RSV Burden and Its Impact on Pediatric Inpatient Bed Occupancy in Belgium: An Analysis of National Hospital Claims Data.
Respiratory syncytial virus (RSV) infections represent a substantial burden on pediatric services during winter. While the morbidity and financial burden of RSV are well studied, less is known about the organizational impact on hospital services (ie, impact on bed capacity and overcrowding and variation across hospitals). Retrospective analysis of the population-wide Belgian Hospital Discharge Data Set for the years 2017 and 2018 (including all hospital sites with pediatric inpatient services), covering all RSV-associated (RSV-related International Classification of Diseases, 10th Version, Clinical Modification diagnoses) inpatient hospitalization by children under 5 years old as well as all-cause acute hospitalizations in pediatric wards. RSV hospitalizations amount to 68.3 hospitalizations per 1000 children less than 1 year and 5.0 per 1000 children 1-4 years of age and are responsible for 20%-40% of occupied beds during the peak period (November-December). The mean bed occupancy rate over the entire year (2018) varies across hospitals from 22.8% to 85.1% and from 30.4% to 95.1% during the peak period. Small-scale pediatric services (<25 beds) are more vulnerable to the volatility of occupancy rates. Forty-six hospital sites have daily occupancy rates above 100% (median of 9 days). Only in 1 of 23 geographically defined hospital networks these high occupancy rates are on the same calendar days. Pediatric services tend to be over-dimensioned to deal with peak activity mainly attributable to RSV. RSV immunization can substantially reduce pediatric capacity requirements. Enhanced collaboration in regional networks is an alternative strategy to deal with peaks and reduce capacity needs.


Pharmaceutical assistance programs for cancer patients in the era of orally administered chemotherapeutics.
Introduction The rising cost of cancer drugs may make treatment unaffordable for some patients. Patients often rely on drug manufacturer-administered Pharmaceutical Assistance Programs (PAPs) to obtain drugs and reduced or no cost. The overall usage of PAPs within cancer care delivery is unknown. Methods We included all cancer patients across an academically affiliated, integrated health system in North Carolina during 2014 ( N = 8591). We identified the subset of patients receiving PAP assistance to afford one or more cancer drugs, in order to calculate the proportion of patients receiving PAP assistance, and the retail value of the assistance. Results Among 8591 cancer patients, 215 unique patients submitted a total of 478 successful PAP requests for cancer drugs. 40% of PAP-utilizing patients were uninsured, 23% had Medicaid coverage, 20% had Medicare coverage, 2% were dual Medicare/Medicaid eligible, and 14% were commercially insured. Among all cancer patients who received medical treatment, 6.0% required PAP assistance, whereas 10.6% receiving an oral agent required PAP assistance. The proportion receiving PAP assistance varied substantially by drug, ranging from <1% of patients (e.g. carboplatin, methotrexate) to 50% of patients (e.g. ponatinib, temsirolimus). The majority of the retail value obtained was for oral agents, including $1,556,575 of imatinib and $1,449,633 of dasatinib, which were the two drugs with the highest aggregate retail value. Conclusions A substantial proportion of cancer patients receive private charitable assistance to obtain standard-of-care treatments. This includes patients with federal and private insurance, suggesting an inability of patients to meet cost-sharing requirements.


Malaria vaccine development.
The malaria parasite life cycle presents several targets for attack, but these different parts of the life cycle are susceptible to different types of host immune response. For example, the sporozoite is most sensitive to immune antibody, while liver stage parasites can be eliminated by cytotoxic T lymphocytes. Attachment of merozoites to erythrocytes, on the other hand, can be blocked by antibody. Convincing experimental evidence shows that completely protective immunity to malaria can be induced. The challenge now is to design recombinant or synthetic vaccines that induce the right types of immune responses to specific life cycle stages. This requires the identification and characterization of B- and T-lymphocyte epitopes expressed by the parasite or by parasitized host cells. These epitopes must be incorporated into a delivery system that maximizes the interaction between the vaccine epitopes and the host immune system. Many epitopes from several parts of the life cycle are already characterized; development of multivalent vaccines, that is, vaccines which contain immunogens from more than one part of the life cycle, is a promising area for research efforts.


Upregulation of TPX2 by STAT3: identification of a novel STAT3 binding site.
TPX2, a protein involved in mitosis, is considered a good marker for actively proliferating tissues, highly expressed in a number of cancer cells. We show the presence of high-affinity binding site for STAT3 in the 5'-flanking region of the Tpx2 gene, which is in vivo bound by activated STAT3. A specific STAT3 peptide inhibitor represses the expression of the Tpx2 gene and inhibits the binding of STAT3 to its consensus sequence in human cell lines where STAT3 is activated. These results indicate that activated STAT3 contributes to the over-expression of Tpx2 through the binding to an enhancer site.


Potentiation of bleomycin cytotoxicity toward cultured mouse cells by hyperthermia and ethanol.
The effects of hyperthermia and ethanol on bleomycin cytotoxicity toward mouse mammary carcinoma FM3A cells in culture were followed in terms of clonal growth in a soft agar medium. Heating the cells at 43 degrees during or before bleomycin treatment sensitized them to bleomycin. Heat treatment after drug treatment was less effective. Bleomycin cytotoxicity was also greatly potentiated by exposing the cells to ethanol at a concentration of more than 4% (v/v) at 37 degrees before or after drug treatment. However, when the cells were exposed to ethanol during bleomycin treatment, the potentiation of cytotoxicity was reduced, and a higher concentration of ethanol was required for inducing equivalent sensitization. The cytoxicity of pepleomycin, a potent antitumor agent derived from bleomycin, was also greatly potentiated by hyperthermia and ethanol. The cellular uptake of 3H-pepleomycin was not increased by either treatment, and thus the increased cytotoxicity was not correlated with a change in membrane permeability to the drug. The sensitizing effect of hyperthermia or ethanol was maximum when the drug treatment was carried out immediately after the cells were exposed to heat or ethanol. Sensitization decreased linearly as the time interval between the two treatments was increased, and after a 6 hr interval the sensitizing effect of either agent disappeared. This recovery from the sensitized state was not inhibited by cycloheximide or by incubation in phosphate-buffered saline instead of complete growth medium, but it was reversibly inhibited at 0 degrees. It is suggested that the modes of action of hyperthermia and ethanol in inducing cell sensitization of bleomycin are similar.


Prolonged analgesia and decreased toxicity with liposomal morphine in a mouse model.
Inadequate control of postoperative pain remains a major clinical problem. A reliable method of providing long-lasting postoperative analgesia with a single dose would be very useful. We synthesized a liposomal morphine formulation and compared it to free morphine with regard to duration of analgesia in the mouse. Analgesia was assessed after intraperitoneal injection using the tail-flick test. The systemic toxicity after administration of liposomal and free morphine was compared. The release rate of morphine from liposomes in vitro was also evaluated. The lethal intraperitoneal dose of free morphine in 50% of mice (LD50) was 400 mg/kg. The maximum safe (non-lethal) dose of free morphine was 130 mg/kg. The highest dose of liposomal morphine administered (1650 mg/kg) did not cause death in any animal. Duration of analgesia was significantly prolonged with the highest dose of liposomal morphine (21.5 +/- 5.3 h) compared to the maximum safe dose of free morphine (3.7 +/- 0.75 h), P < 0.01. In vitro experiments showed a slow release rate of morphine from the liposome depot. Prolonged analgesia and decreased systemic toxicity for liposomal morphine are explained by sustained release of morphine from the liposomal depot. These results suggest that liposomal narcotic formulations may provide prolonged analgesia with single-dose administration.


Short-chain fatty acids and the irritable bowel syndrome: the effect of wheat bran.
Short-chain fatty acids (SCFA) in faeces were examined in 18 patients with the irritable bowel syndrome (IBS) during treatment with wheat bran or placebo. In the placebo period, the patients could be classified in accordance with the faecal concentrations of SCFA into one group with low concentrations (mean, 40 mmol/l; range, 19-77 mmol/l; 10 patients) and another with high concentrations (mean, 168 mmol/l; range, 145-187 mmol/l; 8 patients). The concentrations of SCFA differed (P less than 0.001) in both groups from concentrations found in faeces from a reference group of nine normal individuals (mean, 114 mmol/l; range, 93-155 mmol/l). Patients with low levels of SCFA had lower (P less than 0.001) mean stool mass and longer (P less than 0.05) transit times than those with high concentrations of SCFA in faeces. Ingestion of bran, although a precursor of SCFA, did not change faecal concentrations of SCFA. Abdominal pain, distension, and rumbling were not correlated to low or high concentrations of SCFA in faeces, nor did bran improve these symptoms when compared to placebo. The level of SCFA was rather constant intraindividually and independent of the variability of the daily faecal mass. It is concluded that patients with IBS apparently have continuously abnormal concentrations of SCFA in faeces, either high or low, which are unaffected by the treatment with bran and which hypothetically may be of pathophysiologic importance.


miR-455 inhibits breast cancer cell proliferation through targeting CDK14.
Breast cancer is the most frequently occurring cancer in women worldwide, microRNAs (miRNAs) play critical role in the initiation and progression of breast cancer. Here, we studied the effect of miR-455 on cell proliferation of breast cancer, and found that miR-455 was downregulated in breast cancer tissues and cells. Its overexpression inhibited cell proliferation, whereas its knockdown promoted cell proliferation of breast cancer. We found a Cdc2-related protein kinase CDK14 was the target of miR-455, when the 3'UTR of CDK14 was cloned into luciferase reporter vector and transfected into cells, miR-455 mimic could inhibit the luciferase activity in a dose-dependent manner, miR-455 inhibitor increased the luciferase activity, but the mutant miR-455 mimic couldn't change the luciferase activity, suggesting miR-455 directly bound to the 3'UTR of CDK14. Meanwhile, we also found miR-455 inhibited Cyclin D1 expression and promoted p21 expression, confirming miR-455 inhibited cell proliferation. Double knockdown of miR-455 and CDK14 inhibited the proliferation of breast cancer cell, confirming miR-455 inhibiting cell proliferation by targeting CDK14. Moreover, miR-455 levels were negatively correlated with CDK14 levels in breast cancer tissues. Our finding revealed miR-455 inhibits breast cancer cell proliferation through targeting CDK14, it might be a target for breast cancer therapy.


