Although data in nonrodent species indicate possible effects of fenbendazole on the bone marrow and lymphocyte proliferation and function, little has been reported regarding possible effects on the rodent immune system. The purpose of the current study was to determine the effects of a therapeutic regimen of FBZ on immune parameters in BALB/c mice. Both 9-wk on-off and 5-wk continuous medicated feed protocols were assessed. No significant differences between normal and FBZ diet treated mice were observed in the following parameters: complete blood count, blood chemistry, quantitation of major T and B cell markers in spleen, quantitation of T cell markers in the thymus, spleen cell proliferation to T and B cell mitogens, bone marrow colony-forming cell assays, skin graft rejection, and primary and secondary humoral immune responses. These data indicate that FBZ treatment does not affect many standard broad measures of immune function.


Experiments in Structural DNA Nanotechnology: Arrays and Devices.
In recent years, the chemistry of DNA has expanded from biological systems to nanotechnology. The generalization of the biological processes of reciprocal exchange leads to stable branched motifs that can be used for the construction of DNA-based geometrical and topological objects, arrays and nanomechanical devices. The information in DNA is the basis of life, but it can also be used to control the physical states of a variety of systems, leading ultimately to nanorobotics; these devices include shape-changing, walking and translating machines. We expect ultimately to be able to use the dynamic information-based architectural properties of nucleic acids to be the basis for advanced materials with applications from nanoelectronics to biomedical devices on the nanometer scale.


Bio-assay Guided Isolation of Anti-cancer Compounds from Anthocephalus cadamba Bark.
Anthocephalus cadamba, an important plant in the traditional system of medicine in India, is reported to possess anticancer activity. Guided by bio-assay tests using human colorectal (HCT116) and hepatocellular carcinoma (HepG2) cell lines, it has been shown to contain three active constituents, the triterpenoid saponins 3-O-[α-L-rhamnopyranosyl]-quinovic acid (1) and 3-O-[α-L-rhamnopyranosyl]-quinovic acid 28-O-[β-D-glucopyranosyl] ester (2), and the alkaloid cadambine (3). The structures of the isolated compounds were established using spectroscopic techniques. The isolated compounds demonstrated concentration dependent inhibition of both the cell lines, where compound 3 proved to be the most potent inhibitor of cell line HCT116 (IC50 45 +/- 4 μg/mL) and compound 2 demonstrated maximum inhibitory activity against HepG2 cell line with an IC50 value of 89 +/- 7 μg/mL.


Ethane groups modified DNA nanopores to prolong the dwell time on live cell membranes for transmembrane transport.
Transmembrane transport, mostly relying on biological channels, is crucial for the metabolic processes of live cells including sensing, signaling, cellular communicating and molecular transport. Artificial biomimetic channels offer excellent opportunities for studying the mechanisms of the metabolic processes of live cells and promote the applications of gene transfection, drug delivery, and regulations of cellular communications. DNA nanopores can be designed flexibly and operated easily while maintaining good biocompatibility, offering a good candidate for applications in basic research. However, because of the small size and good biocompatibility of DNA nanopores, it is still difficult to form stable channels on the plasma membrane of live cells by DNA nanopores. As a result, it significantly limits the applications of DNA nanopores in vivo. Thus, in this work, we have constructed ethane-phosphorothioate (PPT) groups modified DNA nanopores (E-DNA nanopores) to simulate biological channels for the transmembrane transport of small molecules. The E-DNA nanopores were found to be more hydrophobic and stable to anchor at the plasma membrane of live cells for a longer time window for subsequent transmembrane transport after the modification of ethane-PPT groups. The membrane-spanning E-DNA nanopores with a longer dwell time window could inspire the design of new DNA nanostructures and expand their biological applications including biosensing and sequencing, construction of artificial cells and regulation of transmembrane transport.


Disease-modifying antirheumatic drugs: gold, penicillamine, antimalarials, and sulfasalazine.
In the assessment of the effects of disease-modifying antirheumatic drugs, three or four clinical measurements supported by the erythrocyte sedimentation rate, and sometimes radiographs, are generally agreed to be correct. Some advocate functional assessments also, or even alternatively. Several studies compared gold, penicillamine, antimalarials, and sulfasalazine either with each other or with placebo, and occasionally with methotrexate. No important differences between the general performance of the four drugs were found. More work was reported on sulfasalazine than on the other three drugs; the data support that it has a place in our armamentarium. Several important contributions concerned strategies of treatment. It is considered that disease-modifying antirheumatic drugs should be used earlier and more aggressively in rheumatoid arthritis. This aspect was perhaps the key note of the 1990 literature on this topic. As part of the new strategies, combination therapy is urged by some rheumatologists, whereas others urge caution on the grounds that we do not yet know enough about the effects of combinations, or by how much the risks of adverse effects are increased in combination.


Clonally expanded mitochondrial DNA deletions within the choroid plexus in multiple sclerosis.
Mitochondrial DNA deletions (∆-mtDNA) have been implicated in the pathogenesis of Alzheimer's disease (AD), multiple sclerosis (MS) and Parkinson's disease (PD), as well as ageing. Clonal expansion of ∆-mtDNA is the process by which a mutant mtDNA molecule increases to high levels within a single cell containing both wild-type and mutant mtDNA. Unlike in AD and PD, the diffuse inflammatory process in MS involves the choroid plexus, and mitochondria are exposed to reactive oxygen and nitrogen species over a prolonged period. We determined the extent of respiratory enzyme deficiency and ∆-mtDNA at a single cell level within choroid plexus epithelial cells in MS as well as in AD, PD and controls. The respiratory enzyme-deficient (lacking complex IV and with intact complex II activity) cells were more prevalent within the choroid plexus in AD, MS and PD compared with controls. The main catalytic subunit of complex IV (subunit-I of cytochrome c oxidase) was lacking in significantly more respiratory enzyme-deficient cells in MS compared with AD, PD and controls. The single cell analysis showed a fourfold increase in the percentage of respiratory enzyme-deficient choroid plexus epithelial cells harbouring clonally expanded ∆-mtDNA in MS. Our findings establish clonal expansion of ∆-mtDNA as a feature relatively more prominent within the choroid plexus epithelium in MS than AD, PD or controls. We propose clonal expansion of ∆-mtDNA as a molecular link between inflammation and part of a delayed cellular energy failure in MS.


Induction and Analysis of Synchronous Meiotic Yeast Cultures.
Meiosis in Saccharomyces cerevisiae can be induced by deprivation of nutrients. Here, we present a protocol for inducing synchronous meiosis in SK1, the most efficient and synchronous yeast strain for meiosis, by exposing SK1 cells to liquid medium that contains potassium acetate as a nonfermentable carbon source and lacks nitrogen. These synchronous meiotic yeast cultures can be subjected to a range of molecular and cytological analyses, making them useful for investigating the genetic and molecular determinants of meiosis.


A complex scenario of tuberculosis transmission is revealed through genetic and epidemiological surveys in Porto.
Tuberculosis (TB) incidence is decreasing worldwide and eradication is becoming plausible. In low-incidence countries, intervention on migrant populations is considered one of the most important strategies for elimination. However, such measures are inappropriate in European areas where TB is largely endemic, such as Porto in Portugal. We aim to understand transmission chains in Porto through a genetic characterization of Mycobacterium tuberculosis strains and through a detailed epidemiological evaluation of cases. We genotyped the M. tuberculosis strains using the MIRU-VNTR system. We performed an evolutionary reconstruction of the genotypes with median networks, used in this context for the first time. TB cases from a period of two years were evaluated combining genetic, epidemiological and georeferencing information. The data reveal a unique complex scenario in Porto where the autochthonous population acts as a genetic reservoir of M. tuberculosis diversity with discreet episodes of transmission, mostly undetected using classical epidemiology alone. Although control policies have been successful in decreasing incidence in Porto, the discerned complexity suggests that, for elimination to be a realistic goal, strategies need to be adjusted and coupled with a continuous genetic characterization of strains and detailed epidemiological evaluation, in order to successfully identify and interrupt transmission chains.


A single dose of enterolactone activates estrogen signaling and regulates expression of circadian clock genes in mice.
Enterolactone (EL) is an enterolignan produced by gut microbiota from dietary plant lignans. Epidemiological and experimental studies suggest that EL and plant lignans may reduce the risk of breast and prostate cancer as well as cardiovascular disease. These effects are thought to at least in part involve modulation of estrogen receptor activity. Surprisingly little is known about the in vivo estrogenicity of EL. In the present study, we investigated the target tissues of EL, the genes affected by EL treatment, and the response kinetics. Following a single dose of EL, luciferase was significantly induced in reproductive and nonreproductive tissues of male and female 3xERE-luciferase mice, indicating estrogen-like activity. Microarray analysis revealed that EL regulated the expression of only 1% of 17β-estradiol target genes in the uterus. The majority of these genes were traditional estrogen target genes, but also members of the circadian signaling pathway were affected. Kinetic analyses showed that EL undergoes rapid phase II metabolism and is efficiently excreted. In vivo imaging demonstrated that the estrogen response followed similar, fast kinetics. We conclude that EL activates estrogen signaling in both male and female mice and that the transient responses may be due to the fast metabolism of the compound. Lastly, EL may represent a link among diet, gut microbiota, and circadian signaling.


[Drug therapy of ankylosing spondylitis].
Ankylosing spondylitis has to be treated with NSAID's in order to influence the inflammatory process and thus reducing pain and slowing down the stiffening of the vertebral column. For this purpose, apart from the NSAID's Sulphasalazine is used. As a symptomatic treatment of pain and muscle spasm simple analgesics and muscle relaxants can be used. In peripheral arthritis a local treatment with intraarticular steroids can be useful, and sometimes a radiosynoviorthesis can stop the inflammation for a longer period. In case of osteoporotic fractures prescription of calcitonin may be considered.


