Effects of antiviral therapy for patients with inflammatory bowel disease and a positive intestinal biopsy for cytomegalovirus.
Cytomegalovirus (CMV) is an opportunistic pathogen; documented tissue involvement of patients with inflammatory bowel disease (IBD) is associated with adverse outcomes. We quantified the density of CMV inclusions in biopsy specimens from patients with IBD and assessed their response to antiviral therapy. In a case-control study, we identified all small bowel and colon biopsy specimens collected from 1111 patients with IBD that had been submitted to the Department of Laboratory Medicine and Pathology, Mayo Clinic, to evaluate for CMV in intestinal tissue from 2005 through 2011. All positive cases were reviewed to confirm the diagnosis of CMV in tissue. We determined the number of viral inclusions in each processed biopsy sample. Biopsy specimens with 5 or more inclusions were considered to have high-grade CMV density. We collected data on response to antiviral therapy and history of surgical resection within 1 year after diagnosis of CMV in tissue. CMV-negative samples (controls) were selected from the same IBD population. Primary outcomes included clinical improvement, hospital admission, time to admission, need for surgical procedures, time to surgery, escalation of therapy, and relapse of CMV infection. In our analysis of the biopsy samples, 68 (6%) were found to contain CMV. Follow-up data and treatment outcomes were available from 50 cases, including 16 patients with high-grade CMV density (all treated) and 34 with low-grade CMV density (20 treated). There was no overall difference in survival, free of surgery, between patients with or without CMV 1 year after diagnosis in tissue. Antiviral treatment improved surgery-free survival outcomes of patients with CMV infection—particularly of patients with high-grade CMV density. Patients with IBD and a high density of CMV inclusions in intestinal biopsy specimens benefit from antiviral therapy. Patients with fewer viral inclusions in biopsy specimens also might benefit, but the severity of the IBD should be the prime consideration in determining treatment strategies.


Diversity of short linear interaction motifs in SARS-CoV-2 nucleocapsid protein.
Short linear motifs (SLiMs) are 3-10 amino acid long binding motifs in intrinsically disordered protein regions (IDRs) that serve as ubiquitous protein-protein interaction modules in eukaryotic cells. Through molecular mimicry, viruses hijack these sequence motifs to control host cellular processes. It is thought that the small size of SLiMs and the high mutation frequencies of viral IDRs allow rapid host adaptation. However, a salient characteristic of RNA viruses, due to high replication errors, is their obligate existence as mutant swarms. Taking advantage of the uniquely large genomic database of SARS-CoV-2, here, we analyze the role of sequence diversity in the presentation of SLiMs, focusing on the highly abundant, multi-functional nucleocapsid protein. We find that motif mimicry is a highly dynamic process that produces an abundance of motifs transiently present in subsets of mutant species. This diversity allows the virus to efficiently explore eukaryotic motifs and evolve the host-virus interface.


Loci-specific differences in blood DNA methylation in HBV-negative populations at risk for hepatocellular carcinoma development.
Late onset of clinical symptoms in hepatocellular carcinoma (HCC) results in late diagnosis and poor disease outcome. Approximately 85% of individuals with HCC have underlying liver cirrhosis. However, not all cirrhotic patients develop cancer. Reliable tools that would distinguish cirrhotic patients who will develop cancer from those who will not are urgently needed. We used the Illumina HumanMethylation450 BeadChip microarray to test whether white blood cell DNA, an easily accessible source of DNA, exhibits site-specific changes in DNA methylation in blood of diagnosed HCC patients (post-diagnostic, 24 cases, 24 controls) and in prospectively collected blood specimens of HCC patients who were cancer-free at blood collection (pre-diagnostic, 21 cases, 21 controls). Out of 22 differentially methylated loci selected for validation by pyrosequencing, 19 loci with neighbouring CpG sites (probes) were confirmed in the pre-diagnostic study group and subjected to verification in a prospective cirrhotic cohort (13 cases, 23 controls). We established for the first time 9 probes that could distinguish HBV-negative cirrhotic patients who subsequently developed HCC from those who stayed cancer-free. These probes were identified within regulatory regions of BARD1, MAGEB3, BRUNOL5, FXYD6, TET1, TSPAN5, DPPA5, KIAA1210, and LSP1. Methylation levels within DPPA5, KIAA1210, and LSP1 were higher in prospective samples from HCC cases vs. cirrhotic controls. The remaining probes were hypomethylated in cases compared with controls. Using blood as a minimally invasive material and pyrosequencing as a straightforward quantitative method, the established probes have potential to be developed into a routine clinical test after validation in larger cohorts.