Height velocity screening: the real world.
To assess the value of the Middlesex-Height Velocity Screening Chart (based on the 50th velocity percentile +/- 1 SD) as a means of screening for abnormal growth in children, the heights of children at seven infant schools were measured on two separate occasions at least 12 months apart. A total of 229 children of mid-age (age of the child between the two measurements) ranging from 4.5 to 6.8 years were measured. Height velocity standard deviation score was calculated for each child by comparison with current Tanner-Whitehouse standards. The subjects were children who had a height recorded during the school entrant medical at least 12 months before the date of the second height measurement. The height velocity standard deviation score calculated for each child was related to the limits set by the Middlesex Screening Chart, and these criteria were used to determine whether or not the child had a 'normal' growth rate. It has been recommended that those falling outside these limits are referred for further assessment. It was found that 95 (41 percent) children fell outside the 'normal' range on the Middlesex chart (66 growing faster and 29 slower), which, if the guidelines were used uncritically, would result in the possible referral of 2500 5-6-year-olds in Sheffield alone. Taking into consideration the additional problems inherent in making two accurate height measurements from which a height velocity could be calculated, and the high number of referrals that would be generated as demonstrated by this study, height velocity screening for growth disorders using current methods is not feasible.


Air analysis in the assessment of fumonisin contamination risk in maize.
In maize-growing areas where fumonisin contamination is endemic, there is an urgent need for novel methods to assess the quality of grain lots before their delivery to common drying and storage collection centres. Aerobiological samples of fungal spores released during harvest were analysed to establish a relationship between fumonisin contamination and the abundance of pathogen propagules collected in the combine harvester using a cyclone and membrane filters. Filter-captured propagules were analysed by direct plating, immunoenzymatic assay of specific Fusarium extracellular polysaccharides and real time polymerase chain reaction of the extracted DNA using fum1, a gene involved in the biosynthesis of fumonisin, as a target. The results showed that time of harvest and environmental conditions strongly influenced the efficiency and performance of the collection system. The data obtained were informative in comparing individual samples collected under similar conditions. The immunoenzymatic assay provided the most reliable data, which improved the ability of a neural network to predict the fumonisin content of lots, when added to agronomic, environmental and phytosanitary data. This is the first attempt to evaluate the Fusarium propagules dispersed during harvesting as a predictive means to assess maize quality. A method based on cyclone/filter capture and immunological detection has been shown to be feasible and to have the potential for the development of a continuous monitoring system, but the prediction capabilities in the present implementation were limited.


Involvement of Normalized Glial Fibrillary Acidic Protein Expression in the Hippocampi in Antidepressant-Like Effects of Xiaoyaosan on Chronically Stressed Mice.
The research has only yielded a partial comprehension of MDD and the mechanisms underlying the antidepressant-like effects of XYS. Therefore, in this study, we aimed to explore the effects of XYS on chronic unpredictable mild stress- (CUMS-) induced changes in the neuronal and the astrocytic markers in the mouse hippocampus. The physical states and depressive-like behaviors in mice with CUMS were recorded. The serum contents of brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) were measured. The protein and mRNA expressions and the immunoreactivities of glial fibrillary acidic protein (GFAP) and neuronal nuclei (NeuN) in mouse hippocampus were detected using a Western blot, qRT-PCR, and immunohistochemical staining, respectively. XYS treatment markedly improved the physical state and depressive-like behaviors in mice subjected to CUMS compared with the model group, and the serum contents of BDNF and GDNF were significantly upregulated. XYS treatment also elevated the protein and mRNA levels, as well as the immunoreactivity of GFAP in the hippocampus. However, CUMS did not influence NeuN expression. In conclusion, these results reveal that chronic administration of XYS elicits antidepressant-like effects in a mouse model of depression and may normalize glial fibrillary acidic protein expression in the hippocampi of mice with CUMS.


[Juvenile malignant lymphomas. 2 Non-Hodgkin lymphomas].
Between 1970 and 1978 33 children with Non-Hodgkin-lymphomas at the age of 2-15 years were treated at the university children's hospital of Jena. 27 patients showed the first appearance of the disease, 6 patients had already been treated in other hospitals and were admitted with relapses. The biopsy material was classified or re-classified after the Kiel-classification. Beside the histological classification the surface markers of the malignant cells of NHL-patients were determined. 20 of 33 children were already in stage IV (Ann-Arbor-classification). Among our patients were 6 lymphoblastic NHL of Brukitt type, 10 of the convoluted cell type and 16 unclassified and one lymphoblastic lymphoma. The main localization of the NHL were mediastinum [15] and the gastrointestinal tract [10]. The therapy consisted of irradiation and chemotherapy (2 protocols) and, in case of an abdominal localization, in the attempt at a radical operation. Patients of stage I and stage II showed a complete remission rate of 50 per cent for 3 years; patients of stages III and IV of 20 per cent only. NHL of the convoluted cell type and of the Burkitt-type proved to have worse three-year-remission rates (16 per cent and 27 per cent) than unclassified lymphoblastic NHL (42 per cent).


[Wegener's granulomatosis and ANCA-associated vasculitis: pathogenic role of autoantigens, autoantibodies, and cytokines].
A real explosion of data is seen concerning the detection of autoantibodies collectively known as antineutrophil cytoplasmic antibodies (ANCA) recognizing granule proteins of granulocytes and monocytes. Distinct fluorescence staining patterns result from their diverse substrate specificities, and can even identify several disease entities mainly in the etiopathogenetically unclear group of primary systemic vasculitides (PSV). The well-documented disease specificity of ANCA recognizing proteinase 3 (PR3), and recent in vitro findings concerning the effect of cytokines on immunocompetent cells including the endothelium are the rationale for the presentation of a new model explaining the development of systemic vasculitis.


The future of measles in highly immunized populations. A modeling approach.
Little is known about how an intensive measles elimination program changes the overall immune status of the population. A computer model was created to study the effect of the measles elimination program in the United States on the number of susceptibles in the population. The simulation reveals that in the prevaccine era, approximately 10.6% of the population was susceptible to measles, most of whom were children less than 10 years of age. With the institution of the measles immunization program, the proportion of susceptibles in the population fell to 3.1% from 1978 through 1981, but then began to rise by approximately 0.1% per year to reach about 10.9% in the year 2050. The susceptibles at this time were distributed evenly throughout all age groups. The model did not consider the potential effect of waning immunity. The results of this study suggest that measles elimination in the United States has been achieved by an effective immunization program aimed at young susceptibles combined with a highly, naturally immunized adult population. However, despite short-term success in eliminating the disease, long-range projections demonstrate that the proportion of susceptibles in the year 2050 may be greater than in the prevaccine era. Present vaccine technology and public health policy must be altered to deal with this eventuality.


Adenine radicals generated in alternating AT duplexes by direct absorption of low-energy UV radiation.
There is increasing evidence that the direct absorption of photons with energies that are lower than the ionization potential of nucleobases may result in oxidative damage to DNA. The present work, which combines nanosecond transient absorption spectroscopy and quantum mechanical calculations, studies this process in alternating adenine-thymine duplexes (AT)n. We show that the one-photon ionization quantum yield of (AT)10 at 266 nm (4.66 eV) is (1.5 ± 0.3) × 10-3. According to our PCM/TD-DFT calculations carried out on model duplexes composed of two base pairs, (AT)1 and (TA)1, simultaneous base pairing and stacking does not induce important changes in the absorption spectra of the adenine radical cation and deprotonated radical. The adenine radicals, thus identified in the time-resolved spectra, disappear with a lifetime of 2.5 ms, giving rise to a reaction product that absorbs at 350 nm. In parallel, the fingerprint of reaction intermediates other than radicals, formed directly from singlet excited states and assigned to AT/TA dimers, is detected at shorter wavelengths. PCM/TD-DFT calculations are carried out to map the pathways leading to such species and to characterize their absorption spectra; we find that, in addition to the path leading to the well-known TA* photoproduct, an AT photo-dimerization path may be operative in duplexes.


New evaluation of reconstructed spatial distribution function from radial distribution functions.
Although three dimensional (3D) solvation structure is much more informative than one dimensional structure, its evaluation is difficult experimentally and theoretically. In our previous Communication [Yokogawa et al., J. Chem. Phys. 123, 211102 (2005)], we proposed a new method to present reconstructed spatial distribution function (RC-SDF) from a set of radial distribution functions (RDFs). In this article, we successfully extended the method more accurately with new basis sets. This new method was applied to two liquid solvation structures, methanol and dimethyl sulfoxide, as examples. Their RC-SDFs evaluated here clearly show that the former solvation structure is well defined while the latter one is broad, which agrees well with the SDFs calculated directly from molecular dynamics simulations. These results indicate that the method can reproduce well these 3D solvation structures in reasonable computational cost.


Hyponatremia and mortality in patients undergoing maintenance hemodialysis: Systematic review and meta-analysis.
This systematic review and meta-analysis examined the relationship between hyponatremia and worse outcomes in patients undergoing maintenance hemodialysis. The MEDLINE, EMBASE, CENTRAL, and Web of Science databases were used to search for relevant articles. The target population was patients on maintenance hemodialysis (those undergoing hemodialysis for ≥60 days). The defined outcomes were death, cardiovascular disease, cognitive decline, and falls. Meta-analysis was performed with a random-effects model of pairwise comparisons of normonatremia and hyponatremia defined for each study, 1-mmol/L increment of sodium analysis, and dose-response analysis using the sodium concentration defined for each study. This study was registered with PROSPERO (registration number CRD42018087667). Thirteen articles were included. The pairwise analysis revealed that the hazard ratio for all-cause mortality was 1.45 (95% confidence interval, 1.31-1.61). The analysis of 1-mmol/L increment of sodium included six studies with a hazard ratio for all-cause mortality of 0.94 (95% confidence interval, 0.91-0.97) for each 1-mmol/L increase in the serum sodium concentration. In the dose-response analysis, assuming a linear relationship, a sodium increment of 1 mmol/L revealed a hazard ratio for all-cause mortality of 0.97 (95% confidence interval, 0.96-0.98). Other outcomes could not be integrated. Hyponatremia is associated with all-cause mortality in patients undergoing maintenance hemodialysis. Healthcare providers should pay special attention to even the slightest indication of hyponatremia.


Pineal Calcification, Melatonin Production, Aging, Associated Health Consequences and Rejuvenation of the Pineal Gland.
The pineal gland is a unique organ that synthesizes melatonin as the signaling molecule of natural photoperiodic environment and as a potent neuronal protective antioxidant. An intact and functional pineal gland is necessary for preserving optimal human health. Unfortunately, this gland has the highest calcification rate among all organs and tissues of the human body. Pineal calcification jeopardizes melatonin's synthetic capacity and is associated with a variety of neuronal diseases. In the current review, we summarized the potential mechanisms of how this process may occur under pathological conditions or during aging. We hypothesized that pineal calcification is an active process and resembles in some respects of bone formation. The mesenchymal stem cells and melatonin participate in this process. Finally, we suggest that preservation of pineal health can be achieved by retarding its premature calcification or even rejuvenating the calcified gland.