Definitive abdominal wall closure using a porcine intestinal submucosa biodegradable membrane in pediatric transplantation.
Abdominal closure in children less than 10 kg following liver or kidney transplantation can be challenging. Excessive pressure attained from a tight closure can result in abdominal compartment syndrome, graft compromise and loss. Concerns over using prosthetic patches are that of infection and dehiscence. We report a series of definitive abdominal wall closure using a biodegradable membrane from porcine intestinal submucosa (Surgisis; Cook Biotech Incorporated, West Lafayette, IN, USA). A prospective collection and follow up of liver and kidney transplant patients weighing less than 10 kg who required abdominal wall augmentation with Surgisis in order to achieve satisfactory closure. There were 10 liver and two renal transplant patients. The average weight of the liver transplant patients was 6.6 kg (5.4-8.5 kg) and the renal 9.8 kg. The average area of Surgisis used was 71.2 cm(2) (25-160 cm(2)) and length of follow up was 15.3 months (1-27 months). Concomitant measures to aid abdominal closure included bilateral recipient nephrectomy for the renal patients and reduction by 33% of the lateral segmental grafts in two liver transplant patients. Delayed closure occurred in all patients except one and the average days to closure from the first surgery was 3.75 days (0-6 days). Following liver transplantation one patient died from multiple organ failure at one month secondary to hemophagocytosis from underlying combined immune deficiency syndrome and one patient with hepatic artery thrombosis was salvaged at re-exploration. There were two wound complications, one patient developed two small sinuses and some skin dehiscence which healed over four months and the second developed a skin sinus following trans-patch liver biopsy which healed in three wk. Both had positive microbial cultures but neither necessitated removal of the graft. There were no incisional hernias. Surgisis is a safe method for facilitating abdominal closure in pediatric transplant patients. It appears to have long-term durability with no incisional hernias on short- and medium-term follow up, and is fairly resistant to infection.


Coinfection with HIV and tropical infectious diseases. II. Helminthic, fungal, bacterial, and viral pathogens.
The morbidity, mortality, and social disruption caused by the human immunodeficiency virus (HIV) pandemic continue to weigh disproportionately on resource-poor regions of the tropics. As a result, the potential for significant epidemiological, biological, and clinical interactions between HIV and other tropical pathogens is great. An overview of the available data on tropical helminths, fungi, bacteria, and viruses is provided here; interactions between HIV and tropical protozoa are covered in a related mini-review in this issue of Clinical Infectious Diseases. Special attention is given to evidence relevant to the hypothesis that helminth coinfection plays a particularly important role in accelerating the pace of HIV pathogenesis in the tropics.


An audit of malaria mortality using the "malaria death investigation form" at United Bulawayo Hospitals, Zimbabwe: 1996-2000.
To identify the main causes of confirmed malaria deaths and assess the validity and the relevance of use of the MDIF in determining areas for improvement of care. A cross sectional study. United Bulawayo Hospitals, a tertiary hospital in Zimbabwe. Patients whose cause of death was malaria using MIDF. Of 470 confirmed cases admitted in UBH during the study period, 53 (11.2%) died and were included in the study. Most deaths occurred in the over 15 years age group (88.6%) with only 3 deaths each in the other groups. All patients were referred or admitted to UBH with complicated and severe malaria; 39(74 %) had more than one complication such as CM and acute renal failure (ARP). Most patients came from or had visited a rural area and did not implement basic prophylactic and therapeutic measures put in place by the NMCP such as early self-medication. Three pregnant women aborted. Guidelines regarding investigations and treatment were not strictly adhered to. Delay in seeking treatment and in referring was generally observed at all levels of the health system. Cases of malaria deaths were found in the city in people who had not travelled to rural area. The MDIF was used in one case only. Malaria mortality accounted for 11 % of confirmed cases. Main causes of death were CM and ARP Parameters contained in the MDIF were those utilised by most authors who have investigated malaria mortality in Africa and there was a similarity in the observations. In view of the information it could provide if properly used, the MDIF is a valid tool for collecting data that the NMCP needs in order to rationalise its strategies at UBH and in other health facilities. Its use should be generalised and compulsory.


[Curcumin induces apoptosis and protective autophagy in human gastric cancer cells with different degree of differentiation].
Objective: To investigate the effect of curcumin on the apoptosis and autophagy of human gastric cancer cells with different degree of differentiation. Methods: Gastric cancer cell lines BGC-823 and MKN-28 were treated with curcumin at different concentrations. The effect of curcumin on cell proliferation was measured by MTT assay. Apoptosis was assessed by flow cytometry. Autophagy status was analyzed by acridine orange staining. The expression levels of apoptotic and autophagy-related proteins were detected by Western blot. Results: The cell viability of BGC-823 and MKN-28 was inhibited by curcumin in a time- and dose-dependent manner. At 48 h after treatment, the IC(50) value of BGC-823 (15.18 μmol/L) was close to that of MKN-28 (15.84 μmol/L), and the difference was not statistically significant (P=0.513). Meanwhile, flow cytometry showed that curcumin induced the apoptosis of gastric cancer cells in a dose-dependent manner. Western blot results showed that the expression of pro-apoptotic proteins bax, active-caspase-3 and active-caspase-9 was significantly increased in BGC-823 and MKN-28 cells, whereas that of the anti-apoptotic protein bcl-2 was strikingly reduced. In addition, the formation of acidic vesicular organelles in cytoplasm, conversion of LC3-Ⅰ to LC3-Ⅱ and increased levels of autophagy-related proteins Beclin1, Atg7 and Atg5-Atg12 were observed in curcumin-treated cells. Moreover, activation of PI3K/Akt/mTOR signaling pathway was also significantly suppressed after curcumin treatment. Blocking autophagy by adding the autophagy inhibitor 3-methyladenine (3-MA) significantly promoted the apoptotic cell death induced by curcumin. Conclusions: Curcumin induces apoptosis and protective autophagy in human gastric cancer cells in vitro. Curcumin combined with autophagy inhibitor may provide a more effective strategy for its clinical application.


Ultrastructural bases for metabolically linked mechanical activity in mitochondria. II. Electron transport-linked ultrastructural transformations in mitochondria.
Isolated mitochondria are capable of undergoing dramatic reversible ultrastructural transformations between a condensed and an orthodox conformation. These two conformations are the extremes in ultrastructural organization between which structually and functionally intact mitochondria transform during reversible respiratory cycles. It has been found that electron transport is required for the condensed-to-orthodox ultrastructural transformation which occurs in mitochondria under State IV conditions, i.e., under conditions in which exogenous substrate is present and ADP is deficient. Inhibition of State IV electron transport at the cyanide-, antimycin A-, or Amytal-sensitive sites in the respiratory chain results in inhibition of this transformation. Resumption of electron transport in initially inhibited mitochondrial systems, initiated by channeling electrons through pathways which bypass the inhibited sites, results in resumption of the ultrastructural transformation. The condensed-to-orthodox transformation is DNP insensitive and, therefore, does not require participation of the coupling enzymes of the energy-transfer pathway. It is concluded that this ultrastructural transformation is manifest by the conversion of the chemical energy of electron transport directly into mechanical work. The reversed ultrastructural transformation, i.e., orthodox-to-condensed, which occurs during ADP-activated State III electron transport, is inhibited by DNP and parallels suppression of acceptor control and oxidative phosphorylation. Mechanochemical ultrastructural transformation as a basis for energy transfer in mitochondria is considered with respect to the results presented.


The role of nicotine and nicotinic mechanisms in neuropsychiatric disease.
Cigarette smoking, and by implication nicotine, may be involved as a negative or positive risk factor in some neuropsychiatric disorders and possibly as a treatment in others. Nicotine exposure may be a negative risk factor for the development of Parkinson's disease, but a positive risk factor for the development of tardive dyskinesia. For Alzheimer's disease and Tourette's syndrome, the role of nicotine exposure is equivocal, however, the role of nicotine as a possible therapeutic agent, alone or in combination, remains an intriguing question. For functional psychiatric disorders, the data are suggestive of a link between tobacco use and at least exacerbation of some disorders. While nicotine exposure is unlikely to be critical in the genesis of these disorders, it may complicate the pharmacological therapeutics and long-term prognosis. Further research is needed to examine the actual importance of tobacco use in behavioural disturbances. The relative importance of central nicotinic mechanisms in normal and disordered human cognition and movement is now beginning to be fully explored.


Middle meningeal artery embolization in the management of chronic subdural hematoma in medically complex pediatric neurosurgical patients: technical note.
Middle meningeal artery (MMA) embolization has gained acceptance as a treatment for chronic subdural hematoma (cSDH) in adult patients but has not been well described in pediatric patients. Standard cSDH treatment has historically consisted of burr hole drainage with or without subdural drain placement. However, due to the high rate of recurrence and frequency of comorbidities within this population, as both pediatric and adult patients with cSDH frequently have concurrent cardiac disease and a need for anticoagulant therapies, MMA embolization has increasingly demonstrated its value as both an adjunctive and primary treatment. In this report, the authors present 3 cases of successful MMA embolization in medically complex children at a single institution. MMA embolization was used as a primary treatment modality and as an adjunctive therapy in the acute setting following surgical hematoma evacuation. Two patients were receiving anticoagulation treatment requiring reversal. Technical considerations specific to the pediatric population as well as those common to both the pediatric and adult populations are addressed. Further work is needed to define the optimal indications and outcomes for MMA embolization in children with cSDH.


Role of P-450 arachidonic acid epoxygenase in the response of cerebral blood flow to glutamate in rats.
Glutamate, a major excitatory neurotransmitter in the brain, has been implicated in the hyperemic response to increases in the activity of neurons, but the mechanism of glutamate-induced dilation of cerebral blood vessels is unknown. Glutamate has been shown to enhance the release of arachidonic acid (AA) in brain tissue and cultured astrocytes. We have previously shown that astrocytes metabolize AA to vasodilator products, epoxyeicostrienoic acids (EETs), and express a P-450 AA epoxygenase, P-450 2C11. We tested the hypothesis that glutamate-induced dilation of cerebral arterioles is mediated in part by changes in the formation and release of EETs by perivascular astrocytes. Primary astrocyte cultures were prepared from 3-day-old rat pups. The cells were labeled with [14C]AA, and the effect of glutamate on the formation of EETs from [14C]AA by cultured astrocytes was studied. The expression of P-450 2C11 protein in the microsomal fractions of cultured astrocytes was assessed by Western blot. In vivo cerebral blood flow measurements were made in adult rats by laser-Doppler flowmetry after administration of glutamate into the subdural space of the rat before and after treatment with miconazole. Glutamate treatment (100 mumol/L for 30 minutes) induced a threefold increase in the formation of EETs from [14C]AA by cultured astrocytes, and the increase was inhibited by miconazole (20 mumol/L), an inhibitor of P-450 AA epoxygenase. Treatment with glutamate (100 mumol/L) for 12 hours increased the expression of P-450 2C11 protein in the microsomal fraction of cultured astrocytes. The response of laser-Doppler cerebral blood flow to administration of glutamate (500 mumol/L) into the subdural space of the rat was significantly attenuated after treatment with miconazole (20 mumol/L for 30 minutes). These findings suggest a role for a P-450 AA epoxygenase in astrocytes in the coupling between the metabolic activity of neurons and regional blood flow in the brain.