Rate distortion optimal bit allocation methods for volumetric data using JPEG 2000.
Computer modeling programs that generate three-dimensional (3-D) data on fine grids are capable of generating very large amounts of information. These data sets, as well as 3-D sensor/measured data sets, are prime candidates for the application of data compression algorithms. A very flexible and powerful compression algorithm for imagery data is the newly released JPEG 2000 standard. JPEG 2000 also has the capability to compress volumetric data, as described in Part 2 of the standard, by treating the 3-D data as separate slices. As a decoder standard, JPEG 2000 does not describe any specific method to allocate bits among the separate slices. This paper proposes two new bit allocation algorithms for accomplishing this task. The first procedure is rate distortion optimal (for mean squared error), and is conceptually similar to postcompression rate distortion optimization used for coding codeblocks within JPEG 2000. The disadvantage of this approach is its high computational complexity. The second bit allocation algorithm, here called the mixed model (MM) approach, mathematically models each slice's rate distortion curve using two distinct regions to get more accurate modeling at low bit rates. These two bit allocation algorithms are applied to a 3-D Meteorological data set. Test results show that the MM approach gives distortion results that are nearly identical to the optimal approach, while significantly reducing computational complexity.


When ESKD complicates cancer screening and cancer treatment.
End-stage kidney disease (ESKD) affects the recommended screening, incidence, treatment, and mortality of cancer. Cancer occurring in a patient with ESKD can influence candidacy for kidney transplantation as well as dialysis decision-making and cancer treatment. Certain cancers are more common among ESKD patients, notably, viral-mediated cancers that are associated with human papilloma or hepatitis viruses, and urothelial cancers associated with analgesic and Balkan nephropathies. Solid tumors are not believed to occur more frequently in ESKD patients. The presence of ESKD may confer a higher risk of post-surgical complications as well as mortality. The cost-effectiveness of cancer screening depends upon individual cancer risk and estimated overall survival. The high mortality associated with ESKD argues against routine cancer screening in dialysis patients. Cancer treatment in ESKD may be complicated by the need to avoid, adjust doses of and/or coordinate the timing of administration of imaging contrast, chemotherapy, and immunotherapy with dialysis treatments. There is a general dearth of information on the treatment of cancer in ESKD patients. These issues will be discussed, and some general guidelines presented based upon the current literature.


Genetics of acute rejection after kidney transplantation.
Treatment of acute rejection (AR) following kidney transplantation has improved in recent years, but there are still limitations to successful outcomes. This review article covers literature in regard to recipient and donor genetics of AR kidney and secondarily of liver allografts. Many candidate gene and some genome-wide association studies (GWASs) have been conducted for AR in kidney transplantation. Genetic associations with AR in kidney and liver are mostly weak, and in most cases, the associations have not been reproducible. A limitation in the study of AR is the lack of sufficiently large populations that account for population stratification to study the AR phenotype which in this era occurs in <10% of transplants. Furthermore, the AR phenotype has been difficult to define and the definitions of classifications have evolved over time. Literature related to the pharmacogenomics of tacrolimus is robust and has been validated in many studies. Associations between gene expression and AR are emerging as markers of outcomes and AR classification. In the future, combinations of pretransplant genotype for AR risk prediction, genotype-based immune suppressant dosing, and pharmacogenomic markers to select AR maintenance or treatment and expression markers from biopsies may provide valuable clinical tools for guiding treatment.


Estimating an EQ-5D-5L Value Set for China.
To estimate a five-level EuroQol five-dimensional questionnaire (EQ-5D-5L) value set for China using the health preferences of residents living in the urban areas of the country. The values of a subset of the EQ-5D-5L-defined health states (n = 86) were elicited using the time trade-off (TTO) technique from a sample of urban residents (n = 1271) recruited from five Chinese cities. In computer-assisted personal interviews, participants each completed 10 TTO tasks. Two additive and two multiplicative regression models were evaluated for their performance in describing the relationship between TTO values and health state characteristics using a cross-validation approach. Final values were generated using the best-performed model and a rescaling method. The 8- and 9-parameter multiplicative models unanimously outperformed the 20-parameter additive model using a random or fixed intercept in predicting values for out-of-sample health states in the cross-validation analysis and their coefficients were estimated with lower standard errors. The prediction accuracies of the two multiplicative models measured by the mean absolute error and the intraclass correlation coefficient were very similar, thus favoring the more parsimonious model. The 8-parameter multiplicative model performed the best in the study and therefore was used to generate the EQ-5D-5L value set for China. We recommend using rescaled values whereby 1 represents the value of instrument-defined full health in economic evaluation of health technologies in China whenever the EQ-5D-5L data are available.


