Sentinel-node biopsy in apparent early stage ovarian cancer: final results of a prospective multicentre study (SELLY).
To evaluate the sensitivity and specificity of sentinel-lymph-node mapping compared with the gold standard of systematic lymphadenectomy in detecting lymph node metastasis in apparent early stage ovarian cancer. Multicenter, prospective, phase II trial, conducted in seven centers from March 2018 to July 2022. Patients with presumed stage I-II epithelial ovarian cancer planned for surgical staging were eligible. Patients received injection of indocyanine green in the infundibulo-pelvic and, when feasible, utero-ovarian ligaments and sentinel lymph node biopsy followed by pelvic and para-aortic lymphadenectomy was performed. Histopathological examination of all nodes was performed including ultra-staging protocol for the sentinel lymph node. 174 patients were enrolled and 169 (97.1 %) received study interventions. 99 (58.6 %) patients had successful mapping of at least one sentinel lymph node and 15 (15.1 %) of them had positive nodes. Of these, 11 of 15 (73.3 %) had a correct identification of the disease in the sentinel lymph node; 7 of 11 (63.6 %) required ultra-staging protocol to detect nodal metastasis. Four (26.7 %) patients with node-positive disease had a negative sentinel-lymph-node (sensitivity 73.3 % and specificity 100.0 %). In a multicenter setting, identifying sentinel-lymph nodes in apparent early stage epithelial ovarian cancer did not reach the expected sensitivity: 1 of 4 patients might have metastatic lymphatic disease unrecognized by sentinel-lymph-node biopsy. Nevertheless, 35.0 % of node positive patients was identified only thanks to ultra-staging protocol on sentinel-lymph-nodes.


Regulation of plant symbiosis receptor kinase through serine and threonine phosphorylation.
We studied the biochemical properties of a plant receptor-like kinase to gain insights into the regulatory mechanism of this largest class of plant kinases. SYMRK (symbiosis receptor kinase) is required for early signal transduction leading to plant root symbioses with nitrogen-fixing rhizobia and phosphate-acquiring arbuscular mycorrhizal fungi. Amino acid substitutions in positions critical for activity of other related kinases cause a nonsymbiotic plant phenotype, suggesting that SYMRK kinase activity is required for symbiosis. SYMRK is capable of intermolecular autophosphorylation. Nonphosphorylated SYMRK is less active than the phosphorylated version, suggesting the phosphorylation status of SYMRK determines its activity. Three Ser/Thr residues were identified as residues required for full kinase activation through targeted mutagenesis. Using quadrupole time-of-flight mass spectrometry analysis, two of these were confirmed to be phosphorylated in vitro. These crucial phosphorylation sites are conserved among various plant receptor-like kinases as well as animal Pelle/interleukin-1 receptor associated kinase. Despite the distinct domain architecture of receptor-like kinases versus Pelle/interleukin-1 receptor associated kinase, our results suggest the existence of conserved activation mechanisms.


Treatment of ras-induced cancers by the F-actin-bundling drug MKT-077.
A rhodacyanine dye called MKT-077 has shown a highly selective toxicity toward several distinct human malignant cell lines, including bladder carcinoma EJ, and has been subjected to clinical trials for cancer therapy. In the pancreatic carcinoma cell line CRL-1420, but not in normal African green monkey kidney cell line CV-1, it is selectively accumulated in mitochondria. However, both the specific oncogenes responsible for its selective toxicity toward cancer cells, and its target proteins in these cancer cells, still remain to be determined. This study was conducted using normal and ras-transformed NIH 3T3 fibroblasts to determine whether oncogenic ras mutants such as v-Ha-ras are responsible for the selective toxicity of MKT-077 and also to identify its targets, using its derivative called "compound 1" as a specific ligand. We have found that v-Ha-ras is responsible for the selective toxicity of MKT-077 in both in vitro and in vivo. Furthermore, we have identified and affinity purified at least two distinct proteins of 45 kD (p45) and 75 kD (p75), which bind MKT-077 in v-Ha-ras-transformed cells but not in parental normal cells. Microsequencing analysis has revealed that the p45 is a mixture of beta- and gamma-actin, whereas the p75 is HSC70, a constitutive member of the Hsp70 heat shock adenosine triphosphatase family, which inactivates the tumor suppressor p53. MKT-077 binds actin directly, bundles actin filaments by cross-linking, and blocks membrane ruffling. Like a few F-actin-bundling proteins such as HS1, alpha-actinin, and vinculin as well as F-actin cappers such as tensin and chaetoglobosin K (CK), the F-actin-bundling drug MKT-077 suppresses ras transformation by blocking membrane ruffling. These findings suggest that other selective F-actin-bundling/capping compounds are also potentially useful for the chemotherapy of ras-associated cancers.


The terpolymer produced by Azotobacter chroococcum 7B: effect of surface properties on cell attachment.
The copolymerization of poly(3-hydroxybutyrate) (PHB) is a promising trend in bioengineering to improve biomedical properties, e.g. biocompatibility, of this biodegradable polymer. We used strain Azotobacter chroococcum 7B, an effective producer of PHB, for biosynthesis of not only homopolymer and its main copolymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHB-HV), but also novel terpolymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate)-poly(ethylene glycol) (PHB-HV-PEG), using sucrose as the primary carbon source and valeric acid and poly(ethylene glycol) 300 (PEG 300) as additional carbon sources. The chemical structure of PHB-HV-PEG was confirmed by (1)H nuclear-magnetic resonance analysis. The physico-chemical properties (molecular weight, crystallinity, hydrophilicity, surface energy) of produced biopolymer, the protein adsorption to the terpolymer, and cell growth on biopolymer films were studied. Despite of low EG-monomers content in bacterial-origin PHB-HV-PEG polymer, the terpolymer demonstrated significant improvement in biocompatibility in vitro in contrast to PHB and PHB-HV polymers, which may be coupled with increased protein adsorption, hydrophilicity and surface roughness of PEG-containing copolymer.


Dynamics of the CapG actin-binding protein in the cell nucleus studied by FRAP and FCS.
FRAP (fluorescence recovery after photobleaching) and FCS (fluorescence correlation spectroscopy) are spectroscopic methods for monitoring the dynamic distribution of proteins inside the nucleus of living cells. As an example we report our studies on the intracellular mobility of the actin-binding protein CapG in live breast cancer cells. This Gelsolin-related protein is a putative oncogene. It appears to be overexpressed especially in metastasizing breast cancer. Furthermore, the CapG protein is known to be involved in the motility control of non-muscle benign cells. Its increased expression triggers an increase in cell motility of benign cells. Thus it can be expected that in cancer cells overexpressing the CapG protein, motility, invasiveness and metastasis might be particularly promoted. Since the nuclear CapG fraction seems to be pivotal to the increase in cell motility, we focused our studies on the CapG mobility in cell nuclei of live breast cancer cells. Using FCS and FRAP we showed that the eGFP-tagged CapG is monomeric and characterized its diffusional properties on the microsecond to minute timescale. This information about the mobility and compartmentalization of CapG might help to provide insight into its function within the cell nucleus and give clues about its altered cellular function in malignant dedifferentiation.


Exploring and validating observations of non-local species in eDNA samples.
The development of DNA-based methods in recent decades has opened the door to numerous new lines of research in the biological sciences. While the speed and accuracy of DNA methodologies are clearly beneficial, the sensitivity of these methods has the adverse effect of increased susceptibility to false positives resulting from contamination in field or lab. Here, we present findings from a metabarcoding study on the diet of and food availability for five insectivorous birds, in which multiple lepidopteran species not known to occur locally were discovered. After describing the pattern of occurrences of these non-local species in the samples, we discuss various potential origins of these sequences. First, we assessed that the taxonomic assignments appeared reliable, and local occurrences of many of the species could be plausibly ruled out. Then, we looked into the possibilities of natural environmental contamination, judging it to be unlikely, albeit impossible to fully falsify. Finally, while dissimilar combinations of non-local species' occurrences across the samples did not initially suggest lab contamination, we found overlap with taxa and sequences handled in the same lab, which was undoubtedly not coincidental. Even so, not all exact sequences were accounted for in these locally conducted studies, nor was it clear if these and other sequences could remain detectable years later. Although the full explanation for the observations of non-local species remains inconclusive, these findings highlight the importance of critical examination of metabarcoding results, and showcase how species-level taxonomic assignments utilizing comprehensive reference libraries may be a tool in detecting potential contamination events, and false positives in general.


Morphometric and biomechanical remodelling following reopening of the obstructed bile duct.
Obstruction in the common bile duct results in duct dilatation, increased wall thickness and stiffening of the wall. Whether these changes are reversible after release of the obstruction has not yet been studied in detail. The aim of this study was to create a porcine model for release of bile duct obstruction and to describe biomechanical and morphometric parameters in a time-dependent design after the reopening. The bile duct of pigs was completely obstructed for one week and then reopened. Eighteen animals were allocated into four groups: unobstructed (group 1, n = 5), one week of obstruction (group 2, n = 5) and one week of obstruction followed by release of the obstruction after one week (group 3, n = 4), and four weeks (group 4, n = 4), respectively. The bile ducts were examined in vitro on the day of termination. One week of obstruction induced pronounced dilatation and wall thickening. Both the diameter and wall thickness decreased after release of the obstruction (p < 0.05) but without reaching the unobstructed level. The biomechanical properties in terms of the circumferential stress-strain relation differed between groups (p < 0.01). Groups 3 and 4 were transposed to the left of the other groups, indicating that they had the stiffest wall. The wall was stiffer in the longitudinal direction when compared to the circumferential direction in all groups (p < 0.05). The collagen amount decreased after releasing the obstruction (p < 0.01). The collagen amount correlated to the wall thickness (r > 0.9 and p < 0.001) and wall thickness-to-radius ratio (r > 0.7 and p < 0.05) in each of the two groups with release of the obstruction.