Comparison of Tight Junction Protein-Related Gene mRNA Expression Levels between Male and Female Gastroesophageal Reflux Disease Patients.
Male predominance has been observed in the erosive reflux disease (ERD), but reverse finding in nonerosive reflux disease (NERD). This suggests sex-specific medicine approach is needed but its mechanism is remained to be elucidated. We aimed to compare clinical characteristics and mRNA expression levels of tight junction-related proteins between male and female gastroesophageal reflux disease (GERD). Sixteen healthy controls, 45 ERD, and 14 NERD patients received upper endoscopies and completed questionnaires. Quantitative real-time polymerase chain reactions of occludin (OCLN), zonal occludens (ZO) 1, claudin-1 (CLDN1) and claudin-4 (CLDN4), and neurokinin 1 receptor (NK1R) were performed in the distal esophageal mucosal specimen. These results were analyzed by sex. Female GERD patients were affected more by reflux symptoms than males. The impairment of overall quality of life was more prominent in female patients with reflux symptoms than male patients (5.6±0.2 vs 4.9±0.6, p=0.009). The levels of OCLN mRNA expression were significantly lower in the male ERD group. On the other hand, those of CLDN1, CLDN4, and NK1R except ZO-1 were significantly higher in the male ERD group. We demonstrated that female ERD/NERD patients were affected more by GERD and male ERD patients showed significant changes of tight junction protein mRNA expression levels.


[Histamine intolerance and pseudoallergy--do they exist?].
Histamine intolerance (HIT) is a controversial syndrome, in which the body is believed to react to histamine released by certain foodstuffs. Symptoms include among other things headache, urticaria and abdominal discomfort. Several fruits, nuts, spices and food additives have been regarded as pseudoallergens. Since there is no objective investigation to prove HIT, the diagnosis is based on symptoms. On the basis of literature, better clinical studies are needed. Because the phenomenon remains unclear, use of the terms histamine intolerance and pseudoallergens is not recommended.


Lactobacillus reuteri NK33 and Bifidobacterium adolescentis NK98 Alleviate Escherichia coli-Induced depression and Gut Dysbiosis in Mice.
Lactobacillus reuteri NK33 (NK33) and Bifidobacterium adolescentis NK98 (NK98) alleviate immobilization stress-induced depression. To understand the gut microbiota-mediated mechanisms of NK33 and NK98 against depression, we examined their effects on Escherichia coli K1 (K1)-induced depression and gut dysbiosis in mice. NK33, NK98, and their mixtures (1:1, 4:1, and 9:1) mitigated K1-induced depression and colitis. NK33 and NK98 additively or synergistically increased BDNF+/NeuN+ cell population and suppressed NF-κB action in the hippocampus. They alleviated gut dysbiosis by reducing the Proteobacteria population and increasing the Clostridia population. These results suggest that NK33 and NK98 may alleviate depression and colitis by ameliorating gut dysbiosis.


EFFECTS OF WHOLE BODY VIBRATION INTERVENTION ON HANDGRIP STRENGTH OF BRAZILIAN HEALTHY SOLDIERS.
Whole body vibration (WBV) exercises have been investigated as an alternative and complementary method to traditional resistance programs for fitness improvements in healthy subjects. Active militaries must have a high fitness level and have to improve some physical abilities to accomplish some specific tasks in Army, as climb ropes, climb walls and pull up. Seven young and healthy soldiers from Brazilian Army were exposed to mechanical vibration in a push-up position, with different frequencies, as 25 (first session), 30 (second session), 35 (third session), 40 (fourth session) and 45 Hz (fifth session). The WBV intervention consisted of five WBV sessions over a consecutive two and a half-weeks period of time. A WBV session consisted of twenty bouts, each one with 10- second vibration, interspersed with 10 seconds of passive rest and 4 minutes pause after the first 10 bouts. Handgrip strength was measured, as proposed by American Society of Hand Therapists - ASHT, before and after the session using 25 Hz of frequency and 45 Hz of frequency (1st and 5th sessions). No significant results were found between groups. Intervention of WBV exercises with 25 and 45 Hz of frequency applied by hands of healthy and young soldiers had no significant results. Further investigations should focus on the efficacy of WBV in the improvement of handgrip strength of young and healthy people.


Nanoscale effects in dendrimer-mediated targeting of neuroinflammation.
Neuroinflammation, mediated by activated microglia and astrocytes, plays a key role in the pathogenesis of many neurological disorders. Systemically-administered dendrimers target neuroinflammation and deliver drugs with significant efficacy, without the need for ligands. Elucidating the nanoscale aspects of targeting neuroinflammation will enable superior nanodevices for eventual translation. Using a rabbit model of cerebral palsy, we studied the in vivo contributions of dendrimer physicochemical properties and disease pathophysiology on dendrimer brain uptake, diffusion, and cell specific localization. Neutral dendrimers move efficiently within the brain parenchyma and rapidly localize in glial cells in regions of injury. Dendrimer uptake is also dependent on the extent of blood-brain-barrier breakdown, glial activation, and disease severity (mild, moderate, or severe), which can lend the dendrimer to be used as an imaging biomarker for disease phenotype. This new understanding of the in vivo mechanism of dendrimer-mediated delivery in a clinically-relevant rabbit model provides greater opportunity for clinical translation of targeted brain injury therapies.


Generation and characterization of antibodies to adhesion-related molecules of retinal pigment epithelial cells.
We have generated antisera to pig retinal pigment epithelial (RPE) cells and to their fractionated membrane proteins. The antisera have been screened functionally by adhesion inhibition assays, and morphologically with immunofluorescence microscopy of cultured cells and frozen sections of the retina. From these sera, we have affinity purified five monospecific antibodies to the RPE surface molecules having molecular weights of 220, 180, 110, 85 and 70 kDa, which effectively inhibit cell-substratum attachment. The 220 and 85 kDa antigens are localized to focal contacts of the cultured cells and are seen in patches in the intact RPE layer in frozen sections of eye tissues. Immunofluorescence microscopy shows that the 180, 110 and 70 kDa antigens localize predominantly to cell-cell junctions. Hence, these antigens may participate in both cell-cell and cell-substratum adhesion in RPE cells. Staining patterns obtained with confocal microscopy of frozen sections of the retina demonstrate that these antibodies are RPE-specific. These adhesion-related molecules may play a role in maintenance of the morphological and functional integrity of RPE.


A loss-of-function NCSTN mutation associated with familial Dowling Degos disease and hidradenitis suppurativa.
Dowling Degos disease (DDD) is a rare autosomal dominant genodermatosis characterized by acquired, slowly progressive reticulated pigmented lesions primarily involving flexural skin areas. Mutations in KRT5, POGLUT-1 and POFUT-1 genes have been associated with DDD, and loss-of-function mutations in PSENEN, a subunit of the gamma-secretase complex, were found in patients presenting with DDD or DDD comorbid with hidradenitis suppurativa (HS). A nonsense mutation in NCSTN, another subunit of the gamma-secretase, was already described in a patient suffering from HS and DDD but whether NCSTN could be considered a novel gene for DDD is still debated. Here, we enrolled a four-generation family with HS and DDD. Through Whole Exome Sequencing (WES) we identified a novel nonsense mutation in the NCSTN gene in all the affected family members. To study the impact of this variant, we isolated outer root sheath cells from patients' hair follicles. We showed that this variant leads to a premature stop codon, activates a nonsense-mediated mRNA decay, and causes NCSTN haploinsufficiency in affected individuals. In fact, cells treated with gentamicin, a readthrough agent, had the NCSTN levels corrected. Moreover, we observed that this haploinsufficiency also affects other subunits of the gamma-secretase complex, possibly causing DDD. Our findings clearly support NCSTN as a novel DDD gene and suggest carefully investigating this co-occurrence in HS patients carrying a mutation in the NCSTN gene.


Selective construction of polycyclic spirooxindoles via a Cu(OTf)2/HOTf-catalyzed domino reaction of o-arylalkynylacetophenones and 3-phenacylideneoxindoles.
Under the combined catalysis of Cu(OTf)2/HOTf, the domino annulation reaction of o-arylalkynyl acetophenones with 3-phenacylideneoxindoles in refluxing acetonitrile selectively afforded functionalized spiro[indoline-3,7'-tetrapheno[7,6-bc]furans] and spiro[indeno[1,2-b]naphtho[2,1-d]furan-7,3'-indolines] depending on the electronic effect of the substituents on both substrates.


Vertebropexy as a semi-rigid ligamentous alternative to lumbar spinal fusion.
To develop ligamentous vertebral stabilization techniques ("vertebropexy") that can be used after microsurgical decompression (intact posterior structures) and midline decompression (removed posterior structures) and to elaborate their biomechanical characteristics. Fifteen spinal segments were biomechanically tested in a stepwise surgical decompression and ligamentous stabilization study. Stabilization was achieved with a gracilis or semitendinosus tendon allograft, which was attached to the spinous process (interspinous vertebropexy) or the laminae (interlaminar vertebropexy) in form of a loop. The specimens were tested (1) in the native state, after (2) microsurgical decompression, (3) interspinous vertebropexy, (4) midline decompression, and (5) interlaminar vertebropexy. In the intact state and after every surgical step, the segments were loaded in flexion-extension (FE), lateral shear (LS), lateral bending (LB), anterior shear (AS) and axial rotation (AR). Interspinous vertebropexy significantly reduced the range of motion (ROM) in all loading scenarios compared to microsurgical decompression: in FE by 70% (p < 0.001), in LS by 22% (p < 0.001), in LB by 8% (p < 0.001) in AS by 12% (p < 0.01) and in AR by 9% (p < 0.001). Interlaminar vertebropexy decreased ROM compared to midline decompression by 70% (p < 0.001) in FE, 18% (p < 0.001) in LS, 11% (p < 0.01) in LB, 7% (p < 0.01) in AS, and 4% (p < 0.01) in AR. Vertebral segment ROM was significantly smaller with the interspinous vertebropexy compared to the interlaminar vertebropexy for all loading scenarios except FE. Both techniques were able to reduce vertebral body segment ROM in FE, LS and LB beyond the native state. Vertebropexy is a new concept of semi-rigid spinal stabilization based on ligamentous reinforcement of the spinal segment. It is able to reduce motion, especially in flexion-extension. Studies are needed to evaluate its clinical application.