Simulated microgravity enhances cerebral artery vasoconstriction and vascular resistance through endothelial nitric oxide mechanism.
Elevations in arterial pressure associated with hypertension, microgravity, and prolonged bed rest alter cerebrovascular autoregulation in humans. Using head-down tail suspension (HDT) in rats to induce cephalic fluid shifts and elevate arterial pressure, this study tested the hypothesis that 2-wk HDT enhances cerebral artery vasoconstriction and that an enhanced vasoconstriction described in vitro will alter regional cerebral blood flow (CBF) and vascular resistance (CVR) during standing and head-up tilt. To test this hypothesis, basal tone and vasoconstrictor responses to increases in transmural pressure, shear stress, and K(+) were determined in vitro in middle cerebral arteries (MCAs) from HDT and control rats. All in vitro measurements were done in the presence and absence of the nitric oxide synthase (NOS) inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME; 10(-5) M) and with endothelium removal. Endothelial NOS (eNOS) mRNA and protein expression levels were measured by RT-PCR and immunoblot, respectively. Regional CBF and CVR were determined with a radiolabeled tracer technique and quantitative autoradiography. Basal tone and all vasoconstrictor responses were greater in MCAs from HDT rats. L-NAME and endothelium removal abolished these differences between groups, and HDT was associated with lower levels of MCA eNOS protein. CBF in select regions was lower and CVR higher during standing and head-up tilt in HDT rats. These results indicate that chronic cephalic fluid shifts enhanced basal tone and vasoconstriction through alterations in the eNOS signaling mechanism. The functional consequence of these vascular alterations with HDT is regional elevations in CVR and corresponding reductions in cerebral perfusion.


A level-set procedure for the design of electromagnetic metamaterials.
Achieving negative permittivity and negative permeability signifies a key topic of research in the design of metamaterials. This paper introduces a level-set based topology optimization method, in which the interface between the vacuum and metal phases is implicitly expressed by the zero-level contour of a higher dimensional level-set function. Following a sensitivity analysis, the optimization maximizes the objective based on the normal direction of the level-set function and induced current flow, thereby generating the desirable patterns of current flow on metal surface. As a benchmark example, the U-shaped structure and its variations are obtained from the level-set topology optimization. Numerical examples demonstrate that both negative permittivity and negative permeability can be attained.


Is the newly described Vietnamese bovid Pseudonovibos spiralis a chamois (genus Rupicapra)?
The "Linh Duong" (Pseudonovibos spiralis), a newly described Vietnamese bovid, has been shown to be related to the Caprinae on the basis of the mitochondrial cytochrome b gene sequencing from horn fragments. To integrate the greatest available biodiversity we performed a new phylogenetic analysis including all genera of Caprinae. Unexpectedly, Pseudonovibos is found to be robustly associated with Rupicapra, and closer to the Alpine than to the Pyrenean chamois. Several clues led us to interpret this result as a carry-over DNA contamination of Pseudonovibos by Rupicapra.


Molecular design of melt-spinnable co-polymers as Si-B-C-N fiber precursors.
Two series of co-polymers with the general formula [B(C2H4SiCH3(NH)x(NCH3)y)3]n, i.e., composed of C2H4SiCH3(NH)x and C2H4SiCH3(NCH3)y (C2H4 = CHCH3, CH2CH2) building blocks in a well defined x : y ratio, have been synthesized by hydroboration of dichloromethylvinylsilane with borane dimethyl sulfide followed by successive reactions with lithium amide and methylamine according to controlled ratios. The role of the chemistry behind their syntheses has been studied in detail by solid-state NMR, FT-IR and elemental analyses. Then, the intimate relationship between the chemistry and the melt-spinnability of these polymers was discussed. By keeping x = 0.50 and increasing y above 0.50, i.e., obtaining methylamine excess, the co-polymers contained more ending groups and especially more tetracoordinated boron, thus allowing tuning very precisely the chemical structure of the preceramic polymer in order to meet the requirements for melt-spinning. The curing treatment under ammonia at 200 °C efficiently rendered the green fibers infusible before their subsequent pyrolysis under nitrogen at 1000 °C to generate Si-B-C-N ceramic fibers. Interestingly, it could be possible to produce also low diameter hollow fibers with relatively high mechanical properties for a further exploration as membrane materials.