Pharmacokinetic properties of intramuscular versus oral syrup paracetamol in Plasmodium falciparum malaria.
Fever is an inherent symptom of malaria in both adults and children. Paracetamol (acetaminophen) is the recommended antipyretic as it is inexpensive, widely available and has a good safety profile, but patients may not be able to take the oral drug reliably. A comparison between the pharmacokinetics of oral syrup and intramuscular paracetamol given to patients with acute falciparum malaria and high body temperature was performed. A randomized, open-label, two-treatment, crossover, pharmacokinetic study of paracetamol dosed orally and intramuscularly was conducted. Twenty-one adult patients with uncomplicated falciparum malaria were randomized to receive a single 600 mg dose of paracetamol either as syrup or intramuscular injection on day 0 followed by a single dose administered by the alternative route on day 1. Paracetamol plasma concentrations were quantified frequently and modelled simultaneously using nonlinear mixed-effects modelling. The final population pharmacokinetic model was used for dose optimization simulations. Relationships between paracetamol concentrations with temperature and parasite half-life were investigated using linear and non-linear regression analyses. The population pharmacokinetic properties of paracetamol were best described by a two-compartment disposition model, with zero-order and first-order absorption for intramuscular and oral syrup administration, respectively. The relative bioavailability of oral syrup was 84.4 % (95 % CI 68.2-95.1 %) compared to intramuscular administration. Dosing simulations showed that 1000 mg of intramuscular or oral syrup administered six-hourly reached therapeutic steady state concentrations for antipyresis, but more favourable concentration-time profiles were achieved with a loading dose of 1500 mg, followed by a 1000 mg maintenance dose. This ensured that maximum therapeutic concentrations were reached rapidly during the first 6 h. No significant relationships between paracetamol concentrations and temperature or parasite half-life were found. Paracetamol plasma concentrations after oral syrup and intramuscular administration in patients with acute falciparum malaria were described successfully by a two-compartment disposition model. Relative oral bioavailability compared to intramuscular dosing was estimated as 84.4 % (95 % CI 68.2-95.1 %). Dosing simulations showed that a loading dose followed by six-hourly dosing intervals reduced the time delay to reach therapeutic drug levels after both routes of administration. The safety and efficacy of loading dose paracetamol antipyretic regimens now needs to be established in larger studies.


Acute effects of cartilage impact.
Posttraumatic arthrosis after joint injury occurs as a result of the disruption of the matrix components and chondrocyte death. Matrix components can be disrupted by mechanical and enzymatic means, and proteoglycan loss also has been shown to occur after cartilage injury. Until recently, chondrocyte death has been thought to occur primarily as a result of necrosis. However, new evidence shows that chondrocyte apoptosis can be stimulated to occur as a result of mechanical injury. The role chondrocyte death plays in the development of posttraumatic arthrosis currently is unknown. The development of in vitro cartilage injury models has made it possible to investigate some of the effects of impact load (acute injury) on cartilage. In vivo models of cartilage injury have made it possible to investigate changes in the integrity of the matrix components and of the chondrocytes in response to injury with time. However, considerably more information regarding this process is necessary before improvements in the prevention and treatment of posttraumatic arthrosis can be developed.


A UV resonance Raman (UVRR) spectroscopic study on the extractable compounds of Scots pine (Pinus sylvestris) wood. Part I: lipophilic compounds.
The wood resin in Scots pine (Pinus sylvestris) stemwood and branch wood were studied using UV resonance Raman (UVRR) spectroscopy. UVRR spectra of the sapwood and heartwood hexane extracts, solid wood samples and model compounds (six resin acids, three fatty acids, a fatty acid ester, sitosterol and sitosterol acetate) were collected using excitation wavelengths of 229, 244 and 257 nm. In addition, visible Raman spectra of the fatty and resin acids were recorded. Resin compositions of heartwood and sapwood hexane extracts were determined using gas chromatography. Raman signals of both conjugated and isolated double bonds of all the model compounds were resonance enhanced by UV excitation. The oleophilic structures showed strong bands in the region of 1660-1630 cm(-1). Distinct structures were enhanced depending on the excitation wavelength. The UVRR spectra of the hexane extracts showed characteristic bands for resin and fatty acids. It was possible to identify certain resin acids from the spectra. UV Raman spectra collected from the solid wood samples containing wood resin showed a band at approximately 1650 cm(-1) due to unsaturated resin components. The Raman signals from extractives in the resin rich branch wood sample gave even more strongly enhanced signals than the aromatic lignin.


Anthropogenic nitrogen pollution inferred by stable isotope records of crustose coralline algae.
Since reef ecosystems can offer intricate habitats for various marine organisms, calcified reefs may contain valuable long-term environmental data. This study investigated stable isotopic composition of marine organisms from the Taoyuan and Linshanbi crustose coralline algae (CCA) reef ecosystems to understand sewage pollution. CCA samples from Taoyuan (Palaeo Xin A: ∼1000 years old and Palaeo G: ∼7000 years old) and Linshanbi (Palaeo L: ∼7000 years old and modern CCA) had significantly lower δ15N values (2.5-5.6 ‰) compared to modern CCA from Taoyuan (10.2 ± 1.2 ‰). Intertidal organisms from the Taoyuan CCA reef also showed higher δ15N values than those from Linshanbi CCA reef, indicating anthropogenic stress in both ecosystems. Long-term pollution monitoring and effective strategies to mitigate sewage pollution are recommended for these CCA reef ecosystems.


Positional nystagmus due to alteration of the specific gravity in the labyrinth.
Horizontal positional nystagmus having characteristics similar to ice water caloric nystagmus was induced by injecting heavy water into the rabbit middle ear cavity or facial nerve through the stylomastoid foramen. Furthermore, with general administration of glycerol to normal subjects, horizontal positional nystagmus as with general administration of heavy water to experimental animals was observed. It is possible that alteration of the specific gravity arising from infiltration of heavy water or glycerol into the labyrinth accounts for the manifestation of positional nystagmus. These studies on positional nystagmus may be one of the means to elucidate the mechanism of caloric nystagmus and to examine the function of the human blood-labyrinth-barrier.


Tyrosol and hydroxytyrosol, two main components of olive oil, protect N2a cells against amyloid-β-induced toxicity. Involvement of the NF-κB signaling.
Alzheimer's disease (AD) is the most common form of dementia. Recently, a number of epidemiological studies have evidence that some dietary factors such as low antioxidants and vitamins intake could increase the risk of AD. In the opposite, diets rich in unsaturated fatty acids, in polyphenols, vitamins and antioxidants were identified as preventive factors. Several studies have reported that adherence to the Mediterranean diet (MeDi) was associated with a reduction in incident of dementia. The beneficial effect of MeDi may be the result of the association of some individual and non-identified food components and high consumption of olive oil. In this study we have investigated the protective effects of two components of olive oil, tyrosol (Tyr) and hydroxytyrosol (OH-Tyr), against Aβ-induced toxicity. In cultured neuroblastoma N2a cells, we found that Aβ(25-35) (100 µg/ml) treatment induced a decrease of glutathione (GSH) and the activation of the transcription factor NF-κB and cell death. Our results demonstrated that the number of cell death decreased when cells were co-treated with Aβ and Tyr or OH-Tyr. However, neither of these phenolic compounds was able to prevent the decrease of GSH induced by H(2)O(2) or Aβ. We found that the increase in the nuclear translocation of the NF-κB subunits after Aβ exposure was attenuated in the presence of Tyr or OH-Tyr. These results identified two individual food components of the MeDi as neuroprotective agent against Aβ and their potential involvement in the beneficial effect of the MeDi for the prevention of AD.


Identification and functional analysis of novel genes expressed in the Anterior Visceral Endoderm.
During early vertebrate development, the correct establishment of the body axes is critical. The anterior pole of the mouse embryo is established when Distal Visceral Endoderm (DVE) cells migrate to form the Anterior Visceral Endoderm (AVE). Symmetrical expression of Lefty1, Cer1 and Dkk1 determines the direction of DVE migration and the future anterior side. In addition to the establishment of the Anterior-Posterior axis, the AVE has also been implicated in anterior neural specification. To better understand the role of the AVE in these processes, we have performed a differential screening using Affymetrix GeneChip technology with AVE cells isolated from cer1P-EGFP transgenic mouse embryos. We found 175 genes which were upregulated in the AVE and 36 genes in the Proximal-posterior sample. Using DAVID software, we characterized the AVE cell population regarding cellular component, molecular function and biological processes. Among the genes that were found to be upregulated in the AVE, several novel genes were identified. Four of these transcripts displaying high-fold change in the AVE were further characterized by in situ hybridization in early stages of development in order to validate the screening. From those four selected genes, one, denominated Adtk1, was chosen to be functionally characterized by targeted inactivation in ES cells. Adtk1 encodes for a serine/threonine kinase. Adtk1 null mutants are smaller and present short limbs due to decreased mineralization, suggesting a potential role in chondrogenesis during limb development. Taken together, these data point to the importance of reporting novel genes present in the AVE.


The DSF type quorum sensing signalling system RpfF/R regulates diverse phenotypes in the opportunistic pathogen Cronobacter.
Several bacterial pathogens produce diffusible signal factor (DSF)-type quorum sensing (QS) signals to control biofilm formation and virulence. Previous work showed that in Burkholderia cenocepacia the RpfFBc/RpfR system is involved in sensing and responding to DSF signals and that this signal/sensor gene pair is highly conserved in several bacterial species including Cronobacter spp. Here we show that C. turicensis LMG 23827(T) possesses a functional RpfF/R system that is involved in the regulation of various phenotypes, including colony morphology, biofilm formation and swarming motility. In vivo experiments using the zebrafish embryo model revealed a role of this regulatory system in virulence of this opportunistic pathogen. We provide evidence that the RpfF/R system modulates the intracellular c-di-GMP level of the organism, an effect that may underpin the alteration in phenotype and thus the regulated phenotypes may be a consequence thereof. This first report on an RpfF/R-type QS system of an organism outside the genus Burkholderia revealed that both the underlying molecular mechanisms as well as the regulated functions show a high degree of conservation.