MolTaut: A Tool for the Rapid Generation of Favorable Tautomer in Aqueous Solution.
Fast and proper treatment of the tautomeric states for drug-like molecules is critical in computer-aided drug discovery since the major tautomer of a molecule determines its pharmacophore features and physical properties. We present MolTaut, a tool for the rapid generation of favorable states of drug-like molecules in water. MolTaut works by enumerating possible tautomeric states with tautomeric transformation rules, ranking tautomers with their relative internal energies and solvation energies calculated by AI-based models, and generating preferred ionization states according to predicted microscopic pKa. Our test shows that the ranking ability of the AI-based tautomer scoring approach is comparable to the DFT method (wB97X/6-31G*//M062X/6-31G*/SMD) from which the AI models try to learn. We find that the substitution effect on tautomeric equilibrium is well predicted by MolTaut, which is helpful in computer-aided ligand design. The source code of MolTaut is freely available to researchers and can be accessed at https://github.com/xundrug/moltaut. To facilitate the usage of MolTaut by medicinal chemists, we made a free web server, which is available at http://moltaut.xundrug.cn. MolTaut is a handy tool for investigating the tautomerization issue in drug discovery.


An unusual cause of chest pain: foreign body in the oesophagus.
This article reports on two patients with unsuspected oesophageal foreign body ingestion, with chest pain as the main symptom. The patients had extensive cardiac evaluation to rule out myocardial ischaemia. Further evaluation showed an impacted oesophageal foreign body. Oesophageal foreign bodies should be considered as factors in chest pain.


Newly Established Genetic System for Functional Analysis of MetSV.
The linear chromosome of the Methanosarcina spherical virus with 10,567 bp exhibits 22 ORFs with mostly unknown functions. Annotation using common tools and databases predicted functions for a few genes like the type B DNA polymerase (MetSVORF07) or the small (MetSVORF15) and major (MetSVORF16) capsid proteins. For verification of assigned functions of additional ORFs, biochemical or genetic approaches were found to be essential. Consequently, we established a genetic system for MetSV by cloning its genome into the E. coli plasmid pCR-XL-2. Comparisons of candidate plasmids with the MetSV reference based on Nanopore sequencing revealed several mutations of yet unknown provenance with an impact on protein-coding sequences. Linear MetSV inserts were generated by BamHI restriction, purified and transformed in Methanosarcina mazei by an optimized liposome-mediated transformation protocol. Analysis of resulting MetSV virions by TEM imaging and infection experiments demonstrated no significant differences between plasmid-born viruses and native MetSV particles regarding their morphology or lytic behavior. The functionality of the genetic system was tested by the generation of a ΔMetSVORF09 mutant that was still infectious. Our genetic system of MetSV, the first functional system for a virus of methanoarchaea, now allows us to obtain deeper insights into MetSV protein functions and virus-host interactions.


Cyclosporin neurotoxicity in cardiac transplant recipients.
Cyclosporin toxicity may produce a wide range of neurological disorders. We report three patients whose neurological problems developed while taking cyclosporin following cardiac transplantation, but resolved rapidly when the drug was discontinued. In two cases, MRI revealed abnormalities which disappeared with clinical recovery. Blood cyclosporin concentrations were not markedly increased, and it is possible that the neurotoxic effects of cyclosporin are mediated by metabolites, through an effect on the blood-brain barrier.


Characterization of the platelet-activating factor acetylhydrolase from human plasma by heterologous expression in Xenopus laevis oocytes.
Platelet-activating factor (PAF) has been implicated as a mediator of inflammation and atherosclerosis. A specific degradative enzyme found in plasma, PAF acetylhydrolase, plays important roles in various pathophysiological events induced by PAF. Human macrophages and Hep G2 cells secrete PAF acetylhydrolase with characteristics identical to the plasma activity. Other investigators reported that apolipoprotein B may possess phospholipase A2 activity, which suggested that apolipoprotein B might be a zymogen for PAF acetylhydrolase. However, while macrophages express PAF acetylhydrolase activity, we did not detect cDNAs for apolipoprotein B in a cDNA library from these cells, indicating that macrophages do not express this protein. In contrast, Hep G2 cells had high levels of cDNA for apolipoprotein B, as expected. We next injected Xenopus laevis oocytes with poly(A)+ RNA extracted from cultured human macrophages and Hep G2 cells. Twenty-five to 50% of Xenopus oocytes injected with poly(A)+ RNA from macrophages or Hep G2 cells secreted a PAF acetylhydrolase activity (1.0-7.8 nmol/ml per h) that also utilized a synthetic oxidized phospholipid as substrate. The activity secreted by poly(A)+ RNA-injected oocytes associated with lipoproteins and transferred between the particles in a pH-dependent manner, much like the plasma activity. These experiments establish that the properties of the enzyme released from poly(A)+ RNA-injected oocytes are identical to those of the plasma form of PAF acetylhydrolase and that the activity detected is not the expression of a domain in apolipoprotein B.


Role of 90-kDa heat shock protein (Hsp 90) and protein degradation in regulating neuronal levels of G protein-coupled receptor kinase 3.
Cellular levels of G protein-coupled receptor kinase (GRK)3 determine the sensitivity of the alpha(2A/B)-adrenoceptor (alpha(2)-AR) to agonist-induced down-regulation. Using human neuroblastoma BE(2)-C cells, this study examines how cellular GRK3 levels are affected by several mechanisms reported to influence stability and degradation of other GRKs. We first examined the interaction between the 90-kDa heat shock protein (Hsp90) and GRK3; Hsp90 reportedly affects the maturation and stability of GRK2. In unstimulated cells, GRK3 coimmunoprecipitates with Hsp90, suggesting a physical interaction. Moreover, when GRK3 protein expression was increased by 24-h epinephrine (EPI) treatment, Hsp90 protein expression increased with a similar but slightly delayed time course. To investigate the influence of Hsp90 on GRK3 protein stability, we determined the effect of the Hsp90 inhibitor geldanamycin (GA) on cellular GRK3 levels. GA eliminated the interaction between Hsp90 with GRK3 and produced a rapid, proteasome-mediated, 70% decrease in GRK3 levels within 24 h. To investigate the influence of Hsp90 on up-regulation of GRK3 expression, we examined the effect of GA on EPI-induced up-regulation. GA reduced the absolute increase in GRK3; however, the percentage of increase in GRK3 by EPI was not significantly different in the absence versus presence of GA (141 +/- 41 versus 94 +/- 12%). Finally, we examined the influence of Ca(2+)-activated proteases on cellular GRK3. Treatment with the calcium ionophore ionomycin produced a rapid decrease in GRK3 levels that was inhibited by the calpain inhibitor calpeptin. In conclusion, several mechanisms influence the degradation of GRK3 and therefore have the potential to affect GPCR signaling by regulating GRK3 levels in neurons.


Cyanobactericidal effect of Streptomyces sp. HJC-D1 on Microcystis auruginosa.
An isolated strain Streptomyces sp. HJC-D1 was applied to inhibit the growth of cyanobacterium Microcystis aeruginosa FACHB-905. The effect of Streptomyces sp. HJC-D1 culture broth on the cell integrity and physiological characteristics of M. aeruginosa FACHB-905 was investigated using the flow cytometry (FCM), enzyme activity and transmission electron microscopy (TEM) methods. Results showed that the growth of M. aeruginosa FACHB-905 was significantly inhibited, and the percentage of live cells depended on the culture broth concentration and exposure time. The activities of antioxidant enzymes including superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) increased with exposure concentration and exposure time, and the significant increase of reactive oxygen species (ROS) led to the disruption of the subcellular structure of M. aeruginosa FACHB-905, and caused the increase of malondialdehyde (MDA). Furthermore, TEM observation suggested the presence of three stages (cell breakage, organelle release and cell death) for the cyanobactericidal process of Streptomyces sp. HJC-D1. Therefore, Streptomyces sp. HJC-D1 not only affected antioxidant enzyme activities and ROS level, but also destroyed the subcellular structure of M. aeruginosa FACHB-905, demonstrating excellent cyanobactericidal properties.


Droplet digital PCR for detection of BRAF V600E mutation in formalin-fixed, paraffin-embedded melanoma tissues: a comparison with Cobas® 4800, Sanger sequencing, and allele-specific PCR.
Cutaneous melanoma has the worst prognosis of all skin cancers. Although emerging targeted therapies, such as B-Raf kinase inhibitor vemurafenib, improve prognosis they require an accurate and sensitive means of detecting the pathogenic BRAF V600E mutation. We compared the sensitivity of four BRAF V600E detection methods in formalin-fixed, paraffin-embedded melanoma biopsies from 87 consecutive melanoma patients with Breslow stage I-V disease (staging based on the depth of tumor of invasion). The methods assessed were the widely used Cobas® 4800 system based on real-time PCR amplification, Sanger sequencing, allele-specific PCR (AS-PCR), and droplet digital PCR (ddPCR). The BRAF V600E mutation was found in 8 (9.2%), 23 (26.4%), 23 (26.4%) and 31 (35.6%) biopsies, respectively. The limit of detection (LoD) was determined by three different methods: Poisson confidence limits, calibration regression and Tzonev's method. Pair-wise agreement between the methods was as follows: Cobas vs. Sanger, P = 0.33; Cobas® 4800 vs. AS-PCR, P = 0.33; Cobas® 4800 vs. ddPCR, P = 0.65; Sanger vs. AS-PCR, P = 1; Sanger vs. ddPCR, P = 0.08; AS-PCR vs. ddPCR, P = 0.06. Multinomial logistic regression was used for predictive modeling of the Breslow-Clark score; ddPCR emerged as the best predictor, the other predictors were mitotic activity, type of malignant melanoma and patient's age. Our results demonstrate that ddPCR is the most sensitive method of detecting the BRAF V600E mutation.


The non-human reservoirs of Ross River virus: a systematic review of the evidence.
Understanding the non-human reservoirs of zoonotic pathogens is critical for effective disease control, but identifying the relative contributions of the various reservoirs of multi-host pathogens is challenging. For Ross River virus (RRV), knowledge of the transmission dynamics, in particular the role of non-human species, is important. In Australia, RRV accounts for the highest number of human mosquito-borne virus infections. The long held dogma that marsupials are better reservoirs than placental mammals, which are better reservoirs than birds, deserves critical review. We present a review of 50 years of evidence on non-human reservoirs of RRV, which includes experimental infection studies, virus isolation studies and serosurveys. We find that whilst marsupials are competent reservoirs of RRV, there is potential for placental mammals and birds to contribute to transmission dynamics. However, the role of these animals as reservoirs of RRV remains unclear due to fragmented evidence and sampling bias. Future investigations of RRV reservoirs should focus on quantifying complex transmission dynamics across environments.