Effect on comfort of administering bubble-humidified or dry oxygen: the Oxyrea non-inferiority randomized study.
The clinical interest of using bubble humidification of oxygen remains controversial. This study was designed to further explore whether delivering dry oxygen instead of bubble-moistened oxygen had an impact on discomfort of ICU patients. This randomized multicenter non-inferiority open trial included patients admitted in intensive care unit and receiving oxygen. Any patient receiving non-humidified oxygen (between 0 and 15 L/min) for less than 2 h could participate in the study. Randomization was stratified based on the flow rate at inclusion (less or more than 4 L/min). Discomfort was assessed 6-8 and 24 h after inclusion using a dedicated 15-item scale (quoted from 0 to 150). Three hundred and fifty-four ICU patients receiving non-humidified oxygen were randomized either in the humidified (HO) (n = 172), using bubble humidifiers, or in the non-humidified (NHO) (n = 182) arms. In modified intention-to-treat analysis at H6-H8, the 15-item score was 26.6 ± 19.4 and 29.8 ± 23.4 in the HO and NHO groups, respectively. The absolute difference between scores in both groups was 3.2 [90% CI 0.0; + 6.5] for a non-inferiority margin of 5.3, meaning that the non-inferiority analysis was not conclusive. This was also true for the subgroups of patients receiving either less or more than 4 L/min of oxygen. At H24, using NHO was not inferior compared to HO in the general population and in the subgroup of patients receiving 4 L/min or less of oxygen. However, for patients receiving more than 4 L/min, a post hoc superiority analysis suggested that patients receiving dry oxygen were less comfortable. Oxygen therapy-related discomfort was low. Dry oxygen could not be demonstrated as non-inferior compared to bubble-moistened oxygen after 6-8 h of oxygen administration. At 24 h, dry oxygen was non-inferior compared to bubble-humidified oxygen for flows below 4 L/min.


CCL2 induces neural stem cell proliferation and neuronal differentiation in Niemann-Pick type C mice.
Niemann-Pick type C disease (NP-C) is a rare and ultimately fatal lysosomal storage disorder with variable neurologic symptoms. Loss of neuronal function and neuronal cell death occur in the NP-C brain, similar to the findings for other neurodegenerative diseases. Targeting of neuronal cells in the brain therefore represents a potential clinical intervention strategy to reduce the rate of disease progression and improve the quality of life. We previously reported that bone marrow stem cells show a neurogenic effect through CCL2 (also known as monocyte chemoattractant protein-1, MCP-1) secretion in the brains of NP-C mice. However, the direct effect of CCL2 on neurogenesis has not been ascertained. Here, to define neurogenic effects of CCL2 in NP-C, we applied human recombinant CCL2 to neural stem cells (NSCs) derived from NP-C mice. CCL2-treated NSCs showed significantly increased capacity for self-renewal, proliferation and neuronal differentiation. Similar results were observed in the subventricular zone of NP-C mice after CCL2 treatment. Furthermore, infusion of CCL2 into the NP-C mouse brain resulted in reduction of neuroinflammation. Taken together, our results demonstrate that CCL2 is a potential new therapeutic agent for NP-C.


Selective reactivation of Epstein-Barr virus-specific cytotoxic T cells by stimulation in vitro with allogeneic virus-transformed HLA-homozygous typing cells.
Epstein-Barr (EB) virus-specific cytotoxic T-cell preparations, produced by stimulation in vitro of peripheral blood lymphocytes with the autologous virus-transformed cell line, are HLA-A and B antigen-restricted and, with some donors, show preferential restriction through one or two of the four relevant antigens of the donor's HLA type. It has now been demonstrated that such EB virus-specific cytotoxic T cells may also be reactivated by stimulation with allogeneic virus-transformed cells provided that there is no mismatch of the HLA-A and B antigens between the responder and stimulator cell donors. In particular, virus-transformed cell lines from HLA-homozygous donors HLA-A and B antigen-matched to one of the haplotypes of an HLA-heterozygous responder were shown to reactivate selectively only those EB virus-specific cytotoxic T cells restricted through the HLA-A and B antigens present on the allogeneic stimulating cells. In addition to confirming the polyclonal nature of the HLA-restricted EB virus-specific cytotoxic T-cell response, this new experimental procedure has allowed the production, and subsequent expansion as cell lines dependent upon T-cell growth factor, of those effector cells restricted through the "nonpreferred" HLA antigens that are poorly represented in the response induced by stimulation with autologous virus-transformed cells.


Taste-Active and Nutritional Components of Thai Native Chicken Meat: A Perspective of Consumer Satisfaction.
The taste-active and nutritional components of Thai native, broilers, black-boned, and spent hen chickens were analyzed. The amounts of tasty amino acids especially glutamic acid were the highest in Thai native chicken. The black-boned chicken had the highest arginine content, related to the least amount of consumer satisfaction. Concerning nutritional quality, choline, and taurine were deemed important for brain function. The black-boned chicken showed the highest choline and taurine contents, unlike that of the spent hens. In contrast, broilers presented the highest betaine content, which might be attributed to their lipid metabolism. L-carnitine content was abundant in black-boned and Thai native chickens. Moreover, the amounts of essential amino acids were high in Thai native chicken. In conclusion, black-boned chicken proved to be an excellent nutritional source for health-conscience consumers, whereas the Thai native chickens were flavourful and delicious.