Small molecule drug activity in melanoma models may be dramatically enhanced with an antibody effector.
Monoclonal antibody (mAb) 38C2 belongs to a group of catalytic antibodies that were generated by reactive immunization and contains a reactive lysine. 38C2 catalyzes aldol and retro-aldol reactions, using an enamine mechanism, and mechanistically mimics natural aldolase enzymes. In addition, mAb 38C2 can be redirected to target integrins alpha(v)beta(3) and alpha(v)beta(5) through the formation of a covalent bond between a beta-diketone derivative of an arginine-glycine-aspartic acid (RGD) peptidomimetic and the reactive lysine residue in the antibody combining site to provide the chemically programmed mAb cp38C2. In this study, we investigated the potential of enhancing the activity of receptor-binding small molecule drug (SCS-873) through antibody conjugation. Using a M21 human melanoma xenograft model in nude mice, cp38C2 inhibited the growth of the tumor by 81%. The chemically programmed antibody was shown to be highly active at a low concentration while SCS-873 alone was ineffective even at dosages 1,000-fold higher than those used for the chemically programmed antibody. In vitro programming of the catalytic antibody was shown to be as effective as in vivo programming. In an experimental metastasis assay, treatment with mAb cp38C2 significantly prolonged overall survival of tumor-bearing severe combined immuno-deficient (SCID) mice when compared to treatment with unprogrammed mAb 38C2, SCS-873 alone or the integrin-specific monoclonal antibody LM609. In vitro, cp38C2 inhibited human and mouse endothelial and human melanoma cell adhesion, migration and invasion. Additionally, cp38C2 inhibited human and mouse endothelial cell proliferation and was active in complement-dependent cytotoxicity assays. These studies establish the potential of chemically programmed monoclonal antibodies as a novel and effective class of immunotherapeutics that combine the merits of traditional small molecule drug design with immunotherapy.


Prevention of corticosteroid induced osteoporosis in inpatients recently discharged from a tertiary teaching hospital.
To determine the medical conditions for which oral corticosteroids are prescribed and to determine the frequency and type of osteoporosis prophylaxis offered to these patients. Medical records of all inpatients for the period March to October 1999 who were documented in pharmacy records as either having received continuous oral steroids for at least 3 months or who had at least 4 courses of oral steroids per year were examined for the following data: age, sex, medical condition for which steroids were required, dose and duration of steroid therapy, whether they were offered bone mineral density (BMD) scans, and whether they were offered drug prophylaxis for steroid induced osteoporosis and the type of drug prophylaxis offered. Followup telephone calls were made to verify patients' use of prophylactic treatment and to validate the chronic use of oral corticosteroids. Use of BMD testing was also validated by comparing the list of patients in this study with the records of bone densitometer units in the area. A total of 189 medical records were examined: 38% were women (n = 72) and 62% were men (n = 117), with an age range of 19-91 years; 73% were taking continuous steroid therapy, the remaining 27% had multiple courses of prednisolone through the year. Steroids were prescribed for respiratory (n = 82, 43%), rheumatological (n = 74, 39%), hematological (n = 16, 8%), dermatological (n = 8, 4%), and gastrointestinal conditions (n = 7, 4%). Chronic obstructive airway disease was the most common respiratory condition for which steroids were prescribed (77, 94%), and polymyalgia rheumatica (36%) and inflammatory arthritis (41%) were the most common rheumatological conditions for which steroids were prescribed. In total, 47% (n = 89) were offered BMD scans while 53% (n = 100) were not. Of the 100 patients not offered BMD scans, 21 (21%) were receiving some form of drug prophylaxis, while 79% of patients were not taking any form of drug prophylaxis. Prophylaxis consisted of calcitriol (64%), alendronate (11%), calcitriol and calcium (7%), calcium alone (7%), alendronate and calcium (3%), etidronate and calcium (2%), alendronate, calcitriol and calcium (1%), alendronate and calcitriol (1%), and hormone replacement therapy (4%). Rheumatologists utilized both BMD testing and prophylactic treatment twice as often in patients taking chronic oral corticosteroid treatment than other specialty physicians. Compared to literature reports, the use of prophylaxis for corticosteroid induced osteoporosis was relatively high at this teaching hospital, with a surprisingly large number of patients receiving this treatment with no monitoring by BMD measurements.


Facilitation of liver cancer SMCC7721 cell aging by sirtuin 4 via inhibiting JAK2/STAT3 signal pathway.
Liver cancer severely threatens public health. Molecular targeted treatment is the further of cancer treatment. The functional role of Sir-related enzymes 4 (sirtuin 4) in treating liver cancer still requires further investigation. This study aimed to elucidate the effect of sirtuin 4 on aging of SMCC7721 liver cancer cell line, to underlying molecular mechanism and potential application in clinics. Adriamycin-induced aging model was established on SMCC7721 liver cancer cell line. Sirtuin 4 over-expression or siRNA plasmid was transfected. Cell aging was measured by β-galactosidase approach. Aging-related proteins P53 and P16 were quantified in Western blot, which also examined activation of Janus kinase 2 (JAK2) signal pathway. CP-690550 was used to suppress JAK2 signal pathway for measuring aging status of SMCC7721 cells. In aged SMCC7721 cells, sirtuin 4 was up-regulated, whilst P53 and P16 protein levels were elevated, in accompanied with JAK2/STAT3 signal pathway. Transfection of sirtuin 4 over-expression plasmid or siRNA increased or decreased sirtuin 4 expression. Adriamycin-induced aging was enhanced or suppressed, accompanied with inhibited or potentiated JAK2 signal pathway in sirtuin 4 up-regulation or down-regulation cells, respectively. The usage of JAK2 signal inhibitor, CP-690550, enhanced Adriamycin-induced cell aging. Sirtuin 4 facilitates Adriamycin-induced aging of SMCC7721 liver cancer cells via inhibiting JAK2/STAT3 signal pathway, thus providing one novel anti-cancer strategy.


Bioartificial liver with whole blood perfusion.
In our series of studies, we have made an effort to develop a bioartificial liver (BAL) system through which whole blood can be perfused as in hemodialysis therapy. In this study, BAL cartridges containing porcine hepatocytes were prepared and perfused in an extracapillary space with human whole blood in vitro. Lidocaine loading tests were performed to evaluate the detoxification ability of the BAL. The clearance value of lidocaine decreased during the initial 6 hours to about 50% of the initial value. After that, it was almost stable until 48 hours. After 48 hours perfusion, thin sections of the hollow fibers containing hepatocytes were prepared and stained with hematoxylin-eosin and immunohistochemically stained for membrane attack complex (MAC). The porcine hepatocytes formed aggregates in the hollow fibers, nuclei in the cells were observed clearly, and MAC was not seen on the porcine hepatocyte aggregation. A hollow fiber that can reject 90% of molecules with molecular weight of 50 kDa effectively protects porcine hepatocytes from humoral immunity. The in vivo assessment of the BAL cartridge was performed using a canine model for 24 hours. No significant hemolysis or thrombus that affected the BAL system and the canine were observed. These results suggest that our BAL system is a promising liver assist device through which patients' whole blood can be perfused.


Pulmonary and esophageal function in lung transplantation: Fundamental principles and clinical application.
The lungs and esophagus have a close anatomical and physiological relationship. Over the years, reflux-induced pulmonary injury has gained wider recognition, but the full effects of pulmonary disease on esophageal function are still unknown. Intrathoracic pressure dynamics potentially affect esophageal function, especially in patients with end-stage lung disease, both obstructive and restrictive. Lung transplantation is the only viable option for patients with end-stage pulmonary disease and has provided us with a unique opportunity to study these effects as transplantation restores the intrathoracic environment. Esophageal and foregut functional testing before and after transplantation provide insights into the pathophysiology of the foregut-pulmonary axis, such as how underlying pulmonary disease and intrathoracic pressure changes affect esophageal physiology. This review summarizes the available literature and shares the research experience of a lung transplant center, covering topics such as pre- and posttransplant foregut function, esophageal motility in lung transplant recipients, immune-mediated mechanisms of graft rejection associated with gastroesophageal reflux, and the role of antireflux surgery in this population.


[Metastasis to the lymph nodes as an index of the effectiveness of preoperative endolymphatic polychemotherapy of cervix cancer].
Ninety eight patients with cervical cancer were subjected to endolymphatic chemotherapy with multiple cytostatic drugs to affect metastatic lymph nodes. A comcentrated mixture of chemotherapeutic drugs (total amount about 20 ml) was injected into plantar lymph vessels, an interval being 10-14 days. The combinations and dosage of the drugs were as follows: thio-TEPA (100-150 mg) or cyclophosphamid (1000-2000 mg)+methotrexate (100 mg)+5-fluorouracil (1000-1500 mg). Subsequently, 76 patients underwent an extended extirpation of the uterus with adnexa, and their lymph-nodes were studied morphologically. It was found that the number of metastases into the lymphnodes was considerably less than it had been expected. Of 45 patients with the disease stage II metastases have been found only in 3 cases (7%). Of 24 patients with cancer stage III the lymphnodes were involved in 4 cases (16.6%). According to the literature data metastatic involvement of the lymphnodes is on an average encountered in 36% for stage II and in 51% - for stage III.


Scapulothoracic dissociation: diagnosis and treatment.
Scapulothoracic dissociation is an important and increasingly common clinical condition resulting from massive traction injury to the anterolateral shoulder girdle with disruption of scapulothoracic articulation. It frequently is accompanied by an acromioclavicular separation, displaced clavicular fracture, or sternoclavicular disruption. Vascular lesions have been reported in 88% of patients and severe neurologic injuries occur in 94% of patients. Many patients have a poor outcome with a flail extremity in 52%, early amputation in 21%, and death in 10%. Early recognition of this injury combined with a logical treatment protocol can help to decrease the substantial morbidity and mortality associated with this condition.


SRIF-sensitive compartmentalization of stored rGH is abolished by hypothyroidism.
Rat adenohypophyses lose immuno- and bioassayable growth hormone in hypothyroidism. We examined whether the somatotroph also loses mechanisms for intracellular hormone compartmentalization during hypothyroidism. A series of identical perifusions was performed using pituitary tissue from thyroidectomized rats before and after thyroxine replacement. Somatostatin (SRIF), (Bu)2cAMP and potassium ion were employed to produce a wide range of hormone release responses. Growth hormone synthesis diminished with hypothyroidism and increased with thyroid hormone replacement. Growth hormone release was therefore expressed as a percent of pituitary content to circumvent effects of variable content. Post-somatostatin rebound release was lost in hypothyroidism: it fell progressively after thyroidectomy (day 7 = 45% of control; day 14 = 11%; day 71 = 3%) and was restored by thyroxine replacement (day 2 = 24%; day 5 = 50%; day 9 = 102%). In conclusion, hypothyroid somatotrophs lose the ability to sequester stored hormone in a SRIF-sensitive compartment. Thyroxine replacement restores that capability. Thus, SRIF-sensitive rGH compartmentalization is thyroid hormone dependent.