Relationships between physical symptoms, emotional distress, and pain appraisal in fibromyalgia: the moderator effect of alexithymia.
Alexithymia is a personality construct that is frequently identified in fibromyalgia (FM). Previous studies have explored the relationship between alexithymia and emotional distress in this disease. Yet, the additional link with factors of pain appraisal is unknown. This study examined the moderating effect of alexithymia in the relationship between emotional distress and pain appraisal in 97 FM women. A control group of 100 healthy women also participated in the study. All participants completed several self-reports about pain experience, sleep quality, impairment, emotional distress, pain appraisal, and alexithymia. FM women showed significantly more difficulty in identifying and describing feelings, but less externally oriented thinking than healthy women. In the clinical group, difficulty in identifying feelings and difficulty in describing feelings significantly correlated with lower sleep quality, higher anxiety and depression, and increased pain catastrophizing and fear of pain. Difficulty in describing feelings significantly correlated with higher pain experience and vigilance to pain. Externally oriented thinking was not correlated with any of the clinical variables. Difficulty in identifying feelings moderated the relationship between anxiety and pain catastrophizing, and difficulty in describing feelings moderated the relationship between anxiety and fear of pain. Implications of the findings for the optimization of care of FM patients are discussed.


p21-activated protein kinase 1 induces the invasion of gastric cancer cells through c-Jun NH2-terminal kinase-mediated activation of matrix metalloproteinase-2.
Gastric cancer (GC) is one of the most common malignancies worldwide. The prognosis of GC is poor, mostly due to widespread metastasis. p21-activated kinase 1 (Pak1), the best characterized member of an evolutionarily conserved family of serine/threonine kinases, plays an important role in the regulation of cell morphogenesis, motility, mitosis and angiogenesis. By qRT-PCR and Gelatin zymograph assay, we demonstrated in the present study that stable overexpression of Pak1 induced matrix metalloproteinase (MMP)-2 mRNA expression and activity in the human MKN45 GC cell line. Conversely, knockdown of endogenous Pak1 expression by small interfering RNA (siRNA) decreased MMP-2 mRNA expression and activity in the MKN45 GC cells. Activation of c-Jun N-terminal kinase (JNK) was required for Pak1-induced upregulation of MMP-2 mRNA level and activity. Moreover, upregulation of MMP-2 by Pak1 via the JNK pathway notably promoted the invasion of MKN45 GC cells. Overexpression of MMP-2 mRNA was once again confirmed to be associated with GC metastasis. In conclusion, our results demonstrated for the first time that Pak1 stimulated MMP-2 mRNA expression and activity in MKN45 GC cells. The JNK signaling pathway was involved in Pak1 modulation of MMP-2, which was important for MKN45 GC cell invasiveness.


Space-charge-limited 2D electron flow between two flat electrodes in a strong magnetic field.
An approximate analytic solution is constructed for the 2D space-charge-limited emission by a cathode surrounded by nonemitting conducting ledges of width Lambda. An essentially exact solution (via conformal mapping) of the electrostatic problem in vacuum is matched to the solution of a linearized problem in the space charge region whose boundaries are sharp due to the presence of a strong magnetic field. The current density growth in a narrow interval near the edges of the cathode depends strongly on Lambda. We obtain an empirical formula for the total current as a function of Lambda which extends to more general cathode geometries.


Fever of unknown origin: a report from China of 208 cases.
Fever of unknown origin (FUO) is a clinical dilemma in western countries and in China. To investigate the causes and prognosis of FUO, 208 patients with FUO admitted to a large university hospital in China were investigated. The final diagnoses established in 158 cases (75.96%) were: infectious disease in 66 cases (31.73%), collagen vascular disease in 46 patients (22.11%), neoplasm in 35 cases (16.83%), and other disease in 11 patients (5.29%). In 66 cases with infectious disease, tuberculosis, septicaemia and typhoid fever were the principal causes. SLE and adult Still's disease were the most important causes among collagen vascular disease. Lymphoma and malignant histiocytosis were mostly associated with FUO among neoplasms. In 50 cases (24.04%), the cause of fever was not found. On discharge from hospital, fever had subsided in 133 cases (63.94%), and had persisted in 63 cases (30.29%); 12 patients (5.87%) died. In China, infectious disease, collagen vascular disease and neoplasm are the main causes of FUO. While most patients recover, there are some differences in the distribution of causes between the West and China, and there are relatively more deaths than in previous reports.


Quantitative analysis of factors affecting intraoperative precision and stability of optoelectronic and electromagnetic tracking systems.
This study aims to provide a quantitative analysis of the factors affecting the actual precision and stability of optoelectronic and electromagnetic tracking systems in computer-aided surgery under real clinical/intraoperative conditions. A "phantom-skull" with five precisely determined reference distances between marker spheres is used for all measurements. Three optoelectronic and one electromagnetic tracking systems are included in this study. The experimental design is divided into three parts: (1) evaluation of serial- and multislice-CT (computed tomography) images of the phantom-skull for the precision of distance measurements by means of navigation software without a digitizer, (2) digitizer measurements under realistic intraoperative conditions with the factors OR-lamp (radiating into the field of view of the digitizer) or/and "handling with ferromagnetic surgical instruments" (in the field of view of the digitizer) and (3) "point-measurements" to analyze the influence of changes in the angle of inclination of the stylus axis. Deviations between reference distances and measured values are statistically investigated by means of analysis of variance. Computerized measurements of distances based on serial-CT data were more precise than based on multislice-CT data. All tracking systems included in this study proved to be considerably less precise under realistic OR conditions when compared to the technical specifications in the manuals of the systems. Changes in the angle of inclination of the stylus axis resulted in deviations of up to 3.40 mm (mean deviations for all systems ranging from 0.49 to 1.42 mm, variances ranging from 0.09 to 1.44 mm), indicating a strong need for improvements of stylus design. The electromagnetic tracking system investigated in this study was not significantly affected by small ferromagnetic surgical instruments.


Immunological effects of the PE/PPE family proteins of Mycobacterium tuberculosis and related vaccines.
Tuberculosis (TB) is a chronic infectious disease caused by Mycobacterium tuberculosis (Mtb), and its incidence and mortality are increasing. The BCG vaccine was developed in the early 20th century. As the most widely administered vaccine in the world, approximately 100 million newborns are vaccinated with BCG every year, which has saved tens of millions of lives. However, due to differences in region and race, the average protective rate of BCG in preventing tuberculosis in children is still not high in some areas. Moreover, because the immune memory induced by BCG will weaken with the increase of age, it is slightly inferior in preventing adult tuberculosis, and BCG revaccination cannot reduce the incidence of tuberculosis again. Research on the mechanism of Mtb and the development of new vaccines against TB are the main strategies for preventing and treating TB. In recent years, Pro-Glu motif-containing (PE) and Pro-Pro-Glu motif-containing (PPE) family proteins have been found to have an increasingly important role in the pathogenesis and chronic protracted infection observed in TB. The development and clinical trials of vaccines based on Mtb antigens are in progress. Herein, we review the immunological effects of PE/PPE proteins and the development of common PE/PPE vaccines.


Lung cancer prevention with (-)-epigallocatechin gallate using monitoring by heterogeneous nuclear ribonucleoprotein B1.
Considering the problems involved in prevention of human lung cancer, growth inhibition of human lung cancer cell line A549 was studied with emphasis on two parameters: green tea polyphenols, such as (-)-epigallocatechin gallate (EGCG) and (-)-epicatechin gallate (ECG); and heterogeneous nuclear ribonucleoprotein B1 (hnRNP B1), a new biomarker of human lung cancer which is highly expressed in the very early stages of human lung cancer. The inhibitory potencies of green tea polyphenols were compared with those of genistein as a control. EGCG or ECG and genistein as a control dose-dependently inhibited the growth of A549 cells, which strongly elevated hnRNP B1 protein, and increased G2/M phase cells associated with induction of apoptotic cells. The results were confirmed by previous evidence with human lung cancer cell line PC-9. Some larger differences in mechanisms of action between green tea polyphenols and genistein were presented. Treatment of A549 cells with EGCG, ECG or genistein significantly inhibited the expression levels of hnRNP B1 mRNA and the elevated levels of hnRNP B1 protein, both of which are constitutively elevated in cancer cells. Furthermore, both EGCG and genistein inhibited the promoter activity of hnRNP A2/B1 gene expression, with IC50 values 29 microM for EGCG and 66 microM for genistein, suggesting the interaction of EGCG or genistein with the transcriptional complex. Looking at our results here, and those of previously reported epidemiological studies with green tea, we discuss the steadily accumulating evidence that clinical trials with green tea extract would be an efficient means of lung cancer prevention.


Association of overexpression of p53 oncoprotein with the state of cell proliferation in oral carcinoma.
p53 is a nuclear phosphoprotein recognised as important in the regulation of normal cell growth and proliferation, the wild-type protein suppressing cell division. Expression of presumptive mutant protein, detected by immunohistochemistry, is used increasingly as a diagnostic and prognostic marker in human neoplasms. A question arises as to whether or not p53 (over)expression in a lesion is any more or less informative than other markers of cell proliferation. Twenty well-differentiated oral squamous cell carcinomas which had earlier been examined for immunoreactivity against a panel of p53 antibodies were examined for the status of cell proliferation--both in islands of invading neoplastic cells and in the non-malignant epithelial margins. The status of epithelial cell proliferation was found to be significantly higher in p53-positive tumours when enumerated by Ki-67 antibody, both within the tumour as well as its margins. This may confer a growth advantage to these neoplasms and reflect a status of inactivated p53 protein, although the actual cause of the rapid proliferation may lie in activation/inactivation of other genes. The PCNA labelling indices, on the other hand, were closely similar in both p53-positive and -negative groups, suggesting that stabilisation of p53 protein does not influence the proliferative advantage in these carcinomas via a deregulation step of PCNA-related gene products.


RACCOONS (PROCYON LOTOR) SHOW HIGHER TRYPANOSOMA CRUZI DETECTION RATES THAN VIRGINIA OPOSSUMS (DIDELPHIS VIRGINIANA) IN SOUTH CAROLINA, USA.
Chagas disease, a significant public health concern in the Americas, is caused by a protozoan parasite, Trypanosoma cruzi. The life cycle of T. cruzi involves kissing bugs (Triatoma spp.) functioning as vectors and mammalian species serving as hosts. Raccoons (Procyon lotor) and opossums (Didelphis virginiana) have been identified as important reservoir species in the life cycle of T. cruzi, but prevalence in both species in the southeastern US is currently understudied. We quantified T. cruzi prevalence in these two key reservoir species across our study area in South Carolina, US, and identified factors that may influence parasite detection. We collected whole blood from 183 raccoons and 126 opossums and used PCR to detect the presence of T. cruzi. We then used generalized linear models with parasite detection status as a binary response variable and predictor variables of land cover, distance to water, sex, season, and species. Our analysis indicated that raccoons experienced significantly higher parasite detection rates than Virginia opossums, with T. cruzi prevalence found to be 26.5% (95% confidence interval [CI], 20.0-33.8) in raccoons and 10.5% (95% CI, 5.51-17.5) in opossums. Overall, our results concur with previous studies, in that T. cruzi is established in reservoir host populations in natural areas of the southeastern US.