Histone acetylation-independent transcription stimulation by a histone chaperone.
Histone chaperones are thought to be important for maintaining the physiological activity of histones; however, their exact roles are not fully understood. The physiological function of template activating factor (TAF)-I, one of the histone chaperones, also remains unclear; however, its biochemical properties have been well studied. By performing microarray analyses, we found that TAF-I stimulates the transcription of a sub-set of genes. The transcription of endogenous genes that was up-regulated by TAF-I was found to be additively stimulated by histone acetylation. On performing an experiment with a cell line containing a model gene integrated into the chromosome, TAF-I was found to stimulate the model gene transcription in a histone chaperone activity-dependent manner additively with histone acetylation. TAF-I bound to the core histones and remodeled the chromatin structure independent of the N-terminal histone tail and its acetylation level in vitro. These results suggest that TAF-I remodel the chromatin structure through its interaction with the core domain of the histones, including the histone fold, and this mechanism is independent of the histone acetylation status.


The effect of stimulus bandwidth on binaural loudness summation.
Binaural loudness summation is an important property of the human auditory system. This paper presents an experimental investigation of how binaural loudness summation varies with stimulus bandwidth. Loudness matches were obtained between dichotic stimuli, with interaural level differences (ILDs) of 2-12 dB, and diotic stimuli. The stimuli were noise bands with seven center frequencies and four bandwidths. Results showed that the loudness of dichotic stimuli increased nonlinearly with ILD, the increase being slightly less with broader bandwidths. There was a bandwidth-dependent difference between the listening tests results and the predictions of Moore and Glasberg's [(2007) J. Acoust. Soc. Am. 121, 1604-1612] loudness model. The size of the difference was, however, small. A characteristic function was derived describing how overall loudness depends on stimulus bandwidth and ILD.


Assessment of Four Serum Biochemical Markers in Elderly Patients with Vascular Dementia after Cerebral Infarction and Their Response to Donepezil and Idebenone.
Objective Our study aimed to explore four serum levels of biochemical markers, including brain-derived neurotrophic factor (BDNF), homocysteine (Hcy), nitric oxide (NO), and γ-interferon (IFN-γ), in elderly patients with vascular dementia (VD) after the cerebral infarction and to elucidate possible connections between them. Method The elderly patients with VD after cerebral infarction admitted in our hospital, and the elderly persons for physical examination from November 2020 to December 2021 were included in this study. The serum levels of BDNF, Hcy, NO, and IFN-γ were compared between the study group and the control group. Results In the study group, the serum levels of Hcy and IFN-γ were significantly higher than that in the control group, whereas significantly lower serum levels of BDNF and NO were found in the study group compared with the control group. After receiving the intervention of donepezil and/or idebenone, the serum levels of Hcy and IFN-γ in group B were significantly lower than that in group A, while the serum levels of BDNF and NO in group B were significantly higher than that in Group A. Conclusion The results of our study showed abnormally expressed serum levels of Hcy, IFN-γ, BDNF, and NO in elderly patients with VD after cerebral infarction which might strongly reflect the severity of VD. Moreover, after intervention of donepezil alone or combined with idebenone, the changes of serum levels of Hcy, IFN-γ, BDNF, and NO may reflect the curative effect of the disease.


Metabolic and endocrine changes associated with undernutrition in the peripubertal gilt.
The effect of feed restriction on circulating metabolites and metabolic and gonadotropic hormones was determined in Large White gilts. From 25 kg BW and 80 d of age, control gilts (CTRL, n = 10) were fed close to ad libitum intake, whereas restricted-fed gilts (REST, n = 10) were offered approximately 50% of the CTRL diet. Estrus was detected with a mature boar. Serial blood samples were collected at approximately 160 and 210 d of age from 0800 to 2000. A meal was supplied at 0900, and an i.v. injection of GnRH (80 ng/kg BW) was given at 1600. The gilts were slaughtered at 220 d of age. No REST and four CTRL gilts exhibited their first estrus between the two series of blood samplings. Feed intake, daily gain, and fat content of the carcass were reduced by 50% in REST gilts (P < .001). Preprandial concentrations of free fatty acids were significantly lower in CTRL gilts at 210 d of age than at 160 d and than in REST gilts at both ages. Within the CTRL group, concentrations were lower (P < .05) at both ages in gilts pubertal before 210 d than in others. After the meal, plasma insulin and urea increased in all gilts, whereas glucose increased and free fatty acids decreased in all except the 210-d CTRL gilts. The afternoon concentration of cortisol was slightly higher (P = .09) in REST than in CTRL gilts. The number of pulses and mean and interpulse concentrations of growth hormone did not show any clear variation with feed level and age.(ABSTRACT TRUNCATED AT 250 WORDS)


Development of rapid and simple experimental and in silico serotyping systems for Citrobacter.
Members of the genus Citrobacter are important opportunistic pathogens responsible for high mortality rate. Therefore, in this study, we aimed to develop efficient and accurate Citrobacter typing schemes for clinical detection and epidemiological surveillance. Using genomic and experimental analyses, we located the O-antigen biosynthesis gene clusters in Citrobacter genome for the first time, and used comparative genomic analyses to reveal the specific genes in different Citrobacter serotypes. Based on the specific genes in O-antigen biosynthesis gene clusters of Citrobacter, we established experimental and in silico serotyping systems for this bacterium. Both serotyping tools are reliable, and our observations are biologically and clinically relevant for understanding and managing Citrobacter infection.


[Down-regulation of Protein Phosphatase 2A Catalytic Subunit Involved in Mitochondria Fission/fusion Dynamics Imbalance and Functional Impairment Induced by Human tau].
Objective To explore whether the downregulation of protein phosphatase 2A catalytic subunit(PP2Ac)involved in the pathogenesis of mitochondria fission/fusion dynamics and functional imbalance induced by human tau accumulation. Methods After cotransfection with mito-dsRed plasmids and pIRES-eGFP-tau40 plasmids 48 hours,the rat primary hippocampal neurons were observed with a laser scanning confocal microscope for their changes in shape and distribution of mitochondria.The expressions of mitochondria fission/fusion protein and PP2Ac and PP2Ab were detected by Western blotting.Furthermore,the shape and distribution of mitochondria of rat primary hippocampal neuron and wild type 293wt cells were assayed 48 hours after co-transfection with siPP2Ac-EGFP plasmids and mito-DsRed plasmids,and the fission/fusion dynamics of 293wt cells was captured with live cell time-lapse imaging after co-transfection with siPP2Ac plasmids and mito-Dendra2 plasmids.After transfection with siPP2Ac plasmids,the relative level of mitochondria fission/fusion protein of 293wt cells was assayed by Western blotting,and mitochondria membrane potential was detected by JC-1 staining,and the cellular viability was measured by CCK8 assay.Finally,the shape and distribution and membrane potential of mitochondria of HEK293 cells with stable transfection of htau40(293htau)were detected after co-transfection with PP2Ac and mito-dsRed plasmids. Results Human tau40 expression decreased distribution of mitochondria and significantly lowered PP2Ac level in primary hippocampal neuron(t=4.814, P=0.0086).Down-regulation of PP2Ac caused mitochondria elongation and perinuclear accumulation in primary hippocampal neuron and 293wt cells;in addition,down-regulation of PP2Ac in 293wt cells significantly increased mitochondria fusion rate(t=2.857, P=0.0074)and the levels of mitochondria fusion protein mitofusin(MFN)1(t=6.768, P=0.0025),MFN2(t=3.121, P=0.0035),and optic atrophy 1(t=3.775, P=0.0199);however,the levels of dynamin-like protein-1 and Fis1 remained unchanged.The down-regulation of PP2Ac in 293wt cells led to the significant decrease in mitochondria membrane potential(t=2.300, P=0.0270)and cell viability(t=6.249, P<0.0001).Finally,up-regulation of PP2Ac attenuated the abnormalities in the shape,distribution and function of mitochondria in the 293htau cells. Conclusion Down-regulation of PP2Ac is involved in the abnormal shape and distribution of mitochondria and its dysfunction induced by human tau40 in rat primary hippocampal neurons and HEK293 cells.


The impact of adverse childhood experiences (ACEs) and combat exposure on mental health conditions among new post-9/11 veterans.
Adverse childhood experiences (ACEs) are early life experiences of abuse and neglect, and observed violence, among others. For military veterans, both ACEs and combat exposure are associated with mental health problems. This study examines the relationship between ACEs and combat exposure on the current mental health in a large sample of recent post-9/11 U.S. veterans. Fifty-nine percent of female and 39% of male veterans reported exposure to 1 ACE, whereas 44% of female and 25% of male veterans were exposed to multiple ACEs. Female veterans were more likely to experience 4 or more ACEs. ACEs were more consistently associated with mental health problems for male veterans than their female peers. For female veterans, exposure to 1 or 2 ACEs did not increase the odds of having any mental health condition, whereas for males, this level of exposure was associated with probable PTSD and anxiety. Combat patrol events were associated with an increase in the likelihood of having a probable mental health problem, with 2 exceptions-combat patrol events were not associated with depression in male veterans and not associated with alcohol misuse in female veterans. Combat was not associated with alcohol misuse. Experiencing a corollary of combat (e.g., accidents, moral injury) was inconsistently associated with the odds of having a probable mental health problem. This study confirms prior studies demonstrating a relationship between ACEs and combat on subsequent mental health problems. Importantly, 2 different types of combat exposure had differential effects on mental health problems. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


Vacuum-ultraviolet (VUV) photoionization of small methanol and methanol-water clusters.
In this work, we report on the vacuum-ultraviolet (VUV) photoionization of small methanol and methanol-water clusters. Clusters of methanol with water are generated via co-expansion of the gas phase constituents in a continuous supersonic jet expansion of methanol and water seeded in Ar. The resulting clusters are investigated by single photon ionization with tunable vacuum-ultraviolet synchrotron radiation and mass analyzed using reflectron mass spectrometry. Protonated methanol clusters of the form (CH3OH)nH(+) (n = 1-12) dominate the mass spectrum below the ionization energy of the methanol monomer. With an increase in water concentration, small amounts of mixed clusters of the form (CH3OH)n(H2O)H(+) (n = 2-11) are detected. The only unprotonated species observed in this work are the methanol monomer and dimer. Appearance energies are obtained from the photoionization efficiency (PIE) curves for CH3OH(+), (CH3OH)2(+), (CH3OH)nH(+) (n = 1-9), and (CH3OH)n(H2O)H(+) (n = 2-9) as a function of photon energy. With an increase in the water content in the molecular beam, there is an enhancement of photoionization intensity for the methanol dimer and protonated methanol monomer at threshold. These results are compared and contrasted to previous experimental observations.