Diabetes mellitus Type 2--the new face of an old lady.
Already 600 years before Christ, type 2 diabetes was known as a disease of elevated blood sugar levels associated with obesity. Since then, it appears, our understanding of the disease has not changed much, aside from the replacement of tasting the patients' urine by the measurement of plasma glucose and glycated haemoglobin levels (HbA1c) for its diagnosis and the discovery of some new drugs. Already, in those old days a physician from India named Sushrut described diabetes mellitus as a disease characterised by the passage of large amounts of urine and its "honey-like" taste and, noteworthy, as a disease that is mainly associated with obesity and a sedentary lifestyle, recommending physical activity as the primary treatment option. Although these milestone observations remain valid, major progress in the underlying pathogenesis of type 2 diabetes has been achieved showing a new face of this old disease and opening doors for novel treatment options. This review will highlight recent pathophysiological aspects of type 2 diabetes, actual diagnostic and treatment guidelines and discuss some possible upcoming new therapeutic strategies.


Usefulness of mindfulness-based cognitive therapy for treating insomnia in patients with anxiety disorders: a pilot study.
The objective of this study was to examine the usefulness of a mindfulness-based cognitive therapy (MBCT) for treating insomnia symptoms in patients with anxiety disorder. Nineteen patients with anxiety disorder were assigned to an 8-week MBCT clinical trial. Participants showed significant improvement in Pittsburgh Sleep Quality Index (Z = -3.46, p = 0.00), Penn State Worry Questionnaire (Z = -3.83, p = 0.00), Ruminative Response Scale (Z = -3.83, p = 0.00), Hamilton Anxiety Rating Scale (Z = -3.73, p = 0.00), and Hamilton Depression Rating Scale scores (Z = -3.06, p = 0.00) at the end of the 8-week program as compared with baseline. Multiple regression analysis showed that baseline Penn State Worry Questionnaire scores were associated with baseline Pittsburgh Sleep Quality Index scores. These findings suggest that MBCT can be effective at relieving insomnia symptoms by reducing worry associated sleep disturbances in patients with anxiety disorder. However, well-designed, randomized, controlled trials are needed to confirm our findings.


Characterization of the katG and inhA genes of isoniazid-resistant clinical isolates of Mycobacterium tuberculosis.
Resistance to isoniazid in Mycobacterium tuberculosis has been associated with mutations in genes encoding the mycobacterial catalase-peroxidase (katG) and the InhA protein (inhA). Among the 26 isoniazid-resistant clinical isolates evaluated in this study, mutations in putative inhA regulatory sequences were identified in 2 catalase-positive isolates, katG gene alterations were detected in 20 strains, and 4 isolates had wild-type katG and inhA genes. Mutations in the katG gene were detected in all 11 catalase-negative isolates: one frameshift insertion, two partial gene deletions, and nine different missense mutations were identified. An arginine-to-leucine substitution at position 463 was detected in nine catalase-positive isolates. However, site-directed mutagenesis experiments demonstrated that the presence of a leucine at codon 463 did not alter the activity of the M. tuberculosis catalase-peroxidase and did not affect the capacity of this enzyme to restore isoniazid susceptibility to isoniazid-resistant, KatG-defective Mycobacterium smegmatis BH1 cells. These studies further support the association between katG and inhA gene mutations and isoniazid resistance in M. tuberculosis, while also suggesting that other undefined mechanisms of isoniazid resistance exist.


Chronotherapeutic strategy: Rhythm monitoring, manipulation and disruption.
Mammalians circadian pacemaker resides in the paired suprachiasmatic nuclei (SCN) and influences a multitude of biological processes, including the sleep-wake rhythm. Clock genes are the genes that control the circadian rhythms in physiology and behavior. 24h rhythm is demonstrated for the function of physiology and the pathophysiology of diseases. The effectiveness and toxicity of many drugs vary depending on dosing time. Such chronopharmacological phenomena are influenced by not only the pharmacodynamics but also pharmacokinetics of medications. The underlying mechanisms are associated with 24h rhythms of biochemical, physiological and behavioral processes under the control of circadian clock. Thus, the knowledge of 24h rhythm in the risk of disease plus evidence of 24h rhythm dependencies of drug pharmacokinetics, effects, and safety constitutes the rationale for pharmacotherapy. Chronotherapy is especially relevant, when the risk and/or intensity of the symptoms of disease vary predictably over time as exemplified by allergic rhinitis, arthritis, asthma, myocardial infarction, congestive heart failure, stroke, and peptic ulcer disease. Morning once-daily administration of corticosteroid tablet medications results in little adrenocortical suppression, while the same daily dose split into four equal administrations to coincide with daily meals and bedtime results in significant hypothalamus-pituitary-adrenal (HPA) axis suppression. However, the drugs for several diseases are still given without regard to the time of day. Identification of a rhythmic marker for selecting dosing time will lead to improved progress and diffusion of chronopharmacotherapy. To monitor the rhythmic marker such as clock genes it may be useful to choose the most appropriate time of day for administration of drugs that may increase their therapeutic effects and/or reduce their side effects. Furthermore, to produce new rhythmicity by manipulating the conditions of living organs by using rhythmic administration of altered feeding schedules or several drugs appears to lead to the new concept of chronopharmacotherapy. Several drugs cause alterations in the 24h rhythms of biochemical, physiological and behavioral processes. The alteration of rhythmicity is sometimes associated with therapeutic effects, or may lead to illness and altered homeostatic regulation. Attention should be paid to the alteration of biological clock and consider it an adverse effect, when it leads to altered regulation of the circadian system which is a serious problem affecting basic functioning of living organisms. One approach to increasing the efficiency of pharmacotherapy is administering drugs at times during which they are best tolerated. From viewpoints of pharmaceutics, the application of biological rhythm to pharmacotherapy may be accomplished by the appropriate timing of conventionally formulated tablets and capsules, and the special drug delivery system to synchronize drug concentrations to rhythms in disease activity. New technology for delivering medications precisely in a time-modulated fashion by bedside or ambulatory pumps is developing to manage human diseases. Therefore, we introduce an overview of the dosing time-dependent alterations in therapeutic outcome and safety of drug. The underlying mechanisms and usefulness are introduced from viewpoint of chronopharmacology and chronotherapy.


The diagnosis and treatment of infertility. One person's philosophic approach.
To justify and at the same time present caveats from one reproductive endocrinologist discussing diagnoses and treatment of various aspects of infertility. Sources of basis of knowledge revealed. Sources for chapter writing includes personal experience, publications (predominantly from the author with some additional sources), national and international meetings, reading of various infertility journals and textbooks. Hopefully, the chapters will be provocative and provide new insights into diagnosis and treatment of infertility. Only areas where the author is most experienced will be discussed. The author promises a non-conventional approach which will lead to lots of controversy.


In silico identification and pharmacological evaluation of novel endocrine disrupting chemicals that act via the ligand-binding domain of the estrogen receptor α.
Endocrine disrupting chemicals (EDCs) pose a significant threat to human health, society, and the environment. Many EDCs elicit their toxic effects through nuclear hormone receptors, like the estrogen receptor α (ERα). In silico models can be used to prioritize chemicals for toxicological evaluation to reduce the amount of costly pharmacological testing and enable early alerts for newly designed compounds. However, many of the current computational models are overly dependent on the chemistry of known modulators and perform poorly for novel chemical scaffolds. Herein we describe the development of computational, three-dimensional multi-conformational pocket-field docking, and chemical-field docking models for the identification of novel EDCs that act via the ligand-binding domain of ERα. These models were highly accurate in the retrospective task of distinguishing known high-affinity ERα modulators from inactive or decoy molecules, with minimal training. To illustrate the utility of the models in prospective in silico compound screening, we screened a database of over 6000 environmental chemicals and evaluated the 24 top-ranked hits in an ERα transcriptional activation assay and a differential scanning fluorimetry-based ERα binding assay. Promisingly, six chemicals displayed ERα agonist activity (32nM-3.98μM) and two chemicals had moderately stabilizing effects on ERα. Two newly identified active compounds were chemically related β-adrenergic receptor (βAR) agonists, dobutamine, and ractopamine (a feed additive that promotes leanness in cattle and poultry), which are the first βAR agonists identified as activators of ERα-mediated gene transcription. This approach can be applied to other receptors implicated in endocrine disruption.


Renal potassium transport: mechanisms and regulation.
The regulation of potassium metabolism involves mechanisms for the appropriate distribution between the intra- and extracellular fluid compartments and for the excretion by the kidney. Clearance and single nephron studies show that renal excretion is determined by regulated potassium secretion and potassium reabsorption, respectively, in principal and intercalated cells of the distal nephron. Measurement of the electrochemical driving forces acting on potassium transport across individual cell membranes and characterization of several ATPases and potassium channels provide insights into the transport and regulation of renal potassium excretion.


The effect of maternal cigarette smoking on the pulmonary function of children and adolescents. Analyses of data from two populations.
To evaluate the role of the use of different analytical techniques in determining differences in the reported effects of passive smoking in children and adolescents, data sets from 2 studies that had previously reported divergent results were subjected to a uniform analytic procedure using a first-order autoregressive model (AR1). Although the populations studied differed in age distribution and the frequency of both personal and maternal smoking, the assumptions of the AR1 model were met by both data sets, and the models developed fit both data sets well. Coefficients for personal smoking from the study-specific models were almost identical. An effect of maternal smoking on FEV1, which has been reported for the East Boston data, again was observed, but no such effect was observed for the Tucson data set. These findings suggest that the differences in reported results are not related simply to model specificity and that real differences in exposure or response to exposure may be occurring in different climatic situations. Such differences need to be taken into account in any assessment of indoor exposure to potentially noxious agents.