Organization of a multifunctional protein in pyrimidine biosynthesis. A domain hypersensitive to proteolysis.
When the multifunctional protein that catalyses the first three steps of pyrimidine biosynthesis in hamster cells is treated with staphylococcal V8 proteinase, a single cleavage takes place. The activities of carbamoyl-phosphate synthetase (EC 220.127.116.11), aspartate carbamoyltransferase (EC 188.8.131.52) and dihydro-orotase (EC 22.214.171.124) and the allosteric inhibition by UTP are unaffected. One fragment, of Mr 182000, has the first and third enzyme activities, whereas the other fragment, of Mr 42000, has aspartate carbamoyltransferase activity and an aggregation site. A similar small fragment is observed in protein digested with low concentrations of trypsin. A similar large fragment is seen after digestion with trypsin and as the predominating form of this protein in certain mutants defective in pyrimidine biosynthesis. These results indicate that a region located adjacent to the aspartate carbamoyltransferase domain is hypersensitive to proteinase action in vitro and may also be sensitive to proteolysis in vivo.


Illumina midi-barcodes: quality proof and applications.
DNA barcoding constitutes a supplemental genetically based characterization tool for the identification of species. Traditionally, the barcodes are generated with a length of 650 bp using standardized Sanger sequencing, but with the introduction of high-throughput sequencing (HTS) methods new opportunities for sequencing are available. To use HTS for barcode collection and identification, the amplification of shorter fragments is preferred. Reference DNA midi-barcodes of wild bees were produced using the Illumina MiSeq as well as the Sanger method. Although DNA midi-barcodes derived from Illumina were comparatively shorter (418 bp), their sequences were coherent to the morphological assignment of species. The Illumina barcodes proved to be effective and dealt better with some general limitations of DNA barcoding.


Differences in the characteristics of families with BRCA1 and BRCA2 mutations in Israel.
Three specific mutations in the BRCA1 (185delAG, 5382insC) and BRCA2 (6174delT) genes have been reported to be of high prevalence in the Jewish Ashkenazi population. We studied the differences in phenotype of families carrying these mutations. All consecutive families found by the CHS Familial Cancer Service to carry one of the three 'Jewish' mutations of the BRCA1/BRCA2 genes were evaluated for phenotypic characteristics. Chi-squared and Student's t-test statistics were employed to study differences in a variety of clinical and demographic parameters. A total of 111 families with 1499 family members were included. Among them 454 cases of cancer (297 in breast/ovary) were reported. Ovarian cancer, but not breast cancer, was detected at a significantly younger age among carriers of 185delT compared with other mutation carriers. In families with 185delAG, 5382insC and 6174delT mutations, breast cancer was found in 20.2, 39.4 and 24.1% of all identified women (born between 1900 and 1975), correspondingly. The corresponding figures for ovarian cancer were 13.9, 6.8 and 4.9%. Families carrying the 5382insC mutation had the highest probability of expressing bilateral breast cancer (38.9% of families, 15.4% of women with cancer, 6.1% of all women in family) and metachronous breast and ovary tumours (22.2, 9.8 and 4.5% correspondingly). Other tumours were reported in 7.9, 9.1 and 12.0% of women and 9.5, 12.9 and 15.8% of men in families with 185delAG, 5382insC, 6174delT, correspondingly. Marked phenotypic differences were found between families carrying different BRCA mutations warranting mutation-specific counselling to families seeking risk-reduction advice. 5382insC emerged as a most aggressive mutation.


[A female patient with a weak, painful thumb].
Here we present the case of a 64-year-old female patient with a painful thumb. This case demonstrates how, with proper physical examination, a correct diagnosis of 'skier's thumb' with concomitant Stener lesion can be made. This condition is a tear of the ulnar collateral ligament (UCL) of the metacarpophalangeal joint with an interposition of the UCL and the aponeurosis adductor pollicis muscle. It is usually treated surgically.


Bronchopulmonary Dysplasia: When the Very Preterm Baby Comes Home.
Many infants with severe bronchopulmonary dysplasia (BPD) can be safely managed with oxygen at home. This review covers criteria for home oxygen therapy, monitoring, and weaning protocols for oxygen therapy in the outpatient setting. Although most infants with BPD are weaned from oxygen within a year, they continue to have pulmonary function abnormalities into adolescence. These infants also require evaluation for pulmonary hypertension, systemic hypertension, and a strong focus on adequate nutritional needs for growth.