Multilocus inference of species trees and DNA barcoding.
The unprecedented amount of data resulting from next-generation sequencing has opened a new era in phylogenetic estimation. Although large datasets should, in theory, increase phylogenetic resolution, massive, multilocus datasets have uncovered a great deal of phylogenetic incongruence among different genomic regions, due both to stochastic error and to the action of different evolutionary process such as incomplete lineage sorting, gene duplication and loss and horizontal gene transfer. This incongruence violates one of the fundamental assumptions of the DNA barcoding approach, which assumes that gene history and species history are identical. In this review, we explain some of the most important challenges we will have to face to reconstruct the history of species, and the advantages and disadvantages of different strategies for the phylogenetic analysis of multilocus data. In particular, we describe the evolutionary events that can generate species tree-gene tree discordance, compare the most popular methods for species tree reconstruction, highlight the challenges we need to face when using them and discuss their potential utility in barcoding. Current barcoding methods sacrifice a great amount of statistical power by only considering one locus, and a transition to multilocus barcodes would not only improve current barcoding methods, but also facilitate an eventual transition to species-tree-based barcoding strategies, which could better accommodate scenarios where the barcode gap is too small or inexistent.This article is part of the themed issue 'From DNA barcodes to biomes'.


Renal uptakes of 99mTc-MAG3, 99mTc-DTPA, and 99mTc-DMSA in rabbits with unilateral ureteral obstruction.
Renal function measurements using (99m)Tc-DTPA and (99m)Tc-MAG(3) dynamic scintigraphs were compared to those obtained using (99m)Tc-DMSA static scintigraphy. Eighteen experimental rabbits were randomly divided into (99m)Tc-DTPA-, (99m)Tc-MAG(3)-, and (99m) Tc-DMSA-injected groups. Experimental unilateral renal damage was induced by ligating a unilateral right ureter in 18 rabbits. Scintigraphic images were obtained at 2 and 5 h after intravenous injection of (99m)Tc-DMSA, or immediately after administration of (99m)Tc-DTPA or (99m)Tc-MAG(3). For the dynamic images using (99m)Tc-DTPA and (99m)Tc-MAG(3), rapid sequential images were obtained every 2 s for 30 images up to 1 min. The three groups presented different relative renal functions between the left normal and the right abnormal kidneys at 1, 2, 3, and 4 weeks post-ligation (p<0.05). However, the between-group comparisons showed no significant differences at any time. These results suggest that dynamic images of (99m)Tc-DTPA and (99m)Tc-MAG(3) can be used to measure the relative renal function in place of the static image of (99m)Tc-DMSA.


Combined therapy of iron chelator and antioxidant completely restores brain dysfunction induced by iron toxicity.
Excessive iron accumulation leads to iron toxicity in the brain; however the underlying mechanism is unclear. We investigated the effects of iron overload induced by high iron-diet consumption on brain mitochondrial function, brain synaptic plasticity and learning and memory. Iron chelator (deferiprone) and antioxidant (n-acetyl cysteine) effects on iron-overload brains were also studied. Male Wistar rats were fed either normal diet or high iron-diet consumption for 12 weeks, after which rats in each diet group were treated with vehicle or deferiprone (50 mg/kg) or n-acetyl cysteine (100 mg/kg) or both for another 4 weeks. High iron-diet consumption caused brain iron accumulation, brain mitochondrial dysfunction, impaired brain synaptic plasticity and cognition, blood-brain-barrier breakdown, and brain apoptosis. Although both iron chelator and antioxidant attenuated these deleterious effects, combined therapy provided more robust results. In conclusion, this is the first study demonstrating that combined iron chelator and anti-oxidant therapy completely restored brain function impaired by iron overload.


Pericardial Effusion During the Perioperative Period for Left Atrial Appendage Closure.
Objectives: To analyze the predictors of pericardial effusion (PE) during the perioperative period of the left atrial appendage closure procedure in our center. Methods: A total of 624 consecutive patients with non-valvular AF undergoing LAAC from May 2014 to October 2019 were involved in this study. Patients were divided into groups depending on whether they showed no PE, intraoperative PE or postoperative PE. We analyzed the predictors of PE during the perioperative period of the LAAC procedure. Results: (1) Of the 624 patients in our population (age 68.2 ± 9.1 years, 63% male, CHA2DS2-VASc score 3.4 ± 1.6), 30 patients experienced PE in the perioperative period, including 10 intraoperative PE and 20 postoperative PE. (2) A total of 26 (86.6%) patients had mild PE. 4 (13.4%) patients had pericardial tamponade, 2 (6.7%) of which were intraoperative, and the other 2 (6.7%) postoperative. (3) Significant differences were measured in relation to female sex, intraoperative time, combined procedures, changes in sinus rhythm, device retrieval times and duration of hospitalization between 2 groups (no PE occurred, intraoperative PE), P values were 0.039, 0.024, 0.004, 0.015, 0.003 and 0.039. Conclusions: Female sex, paroxysmal AF, changing in sinus rhythm, device retrieval times and intraoperative time all had a positive association with PE during the perioperative period.


A KV2.1 gating modifier binding assay suitable for high throughput screening.
Gating modifier peptides alter gating of voltage-gated potassium (KV) channels by binding to the voltage sensor paddle and changing the energetics of channel opening. Since the voltage sensor paddle is a modular motif with low sequence similarity across families, targeting of this region should yield highly specific channel modifiers. To test this idea, we developed a binding assay with the KV2.1 gating modifier, GxTX-1E. Monoiodotyrosine-GxTX-1E (125I-GxTX-1E) binds with high affinity (IC50 = 4 nM) to CHO cells stably expressing hKV2.1 channels, but not to CHO cells expressing Maxi-K channels. Binding of 125I-GxTX-1E to KV2.1 channels is inhibited by another KV2.1 gating modifier, stromatoxin (IC50 = 30 nM), but is not affected by iberiotoxin or charybdotoxin, pore blocking peptides of other types of potassium channels, or by ProTx-II, a selective gating modifier peptide of the voltage-gated sodium channel NaV1.7. Specific 125I-GxTX-1E binding is not detectable when CHO-KV2.1 cells are placed in high external potassium, suggesting that depolarization favors dissociation of the peptide. The binding assay was adapted to a 384-well format, allowing high throughput screening of large compound libraries. Interestingly, we discovered that compounds related to PAC, a di-substituted cyclohexyl KV channel blocker, displayed inhibitory binding activity. These data establish the feasibility of screening large libraries of compounds in an assay that monitors the displacement of a gating modifier from the channel's voltage sensor. Future screens using this approach will ultimately test whether the voltage sensor of KV channels can be selectively targeted by small molecules to modify channel function.


NUP214 in Leukemia: It's More than Transport.
NUP214 is a component of the nuclear pore complex (NPC) with a key role in protein and mRNA nuclear export. Chromosomal translocations involving the NUP214 locus are recurrent in acute leukemia and frequently fuse the C-terminal region of NUP214 with SET and DEK, two chromatin remodeling proteins with roles in transcription regulation. SET-NUP214 and DEK-NUP214 fusion proteins disrupt protein nuclear export by inhibition of the nuclear export receptor CRM1, which results in the aberrant accumulation of CRM1 protein cargoes in the nucleus. SET-NUP214 is primarily associated with acute lymphoblastic leukemia (ALL), whereas DEK-NUP214 exclusively results in acute myeloid leukemia (AML), indicating different leukemogenic driver mechanisms. Secondary mutations in leukemic blasts may contribute to the different leukemia outcomes. Additional layers of complexity arise from the respective functions of SET and DEK in transcription regulation and chromatin remodeling, which may drive malignant hematopoietic transformation more towards ALL or AML. Another, less frequent fusion protein involving the C terminus of NUP214 results in the sequestosome-1 (SQSTM1)-NUP214 chimera, which was detected in ALL. SQSTM1 is a ubiquitin-binding protein required for proper autophagy induction, linking the NUP214 fusion protein to yet another cellular mechanism. The scope of this review is to summarize the general features of NUP214-related leukemia and discuss how distinct chromosomal translocation partners can influence the cellular effects of NUP214 fusion proteins in leukemia.


Hypoxia in static and dynamic 3D culture systems for tissue engineering of bone.
Tissue engineering of sizeable cell-scaffold constructs is limited by gradients in tissue quality from the periphery toward the center. Because homogenous delivery of oxygen to three-dimensional (3D) cell cultures remains an unsolved challenge, we hypothesized that uneven oxygen supply may impede uniform cellular growth on scaffolds. In this study we challenged static and dynamic 3D culture systems designed for bone tissue engineering applications with a well-growing subclone of MC3T3-E1 preosteoblasts and continuously measured the oxygen concentrations in the center of cell-seeded scaffolds and in the surrounding medium. After as little as 5 days in static culture, central oxygen concentrations dropped to 0%. Subsequently, cells died in central regions of the scaffold but not in its periphery, where oxygen levels were approximately 4%. The use of perfusion bioreactors successfully prevented cell death, yet central oxygen concentrations did not rise above 4%. We conclude that 3D culture in vitro is associated with relevant oxygen gradients, which can be the cause of inhomogeneous tissue quality. Perfusion bioreactors prevent cell death but they do not entirely eliminate 3D culture-associated oxygen gradients. Therefore, we advise continuous oxygen monitoring of 3D culture systems to ensure tissue quality throughout engineered constructs.