Photoreceptor discs form through peripherin-dependent suppression of ciliary ectosome release.
The primary cilium is a highly conserved organelle housing specialized molecules responsible for receiving and processing extracellular signals. A recently discovered property shared across many cilia is the ability to release small vesicles called ectosomes, which are used for exchanging protein and genetic material among cells. In this study, we report a novel role for ciliary ectosomes in building the elaborate photoreceptor outer segment filled with hundreds of tightly packed "disc" membranes. We demonstrate that the photoreceptor cilium has an innate ability to release massive amounts of ectosomes. However, this process is suppressed by the disc-specific protein peripherin, which enables retained ectosomes to be morphed into discs. This new function of peripherin is performed independently from its well-established role in maintaining the high curvature of disc edges, and each function is fulfilled by a separate part of peripherin's molecule. Our findings explain how the outer segment structure evolved from the primary cilium to provide photoreceptor cells with vast membrane surfaces for efficient light capture.


The role of settling velocity formulation in the determination of gully pot trapping efficiency: comparison between analytical and experimental data.
Roadside gully pots are the connecting points between surface runoff and the underground drainage network; therefore they can be considered as the most superficial component of urban drainage systems. Gully pots are supposed to trap particulate matter washed off the catchment surface, but also to collect and convey stormwater into the network. The continuous accumulation of particulate matter results in a progressive loss of the gully pot hydraulic conveyance, thereby increasing the probability of urban flooding during rainstorm events. This study has therefore the objective to determine which variables influence the gully pot capability of retaining solids (efficiency), both experimentally and analytically. Several laboratory tests have been performed on a simple plastic gully pot, with different inflow rates and using both mono and hetero-disperse particle samples. Particular attention has been given to the influence exerted by the way particle settling velocity is expressed: efficiency has been analytically determined by means of multiple settling velocity formulas proposed by various authors and eventually compared to experimental data. Results deriving from the adoption of each single settling velocity formula have been extensively analysed, showing fairly different outcomes.


Identifying Molecular Biomarkers for Diseases With Machine Learning Based on Integrative Omics.
Molecular biomarkers are certain molecules or set of molecules that can be of help for diagnosis or prognosis of diseases or disorders. In the past decades, thanks to the advances in high-throughput technologies, a huge amount of molecular 'omics' data, e.g., transcriptomics and proteomics, have been accumulated. The availability of these omics data makes it possible to screen biomarkers for diseases or disorders. Accordingly, a number of computational approaches have been developed to identify biomarkers by exploring the omics data. In this review, we present a comprehensive survey on the recent progress of identification of molecular biomarkers with machine learning approaches. Specifically, we categorize the machine learning approaches into supervised, un-supervised and recommendation approaches, where the biomarkers including single genes, gene sets and small gene networks. In addition, we further discuss potential problems underlying bio-medical data that may pose challenges for machine learning, and provide possible directions for future biomarker identification.


Within-clutch variation in yolk testosterone as an adaptive maternal effect to modulate avian sibling competition: evidence from a comparative study.
In many species, embryos are exposed to maternal hormones in utero, in the egg, or in the seed. In birds, mothers deposit substantial testosterone into their eggs, which enhances competitive ability of offspring. These maternal testosterone concentrations vary systematically within clutches in different patterns and may enable mothers to adaptively fine-tune competitive hierarchies within broods. We performed a comparative analysis to investigate this hypothesis using a broad set of avian species. We expected species with small size differences among siblings (arising from small hatching asynchrony or slow growth rates) to aim for survival of the whole brood in good years and therefore compensate last-hatching eggs with relatively more testosterone. We expected species with large size differences among siblings (large hatching asynchrony or fast growth rates) to produce surplus young as insurance against failed offspring and to facilitate elimination of redundant surplus young by bestowing last-hatching eggs with relatively less testosterone. As predicted, we found that maternal testosterone compensation to last-hatching eggs is stronger when size differences among siblings become smaller. Maternal testosterone compensation to last-hatching eggs also correlated negatively with hatching asynchrony and growth rates. These findings provide evidence for correlated evolution of several maternal effects that together support different maternal reproductive strategies.


[The enzymatic system of phospholipid deacylation-reacylation in rat liver lysosomal membranes].
It has been shown for the first time that lysosomal (tritosomal) membranes of rat liver contain enzymes that are responsible for the deacylation-reacylation of phospholipids; their activity optimum lies at pH 7.0. Deacylation of lysosomal membrane phospholipids is controlled by a cascade of enzymatic reactions involving Ca2(+)-dependent phospholipase A1 which exhibits the maximal activity at 2.5 mM Ca2+ and at neutral values of pH, as well as lysophospholipase. Reacylation of lyso-derivatives of phospholipids is catalyzed by Mg2(+)-activated oleoyl-CoA:lysophosphatidylcholine acyltransferase having an activity optimum at pH 7.2.


Application of artificial intelligence to the electrocardiogram.
Artificial intelligence (AI) has given the electrocardiogram (ECG) and clinicians reading them super-human diagnostic abilities. Trained without hard-coded rules by finding often subclinical patterns in huge datasets, AI transforms the ECG, a ubiquitous, non-invasive cardiac test that is integrated into practice workflows, into a screening tool and predictor of cardiac and non-cardiac diseases, often in asymptomatic individuals. This review describes the mathematical background behind supervised AI algorithms, and discusses selected AI ECG cardiac screening algorithms including those for the detection of left ventricular dysfunction, episodic atrial fibrillation from a tracing recorded during normal sinus rhythm, and other structural and valvular diseases. The ability to learn from big data sets, without the need to understand the biological mechanism, has created opportunities for detecting non-cardiac diseases as COVID-19 and introduced challenges with regards to data privacy. Like all medical tests, the AI ECG must be carefully vetted and validated in real-world clinical environments. Finally, with mobile form factors that allow acquisition of medical-grade ECGs from smartphones and wearables, the use of AI may enable massive scalability to democratize healthcare.


Navigating Undiagnosed Dissociative Identity Disorder in the Inpatient Setting: A Case Report.
This case illustrates previously undiagnosed dissociative identity disorder (DID) in a middle-aged female with extensive childhood trauma, who was high functioning prior to a trigger that caused a reemergence of her symptoms. The trigger sparked a dissociative state, attempted suicide, and subsequent inpatient psychiatric hospitalization. Practitioners should include in their differential and screen for undiagnosed DID in patients with episodic psychiatric hospitalizations refractory to the standard treatments for previously diagnosed mental illnesses. Case study. During hospitalization, the diagnosis of DID became apparent and treatment included low-dose risperidone, mirtazapine, sertraline, unconditional positive regard, normalization of her dissociative states in an attempt to decrease her anxiety during treatment, and documentation for the patient via written notes following interviews. These methods helped her come to terms with the diagnosis and allowed the treatment team to teach her coping skills to lessen the impact of dissociative states following discharge.


Recognition of familiar melodies by adult cochlear implant recipients and normal-hearing adults.
The purpose of this study was to compare melody recognition and pitch perception of adult cochlear implant recipients and normal-hearing adults and to identify factors that influence the ability of implant users to recognize familiar melodies. Forty-nine experienced cochlear implant recipients and 18 normal-hearing adults were tested on familiar melody recognition. The normal-hearing adults were significantly (p < 0.0001) more accurate than implant recipients. Implant recipients showed considerable variability in perception of complex tones and pure tones. There were significant negative correlations between melody recognition, age at the time of testing, length of profound deafness and complex-tone perception, and significant positive relations between melody recognition and speech recognition scores.


Nature-Based Resilience: Experiences of Five Cities from South Asia.
As in many other parts of the world, the urban areas of the South Asian region are increasingly expanding. While cities today are the heart of commercial, technological and social development, they are also vulnerable to a variety of natural and anthropogenic threats. The complex urban infrastructure, and the ever-expanding population in cities, exacerbate the impacts of climate change and increase the risk of natural hazards. Throughout history, various hydrological disasters including floods, tidal surges, and droughts, and non-hydrological disasters such as earthquakes, landslides, and storms have led to catastrophic social, economic and environmental impacts in numerous South Asian cities. Disaster risk reduction is therefore central to ensure sustainability in urban areas. Although Nature-based Solutions (NbS) are identified as a promising strategy to reduce risk and increase resilience, there appears to be a lack of evidence-based approaches. NbS are measures that can be practiced to obtain benefits of nature for the environmental and community development through conserving, managing, and restoring ecosystems. Against this backdrop, the South Asian cities provide opportunities to evaluate capacities for achieving Nature-based Resilience (NbR) through NbS. This study documents insights from five cities of five different countries of the South Asian region which are subjected to a wide array of disasters: Barishal (Bangladesh), Phuentsholing (Bhutan), Gurugram (India), Kathmandu (Nepal), and Colombo (Sri Lanka). The primary objective of this study is to provide evidence on how NbS are being practiced. Thus, some success stories in cities under consideration are highlighted: restoration of natural canals through integrated development plans and community participation (Barishal), concepts of Gross National Happiness (GNH) and minimal nature interventions (Phuentsholing), "Greening cities'' including eco-corridors, vegetation belts, biodiversity parks (Gurugram), proper land use planning aims at different disasters (Kathmandu), and wetland restoration and management with multiple benefits (Colombo). These cases could therefore, act as a "proxy" for learning from each other to prepare for and recover from future disasters while building NbR.


Description of Cobetia amphilecti sp. nov., Cobetia litoralis sp. nov. and Cobetia pacifica sp. nov., classification of Halomonas halodurans as a later heterotypic synonym of Cobetia marina and emended descriptions of the genus Cobetia and Cobetia marina.
A group of five Gram-negative, aerobic, halotolerant, non-pigmented bacteria isolated from shallow sediment samples and invertebrate specimens collected from the Gulf of Alaska and the Sea of Japan was subjected to taxonomic study. On the basis of 16S rRNA gene sequence analysis, the novel isolates were affiliated to the genus Cobetia, sharing the highest sequence similarity of 99.3-99.9 % with Cobetia marina DSM 4741(T). DNA-DNA hybridization experiments between and among the novel strains and C. marina DSM 4741(T) and Cobetia crustatorum JCM 15644(T) revealed that the five strains represent three separate genospecies, which could be differentiated in their morphological, physiological and biochemical characteristics. Halomonas halodurans NBRC 15607(T) was included in this study as it has recently been reported to exhibit high 16S rRNA gene sequence similarity to C. marina DSM 4741(T), and it showed a high DNA relatedness value of 96 % with C. marina DSM 4741(T), indicating that they belong to the same species. On the basis of phylogenetic analysis, DNA-DNA hybridization and phenotypic characterization, three novel species are proposed, named Cobetia amphilecti sp. nov. (type strain KMM 1561(T) = NRIC 0815(T) = CCUG 49560(T)), Cobetia litoralis sp. nov. (type strain KMM 3880(T) =NRIC 0814(T) =CCUG 49563(T)) and Cobetia pacifica sp. nov. (type strain KMM 3879(T) = NRIC 0813(T) = CCUG 49562(T)). It is also proposed that Halomonas halodurans is a later heterotypic synonym of Cobetia marina, and emended descriptions of the genus Cobetia and the species Cobetia marina are provided.