[Emergency colonic surgery: analysis of risk factors predicting morbidity and mortality].
The aim of the present study was to identify risk factors for morbidity and mortality in patients submitted to emergency colonic surgery. Between 1997 and 2008 157 patients, 106 of whom affected by colon cancer (67.5%) and 51 by benign disease (32.5%), were treated. The risk factors for morbidity and mortality were evaluated by univariate and multivariate analysis considering clinical and demographic data. The overall 30-day morbidity and mortality rates were 19.1% (30 patients) and 12.7% (20 patients), respectively. Among patients affected by cancer the mortality rate was 15% (16 patients) and the morbidity rate 23.6% (25 patients), while among the patients with benign disease the mortality rate was 7.8% (4 patients) and the morbidity rate 9.8% (5 patients). No postoperative surgical complications were noted. The strongest risk factors for early death were postoperative medical complications such as cardiopulmonary, renal, thrombo-embolic and infectious complications. The results of the univariate analysis showed that advanced age, neoplastic disease, advanced stage of cancer and associated medical disease prior to surgery play a role as risk factors for morbidity and mortality. In the multivariate analysis only the presence of associated medical disease proved to be a significant independent predictor of outcome. Emergency surgery for both neoplastic and benign colonic disease is still associated with an increased risk of death. Although restorative colectomy should be regarded as the first choice procedure in the emergency setting, Hartmann's procedure is still widely used in high-risk patients.


The PTPN22 620W allele confers susceptibility to systemic sclerosis: findings of a large case-control study of European Caucasians and a meta-analysis.
To determine whether genetic variants of the PTPN22 gene, including the R620W (1858C>T) missense single-nucleotide polymorphism (SNP), are associated with systemic sclerosis (SSc). Since PTPN22 is involved in multiple autoimmune diseases, we also examined the occurrence of a concomitant autoimmune disease. We then conducted a meta-analysis of the most recent studies of SSc in order to verify the association or lack of association between the PTPN22 1858C>T variant and SSc. Seven PTPN22 SNPs were analyzed in a French Caucasian cohort of 659 SSc patients and 504 healthy controls. All SSc patient sera were tested for the presence of autoantibodies against topoisomerase I (anti-topo I) and for anticentromere antibodies (ACAs). The co-occurrence of an autoimmune disease was observed in 22% of the 416 SSc patients who were exhaustively screened. In 33 of the 416 patients (8%), the concomitant autoimmune disease was known to be associated with PTPN22 1858T; these patients were excluded prior to analysis. No association was detected for any of the SNPs tested. PTPN22 haplotype analysis identified a strong association between SSc and the presence of a risk haplotype carrying the 1858T allele (P = 1.52 x 10(-7)) and a protective haplotype carrying the 1858C allele (P = 2.20 x 10(-16)) in our French Caucasian population. The meta-analysis provided evidence that the PTPN22 1858T allele is involved in the genetic susceptibility to SSc in Caucasian (P = 8.39 x 10(-3), OR 1.08 [95% CI 1.02-1.15]) and mixed (P = 3.11 x 10(-3), OR 1.09 [95% CI 1.04-1.16]) populations, particularly in the anti-topo I-positive subset. Our results indicate that PTPN22, a shared genetic factor of multiple autoimmune diseases, also contributes to the genetic background of SSc.


Ultrastructural and morphometric studies of normal rat hepatocytes.
To investigate the arrangement of organelles in normal rat liver hepatocytes, we examined liver tissues by electron microscopy. We also analyzed hepatocyte organelles by morphometry using the NIH image software. The smooth ER area appears as a network of branching tubules and contains many glycogen rosettes. The rough ER area presents lamellar plates and contains free polyribosomes. The Golgi complex area comprises the parallel smooth surface of associated vesicles and contains secretory granules, lysosomes and a few glycogens. Ratio of the cross-sectional area of hepatocytes shows the smooth ER area (60 +/- SD 1.5%), the rough ER area (35 +/- SD 1.4%) and the Golgi complex area (4 +/- SD 1.7%) of the total cytoplasm. The rough ER shows a dendrite pattern in a hepatocyte. Many mitochondria are spread between the smooth ER and the rough ER area. The smooth ER directly communicates with the rough ER, the Golgi complex cisternae at a forming face and the nuclear envelope. Anastomosing tubules with polyribosomes between the smooth ER and rough ER, between smooth ER and the nuclear envelope frequently. These results provide a basics at the cellular level for better understanding the experimentally induced changes and for improving the pathological hepatic diagnosis.


Treatment of severe or resistant corticosteroid-responsive dermatoses with Diprolene cream.
In an open study Diprolene Cream was administered to fifty patients with severe or resistant psoriasis, atopic dermatitis or other corticosteroid-responsive dermatoses. For a period of 14 days, patients applied 3.5 grams of medication twice daily. Efficacy was evaluated in forty-six patients, while tolerance and safety were evaluated in fifty patients. Therapeutic response in forty-one of forty-six patients was noted by treatment Day 3. By Day 14, mean total severity score of signs and symptoms in twenty-one patients with psoriasis was reduced by 88%. Similarly, in twenty-five patients with other severe or resistant corticosteroid-responsive dermatoses, a 99% decrease in the mean total severity score of signs and symptoms was evident. Of the patients evaluable for tolerance and safety, twenty-four of fifty reported transient, mild to moderate burning that occurred most often upon application of the study preparation. Therapy was not discontinued in any of these patients, and none required additional treatment for the local reaction. Morning plasma cortisol levels in monitored patients remained within normal limits throughout the study.


Technical note: how to use Winbugs to draw inferences in animal models.
This paper deals with Bayesian inferences of animal models using Gibbs sampling. First, we suggest a general and efficient method for updating additive genetic effects, in which the computational cost is independent of the pedigree depth and increases linearly only with the size of the pedigree. Second, we show how this approach can be used to draw inferences from a wide range of animal models using the computer package Winbugs. Finally, we illustrate the approach in a simulation study, in which the data are generated and analyzed using Winbugs according to a linear model with i.i.d errors having Student's t distributions. In conclusion, Winbugs can be used to make inferences in small-sized, quantitative, genetic data sets applying a wide range of animal models that are not yet standard in the animal breeding literature.


Repression of HMGA2 gene expression by human cytomegalovirus involves the IE2 86-kilodalton protein and is necessary for efficient viral replication and inhibition of cyclin A transcription.
Human cytomegalovirus (HCMV) infection results in dysregulation of several cell cycle genes, including inhibition of cyclin A transcription. In this work, we examine the effect of the HCMV infection on expression of the high-mobility group A2 (HMGA2) gene, which encodes an architectural transcription factor that is involved in cyclin A promoter activation. We find that expression of HMGA2 RNA is repressed in infected cells. To determine whether repression of HMGA2 is directly related to the inhibition of cyclin A expression and impacts on the progression of the infection, we constructed an HCMV recombinant that expressed HMGA2. In cells infected with the recombinant virus, cyclin A mRNA and protein are induced, and there is a significant delay in viral early gene expression and DNA replication. To determine the mechanism of HMGA2 repression, we used recombinant viruses that expressed either no IE1 72-kDa protein (CR208) or greatly reduced levels of IE2 86-kDa (IE2 86) protein (IE2 86DeltaSX-EGFP). At a high multiplicity of infection, the IE1 deletion mutant is comparable to the wild type with respect to inhibition of HMGA2. In contrast, the IE2 86DeltaSX-EGFP mutant does not significantly repress HMGA2 expression, suggesting that IE2 86 is involved in the regulation of this gene. Cyclin A expression is also induced in cells infected with this mutant virus. Since HMGA2 is important for cell proliferation and differentiation, particularly during embryogenesis, it is possible that the repression of HMGA2 expression during fetal development could contribute to the specific birth defects in HCMV-infected neonates.


Associations of serum uric acid with cardiovascular events and mortality in moderate chronic kidney disease.
It is unclear whether the presence of kidney disease modifies the associations of uric acid with cardiovascular events and death. In the limited access, public use Atherosclerosis Risk In Communities (ARIC) database, associations of serum uric acid levels with cardiovascular events and death were analysed using a parametric proportional hazards model and the modification of these associations by the presence of CKD was assessed using a likelihood ratio test. Of the 15 366 ARIC participants included in this analysis, 461 had CKD (eGFR <60 ml/min/1.73 m(2)). In both non-CKD and CKD sub-groups, participants with hyperuricaemia (> or = 7 mg/dl in men and > or = 6 mg/dl in women) compared to those with normal serum uric acid levels had higher waist circumference and fasting serum insulin levels. In the entire cohort, in a multivariate parametric proportional hazards model, each mg/dl increase in serum uric acid was associated with an increased hazard of cardiovascular events (HR 1.09, 95% CI 1.05-1.12) and death. A multiplicative interaction term of serum uric acid and CKD when added to the above models was significant (P < 0.001). The likelihood ratio test of the models with and without the interaction term was also significant (P < 0.001). In the non-CKD population, a multivariate analysis after adjusting for comorbidities and metabolic syndrome showed a significant association between hyperuricaemia and mortality (HR 1.18, 95% CI 1.04-1.33) but not for cardiovascular events (HR 1.07, 95% CI 0.96-1.19). In the CKD population, the association was not significant for both mortality and cardiovascular events. We conclude that hyperuricaemia is associated with insulin resistance and mortality in the non-CKD population. The presence of CKD attenuates the associations of uric acid with mortality. Interventional studies are warranted to establish the biological role of hyperuricaemia in mortality in non-CKD and CKD populations.


Determinants of complete remission induction and maintenance in chemotherapy with or without irradiation of small cell lung cancer.
The possible influence of pretreatment patient characteristics upon the probabilities of complete remission (CR) induction and maintenance was investigated in a series of 815 nonresected patients with small cell lung cancer. All patients underwent pretreatment staging which enabled allocation of 391 patients to trials for limited stage disease and 424 patients to trials for extensive disease. Three controlled trials for each disease stage were conducted between 1973 and 1981. All therapeutic regimens consisted of combinations of between three and six agents (lomustine, cyclophosphamide, methotrexate, vincristine, doxorubicin, etoposide) with or without irradiation. Thirty-five % of the limited stage patients and 18% of the extensive stage patients were alive and had achieved a complete remission 16 weeks after initiation of the treatment, i.e., after four cycles of chemotherapy. Relationships between pretreatment characteristics and the probability to pass this benchmark were examined by logistic regression analysis. The probability of CR was negatively related to increased serum lactate dehydrogenase and male sex in both disease stages. Pretreatment anemia (less than 12 g/liter) and poor performance status were associated with a reduced CR rate in limited and extensive stage disease, respectively. Factors related to the maintenance of complete remission were subsequently examined in the 211 complete responders by use of Cox's regression analysis. Complete responders with extensive disease prior to treatment had greater cumulative risk of relapse than those with limited disease (P less than 0.01). Hyponatremia had a significant negative influence on the remission duration in limited disease while age greater than 60 years and bone marrow metastases had significantly negative influence in extensive disease. Using the models it was possible to identify subgroups of patients with CR rates ranging from 5 to 55% and to stratify complete responders according to estimated risks of subsequent relapse.