Pathophysiological aspects of malignant brain tumors studied with positron emission tomography.
To further understand the control of brain tumor fluid balance and pH, the following studies were undertaken. The transport of a water soluble molecule across the brain and tumor capillary endothelium was studied during glucocorticoid and radiation treatment. The brain and brain-tumor acidity (pH) was evaluated as a single measurement in patients receiving a low maintenance dose of glucocorticoid. Transport changes and pH were measured in 61 patients with cerebral tumors using 82Rubidium (82Rb) and 11C-Dimethyloxa-zolidindione (11C-DMO), respectively, and Positron Emission Tomography (PET). Supplementary studies of tumor and contralateral brain blood flow and blood volume using the C15O2/PET and C15O/PET technique, respectively, were included to validate the 82Rb/PET model and obtain further information. A total of 125 PET scans were performed. Supplementary studies were undertaken to estimate delay of blood registration and form distribution of arterial blood isotope activity curves. Blood-to-tumor barrier transport was outlined at baseline and at 6 and 24 hours after the start of glucocorticoid treatment, finding a significant decrease in the transport. Radiation treatment (2-6 gray) did not alter the blood-to-tumor barrier transport when restudied within one hour in patients receiving glucocorticoid. In accordance with others, we observed pH values in gray and white matter in the range of 6.74-7.09 and 6.77-7.03 respectively. The pH in brain tumors was as high as 6.88-7.26, suggesting that tumors are more alkalotic than the normal brain. The permeability surface area product and the permeability coefficient were determined from the 82Rb/PET transport and C15O2/PET flow studies. Baseline permeability values were comparable to the literature values both for 82Rb and potassium. No difference in tissue blood volume was seen between 82Rb/PET and C15O/PET models and was of the same magnitude in the tumor and the contralateral tissue. The pH and fluid control in human brain tumors are perceived as metabolically controlled rather than, as previously believed, a result of simple passive exchange of alkalotic or osmotic active molecules between plasma and tumor interstitial space. Aspects of tumor alkalosis, tumor edema production, glucocorticoid edema clearance, and relationship between the anti-edema effect of glucocorticoid and the shown transport changes to 82Rb will be reviewed in the light of metabolic control mechanisms.


Blood pressure in professional male football players in Norway.
Scarce data exist on blood pressure (BP) and secondary effects in young athletes. Hence, we aimed to study prevalence of high BP (≥140/90 mmHg) and the association to arterial compliance, left ventricle mass and left atrium volume. A Norwegian preparticipation cardiac screening of male professional football players enrolled 493 white European, 47 matched controls, 49 black and 53 players of other ethnicity. BP was measured as a mean of two measurements. Height and weight were self-reported, and body surface area (BSA) was calculated. The echocardiographic parameters were indexed to BSA. Heart rates (HRs) by electrocardiography and pulse pressure (PP) were considered as surrogates for sympathetic activity. Arterial compliance was calculated as stroke volume(BSA)/PP. The players mean age was 25 years (18-38) and mean BP 122/69 ± 11/8 mmHg. There were no significant differences in prevalence of hypertension between all players, 39 (7%), and controls, four (9%), or between white, 32 (7%), and black, five (10%), players. There was a significant positive linear relationship between BP and left ventricle mass(BSA), left atrium volume(BSA), stroke volume(BSA), HR and PP, and negative relationship to arterial compliance(BSA). Although the prevalence of high BP in professional football players was low, our data indicate a novel association between elevated BP and reduced arterial compliance, increased left ventricle mass and left atrium volume even in young athletes. This emphasizes closer focus on BP measurements and standardized follow-up after preparticipation screening of athletes.


Bilateral ectopic ureters with bladder agenesis.
Agenesis of Urinary bladder is an extremely rare congenital genitourinary anomaly and is associated with other severe malformations that are incompatible with life. About 60 cases and 22 live births only have been reported in English literature. We report an eight year old girl with triad of complete agenesis of bladder and urethra, solitary functioning left kidney and an ectopic ureter opening into the vagina. The management of the child along with embryo-pathogenesis and literature review is discussed.


Iron Pharmacokinetics in Women with Iron Deficiency Anaemia Following A Single Oral Dose of a Novel Formulation of Tardyferon (Prolonged Release Ferrous Sulphate).
Numerous iron-containing preparations are available on the market; these vary in dosage, salt, chemical state of iron (ferric or ferrous) and in the iron delivery process (immediate or prolonged-release). Tardyferon® is a prolonged-release tablet containing 80 mg ferrous sulphate. The formulation has recently been modified; changes to the excipients which constitute the inert formulation matrix have allowed a decrease in tablet size for easier swallowing. The aim of this multicenter open-label study was to characterize the serum pharmacokinetics of iron in non-pregnant women aged 23-45 years with iron deficiency anaemia (IDA) following single oral administration of 160 mg Tardyferon® under fasting conditions. Blood samples were collected from the 29 participants before dosing and until 24 h post-dosing. Serum iron concentrations were determined using a routine colorimetric analytical method; pharmacokinetic parameters were derived using a non-compartmental approach. In these patients, median time to maximum serum concentrations (Tmax) was 4 h. Serum profiles were consistent with prolonged release; iron levels were elevated up to 12 h after dosing, with mean C12h still more than 7 times higher than baseline (CT0), and mean C2h and C8h representing 69.7% and 81.9% of the Cmax, respectively. In vitro dissolution testing performed on the clinical batch also demonstrated prolonged release of iron from this formulation. A single oral dose of 160 mg Tardyferon® administered under fasting conditions to this target population resulted in a long-lasting release of iron in the gastrointestinal tract, leading to optimal iron absorption. Moreover, Tardyferon® was well tolerated.


[Increased expressions of substance P and neurokinin/tachykinin receptor 1 in eosinophils of patients with psoriasis].
Objective To investigate the expressions of substance P (SP) and its receptor neurokinin/tachykinin receptor 1 (NK1R) in peripheral blood eosinophils of patients with psoriasis. Methods The levels of SP and NK1R in the peripheral blood of both patients with psoriasis and healthy people were detected by flow cytometry. This method was again used to detect the levels of SP and NK1R in the peripheral blood eosinophils of patients with psoriasis after stimulated with the crude extracts of Artemisia pollen, dust mite and Platanus pollen (all at concentrations of 0.1 and 1.0 μg/mL). Results Compared with the healthy controls, the percentages of SP+ and NK1R+ eosinophils in psoriasis patients increased up to 2.7 and 0.5 folds, respectively. Moreover, the mean fluorescence intensity (MFI) of SP+ and NK1R+ eosinophils of psoriasis patients were elevated by 1.5 and 0.2 folds, respectively. The percentage of SP+ eosinophils in psoriasis were down-regulated by 60% after the stimulation with Platanus pollen extract (1 μg/mL), while 0.1 μg/mL Platanus pollen extract induced a 0.6-fold increase in the percentage of NK1R+ eosinophis. Conclusion The expressions of SP and NK1R are up-regulated in peripheral blood eosinophils of patients with psoriasis.


Anticoagulation and hemostasis in cardiac surgical patients.
Optimizing anticoagulation and hemostasis during cardiopulmonary bypass and perioperatively helps to ensure the best possible clinical outcome. This article reviews the pharmacology of unfractionated and low-molecular weight heparin, aprotinin, desmopressin, dextran, antiplatelet agents, warfarin, and direct thrombin inhibitors. Their use is discussed in the context of coronary artery surgery, valvular surgery, and mechanical cardiac support devices, as well as in the management of acute ischemic syndromes, atrial fibrillation, and prevention and treatment of venous thromboembolism. Progress in the development and utilization of these anticoagulants and antiplatelet agents has supported the major advances that have been achieved in cardiac surgery.


The role of constant region carbohydrate in the assembly and secretion of human IgD and IgA1.
Immunoglobulins are glycoproteins, containing N- linked carbohydrates in the heavy chain constant regions of all isotypes and O-linked carbohydrates in the hinge regions of human IgA1 and IgD. A previous study showed that IgD synthesized in the presence of tunicamycin and lacking the three N-linked glycans on the heavy chain was not secreted (Shin, S. U., Wei, D. F., Amin, A. R., Thorbecke, G. J., and Morrison, S. L. (1992) Hum. Antibodies 3, 65-74). The contribution of each of the carbohydrates in the Fc of IgD to assembly and secretion was now analyzed by eliminating the carbohydrate addition sequence, Asn-X-Ser/Thr, through site-directed mutagenesis. Only the carbohydrate nearest the sole disulfide bond between heavy chains, which remained high mannose and appeared to be buried within the folded molecule, was found to be essential for secretion. When IgD lacked that glycan, assembly reached only the heavy/light chain half-molecule stage, and heavy chains were held inside the endoplasmic reticulum. Using benzyl 2-acetamido-2-deoxy-alpha-d-galactopyranoside (BADG) to inhibit complete O-linked glycosylation, we found that IgA1 and IgD with incomplete hinge carbohydrates were assembled and secreted from cells. Thus, one N-linked glycan plays a structural role in IgD and is required for proper assembly and secretion, but the O-linked carbohydrates in the hinge of IgD and IgA1 are not required for folding and export.


Blunt Traumatic Tracheobronchial Injury: a Clinical Pathway.
Motor vehicle collisions (MVC) are a major burden on healthcare systems. Saudi Arabia is one of the countries with a high mortality rate of MVC. Blunt tracheobronchial injuries are rare; however, it is a catastrophic event that requires a high center of care. Lack of experience and advanced faculty prompt early stabilization and transfer of the victim for advanced care. Due to the uncertainty of management of these injuries, we would like to share our experience in dealing with such injuries. To address the difficulties in initial management and transfer of patient with blunt traumatic tracheobronchial injuries. This is a single-center retrospective case-series study including patients admitted as cases of trauma including all age groups with blunt acute tracheobronchial injuries confirmed by imaging or bronchoscope. In our study, four patients with tracheobronchial injuries were identified, and a retrospective analysis was performed. Two of the males and one of the females are adults, while the other two are pediatrics. Two of them have a right main bronchial injury and the other two have a left main bronchial injury. Posterolateral thoracotomy and bronchial anastomosis were performed on all four patients and were followed up. In Saudi Arabia, blunt trauma is a prevalent type of injury, although tracheobronchial injuries are uncommon. In the event of trauma, a high index of suspicion of tracheobronchial injuries in a high mechanism injury warrants prompt treatment. Due to a lack of experienced and specialized hands in this field, management may be delayed, and eventually lead to unfavorable outcomes, hence we thought of a guide to facilitate the decision-making.