Secondary structure-dependent evolution of Cymbidium ringspot virus defective interfering RNA.
Mutational analysis of defective interfering (DI) RNAs of Cymbidium ringspot virus (CymRSV) was used to study the mechanism of DI RNA evolution. It was shown that a highly base-paired structure in the 3' region of the longer DI RNA directed the formation of smaller DI RNA molecules. Mutations which increased the stability of the computer-predicted, highly structured 3' region of the longest DI RNA of CymRSV significantly enhanced the generation and accumulation of the smaller derivatives. Sequence analysis of smaller progeny molecules revealed that the highly base-paired region was deleted from the precursor DI RNA. Moreover, sites of recombination were found in other regions of the DI RNA progenies due to transposition of the highly base-paired structure. It is likely that the deletion event was structure- and not sequence-specific, and operated when a foreign sequence containing a 37-nt-long base-paired stem was inserted at the appropriate position of DI RNA.


Identification of minority ion-cyclotron emission during radio frequency heating in the JET tokamak
First measurements and identification of minority ion-cyclotron emission (MICE) during ICRF (H)D minority heating in the JET tokamak are presented. An inner wall radiofrequency (rf) probe shows the new single MICE spectral line, down-shifted from the heating frequency and appearing approximately 400 ms after the ICRH switch-on. The line is narrow (Deltaomega/omega approximately 0.04), characterized by the ion-cyclotron frequency of minority protons in the outer-edge midplane plasma and is observed irrespective of whether single or multifrequency ICRH is applied. The observations are consistent with the classical evolution and population of the plasma edge with approximately 3 MeV ICRH protons on orbits near the outboard limiters. Particle loss and energy filtering contribute to a local non-Maxwellian energetic ion distribution, which is susceptible to ion-cyclotron instability.


A Molecular Epidemiological Investigation of Carriage of the Adeno-Associated Virus in Murine Rodents and House Shrews in China.
To investigate the prevalence of the adeno-associated virus (AAV) in murine rodents and house shrews in 4 provinces of China. A total of 469 murine rodents and 19 house shrews were captured between May 2015 and May 2017. Cap gene of AAV sequences was obtained to evaluate the genetic characteristics of rat AAV. Rat AAVs were found in 54.7% (267/488) of throat swabs, 14.3% (70/488) of fecal samples, and 18.4% (41/223) of serum samples. Rat AAVs were detected in 3 species of murine rodents including Rattus norvegicus (34.8%), R. tanezumi (43.0%), and R. losea (2.3%), and house shrews (Suncus murinus) (26.1%) from the selected sampling sites. Fourteen near-full-length Cap gene sequences, ranging in length from 2,156 to 2,169 nt, were isolated from the fecal samples of R. norvegicus and R. tanezumi. These 14 sequences shared a high identity of 97.4% at the nucleotide level and 99.1% at the amino acid level. Phylogenetic analysis showed that the rat AAV formed a distinct clade, distinguishable from the AAV discovered in humans and in other animals. A high prevalence of rat AAV that was highly conserved within the Cap gene was found in 3 common murine rodents and house shrews in China.


Novel variants in KAT6B spectrum of disorders expand our knowledge of clinical manifestations and molecular mechanisms.
The phenotypic variability associated with pathogenic variants in Lysine Acetyltransferase 6B (KAT6B, a.k.a. MORF, MYST4) results in several interrelated syndromes including Say-Barber-Biesecker-Young-Simpson Syndrome and Genitopatellar Syndrome. Here we present 20 new cases representing 10 novel KAT6B variants. These patients exhibit a range of clinical phenotypes including intellectual disability, mobility and language difficulties, craniofacial dysmorphology, and skeletal anomalies. Given the range of features previously described for KAT6B-related syndromes, we have identified additional phenotypes including concern for keratoconus, sensitivity to light or noise, recurring infections, and fractures in greater numbers than previously reported. We surveyed clinicians to qualitatively assess the ways families engage with genetic counselors upon diagnosis. We found that 56% (10/18) of individuals receive diagnoses before the age of 2 years (median age = 1.96 years), making it challenging to address future complications with limited accessible information and vast phenotypic severity. We used CRISPR to introduce truncating variants into the KAT6B gene in model cell lines and performed chromatin accessibility and transcriptome sequencing to identify key dysregulated pathways. This study expands the clinical spectrum and addresses the challenges to management and genetic counseling for patients with KAT6B-related disorders.


Novel biological therapies for the treatment of diabetic foot ulcers.
The number of people with diabetes mellitus (DM) is estimated to exceed 640 million by the year 2040. Diabetic foot ulcer (DFU) is a debilitating illness that affects more than 2% of DM patients. DFU is caused by DM-induced neural and vascular lesions leading to a reduced sensation and microcirculation. The increase in the prevalence of DFU has prompted researchers to find new therapies for the management of DFU. Areas covered: This review presents the current status of novel biological therapies used in the treatment of DFU. Literature information and data analysis were collected from PubMed, the website of the American Diabetes Association, and ClinicalTrials.gov. The keywords used in the search were: DM, DFU, complications of DM. Expert opinion: Many biological agents have been investigated in a bid to find an effective therapy for DFU. These include growth factors (platelet-derived growth factor, vascular endothelial growth factor etc), stem cells (epithelial progenitor-, adipose-derived stem cells etc), anti-diabetic drugs (insulin, exendin-4), herbs, urokinase, dalteparin, statins and bio-agents such as acid peptide matrix. Biological agents that can reduce hyperglycaemia, increase sensation, microcirculation and oxygenation and repair lost tissue are the most ideal for the treatment of DFU.


Mutations in RASA1 and GDF2 identified in patients with clinical features of hereditary hemorrhagic telangiectasia.
Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant vascular disorder caused by mutations in ENG, ACVRL1 and SMAD4, which function in regulating the transforming growth factor beta and bone morphogenetic protein signaling pathways. Symptoms of HHT can be present in individuals who test negative for mutations in these three genes indicating other genes may be involved. In this study, we tested for mutations in two genes, RASA1 and GDF2, which were recently reported to be involved in vascular disorders. To determine whether RASA1 and GDF2 have phenotypic overlap with HHT and should be included in diagnostic testing, we developed a next-generation sequencing assay to detect mutations in 93 unrelated individuals who previously tested negative for mutations in ENG, ACVRL1 and SMAD4, but were clinically suspected to have HHT. Pathogenic mutations in RASA1 were identified in two samples (2.15%) and a variant of unknown significance in GDF2 was detected in one sample. All three individuals experienced epistaxis with dermal lesions described in medical records as telangiectases. These results indicate that the inclusion of RASA1 and GDF2 screening in individuals suspected to have HHT will increase the detection rate and aid clinicians in making an accurate diagnosis.


Helping may be Harming: unintended negative consequences of providing social support.
While social support is generally considered a helpful resource for employees, it can also serve as a job stressor. Unhelpful workplace social support (UWSS) is any action taken by a supervisor and/or colleague that the recipient believes was intended to benefit him or her but is perceived as unhelpful or harmful. Two studies, one qualitative and one quantitative, identified types of UWSS and demonstrated that unhelpful support can operate as a job stressor in relating to strains. In Study 1, critical incidents were collected from 116 employees, and a content analysis revealed 11 distinct categories of UWSS. In Study 2, the taxonomy of UWSS was further refined using quantitative methods. Results of two samples (176 diverse employees and 496 registered nurses) demonstrate that UWSS is associated with higher job-related negative affect, lower competence-based self-esteem, lower coworker satisfaction, higher work-related burnout, higher organisational frustration, and more physical symptoms (e.g. headache, nausea, and fatigue) among recipients. Together, the studies demonstrate that unhelpful workplace social support is a meaningful job stressor worthy of further investigation.


Caring to death: the murder of patients by nurses.
Beyond the initial 'shock-horror' reaction in the mass media, little attention is paid by nurses or the public to nurses who murder patients. This study used discursive inquiry to uncover social constructions of this phenomenon and their implications for the definition and treatment of such murders. The mass media and professional literature were searched for commentary on cases of nurses who had been convicted of murder between 1980 and 2006. The retrieved texts were subjected to discursive analysis. Discursive constructions included the profile of murderous nurses; types of murders; contexts in which murder occurs; factors that aid detection and apprehension; legal processes and punishment; and reactions of the public, profession, regulators and families. The findings imply that murder of a patient by a nurse might occur in any setting in which nurses care for vulnerable patients--the old, the young, the sick and the disabled. Trust in nurses assists a nurse to murder. Nurses have a responsibility to understand how their workplaces can form crucibles in which murder can take place. The profession needs to acknowledge the possibility of nurses who murder patients and to commence a discussion about what might be done to limit the harm they do.


The pollen S-determinant in Papaver: comparisons with known plant receptors and protein ligand partners.
Cell-cell communication is vital to multicellular organisms and much of it is controlled by the interactions of secreted protein ligands (or other molecules) with cell surface receptors. In plants, receptor-ligand interactions are known to control phenomena as diverse as floral abscission, shoot apical meristem maintenance, wound response, and self-incompatibility (SI). SI, in which 'self' (incompatible) pollen is rejected, is a classic cell-cell recognition system. Genetic control of SI is maintained by an S-locus, in which male (pollen) and female (pistil) S-determinants are encoded. In Papaver rhoeas, PrsS proteins encoded by the pistil S-determinant interact with incompatible pollen to effect inhibition of pollen growth via a Ca(2+)-dependent signalling network, resulting in programmed cell death of 'self' pollen. Recent studies are described here that identified and characterized the pollen S-determinant of SI in P. rhoeas. Cloning of three alleles of a highly polymorphic pollen-expressed gene, PrpS, which is linked to pistil-expressed PrsS revealed that PrpS encodes a novel approximately 20 kDa transmembrane protein. Use of antisense oligodeoxynucleotides provided data showing that PrpS functions in SI and is the pollen S-determinant. Identification of PrpS represents a milestone in the SI field. The nature of PrpS suggests that it belongs to a novel class of 'receptor' proteins. This opens up new questions about plant 'receptor'-ligand pairs, and PrpS-PrsS have been examined in the light of what is known about other receptors and their protein-ligand pairs in plants.