Quercetin but not quercitrin ameliorates tumor necrosis factor-alpha-induced insulin resistance in C2C12 skeletal muscle cells.
Skeletal muscle is a major site for glucose metabolism and its injury by cytokines can induce insulin resistance leading to type 2 diabetes. It has been suggested that quercetin may act as an anti-diabetic agent, however, the effects of quercetin on insulin resistance in skeletal muscle remain unknown. We aimed to investigate the role of quercetin and its glycoside, quercitrin in tumor necrosis factor-alpha (TNF-α) induced C2C12 skeletal muscle cell impairment. Quercetin, but not quercitrin moderately attenuated the effects of TNF-α and enhanced the basal and insulin stimulated uptake of glucose in a dose-dependent manner via the activation of the protein kinase B (Akt) and AMP-activated protein kinase (AMPK) pathways. Furthermore, the underlying mechanism also involved the suppression of nuclear factor-κB (NF-κB) signaling and the nitric oxide (NO)/inducible nitric oxide synthase (iNOS) system, downstream of AMPK transduction. In summary, quercetin exhibited its effect of improving glucose uptake and insulin sensitivity in skeletal muscle cells via the two independent signaling pathways of Akt and AMPK, and can be developed as a potential anti-diabetic agent.


Presence of Streptococcus dentisani in the dental plaque of children from different Colombian cities.
Streptococcus dentisani has been identified as an oral cavity probiotic due to its beneficial characteristics. One of its beneficial features is the production of bacteriocins, which inhibit the growth of cariogenic bacteria, and another is its buffering capacity through the production of ammonium from arginine. The purpose of this study was to determine the presence of S. dentisani in the dental plaque of Colombian children and whether the presence of this bacterium is related to oral health and other conditions. Dental plaque and information on diet and oral hygiene habits were collected from children between 6 and 12 years of age from four Colombian cities, divided into caries-free children (International Caries Detection and Assessment System [ICDAS] 0, Decayed Missing Filled Teeth index [DMFT] 0), children with ICDAS 1 and 2, and children with ICDAS >3. Plaque DNA was extracted and quantified, and real-time polymerase chain reaction was performed using specific primers. This bacterium was identified in all samples, with a median of 0.46 cells/ng DNA (interquartile range [IQR] 0.13-1.02), without finding significant differences between the groups (P > 0.05). In caries-free children, a median of 0.45 cells/ng DNA (IQR 0.14-1.23) was found. In children with ICDAS 1 and 2, the median was 0.49 cells/ng DNA (IQR 0.11-0.97), and in children with ICDAS >3, the median was 0.35 cells/ng DNA (IQR 0.12-1.07). However, statistically significant differences were found in the origin of children (P < 0.01), the use of fluoride-containing products (P < 0.01), and the frequency of food intake (P < 0.05). In conclusion, the presence of S. dentisani was quantified in children from four Colombian cities, without finding significant differences in oral health status. Nevertheless, three conditions showed a possible relationship with S. dentisani.


Focal segmental glomerulosclerosis and kidney transplantation.
The recurrence rate of focal segmental glomerulosclerosis after kidney transplantation is ranging between 20% and 40%. Focal segmental glomerulosclerosis is associated with poor graft survival. In this review, the etiology, pathogenesis, clinicopathological features, risk factors of recurrence, and updated lines of management are discussed.


Sexuality of adolescent girls born with HIV in Senegal: an anthropological analysis.
In Senegal, the dominant social norm upholds virginity before marriage and edifies abstinence for adolescents as a cardinal moral value. Currently, sex outside of marriage remains socially condemned. The onset of sex for adolescent girls born with HIV in Senegal brings up several challenges. In Dakar, initiatives, especially through digital applications, are being developed to support these young people. These programs are much rarer in rural settings. A study conducted in 2021 explored how adolescent girls born with HIV who live outside of Dakar experience sexuality, what socio-health constraints they face, and what support they receive from the healthcare system. An anthropological study titled 'Treatment Failure among Children and Adolescents Living with HIV in Senegal, Outside Dakar' (ETEA-VIH, ANRS 12421) was conducted in 2021 in 14 regional hospitals and health centers. Semi-structured interviews were conducted with 87 HIV-positive children and adolescents, 95 parents/guardians, and 47 health care workers. Adolescent girls' onset of sexuality was specifically analyzed for 40 adolescent girls age 12-19 years old. Generally, parents feign oblivion about their children's sexual lives. Mothers dread a pregnancy out of marriage because they are responsible for overseeing sex education and would be 'blamed' for the transgression. The occurrence of an unintended pregnancy can lead to exclusion from the family and a risk of transmitting HIV to the child due to the lack of medical and social support. HIV remains a stigmatizing disease that families keep secret. The risk of disclosure is a major concern. Despite sexual and reproductive health (SRH) programs, most healthcare workers are reluctant to discuss sexuality or to offer contraception to adolescent girls. Information spaces have been set up in some regional hospitals by associations trained in SRH. They are rarer in health centers. Accessibility to digital applications and discussion forums is limited due to the lack of smartphones and Internet access. In rural settings, HIV-positive adolescent girls are confronted with the silence that surrounds sexuality and HIV. An individualized approach and confidential access to contraception should be prioritized to support them with assistance from PLHIV associations.


Evaluation and Application of Population Pharmacokinetic Models for Identifying Delayed Methotrexate Elimination in Patients With Primary Central Nervous System Lymphoma.
Objective: Several population pharmacokinetic (popPK) models have been developed to determine the sources of methotrexate (MTX) PK variability. It remains unknown if these published models are precise enough for use or if a new model needs to be built. The aims of this study were to 1) assess the predictability of published models and 2) analyze the potential risk factors for delayed MTX elimination. Methods: A total of 1458 MTX plasma concentrations, including 377 courses (1-17 per patient), were collected from 77 patients who were receiving high-dose MTX for the treatment of primary central nervous system lymphoma in Huashan Hospital. PopPK analysis was performed using the NONMEM® software package. Previously published popPK models were selected and rebuilt. A new popPK model was then constructed to screen potential covariates using a stepwise approach. The covariates were included based on the combination of theoretical mechanisms and data properties. Goodness-of-fit plots, bootstrap, and prediction- and simulation-based diagnostics were used to determine the stability and predictive performance of both the published and newly built models. Monte Carlo simulations were conducted to qualify the influence of risk factors on the incidence of delayed elimination. Results: Among the eight evaluated published models, none presented acceptable values of bias or inaccuracy. A two-compartment model was employed in the newly built model to describe the PK of MTX. The estimated mean clearance (CL/F) was 4.91 L h-1 (relative standard error: 3.7%). Creatinine clearance, albumin, and age were identified as covariates of MTX CL/F. The median and median absolute prediction errors of the final model were -10.2 and 36.4%, respectively. Results of goodness-of-fit plots, bootstrap, and prediction-corrected visual predictive checks indicated the high predictability of the final model. Conclusions: Current published models are not sufficiently reliable for cross-center use. The elderly patients and those with renal dysfunction, hypoalbuminemia are at higher risk of delayed elimination.


XPD Polymorphisms and Risk of Squamous Cell Carcinoma of the Head and Neck in a Korean Sample.
XPD is a major player in nucleotide excision repair, which is one of the basic pathways of DNA repair. The objective of this study was to investigate the association of XPD single nucleotide polymorphisms (SNPs) and the risk of squamous cell carcinoma of the head and neck (SCCHN) in Koreans. We performed XPD +23591G>A and +35931A>C genotyping in 290 SCCHN patients and 358 controls. The frequencies of the XPD +23591G>A (GG/GA/AA) genotypes were 89.0%/11.0%/0% in the patients and 90.3%/8.8%/0.9% in the controls, respectively. The odds ratio (OR) of the XPD +23591 GA genotype was 1.94 (0.92 to 4.08) in reference to the GG genotype. The frequencies of the XPD +35931A>C (AA/AC/CC) genotypes were 86.9%/12.0%/1.1% in the patients and 85.6%/13.8%/0.6% in the controls, respectively. The OR of the XPD +35931 AC and CC genotypes were 0.98 (0.51 to 1.88) and 2.68 (0.71 to 10.1), respectively, in reference to the AA genotype. On the subgroup analyses according to the smoking and drinking statuses, the SNPs and haplotypes of XPD showed no statistically significant association with the risk of SCCHN. The results of this study suggest that the XPD +23591G>A and +35931A>C SNPs are not associated with the risk of SCCHN in Koreans; however, a further study with a larger number of subjects is necessary to verify this conclusion.


Sex, Organ, and Breed Differences in the mRNA Expression of Drug Transporters in the Liver and Kidney of Pigs.
Domestic pigs are attractive as an animal model for humans because of their anatomical and physiological similarities to humans. In this study, sex, organ, and breed differences in the mRNA expression of drug transporters such as breast cancer resistance protein (BCRP), multidrug resistance protein 1 (MDR1), multidrug resistance associated protein 2 (MRP2), organic anion porting polypeptide 1B3 (OATP1B3), organic anion transporters (OAT1, OAT2, and OAT3), and organic cation transporters (OCT1 and OCT2) were examined by RT-PCR in the liver and kidney of 5-month-old Meishan and Landrace pigs. No sex differences in the amount of BCRP mRNA were observed in both breeds. In Meishan pigs, sex differences (male < female) in the mRNA amounts of MDR1, OATP1B3, and OCT1 were observed in the liver. Similarly, sex differences in the mRNA amounts of MRP2, OAT1, OAT2, OAT3, and OCT2 were observed in the kidney of Meishan pigs: male > female for MRP2, OAT3, and OCT2, and male < female for OAT1 and OAT2. However, no such sex differences were observed in Landrace pigs. In addition, regardless of breed, hepatic OAT1, OAT3, and OCT2 mRNAs and renal OATP1B3 mRNA were not detected. Thus, organ and breed differences in the expression of drug transporters suggest the existence of genetically controlled organ-selective factors. Furthermore, additional experiments in castrated and/or testosterone propionate-treated pigs strongly suggested that sex and breed differences in the gene expression of drug transporters, especially hepatic OCT1 and renal OAT1, were primarily due to the difference in serum testosterone concentration.