The influence of hatha yoga on stress, anxiety, and suppression: A randomized controlled trial.
Engaging in yoga may mitigate stress and anxiety in individuals while potentially enhancing one's capacity to manage distractions. Our research aimed to explore the relation between these two outcomes: Can an eight-week yoga program foster distraction suppression, thereby reducing stress and discomfort? To answer this question, we used Hatha Yoga, the most commonly practiced form of yoga. We tested if the intervention improved participants' ability to suppress distractions and selectively decrease self-reported stress and stress reactivity. In Addition, we investigated whether such an intervention would increase participants' mindfulness. Our study included 98 healthy yoga novices between 18 and 40 years who were randomly assigned to either an experimental or a waitlist condition, with each participant completing pre- and post-intervention assessments, including questionnaires, as well as electrophysiological and behavioral measures. After eight weeks of yoga practice, significant reductions in self-reported stress and stress reactivity levels, as well as increased mindfulness, were observed among those participating in the intervention relative to those in the waitlist control group. There were, however, no significant changes in state or trait anxiety due to the intervention. Changes in stress measures could not be explained by changes in participants' ability to suppress distractors, which was not affected by the intervention. Overall, our findings suggest that regular participation in Hatha Yoga can improve mental health outcomes without impacting cognitive functioning directly related to distractor suppression. CLINICAL TRIAL REGISTRATION NUMBER: NCT05232422.


Influence of benzene exposure, fat content, and their interactions on erythroid-related hematologic parameters in petrochemical workers: a cross-sectional study.
Ubiquitously distributed benzene is a known hematotoxin. Increasing evidence has suggested that erythroid-related hematologic parameters may be sensitive to benzene exposure. Fat content, which is also closely associated with erythroid-related hematologic parameters, may affect the distribution and/or metabolism of benzene, and eventually benzene-induced toxicity. To explore the influence of benzene exposure, fat content, and their interactions on erythroid-related hematologic parameters, we recruited 1669 petrochemical workers and measured their urinary S-phenylmercapturic acid (SPMA) concentration and erythroid-related hematological parameters. Indices for fat content included body fat percentage (BF%), plasma total cholesterol (TC) and triglycerides (TG), and occurrence of fatty liver. The dose-response curve revealed U-shaped nonlinear relationships of SPMA with hematocrit (HCT) and mean corpuscular hemoglobin concentration (MCHC) (P-overall < 0.001, and P-nonlinear < 0.015), as well as positive linear associations and r-shaped nonlinear relationships of continuous fat content indices with erythroid-related hematological parameters (P-overall ≤0.005). We also observed modification effects of fat content on the associations between benzene exposure and erythroid-related hematological parameters, with workers of lower or higher BF% and TG more sensitive to benzene-induced elevation of MCHC (Pinteraction = 0.021) and benzene-induced decrease of HCT (Pinteraction = 0.050), respectively. We also found that some erythroid-related hematologic parameters differed between subgroups of workers with different SPMA levels and fat content combination. Our study suggested that benzene exposure, fat content, and their interactions may affect erythroid-related hematological parameters in petrochemical workers in a complex manner that are worthy of further investigation.


Suicide and Attempted Suicide in Jhenaidah District, Bangladesh, 2010-2018.
Background: Suicide and attempted suicide are a serious but under-explored public health problem in Bangladesh. Survey estimates suggest that Jhenaidah District, one of the 64 districts that make up Bangladesh, is one of the highest suicide-prone regions in Bangladesh. Relatively little is known about the magnitude of suicide attempts in the district. Aims: This article describes the incidence of suicide and suicide attempts for Jhenaidah, Bangladesh for the period 2010-2018. Method: Primary descriptive analysis was performed on routine data collected by a Bangladesh-based nongovernment organization (NGO): Societies for Voluntary Activities (SOVA). Results: A total of 22,675 suicide attempts and 3,152 suicides occurred in the district. The rate of suicide attempts was found to be 136.35/100,000 and the suicide rate was found to be 20.6/100,000 in Jhenaidah. The subdistrict Sadar had the highest incidence of suicide attempt (38.09%) and suicide (33.47%). Poisoning was the most common method of suicide attempt for both males (77.07%) and females (77%). Limitations: Many cases of suicide attempts and suicide are unreported in Bangladesh owing to stigmatization; only reported cases form part of this investigation. Conclusion: Jhenaidah has very high rates of suicide and suicide attempts that surpass the global and Bangladesh averages. Although females demonstrate higher suicide rates, male suicide rates have gradually increased over the study period. Future studies are called for to better understand the local patterns and dynamics of fatal and nonfatal suicidal behaviors. Developing a sub-district-, district-, and national-level suicide prevention strategy ought to be considered a priority.