Maximal proton secretory rate of rat distal tubules is higher during chronic metabolic alkalosis.
In vivo microperfusion studies show augmented proton secretion in the distal tubule of rats with chronic metabolic alkalosis. The present studies used the same technique to determine whether this augmented proton secretion is due predominantly to an increase in substrate affinity or alternatively to a predominant increase in the maximal proton secretory rate. Surface distal tubules of alkalotic and control rats were microperfused in vivo with solutions containing increasing concentrations of HCO3. Proton secretion was determined as the difference between measured net HCO3 reabsorption and passive HCO3 transport calculated by use of the permeability derived from perfusing with a HCO3-free solution. Proton secretion was expressed as a function of the initial luminal HCO3 concentration and was assumed to follow saturable Michaelis-Menten kinetics. Alkalotic animals had a significantly higher Km (33.9 vs. 21.6 mM, P less than 0.03) and Vmax (223.8 vs. 99.1 pmol.mm-1.min-1, P less than 0.001) compared with control. These data are consistent with the augmented proton secretion in the distal tubule of alkalotic animals as predominantly due to an increased maximal proton secretory rate rather than to increased substrate affinity.


An unusual case of seronegative, 16S PCR positive Brucella infection.
Brucella is a zoonotic infection commonly diagnosed by isolation of the organism from blood culture or positive serological testing. It is an uncommon cause of a pyrexia of unknown origin in the United Kingdom. We describe the case of a 14-year-old girl with no history of travel who presented with pyrexia, weight loss, arthralgia, multiple splenic abscesses and a subsequent pleural effusion, the latter of which isolated a Brucella species on 16S rRNA PCR. The patient responded well to initiation of treatment for brucellosis and on repeat imaging, after 3 months, the splenic abscesses had resolved. This unique case demonstrates uncommon complications of brucellosis and the challenges of diagnosing the organism, the latter of which can be alleviated by the utilization of molecularbased technologies. This patient had a negative serology result for brucellosis, which highlights the need to interpret serology results with caution in non-endemic regions for brucellosis.


A new polymer gel for magnetic resonance imaging (MRI) radiation dosimetry.
New composition polymer gels, the N-vinylpyrrolidone argon (VIPAR) gels, were developed and investigated as MRI dosimeters. VIPAR gels were irradiated in the dose range of 0-12 Gy by a 6 MV x-ray linear accelerator and MR-scanned in a 1.5 T magnetic resonance imager. A linear relationship was found between absorbed dose and spin spin relaxation rate R2. The dose sensitivity was found to be approximately 0.1 s(-1) Gy(-1) for a gel composition of 4% w/w in N-vinylpyrrolidone, 4% w/w in N,N'-methylene-bisacrylamide, 5% w/w in gelatine type A and 87% w/w in water. This dose sensitivity was stable with time and did not deteriorate even when a boost radiation dose of 2.5 Gy was applied 15 days after the first irradiation. Good reproducibility of these results was observed when a new batch of gels was produced and used for corresponding measurements and analysis.


[The federal politics of basic sanitation and the initiatives of participation, mobilization, social control, health and environmental education].
The purpose of this article is to accomplish a critical analysis of two governmental important programs in health and environmental education - Health Education and Social Mobilization Program (PESMS) and Environmental Education and Sanitation Social Mobilization Program (PEAMSS), aiming at stimulate participative educational actions and social mobilization in sanitation projects. The methodology was based on reading and analysis of documents and observation in Workshops, Meetings, Seminars, Conventions, Congresses and Interviews. The authors describe the process of Program creation - PESMS and PEAMSS. They promoted a reflection and thought about Participation, Mobilization, Social Control, Health Education and Environmental Education. They also made considerations about the difficulties, facilities, advances and challenges in the implantation and implementation of PESMS and PEAMSS in the fundament for the realization of the public services of basic sanitation. They conclude that the creation of conditions by means of initiatives of Participation, Mobilization, Social Control, Health Education and Environmental Education become necessary for the development of Federal Policies of Basic Sanitation.


From job stress to intention to leave among hospital nurses: A structural equation modelling approach.
The aim of this study was to examine the structural relationships linking job stress to leaving intentions through job satisfaction, depressed mood and stress adaptation among hospital nurses. High turnover among nurses is a global concern. Structural relationships linking job stress to leaving intentions have not been thoroughly examined. Two nationwide cross-sectional surveys of full-time hospital staff in 2011 and 2014. The study participants were 26,945 and 19,386 full-time clinical nurses in 2011 and 2014 respectively. Structural equation modelling was used to examine the interrelationships among the study variables based on the hypothesized model. We used cross-validation procedures to ensure the stability and validity of the model in the two samples. There were five main paths from job stress to intention to leave the hospital. In addition to the direct path, job stress directly affected job satisfaction and depressed mood, which in turn affected intention to leave the hospital. Stress adaptation mitigated the effects of job stress on job satisfaction and depressed mood, which led to intention to leave the hospital. Intention to leave the hospital preceded intention to leave the profession. Those variables explained about 55% of the variance in intention to leave the profession in both years. The model fit was good for both samples, suggesting validity of the model. Strategies to decrease turnover intentions among nurses could focus on creating a less stressful work environment, increasing job satisfaction and stress adaptation and decreasing depressed mood. Hospitals should cooperate in this issue to decrease nurse turnover.


Expression profiling of nuclear receptors in human and mouse embryonic stem cells.
Nuclear receptors (NRs) regulate gene expression in essential biological processes including differentiation and development. Here we report the systematic profiling of NRs in human and mouse embryonic stem cell (ESC) lines and during their early differentiation into embryoid bodies. Expression of the 48 human and mouse NRs was assessed by quantitative real-time PCR. In general, expression of NRs between the two human cell lines was highly concordant, whereas in contrast, expression of NRs between human and mouse ESCs differed significantly. In particular, a number of NRs that have been implicated previously as crucial regulators of mouse ESC biology, including ERRbeta, DAX-1, and LRH-1, exhibited diametric patterns of expression, suggesting they may have distinct species-specific functions. Taken together, these results highlight the complexity of the transcriptional hierarchy that exists between species and governs early development. These data should provide a unique resource for further exploration of the species-specific roles of NRs in ESC self-renewal and differentiation.


Reactive oxygen species-mediated homologous downregulation of angiotensin II type 1 receptor mRNA by angiotensin II.
Recent studies suggest a crucial role of reactive oxygen species (ROS) for the signaling of angiotensin (Ang) II through Ang II type 1 receptor (AT(1)-R). However, the role of ROS in the regulation of AT(1)-R expression has not been explored. In this study, we examined the effect of an antioxidant on the homologous downregulation of AT(1)-R by Ang II. Ang II (10(-6) mol/L) decreased AT(1)-R mRNA with a peak suppression at 6 hours of stimulation in rat aortic vascular smooth muscle cells. Preincubation of vascular smooth muscle cells with N:-acetylcysteine (NAC), a potent antioxidant, almost completely inhibited the Ang II-induced downregulation of AT(1)-R mRNA. The effect of NAC was due to stabilization of the AT(1)-R mRNA that was destabilized by Ang II. The Ang II-induced AT(1)-R mRNA downregulation was also blocked by PD98059, an extracellular signal-regulated protein kinase (ERK) kinase inhibitor. Ang II-induced ERK activation was inhibited by NAC as well as by PD98059. Exogenous H(2)O(2) also suppressed AT(1)-R mRNA. These results suggest that the production of ROS and the activation of ERK are critical for the downregulation of AT(1)-R mRNA. The generation of ROS through stimulation of AT(1)-R not only mediates signaling of Ang II but also may play a crucial role in the adaptation process of AT(1)-R to the sustained stimulation of Ang II.


Ultrasonic Deposition of Carbon Nanotubes on Polycrystalline Cubic Boron Nitride Composites.
Polycrystalline cubic boron nitride (PcBN) are super-hard materials with high hardness and excellent abrasive resistance, widely used in cutting tools for precision machining of automotive and aerospace parts; however, their brittle properties make them prone to premature failure. Coatings are often applied to PcBN to extend their range of applicability and durability. Conventional coating methods are limited to the thickness range of a few hundred nanometres, poor adhesion to the substrate, and limited stability under ambient conditions. To further the properties of PcBN composites, in this paper, we explore the use of ultrasonic bonding to apply thick coatings (30-80 μm) on PcBN cutting tools. For the first time, a multi-walled carbon nanotube (MWCNT) powder is preplaced on a PcBN substrate to allow an unconventional coating technique to take place. The effects of ultrasonic bonding parameters on the change of mechanical properties of the coated tools are investigated through scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), micro-hardness analyses, and white light interferometry. The structure of the carbon nanotubes is investigated through transmission electron microscopy (pre coating) and cross-section of the bonded MWCNTs is studied via focused ion beam milling and SEM to evaluate the bonding between the multi-walled nanotubes. Optimum processing windows (i.e., bonding speed, energy, and pressure) are discovered for coating MWCNTs on PcBN. Focus ion beam milling analyses revealed a relationship between consolidation parameters and porosity of MW(pCNT) bonds. The proposed method paves the way for the novel design of functional coatings with attunable properties (i.e., thickness and hardness) and therefore improved productivity in the machining of aerospace and automotive parts.


Metabolomic profiling, biological evaluation of Aspergillus awamori, the river Nile-derived fungus using epigenetic and OSMAC approaches.
LC-HRMS-based metabolomics approach was applied to the river Nile-derived fungus Aspergillus awamori after its fermentation on four different media and using four epigenetic modifiers as elicitors. Thereafter, a comprehensive multivariate statistical analysis such as PCA, PLS-DA and OPLS-DA were employed to explain the generated metabolomic data (1587 features). PCA showed that the fungus displayed a unique chemical profile in each medium or elicitor. Additionally, PLS-DA results revealed the upregulated metabolites under each of these conditions. Results indicated that both rice and malt dextrose agar were recognized as the best media in terms of secondary metabolites diversity and showed better profiles than the four applied epigenetic modifiers, of which nicotinamide was the best secondary metabolite elicitor. Testing the antibacterial and cytotoxic effects of all A. awamori-derived extracts revealed that using epigenetic modifiers can induce antimicrobial metabolites against S. aureus and E. coli, whereas using rice, malt dextrose or nicotinamide can induce groups of cytotoxic metabolites. OPLS-DA results assisted in the putative identification of the induced metabolites that could be responsible for these observed inhibitory activities. This study highlighted how powerful the OSMAC approach in maximizing of the chemical diversity of a single organism. Furthermore, it revealed the power of metabolomics in tracing, profiling and categorizing such chemical diversity and even targeting the possible bioactive candidates which require further scaling up studies in the future.