Rhenium(IV)-copper(II) heterobimetallic complexes with a bridge malonato ligand. Synthesis, crystal structure, and magnetic properties.
The Re(IV) complex [ReCl4(mal)]2-, in the form of two slightly different salts, (AsPh4)1.5(HNEt3)0.5[ReCl4(mal)] (1a) and (AsPh4)(HNEt3)[ReCl4(mal)] (1b), and the Re(IV)-Cu(II) bimetallic complexes [ReCl4(mu-mal)Cu(phen)2].CH3CN (2), [ReCl4(mu-mal)Cu(bpy)2] (3), and [ReCl4(mu-mal)Cu(terpy)] (4) (mal=malonate dianion, AsPh4=tetraphenylarsonium cation, HNEt3=triethylammonium cation, phen=1,10-phenanthroline, bpy=2,2'-bipyridine and terpy=2,2':6',2' '-terpyridine) have been synthesized and the structures of 1a, 1b, 2, and 3 determined by single-crystal X-ray diffraction. The structures of 1a and 1b are made up of discrete [ReCl4(mal)]2- anions and AsPh4+ and HNEt3+ cations, held together by electrostatic forces and hydrogen bonds. The Re(IV) atom is surrounded by four chloride anions and a bidentate malonate group, in a distorted octahedral environment. The structure of 2 consist of neutral dinuclear units [ReCl4(mu-mal)Cu(phen)2], with the metal ions united through a bridge carboxilato. The environment of Re(IV) is nearly identical to that in the mononuclear complex, and Cu(II) is five coordinate, being surrounded by four nitrogen atoms of two bidentate phen ligands and one oxygen atom of the malonato ligand. In 3, there are also dinuclear units, [ReCl4(mu-mal)Cu(bpy)2], but the Cu(II) ions complete a distorted octahedral coordination by binding with the free malonato oxygen atom of a neighbor unit, resulting in an infinite chain. The magnetic properties of 1-4 were also investigated in the temperature range 2.0-300 K. The magnetic behavior of 1a and 1b is as expected for a Re(IV) complex with a large value of the zero-field splitting (2D ca. 110 cm(-1)). For the bimetallic complexes, the magnetic coupling between Re(IV) and Cu(II) is antiferromagnetic in 2 (J=-0.39 cm(-1)), ferromagnetic in 4 (J=+1.51 cm(-1)), and nearly negligible in 3 (J=-0.09 cm(-1)).


Nuclear quiescence and histone hyper-acetylation jointly improve protamine-mediated nuclear remodeling in sheep fibroblasts.
Recently we have demonstrated the possibility to replace histones with protamine, through the heterologous expression of human protamine 1 (hPrm1) gene in sheep fibroblasts. Here we have optimized protaminization of somatic nucleus by adjusting the best concentration and exposure time to trichostatin A (TSA) in serum-starved fibroblasts (nuclear quiescence), before expressing Prm1 gene. To stop cell proliferation, we starved cells in 0.5% FBS in MEM ("starved"-ST group), whereas in the Control group (CTR) the cells were cultured in 10% FBS in MEM. To find the most effective TSA concentration, we treated the cells with increasing concentrations of TSA in MEM + 10% FBS. Our results show that combination of cell culture conditions in 50 nM TSA, is more effective in terminating cell proliferation than ST and CTR groups (respectively 8%, 17.8% and 90.2% p<0.0001). Moreover, nuclear quiescence marker genes expression (Dicer1, Smarca 2, Ezh1 and Ddx39) confirmed that our culture conditions kept the cells in a nuclear quiescent state. Finally, ST and 50 nM TSA jointly increased the number of spermatid-like cell (39.4%) at higher rate compared to 25 nM TSA (20.4%, p<0.05) and 100 nM TSA (13.7%, p<0.05). To conclude, we have demonstrated that nuclear quiescence in ST cells and the open nuclear structure conferred by TSA resulted in an improved Prm1-mediated conversion of somatic nuclei into spermatid-like structures. This finding might improve nuclear reprogramming of somatic cells following nuclear transfer.


Induced plasmid-genome rearrangements in Rhizobium japonicum.
The P group resistance plasmids RP1 and RP4 were introduced into Rhizobium japonicum by polyethylene-glycol-induced transformation of spheroplasts. After cell wall regeneration, transformants were recovered by selecting for plasmid determinants. Plant nodulation, nitrogen fixation, serological, and bacterial genetics studies revealed that the transformants were derived from the parental strains and possessed the introduced plasmid genetic markers. Agarose gel electrophoresis, restriction enzyme analysis, and DNA hybridization studies showed that many of the transformant strains had undergone genome rearrangements. In the RP1 transformants, chromosomal DNA was found to have transposed into a large indigenous plasmid of R. japonicum, producing an even larger plasmid, and the introduced R plasmid DNA was found to be chromosomally integrated rather than replicating autonomously or integrated into the endogenous plasmid. Seemingly, a similar section of chromosomal DNA was involved in all the genomic rearrangements observed in the R. japonicum RP1 and RP4 transformant strains.


[Investigation of Leishmania RNA Virus 2 in Leishmania major and Leishmania tropica Strains Isolated from Cutaneous Leishmaniasis Patients in Türkiye].
Leishmania RNA virus (LRV) is a double-stranded RNA (dsRNA) virus that is thought to contribute to the severe inflammatory response of the causative Leishmania parasite in the mammalian host by being present in many isolates of Leishmania spp. In our study, it was aimed to obtain data on the presence of Leishmania RNA Virus 2 (LRV2), which is thought to cause a change in the clinical course of leishmaniasis, in Leishmania major and Leishmania tropica isolates isolated from cutaneous leishmaniasis (CL) patients in Türkiye. Leishmania strains stored in liquid nitrogen tank by cryopreservation in Manisa Celal Bayar University Faculty of Medicine Parasite Bank were resuscitated under suitable conditions and cultivated in NNN and RPMI-1640 media. Then, the isolates were allowed to enter the logarithmic phase in a 26ºC incubator and DNA isolations were made using the "High Pure PCR Template Preparation Kit". Real-time polymerase chain reaction (Rt-PCR) melting analyzes were applied to the DNAs obtained by using primers and probes specific to the internal transcribed spacer-1 (ITS-1) gene region of Leishmania. After RNA isolation from promastigote suspension, cDNA synthesis was performed by reverse transcription. After gel electrophoresis with PCR amplification products, dsRNA band formation was evaluated in terms of LRV2 positivity under ultraviolet light. Among the 20 examined Leishmania spp. isolates (10 L.tropica and 10 L.major), four (three L.tropica, one L.major) were found to be positive for the presence of LRV2. Although the mechanism of LRV in recent studies has not been fully understood, it is known that it exacerbates the clinic of the disease and even has an effect on the formation of drug resistance by the parasite. It is important to obtain data on the presence of LRV in our country and to contribute to various clinical, drug development, prevalence studies, diagnosis and treatment of the disease in the future.


Portable fluorescence-based microRNA detection system based on isothermal signal amplification technology.
MicroRNAs (miRNAs) diagnostics can be useful for diagnosing or confirming miRNA abundance and are used in screening tests and to assess changes in miRNAs in vivo. At present, the use of traditional nucleic acid amplification assays to detect miRNAs has been limited in laboratory environment because of the time, equipment, and technical expertise required to perform these assays. A specialized, rapid affordable miRNA detection system is necessary when there are limited resources or point-of-care testing needs. We designed a portable and affordable fluorescence-based miRNA detection system based on isothermal signal amplification technology, using SYBR Green II as a fluorescent dye. To reduce costs, we chose LED as a light source and designed the corresponding optical path for LED. The portable detection system shows results consistent with those by real-time PCR, and can be used to detect miR-183 with a limit of detection of approximately 2 fmol. We used the system to detect miR-183 in tissues and blood from patients with hepatocellular carcinoma (HCC). The results from the portable detection device were compared with those from clinical trials and indicated that the miR-183 fluorescence signal could successfully identify HCC and provide information related to cancer progression.


[Professional confidentiality and the best interests of the child].
Sharing secret information raises many questions. In child protection in particular, the obligation to remain silent and conditions of information sharing are associated with the concept of the best interests of the child. The latter is based on a set of criteria of which case-by-case evaluation remains the responsibility of professionals and requires a framework for decision-making procedures, which must be collegial.


Life Interrupted: Family Routines Buffer Stress during the COVID-19 Pandemic.
Adoption of certain behavioral and social routines that organize and structure the home environment may help families navigate the challenges presented by the COVID-19 pandemic. The current cross-sectional study aimed to assess family routines prior to and during the COVID-19 pandemic and examine associations with individual and family well-being. Using a national sample, 300 caregivers of children ages 6-18 were surveyed using Amazon Mechanical Turk platform during the first three months of COVID-19 pandemic in the United States. Caregivers reported on family demographics, COVID-19-related stress, engagement in family routines (prior to and during the COVID-19 pandemic), stress mindset, self-efficacy, and family resiliency. Overall, families reported engaging in fewer routines during the COVID-19 pandemic compared to prior to the pandemic. COVID-19-related stress was highest in low-income families, families of healthcare workers, and among caregivers who had experienced the COVID-19 virus. Moreover, COVID-19-related stress was negatively related to self-efficacy, positively related to an enhancing stress mindset, and negatively related to family resilience. Engagement in family routines buffered relations between COVID-19-related stress and family resilience, such that COVID-19-related stress was not associated with lower family resilience among families that engaged in high levels of family routines. Results suggest that family routines were challenging to maintain in the context of the COVID-19 pandemic, but were associated with better individual and family well-being during this period of acute health, economic, and social stress.


HNK1 and Sox10 are present during repair of mandibular bone defects.
Neural crest cells possess characteristics of stem cells including plasticity and ability to differentiate into various cell types. HNK1 and Sox10 are markers of neural crest cell progenitors that have been demonstrated in osteoblasts during osteogenesis of the maxilla and mandible. We investigated the presence of Sox10 and HNK1 during regeneration of mandibular bone defects. Defects were created in mandibles of rats. Samples of these defects were collected at 7, 14 and 28 days post-surgery; bone regeneration was observed during this period. Immunohistochemical analysis revealed expression of HNK1 and Sox10 in osteoblasts, osteocytes and osteogenic cells, whereas osteoclasts were unstained. HNK1 expression was increased in osteoblasts and osteocytes over time and SOX10 expression was found in osteoblasts and osteogenic cells at 7, 14 and 28 days post-surgery. HNK1 and Sox10 are present in osteoblasts, osteocytes and osteogenic cells during mandible bone regeneration.