[The analgesic and anti-stress effects of middle-molecule peptides in a normal state and in thermal burns].
Fractions of medium-weight molecular peptides (MWP) were isolated from blood of intact dogs and those with burns, which possessed the property of increasing the latency of the pain reaction in mice by 23-28% after intravenous infusion. The analgesic properties of MWP change essentially in the first 24 hours after the burn. The dose dependences of the antinociceptive effect of MWP have a marked non-linear character. The studied MWP fractions also prevent involution of the thymus induced by 9-hour immobilization stress. An MWP fraction was isolated from the blood of dogs 12 hours after a burn; its intravenous infusion led to reduction of the thymus equal to that caused by 9-hour immobilization. The relationship of adaptational and damaging effects of MWP in thermal burns in discussed.


Detailed distribution of NK012, an SN-38-incorporating micelle, in the liver and its potent antitumor effects in mice bearing liver metastases.
To clarify and compare the antitumor effects and specific biodistribution of NK012, an SN-38-incorporating polymeric micelle, in mice bearing multiple liver metastases of human colon cancer HT-29 cells with irinotecan hydrochloride (CPT-11). The maximum tolerable dose of NK012 (30 mg/kg) or CPT-11 (66.7 mg/kg) was i.v. administered three times every 4 days to mice bearing metastases to the liver colonized 7 days after the portal administration of HT-29 cells (n = 6). In vivo antitumor effects were evaluated by bioluminescence imaging and histopathologic examination. Drug biodistribution was analyzed by high-performance liquid chromatography and fluorescence microscopy (n = 3). NK012 eradicated the liver metastases and produced a significant longer survival rate than CPT-11 (P = 0.0006). High-performance liquid chromatography showed the prolonged distribution of NK012 and free SN-38 released from NK012 in the tumors, liver, and spleen for weeks after NK012 administration. On the other hand, the accumulation levels of CPT-11 and free SN-38 converted from CPT-11 rapidly decreased within 1 day after CPT-11 administration. In the liver metastases, fluorescence microscopy and immunohistochemistry showed that administered NK012 was distributed mainly adjacent to tumor vessels after 1 day. As for the normal liver, NK012 was distributed in Kupffer cells instead of hepatocytes for at least 7 days after administration. This study suggests that NK012 is strongly effective against liver metastases and does not damage the liver despite the long retention time of NK012 in Kupffer cells.


Identification of MUC5B, MUC5AC and small amounts of MUC2 mucins in cystic fibrosis airway secretions.
To investigate the genetic identities of the mucins secreted in cystic fibrosis (CF) airways, sputum was collected from five individuals. Samples were separated into gel and sol phases by high-speed centrifugation and the gel phase was extracted in 6 M guanidinium chloride. The 'insoluble' residue remaining after extraction of the gel phase was brought into solution by reduction/alkylation. Density-gradient centrifugation in CsCl revealed polydisperse distributions of sialic acid-containing mucins in the gel phase, insoluble residue and sol phase fractions and the degree of variation between the different individuals was low. Antibodies recognizing MUC5AC and MUC5B identified these mucins in each of the fractions. MUC2, however, was present only as a component of the insoluble residue from the gel which accounted for less than 4% by mass of the total mucins. MUC5B and MUC5AC from the gel phase were large oligomeric species composed of disulphide-bond linked subunits and MUC5B was present as two populations with different charge densities which are likely to correspond to MUC5B 'glycoforms'. The sol phase contained, in addition to MUC5AC and MUC5B, mainly smaller mucins which did not react with the antisera and which were probably degraded. MUC5AC appeared to be enriched in the sol, suggesting that this mucin may be more susceptible to proteolytic degradation than MUC5B. The mucins present in sputum remained broadly similar during acute exacerbation and following antibiotic treatment, although the relative amount of an acidic MUC5B glycoform was decreased during infection.


Genetic and functional analysis of full-length human immunodeficiency virus type 1 env genes derived from brain and blood of patients with AIDS.
The genetic evolution of human immunodeficiency virus type 1 (HIV-1) in the brain is distinct from that in lymphoid tissues, indicating tissue-specific compartmentalization of the virus. Few primary HIV-1 envelope glycoproteins (Envs) from uncultured brain tissues have been biologically well characterized. In this study, we analyzed 37 full-length env genes from uncultured brain biopsy and blood samples from four patients with AIDS. Phylogenetic analysis of intrapatient sequence sets showed distinct clustering of brain relative to blood env sequences. However, no brain-specific signature sequence was identified. Furthermore, there was no significant difference in the number or positions of N-linked glycosylation sites between brain and blood env sequences. The patterns of coreceptor usage were heterogeneous, with no clear distinction between brain and blood env clones. Nine Envs used CCR5 as a coreceptor, one used CXCR4, and two used both CCR5 and CXCR4 in cell-to-cell fusion assays. Eight Envs could also use CCR3, CCR8, GPR15, STRL33, Apj, and/or GPR1, but these coreceptors did not play a major role in virus entry into microglia. Recognition of epitopes by the 2F5, T30, AG10H9, F105, 17b, and C11 monoclonal antibodies varied among env clones, reflecting genetic and conformational heterogeneity. Envs from two patients contained 28 to 32 N-glycosylation sites in gp120, compared to around 25 in lab strains and well-characterized primary isolates. These results suggest that HIV-1 Envs in brain cannot be distinguished from those in blood on the basis of coreceptor usage or the number or positions of N-glycosylation sites, indicating that other properties underlie neurotropism. The study also demonstrates characteristics of primary HIV-1 Envs from uncultured tissues and implies that Env variants that are glycosylated more extensively than lab strains and well-characterized primary isolates should be considered during development of vaccines and neutralizing antibodies.