Longitudinal Assessment of Bone Mineral Density and Body Composition in Competitive Cyclists.
Baker, BS and Reiser II, RF. A longitudinal assessment of bone mineral density and body composition in competitive cyclists. J Strength Cond Res 31(11): 2969-2976, 2017-Competitive cycling has been associated with low bone mineral density (BMD); however, BMD is a multifaceted issue. The purpose of this study was to investigate how age (18-49 years), sex, USA Cycling Category (elite-4), and racing type (road and multiple bikes), influenced body composition across a season in competitive cyclists. February marked the preseason, where 42 participants (22 males, 20 females) completed a health history and cycling questionnaire, 4-day dietary log and a dual-energy X-ray absorptiometry scan, and repeated the measures 180 ± 11 days later. Preseason BMD and Z-Scores were within healthy ranges and similar between sexes, age groups, competition levels and racing-type groups (p ≥ 0.053). Age was significantly correlated with whole group BMD (r = 0.309; p = 0.047). Postseason analysis revealed very encouraging findings as no significant changes in BMD or Z-Score were observed in any group (p ≥ 0.067). A significant main effect for time was found in all groups as lean mass (LM) decreased and fat mass increased across the season (p ≤ 0.001). Additional analysis showed a significant time × group interaction as cat. 1 riders decreased body mass and body mass index, whereas cat. 4 riders responded in the opposite direction (p ≤ 0.037). Postseason correlations highlighted significant positive relationships between BMD and age, LM, and Kcal ingested (r ≥ 0.309; p ≤ 0.047). The only significant negative correlate of BMD was percent body fat (r = -0.359; p = 0.020). Armed with this information, cyclists and coaches should aim to prioritize balance between body mass and caloric intake while meeting the demands of training to minimize risk of cycling related low bone mass.


Potential predictors for progression of moyamoya disease: A systematic review and meta-analysis.
The progress of Moyamoya disease (MMD) is often accompanied by the occurrence of new ischemia or hemorrhagic events, which was difficult to predict. This systematic review and meta-analysis aimed to identify predictors for progression in MMD patients. We searched PubMed, Web of Science, Cochrane Library, and Embase databases up to December 10th, 2022 for randomized controlled trials, case-control studies, or cohort studies reporting predictors of disease progression in MMD patients. The results of each predictor were pooled by meta-analysis and further analyzed by subgroup analysis for predictors of unilateral to bilateral progression of MMD. A total of 842 patients from 12 studies were included. The estimated pooled means indicated lower age (standard mean difference [SMD]: -0.29, 95% confidence interval [CI]: -0.55 to -0.03; P = 0.03), family history (odds ratio [OR] 3.97, 95% CI: 1.96 to 8.03; P < 0.001) and contralateral abnormality (OR 3.95, 95% CI: 1.10 to 14.20; P = 0.04) were associated with progression in MMD patients. Subgroup analyses indicated that the same three factors were associated with the progression of unilateral to bilateral MMD. This meta-analysis revealed that lower age, family history and contralateral abnormality were associated with progression in MMD patients. The same three factors are associated with the progression of unilateral to bilateral MMD. Further studies are needed to validate our results.


[The transcriptional regulation of the TCA1 elements and their effects on morphogenesis of Candida albicans].
We have cloned two copies of retrotransposon-like elements Tca1-1 and Tca1-2 (Transposon Candida albicans) from Candida albicans strain SC5314. Comparing the restriction maps of the two genomic fragments, similar patterns were observed. The two composite elements were transcribed into an approximately unit length 6 kb RNA. The expression of this transcript was greatly increased when cells were grown at 25 degrees C versus 37 degrees C. Deletion of the partial or complete intervening region of the Tca1-1 element causes the morphogenetic variation of the cells. The morphogenetic variation is in cooperation with the expression of the URA3 gene.


Gastric and non-gastric marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue: A single-center experience.
Extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) is a distinct lymphoma with specific clinical and pathological features that occurs in diverse anatomic locations. We conducted this retrospective study to demonstrate our experience in patients with MALT lymphomas and compare our results with the literature. We studied 23 patients with histologically confirmed diagnosis of MALT lymphomas (12 with gastric, 11 with non-gastric localization) treated during the past 13 years. The female/male ratio of patients was 15/8 with a median age at presentation of 56 years (range 27-88 years). 16 patients (70%) with stage I and II, 7 patients (30%) with stage III and IV were admitted. At presentation 93% of patients had good performance status (ECOG<2) and 5 (22%) had B-symptoms. There was no difference between gastric and non-gastric MALT lymphomas when compared with sex, age, ECOG performance status, stage of the disease. Patients were treated with different treatment modality; H. Pylori eradication only (8.6%), radiotherapy only (21.7%), surgery alone (4.%) or followed by radiotherapy (26%) or chemotherapy (21.7%). All the patients are alive with a median 33 months (range 8-153 months) of follow-up and the 5- year PFS in gastric lymphoma and non-gastric lymphoma were 86% and 84% respectively with no statistical difference (p=0.5). Because of the indolent course the prognosis of MALT lymphoma was good regardless of the treatment modalities. The treatment choice should be patient-tailored, taking into account the site, stage, age and other clinical characteristic of patient.


The application of an information support robot to reduce agitation in an older adult with Alzheimer's disease living alone in a community dwelling: a case study.
Agitation is a common behaviour of Alzheimer's Disease (AD). Although healing robots have previously been used for alleviating agitation for people with dementia in hospitals and other healthcare facilities, its effects remain unclear whether an information-support robot capable of disseminating information with user needs may have similar effects when it is applied in community dwellings. This study was to investigate the effects of information-support robots for reducing agitation in older people with AD residing alone in community-dwellings. One participant with AD who lived alone in a community-dwelling was recruited. Participant's frequencies, durations, and time courses of four patterns of agitation at home at baseline and during intervention were analysed. In this study, agitation was defined as: (a) restlessness going to the corridor (RLtoC), (b) restlessness going to the toilet (RLtoT), (c) restlessness going to the entrance (RLtoE), and (d) restlessness going to multiple places (RLtoMP). In the intervention phase, the information-support robot disseminated daily schedule or date information to the participant. RLtoC and RLtoMP were significantly reduced in frequency during the intervention. The durations of each episode of RLtoT and RLtoMP in the intervention were slightly reduced than those at the baseline. There were no differences in time courses of episodes of RLtoC and RLtoMP after the intervention. This case study showed that information-support robot was useful to alleviate agitation of an older adult with AD living alone in a community-dwelling. Further study is warranted.


Circadian periodicity of intestinal Na+/glucose cotransporter 1 mRNA levels is transcriptionally regulated.
Intestinal expression of the high affinity Na+/glucose cotransporter 1 (SGLT1), which absorbs dietary glucose and galactose, exhibits both circadian periodicity in its activity and induction by dietary carbohydrate. Because the daily variation in SGLT1 activity is established by the feeding schedule (whether ad libitum or imposed) and persists in the absence of food, this variation has been described as anticipatory. To delineate the mechanisms regulating SGLT1, its expression was examined in rats maintained in a 12-h photoperiod with free access to chow. SGLT1 mRNA levels varied significantly, with the maximum abundance occurring near the onset of dark and the minimum near the onset of light. The SGLT1 transcription rate was 7-fold higher in the morning (1000-1100 h) than in the afternoon (1600-1700 h). An element for hepatocyte nuclear factor 1 (HNF-1) was identified in the SGLT1 promoter that formed different complexes with small intestinal nuclear extracts, depending on the time when the source animal was killed. Serological tests indicated that HNF-1alpha was present in complexes throughout the day, while HNF-1beta binding exhibited circadian periodicity. We propose that exchange of HNF-1 dimerization partners contributes to circadian changes in SGLT1 transcription. Because SGLT1 mRNA levels also varied in rhesus monkeys (offset by approximately one-half day from rats), a similar mechanism appears to be present in primates.


[The prevalence of diabetes mellitus in children of different regions of Ukraine].
Prevalence of diabetes mellitus among the children of Ukraine increased by 7% during the period from 1985 to 1992. The incidence that are revealed each year does not change. Different regions are significantly distinct from each other. The high and stable level of this disease incidence in Cherkassy Province, including cases revealed for the first time, two times exceeds these indices for Ukraine that attracts considerable interest.


Evaluation of Serratus Anterior Plane Block for Pain Relief in Patients Undergoing MIDCAB Surgery.
The minimally invasive direct coronary artery bypass (MIDCAB) surgery is associated with severe chest pain in the first 2 to 3 postoperative days; this may delay the patient recovery. In this randomized controlled trial we evaluated the role of serratus anterior plane (SAP) block for postoperative pain relief in patients undergoing MIDCAB surgery. Patients undergoing MIDCAB surgery were randomized into 2 groups of 25 each; SAP group received 20 mL of 0.2% ropivacaine with 1 μg/mL fentanyl as bolus followed by infusion at 8 mL/h; control group received saline for both bolus and infusion. Primary outcome measure was postoperative pain when supine, and during deep inspiration, coughing, and patient movement; secondary outcome measures were requirement of postoperative intravenous fentanyl and opioid-related side effects. All patients were followed at 6-hourly intervals for 48 hours in the postoperative period. Results were analyzed by the Student's t -test, χ 2 test, Mann-Whitney U-test and Kruskall-Wallis test. A P-value <0.05 was considered significant. The 2 groups were similar with respect to patient characteristics. Static and dynamic pain visual analog scale scores were significantly reduced in the SAP group as compared to the control group at most of the time points of assessment (P < 0.05). Patient-controlled fentanyl requirements were reduced in the SAP group as compared to control group on the second postoperative day (P < 0.05). SAP block reduced the postoperative pain scores and opioid requirements in patients undergoing MIDCAB surgery.


Behavioural changes and occlusal splints are effective in the management of masticatory myofascial pain: a short-term evaluation.
The aim of this research was to test the hypothesis that treatment with intra-oral appliances with different occlusal designs was beneficial in the management of pain of masticatory muscles compared with a control group. A total of 51 patients were analysed according to the research diagnostic criteria for temporomandibular disorders (RDC/TMD) to obtain the diagnosis of masticatory myofascial pain (MMP). The sample was then randomly divided into three groups: group I (n = 21) wore a full coverage acrylic stabilisation occlusal splint; group II (n = 16) wore an anterior device nociceptive trigeminal inhibitory (NTI) system; and group III (n = 14) only received counselling for behavioural changes and self-care (the control group). The first two groups also received counselling. Follow-ups were performed after 2 and 6 weeks and 3 months. In these sessions, patients were evaluated by means of a visual analogue scale (VAS) and pressure pain threshold (PPT) of the masticatory muscles. Possible adverse effects were also recorded, such as discomfort while using the appliance and occlusal changes. The results were analysed with Kruskal-Wallis, anova, Tukey's and Friedman tests, with a significance level of 5%. Group I showed improvement in the reported pain at the first follow-up (2 weeks), whereas for groups II and III, this progress was detected only after 6 weeks and 3 months, respectively. The PPT values did not change significantly. It was concluded that behavioural changes are effective in the management of pain in MMP patients. However, the simultaneous use of occlusal devices appears to produce an earlier improvement.