Lymph Node Metastasis and Extrathyroidal Extension in Papillary Thyroid Microcarcinoma in Cyprus: Suspicious Subcentimeter Nodules Should Undergo FNA When Multifocality is Suspected.
To determine the prevalence of lymph node (LN) metastasis and extrathyroidal extension (ETE) in patients with papillary thyroid microcarcinoma (PTMC) in Cyprus and to evaluate the role of preoperative ultrasound (U/S) examination. A retrospective study of 102 patients who underwent thyroidectomy for PTMC in a 2-year period. Preoperatively, all patients had a thyroid and neck U/S examination with LN mapping. Tumor size according to the largest diameter, number of foci, LN metastasis, and ETE data was collected from the histopathological report and was compared to the preoperative U/S reports. LN metastasis was present in 23.5% of patients. 15.7% had central, 3.9% had lateral, and 3.9% had both central and lateral LN metastasis. ETE was present in 27.5% of patients. 21.6% had multifocal disease, and in this group, 40.9% had LN metastasis and 36.4% had ETE. Multifocality (p = 0.03), size of tumor (p = 0.05), and ETE (p ≤ 0.001) were significantly associated with LN metastasis. The prevalence of LN metastasis in multifocal PTMC ≤5 mm was the same with multifocal PTMC >5 mm. The preoperative U/S sensitivity for the suspicious lateral neck and central LN was 100%, and the specificity was 100%. The preoperative U/S sensitivity for nodules suspicious for ETE was 53.6%, and the specificity was 100%. The presence of LN metastasis and ETE in our PTMC patients in Cyprus is frequent. Neck U/S mapping is a highly reliable and accurate tool in identifying metastatic nodes. LN metastasis is associated with ETE and multifocality. Suspicious subcentimeter nodules should undergo FNA irrespective of size when multifocality is suspected.


The lived experience of visual creative expression for young adult cancer survivors.
Engaging in visual creative expression individually and in a therapeutic setting can be a beneficial experience for cancer survivors; however, most research in this field has been conducted with older adults. The current study aimed to address this gap by utilising van Manen's hermeneutic phenomenology to answer the following question: 'What is the lived experience and meaning of visual creative expression for young adult cancer survivors?' Seven young adults, diagnosed with cancer between the ages of 18 and 35, were interviewed about creative expression experiences, which they engaged in individually and/or in a therapeutic setting. Data analysis included a thematic reflection, guided existential reflection, and a process of writing and rewriting. Two superordinate themes were identified: increased self-understanding and a healing experience. Seven subthemes were also identified and included the following: being in the flow, allowing the body to express itself, renegotiating control, changing one's environment, being seen, respect for art as a separate entity and giving back. Findings suggest that visual creative expression can be a meaningful experience for young adult cancer survivors, and that this experience espouses both similarities and differences from experiences of older adult survivors. Recommendations are made for future research, in addition to implications for practitioners.


Association between mannose-binding lectin deficiency and septic shock following acute pyelonephritis due to Escherichia coli.
Structural and promoter MBL2 gene polymorphisms responsible for low MBL levels are associated with increased risk of infection. The objective of this study was to assess the possible association between polymorphisms of the MBL2 gene and the incidence of septic shock and bacteremia in patients with acute pyelonephritis due to Escherichia coli. The study included 62 female patients with acute pyelonephritis due to E. coli who required hospital admission, as well as 133 healthy control subjects. Six single-nucleotide polymorphisms (-550 G/C, -221 C/G, +4 C/T, codon 52 CGT/TGT, codon 54 GGC/GAC, and codon 57 GGA/GAA) in the MBL2 gene were genotyped by using a sequence-based typing technique. No significant differences were observed in the frequencies for low-expression MBL2 genotypes (O/O and LXA/O) between patients with acute pyelonephritis and healthy controls. Patients with acute pyelonephritis and septic shock had a higher incidence of low-expression MBL2 genotypes than patients with acute pyelonephritis without septic shock (odds ratio = 9.019, 95% confidence interval = 1.23 to 65.93; P = 0.03). No association was found between bacteremic acute pyelonephritis and low-expression MBL2 genotypes. We found that low-expression MBL2 genotypes predispose to septic shock but not to bacteremia in patients with E. coli-induced acute pyelonephritis. Determination of MBL2 polymorphisms could be useful for assessing the risk of septic shock in women undergoing acute pyelonephritis.


An immunohistochemical study of temperature-related changes in galanin and nitric oxide synthase immunoreactivity in the hypothalamus of the toad.
Galanin (GAL) and nitric oxide synthase (NOS) have been implicated in the control of thermogenesis in mammals. An experimental protocol was designed to determine whether or not the expression of these molecules in the hypothalamus of the toad could be related to environmental temperature changes. Exposure of the animals to low temperature increased the number and intensity of NOS-positive neurons in the magnocellular hypothalamic region, in contrast to a weak immunoreactivity observed in control animals kept in a natural environment at a spring-summer temperature (23-27 degrees C). Also a significantly higher number of GAL-immunoreactive (-IR) cells was observed in the preoptic area as compared to that observed in controls, while no difference in the intensity of GAL immunostaining intensity was detected. These results show a temperature-related expression of GAL and NOS in the hypothalamus and preoptic area of the toad. The results suggest a possible role of GAL and NOS in the regulation of hibernation in these animals.


Internal hernia of the stomach through the lateral space of the colostomy: a case report.
An 81-year-old woman had undergone laparoscopic abdominoperineal resection for rectal cancer. A permanent colostomy was created through an intraperitoneal route. Three months after the surgery, the patient presented with lower abdominal pain and vomiting. Computed tomography showed gastric incarceration through the lateral space of the lifted sigmoid colostomy. Although the herniated stomach was reduced by nasogastric tube decompression, the patient experienced a recurrence of gastric hernia shortly thereafter. A laparoscopic operation was performed, and a new colostomy was constructed through an extraperitoneal route. The patient had no hernia recurrence during the 20 months of follow-up after the operation. Gastric internal hernia associated with colostomy can occur as a rare complication. Although reduction of the incarcerated stomach is possible by nasogastric tube decompression, surgical repair of the hernia may be the optimal management to prevent recurrence.


Effect of volunteering on risk factors for cardiovascular disease in adolescents: a randomized controlled trial.
The idea that individuals who help others incur health benefits themselves suggests a novel approach to improving health while simultaneously promoting greater civic orientation in our society. The present study is the first experimental trial, to our knowledge, of whether regular volunteering can reduce cardiovascular risk factors in adolescents. To test a novel intervention that assigned adolescents to volunteer with elementary school-aged children as a means of improving adolescents' cardiovascular risk profiles. Randomized controlled trial, with measurements taken at baseline and 4 months later (postintervention). Urban public high school in western Canada. One hundred six 10th-grade high school students who were fluent in English and free of chronic illnesses. Weekly volunteering with elementary school-aged children for 2 months vs wait-list control group. Cardiovascular risk markers of C-reactive protein level, interleukin 6 level, total cholesterol level, and body mass index. No statistically significant group differences were found at baseline. Postintervention, adolescents in the intervention group showed significantly lower interleukin 6 levels (log10 mean difference, 0.13; 95% CI, 0.004 to 0.251), cholesterol levels (log10 mean difference, 0.03; 95% CI, 0.003 to 0.059), and body mass index (mean difference, 0.39; 95% CI, 0.07 to 0.71) compared with adolescents in the control group. Effects for C-reactive protein level were marginal (log10 mean difference, 0.13; 95% CI, -0.011 to 0.275). Preliminary analyses within the intervention group suggest that those who increased the most in empathy and altruistic behaviors, and who decreased the most in negative mood, also showed the greatest decreases in cardiovascular risk over time. Adolescents who volunteer to help others also benefit themselves, suggesting a novel way to improve health. TRIAL REGISTRATION clinicaltrials.gov Identifier: NCT01698034.


A patchy continuum? Stream processes show varied responses to patch- and continuum-based analyses.
Many conceptual syntheses in ecology and evolution are undergirded by either a patch- or continuum-based model. Examples include gradualism and punctuated equilibrium in evolution, and edge effects and the theory of island biogeography in ecology. In this study, we sought to determine how patch- or continuum-based analyses could explain variation in concentrations of stream macronutrients and system metabolism, represented by measures of productivity and respiration rates, at the watershed scale across the Kanawha River Basin, USA. Using Strahler stream order (SSO; continuum) and functional process zone (FPZ; patch) as factors, we produced statistical models for each variable and compared model performance using likelihood ratio tests. Only one nutrient (i.e., PO43- ) responded better to patch-based analysis. Both models were significantly better than a null model for ecosystem respiration; however, neither outperformed the other. Importantly, in most cases, a combination model, including both SSO and FPZ, best described observed variation in the system. Our findings suggest that several patch- and continuum-based processes may simultaneously influence the concentration of macronutrients and system metabolism. Nutrient spiral- ing along a continuum and the patch mosaic of land cover may both alter macronutrients, for example. Similarly, increases in temperature and discharge associated with increasing SSO, as well as the differences in light availability and channel morphology associated with different FPZs, may influence system metabolism. For these reasons, we recommend a combination of patch- and continuum-based analyses when modeling, analyzing, and interpreting patterns in stream ecosystem parameters.


Sample matrix effects on measured carbon and oxygen isotope ratios during continuous-flow isotope-ratio mass spectrometry.
Continuous-flow isotope-ratio mass spectrometry (CF-IRMS) is frequently used to analyze CO2 found in media such as air, breath, and soil pore space gas with the aid of a sample preparation and transfer device such as a Gasbench II. This study investigated the effect that matrices other than helium (He) have on the measured δ(13)C and δ(18)O isotope ratios of CO2. Identical CO2 was added to sample vials with matrices of pure He, pure N2, or a 21:79 mixture of O2/N2 and analyzed by a ThermoFinnigan Delta(Plus) XP isotope-ratio mass spectrometer coupled to a ThermoFinnigan Gasbench II. Variables such as CO2 concentration, sample analysis sequence, and sample matrix removal ('blanking') through manipulation of an injection and dilution open split were tested to identify systematic isotope ratio offsets between the different matrix types. The process of blanking induced a δ(13)C and δ(18)O offset of ≤0.2‰ between otherwise identical populations of CO2 samples in He. The (13)C/(12)C and (18)O/(16)O isotope ratios of CO2 sampled from pure N2 or a mixture of O2/N2 were found to be within 0.1 to 0.2‰ of those of an identical CO2 sampled from a He matrix when N2 or O2/N2 was removed prior to transport to the mass spectrometer. The measured oxygen isotope ratios of CO2 sampled from N2 and O2/N2 varied by as much as 0.6‰ and 4‰, respectively, if matrix gas was not removed prior to ionization. Sampling CO2 from matrices similar to air does not significantly affect the measured (13)C/(12)C and (18)O/(16)O isotope ratios of CO2 when a gas-handling procedure that includes the removal of matrix gas is utilized. This procedure is much preferable to introducing matrix gas and potentially isobaric interference to the ion source.