Maximum tolerable dose of cyclophosphamide and azathioprine in Pakistani patients with primary renal disease.
The immunosuppressive regimens, at present, mainly rely on western guidelines that were derived from studies conducted in western populations. No such study exists for South Asian population, which is home to almost two billion people different in both genetics and environment from west. Locally derived thresholds for side effects markedly different from western figures may warrant re-adjustment of current local immunosuppressive regimens that are at present based largely on western guidelines. In order to define optimum dose for Cyclophosphamide (CYC) and Azathioprine (AZA) based immunosuppressive therapy, we conducted this study to find out maximum tolerable doses of azathioprine (AZA) and cyclophosphamide (CYC) beyond which neutropenia and thrombocytepenia are most likely to occur in patients with primary renal pathology. Patients with systemic vasculitis and idiopathic glomerulonephritis who were on CYC and AZA were identified through review of medical records at a tertiary care hospital in Pakistan (The Aga Khan University Hospital, Karachi). Patients were categorized under three principal diagnosis i.e. systemic lupus erythematosus (SLE), primary (idiopathic) glomerulonephritis (GN) and Wegener's granulomatosis (WG). The Receiver Operating Curve (ROC) was used to calculate the maximum tolerable dose for both CYC and AZA. We identified 94 patients aged 6-82 years (median 44.5 years) with primary renal disease (Wegener's granulomatosis n=13, Systemic lupus erythematosis n=62 and idiopathic glomerulonephritis n=19) who received CYC or AZA. Of these 94 patients, 36.2% (n=34) received CYC and 63.8% (n=60) received AZA. The mean dose of CYC was 1.54 +/- 0.50 mg/kg of body weight (range: 0.77-2.93). The mean dose of AZA was 1.64 +/- 0.59 mg/kg of body weight (range: 0.47-2.97). The maximum tolerable doses calculated for CYC and AZA were 1.25 mg/kg and 1.30 mg/kg of body weight respectively. The maximum tolerable dose for CYC and AZA among males could not be calculated, because of insufficient number of patients who developed neutropenia and thrombocytopenia. The maximum tolerable doses for CYC and AZA among females were 1.34 mg/kg and 1.03 mg/kg of body weight respectively. Also we found out that AZA was relatively more likely to cause neutropenia and thrombocytopenia (p=0.07). We thereby recommend that CYC should be initiated at a dose no more than 1 mg/kg of body weight and AZA at an initial dose of 0.75-1.0 mg/kg of body weight. The dose may be adjusted later on the basis of clinical response and laboratory reports.


[Prenatal diagnosis of placenta acretta by ultrasound and its histopathological association].
Anomalous adhesions of the placenta, known as placenta accreta and its variants, are the cause of obstetric hemorrhages that put the pregnant woman at risk. Accretism is strongly associated with a history of uterine surgery (cesarean section, myomectomy, curettage), as well as ultrasonographic signs, such as the presence and size of placental lacunae, loss of the placenta/bladder interface, location on the anterior face of the placenta, and presence of Doppler flow; these markers can be assessed by prenatal ultrasound. To analyze the association of prenatal diagnosis of placenta accreta by ultrasound with the histopathological result using the Tovbin index. Observational, cross-sectional and analytical study. 63 patients who had placenta accreta data by ultrasound measured with the Tovbin index and by means of the histopathological result obtained from the platform of the Mexican Institute for Social Security (IMSS) were included. The association between the two studies with the presence of placenta accreta was analyzed. 63 patients were analyzed; the Tovbin index was positive in 89% of the patients with a diagnosis of placenta accreta confirmed by histopathology. Both the Tovbin index and the histopathology report showed a statistically significant association with a p value of 0.04 for the diagnosis of placenta accreta. The Tovbin index as an ultrasonographic prenatal diagnosis of placenta accreta has a statistically significant association with histopathology diagnosis.


Citizen science networks in natural history and the collective validation of biodiversity data.
Biodiversity data are in increasing demand to inform policy and management. A substantial portion of these data is generated in citizen science networks. To ensure the quality of biodiversity data, standards and criteria for validation have been put in place. We used interviews and document analysis from the United Kingdom and The Netherlands to examine how data validation serves as a point of connection between the diverse people and practices in natural history citizen science networks. We found that rather than a unidirectional imposition of standards, validation was performed collectively. Specifically, it was enacted in ongoing circulations of biodiversity records between recorders and validators as they jointly negotiated the biodiversity that was observed and the validity of the records. These collective validation practices contributed to the citizen science character or natural history networks and tied these networks together. However, when biodiversity records were included in biodiversity-information initiatives on different policy levels and scales, the circulation of records diminished. These initiatives took on a more extractive mode of data use. Validation ceased to be collective with important consequences for the natural history networks involved and citizen science more generally.


Gender Differences in the Combined Effects of Cardiovascular Disease and Osteoarthritis on Progression to Functional Impairment in Older Mexican Americans.
Comorbidity (COM) is an important issue in aging. Cardiovascular disease (CVD) and osteoarthritis separately and together may modify the trajectories of functional decline. This analysis examines whether specific and unrelated COMs influence functional change differently and vary by gender. A cohort study of 1,789 (aged 60 years and older) Mexican Americans was followed annually for up to 10 years. We created four groups of COM (CVD alone, lower body osteoarthritis alone [OA], neither, or both). We employed mixed effects Poisson models with Instrumental Activities of Daily Living (IADL) as the outcome. We tested whether the association between COM and decline in functional status differed by gender. IADL impairments in those with CVD, OA, or both were significantly higher at baseline and increased more rapidly over time compared to those with neither condition. Compared to women with no COM, the number of IADL impairments in women with CVD alone were 1.36 times greater, with OA were 1.35 times greater, and both conditions were 1.26 times greater. Compared to men with no COM, IADL impairments in men with CVD alone were 1.15 times greater, OA alone were 1.12 times greater, and both were 1.26 times greater. Over time, the influence of COM on functional decline differs by specific combinations of COM and by gender. Aggregate COM scales obscure the biological and temporal heterogeneity in the effects of COM. Time-dependent-specific COMs better assess the development of impairment. Women experience a higher burden of functional impairment due to COM than men.


Subdiffraction resolution in total internal reflection fluorescence microscopy with a grating substrate.
We propose a fluorescence surface imaging system that presents a power of resolution beyond that of the diffraction limit without resorting to saturation effects or probe scanning. This is achieved by depositing the sample on an optimized periodically nanostructured substrate in a standard total internal reflection fluorescence microscope. The grating generates a high-spatial-frequency light grid that can be moved throughout the sample by changing the incident angle. An appropriate reconstruction procedure permits one to recover the fluorescence amplitude from the images obtained for various incidences. Simulations of this imaging system show that the resolution is not limited by diffraction but by the period of the grating.


Microsatellite typing of clinical and environmental Cryptococcus neoformans var. grubii isolates from Cuba shows multiple genetic lineages.
Human cryptococcal infections have been associated with bird droppings as a likely source of infection. Studies toward the local and global epidemiology of Cryptococcus spp. have been hampered by the lack of rapid, discriminatory, and exchangeable molecular typing methods. We selected nine microsatellite markers for high-resolution fingerprinting from the genome of C. neoformans var. grubii. This panel of markers was applied to a collection of clinical (n = 122) and environmental (n = 68; from pigeon guano) C. neoformans var. grubii isolates from Cuba. All markers proved to be polymorphic. The average number of alleles per marker was 9 (range 5-51). A total of 104 genotypes could be distinguished. The discriminatory power of this panel of markers was 0.993. Multiple clusters of related genotypes could be discriminated that differed in only one or two microsatellite markers. These clusters were assigned as microsatellite complexes. The majority of environmental isolates (>70%) fell into 1 microsatellite complex containing only few clinical isolates (49 environmental versus 2 clinical). Clinical isolates were segregated over multiple microsatellite complexes. A large genotypic variation exists in C. neoformans var. grubii. The genotypic segregation between clinical and environmental isolates from pigeon guano suggests additional source(s) of human cryptococcal infections. The selected panel of microsatellite markers is an excellent tool to study the epidemiology of C. neoformans var. grubii.


Determinants of diabetes in Bangladesh using two approaches: an analysis of the Demographic and Health Survey 2011.
This cross-sectional study analysed data from the Bangladesh Demographic and Health Survey 2011 to investigate factors associated with diabetes in Bangladesh. Data were analysed using logistic and log-binomial regressions to estimate odds ratios (ORs) and prevalence ratios (PRs), respectively. Among the 7544 respondents aged ≥35 years, the estimated prevalence of diabetes was 11.0%. In the adjusted analysis, survey participants in the age group 55-64 years (adjusted PR [APR]: 1.8, 95% Confidence Interval (CI): 1.4, 2.2; adjusted OR [AOR]: 1.9, 95% CI: 1.5, 2.5) and those with at least secondary education level (APR: 1.3, 95% CI: 1.0, 1.6; AOR: 1.3, 95% CI: 1.0, 1.7) were more likely to have diabetes than those in the age group 35-44 years and those with no education. Furthermore, respondents living in Khulna (APR: 0.5, 95% CI: 0.4, 0.6; AOR: 0.4, 95% CI: 0.3, 0.6) were less likely to have diabetes than people living in Barisal. While adjusted estimates of PR and OR were similar in terms of significance of association, the magnitude of the point estimate was attenuated in PR compared with the OR. Nevertheless, the measured factors still had a significant association with diabetes in Bangladesh. The results of this study suggest that Bangladeshi adults would benefit from increased education on, and awareness of, the risk factors for diabetes. Focused public health intervention should target these high-risk populations.