Human CST Prefers G-Rich but Not Necessarily Telomeric Sequences.
The human CST (CTC1-STN1-TEN1) heterotrimeric complex plays roles in both telomere maintenance and DNA replication through its ability to interact with single-stranded DNA (ssDNA) of a variety of sequences. The precise sequence specificity required to execute these functions is unknown. Telomere-binding proteins have been shown to specifically recognize key telomeric sequence motifs within ssDNA while accommodating nonspecifically recognized sequences through conformationally plastic interfaces. To better understand the role CST plays in these processes, we have produced a highly purified heterotrimer and elucidated the sequence requirements for CST recognition of ssDNA in vitro. CST discriminates against random sequence and binds a minimal ssDNA comprised of three repeats of telomeric sequence. Replacement of individual nucleotides with their complement reveals that guanines are specifically recognized in a largely additive fashion and that specificity is distributed uniformly throughout the ligand. Unexpectedly, adenosines are also well tolerated at these sites, but cytosines are disfavored. Furthermore, sequences unrelated to the telomere repeat, yet still G-rich, bind CST well. Thus, CST is not inherently telomere-specific, but rather is a G-rich sequence binder. This biochemical activity is reminiscent of the yeast t-RPA and Tetrahymena thermophila CST complexes and is consistent with roles at G-rich sites throughout the genome.


Clinical Peptidomics: Advances in Instrumentation, Analyses, and Applications.
Extensive effort has been devoted to the discovery, development, and validation of biomarkers for early disease diagnosis and prognosis as well as rapid evaluation of the response to therapeutic interventions. Genomic and transcriptomic profiling are well-established means to identify disease-associated biomarkers. However, analysis of disease-associated peptidomes can also identify novel peptide biomarkers or signatures that provide sensitive and specific diagnostic and prognostic information for specific malignant, chronic, and infectious diseases. Growing evidence also suggests that peptidomic changes in liquid biopsies may more effectively detect changes in disease pathophysiology than other molecular methods. Knowledge gained from peptide-based diagnostic, therapeutic, and imaging approaches has led to promising new theranostic applications that can increase their bioavailability in target tissues at reduced doses to decrease side effects and improve treatment responses. However, despite major advances, multiple factors can still affect the utility of peptidomic data. This review summarizes several remaining challenges that affect peptide biomarker discovery and their use as diagnostics, with a focus on technological advances that can improve the detection, identification, and monitoring of peptide biomarkers for personalized medicine.


Biology of colon cancer: an overview.
The altered growth characteristics of neoplastic cells have recently been associated with changes in membrane glycoproteins present on the cell surface. Since the carbohydrate moieties of surface membrane glycoproteins are asymmetrically located on the external cell surface, these glycoconjugates are likely candidates for providing cell surfaces with many of their biological properties. Using specific external cell surface labeling techniques, we have broadened our investigation of tumor cell surface glycoconjugates to include studies on cultured human epithelial cells from fetal intestine and from colonic carcinoma. We have isolated by affinity chromatography and gel filtration and integral membrane glycoprotein, termed Galactoprotein I, from a cultured human colonic adenocarcinoma cell line, which appears to be identical in many respects to CEA. Further examination of cell surface glycoproteins regarding quantitative and qualitative alterations and topographical redistribution should provide an insight into the biological aspects of tumor development.


Serum vascular endothelial growth factor concentrations and ovarian stromal blood flow are increased in women with polycystic ovaries.
The aim of this study was to determine basal serum vascular endothelial growth factor (VEGF) concentrations and Doppler blood flow changes within the ovarian stroma of women with polycystic ovaries (PCO) and women with normal ovaries. Pulsed and colour Doppler blood flows within the ovarian stroma were recorded, and serum VEGF concentrations measured, in the early follicular phase (days 2-3 of a menstrual cycle) in 60 women undergoing ovarian stimulation for in-vitro fertilization. 36 women had normal ovaries, 14 women had PCO as seen on pelvic ultrasound examination and 10 had polycystic ovarian syndrome (PCOS). Mean+/-SD serum VEGF concentrations were significantly higher (P < 0.001) in women with PCO and PCOS (3.4+/-0.7 and 3.2+/-0.66 ng/ml respectively) compared with women with normal ovaries (2.3+/-0.5 ng/ml). Mean peak systolic blood flow velocity (PSV) and time-averaged maximum flow velocity (TAMXV) were significantly higher (P < 0.001) in women with PCO and PCOS compared with women with normal ovaries. The mean PSV were 15+/-4 and 16+/-4 cm/s in women with PCO and PCOS respectively, compared with 9+/-2 cm/s in women with normal ovaries. The TAMXV were 9+/-3 and 11+/-3 cm/s in women with PCO and PCOS respectively compared with women with normal ovaries (5.8+/-1.5 cm/s). Serum VEGF concentrations were positively correlated with PSV (r=0.44, P=0.001) and TAMXV (r=0.45, P < 0.000) in all three groups of women. Higher serum concentrations of VEGF in women with PCO and PCOS may relate to the increased vascularity that underlies the increased blood flow demonstrated by Doppler blood flow velocity measurements in these women. The results may explain the higher risk of ovarian hyperstimulation syndrome in programmes of ovarian stimulation in patients with PCO compared with those with normal ovaries.


[Expression of mucin 15 in hepatocellular carcinoma and its clinical implications].
To investigate the expression of mucin 15 (MUC15) in hepatocellular carcinoma and explore its association with the prognosis of patients. The expression of MUC15 was detected by quantitative RT-PCR and Western blotting in liver cell line L02, liver cancer cell lines HepG2, MHCC-97H, and SMMC-7721, and in 122 HCC and corresponding adjacent non-tumor liver tissues. The association of MUC15 expression in HCC tissues with the clinical parameters and the patients' survival was analyzed. The 1iver cell line L02 showed significantly higher MUC15 expression level than the liver cancer cell lines HepG2, MHCC-97H, and SMMC-7721 (P<0.05). The expression level of MUC15 was markedly lower in the HCC tissues than in the adjacent non-tumor liver tissues (P<0.05). MUC15 expression in the HCC tissues was significantly correlated with the tumor TNM stage, intrahepatic or lymphatic metastasis, portal vein thrombosis and tumor differentiation (P<0.05). Kaplan-Meier survival analysis suggested that a low MUC15 expression was associated with a poor clinical prognosis of the patients. The expression of MUC15 is correlated with the clinicopathological features and prognosis of HCC patients, and may potentially serve as a novel prognostic marker for HCC.


Pulmonary atresia with intact ventricular septum: range of morphology in a population-based study.
We describe the morphologic variability in pulmonary atresia with intact ventricular septum (PAIVS) within a population-based study. An uncommon disease, PAIVS shows considerable morphologic heterogeneity. Clinical reports, based mostly on small samples of patients, may not reflect the true spectrum of pathology of this condition. We have studied the entire range of morphology in a prospective population-based study of patients over a five-year period (1991 to 1995). METHODS; As part of the United Kingdom and Ireland Collaborative Study of PAIVS, all 18 pediatric cardiac centers were visited by a single investigator. Morphologic features of each case were determined by direct review of the echocardiograms and angiocardiograms, from surgical and autopsy reports, and by review of pathology specimens where available. Among 183 live-born infants, atresia was valvar (membranous) in 74.7% and muscular in 25.3%. Muscular obliteration of the apical trabecular cavity, and in some cases its infundibulum, resulted in "bipartite" right ventricle (RV) in 33.6%, and a "unipartite" chamber in 7.7%. The remaining 58.7% had "tripartite" morphology. Coronary arterial abnormalities were identified in 45.8%, including arterial stenoses, interruptions and ectasia in 7.6%. Ebstein's malformation coexisted in 18 patients. Median tricuspid valvar size and RV inlet Z-scores were -5.2 and -5.1, respectively. This study provides unique data on the diverse pathology of PAIVS in an unselected population. This will help determine if published reports reflect the true spectrum of pathology of the condition.


Overriding imatinib resistance with a novel ABL kinase inhibitor.
Resistance to the ABL kinase inhibitor imatinib (STI571 or Gleevec) in chronic myeloid leukemia (CML) occurs through selection for tumor cells harboring BCR-ABL kinase domain point mutations that interfere with drug binding. Crystallographic studies predict that most imatinib-resistant mutants should remain sensitive to inhibitors that bind ABL with less stringent conformational requirements. BMS-354825 is an orally bioavailable ABL kinase inhibitor with two-log increased potency relative to imatinib that retains activity against 14 of 15 imatinib-resistant BCR-ABL mutants. BMS-354825 prolongs survival of mice with BCR-ABL-driven disease and inhibits proliferation of BCR-ABL-positive bone marrow progenitor cells from patients with imatinib-sensitive and imatinib-resistant CML. These data illustrate how molecular insight into kinase inhibitor resistance can guide the design of second-generation targeted therapies.


Effects of acute plasma volume expansion on renal perfusion, filtration, and oxygenation after cardiac surgery: a randomized study on crystalloid vs colloid.
In the present randomized study, we evaluated the differential effects of a colloid and a crystalloid fluid on renal oxygen delivery (RD(O2)), glomerular filtration (GFR), renal oxygen consumption ((RV(O2))), and the renal oxygen supply-demand relationship (i.e., renal oxygenation) after cardiac surgery with cardiopulmonary bypass. Thirty patients with normal preoperative renal function, undergoing uncomplicated cardiac surgery, were studied in the intensive care unit in the early postoperative period. Patients were randomized to receive a bolus dose of either a crystalloid (Ringers-acetate 20 ml kg(-1), n=15) or a colloid solution (Venofundin) 10 ml kg(-1), n=15). Systemic haemodynamics were measured via a pulmonary artery catheter. Renal blood flow and GFR were measured by the renal vein retrograde thermodilution technique and by renal extraction of 51Cr-EDTA (=filtration fraction). Arterial and renal vein blood samples were obtained for measurements of renal oxygen delivery (RD(O2)) and RV(O2). Renal oxygenation was estimated from the renal oxygen extraction. Despite an increase in cardiac index and renal blood flow with both fluids, neither of the fluids improved RD(O2), because they both induced haemodilution. The GFR increased in the crystalloid (28%) but not in the colloid group. The crystalloid increased the filtration fraction (24%) and renal oxygen extraction (23%), indicating that the increase in GFR, the major determinant of RV(O2), was not matched by a proportional increase in RD(O2). Neither the colloid nor the crystalloid improved RD(O2) when used for postoperative plasma volume expansion. The crystalloid-induced increase in GFR was associated with impaired renal oxygenation, which was not seen with the colloid. NCT01729364.