Persistent sex-by-environment effects on offspring fitness and sex-ratio adjustment in a wild bird population.
A major component of sex-allocation theory, the Trivers-Willard model (TWM), posits that sons and daughters are differentially affected by variation in the rearing environment. In many species, the amount of parental care received is expected to have differing effects on the fitness of males and females. When this occurs, the TWM predicts that selection should favour adjustment of the offspring sex ratio in relation to the expected fitness return from offspring. However, evidence for sex-by-environment effects is mixed, and little is known about the adaptive significance of producing either sex. Here, we test whether offspring sex ratios vary according to predictions of the TWM in the house wren (Troglodytes aedon, Vieillot). We also test the assumption of a sex-by-environment effect on offspring using two experiments, one in which we manipulated age differences among nestlings within broods, and another in which we held nestling age constant but manipulated brood size. As predicted, females with high investment ability overproduced sons relative to those with lower ability. Males were also overproduced early within breeding seasons. In our experiments, the body mass of sons was more strongly affected by the sibling-competitive environment and resource availability than that of daughters: males grew heavier than females when reared in good conditions but were lighter than females when in poor conditions. Parents rearing broods with 1:1 sex ratios were more productive than parents rearing broods biased more strongly towards sons or daughters, suggesting that selection favours the production of mixed-sex broods. However, differences in the condition of offspring as neonates persisted to adulthood, and their reproductive success as adults varied with the body mass of sons, but not daughters, prior to independence from parental care. Thus, selection should favour slight but predictable variations in the sex ratio in relation to the quality of offspring that parents are able to produce. Offspring sex interacts with the neonatal environment to influence offspring fitness, thus favouring sex-ratio adjustment by parents. However, increased sensitivity of males to environmental conditions, such as sibling rivalry and resource availability, reduces the fitness returns from highly male-biased broods.


Single-cell analyses demonstrate that a heme-GATA1 feedback loop regulates red cell differentiation.
Erythropoiesis is the complex, dynamic, and tightly regulated process that generates all mature red blood cells. To understand this process, we mapped the developmental trajectories of progenitors from wild-type, erythropoietin-treated, and Flvcr1-deleted mice at single-cell resolution. Importantly, we linked the quantity of each cell's surface proteins to its total transcriptome, which is a novel method. Deletion of Flvcr1 results in high levels of intracellular heme, allowing us to identify heme-regulated circuitry. Our studies demonstrate that in early erythroid cells (CD71+Ter119neg-lo), heme increases ribosomal protein transcripts, suggesting that heme, in addition to upregulating globin transcription and translation, guarantees ample ribosomes for globin synthesis. In later erythroid cells (CD71+Ter119lo-hi), heme decreases GATA1, GATA1-target gene, and mitotic spindle gene expression. These changes occur quickly. For example, in confirmatory studies using human marrow erythroid cells, ribosomal protein transcripts and proteins increase, and GATA1 transcript and protein decrease, within 15 to 30 minutes of amplifying endogenous heme synthesis with aminolevulinic acid. Because GATA1 initiates heme synthesis, GATA1 and heme together direct red cell maturation, and heme stops GATA1 synthesis, our observations reveal a GATA1-heme autoregulatory loop and implicate GATA1 and heme as the comaster regulators of the normal erythroid differentiation program. In addition, as excessive heme could amplify ribosomal protein imbalance, prematurely lower GATA1, and impede mitosis, these data may help explain the ineffective (early termination of) erythropoiesis in Diamond Blackfan anemia and del(5q) myelodysplasia, disorders with excessive heme in colony-forming unit-erythroid/proerythroblasts, explain why these anemias are macrocytic, and show why children with GATA1 mutations have DBA-like clinical phenotypes.


Heterologous booster vaccines reduce severity and mortality in COVID-19 during BA.2 and BA.4/BA.5 omicron predominance in Thailand.
The COVID-19 pandemic has evolved quickly, with variants of concern resulting in the need to offer booster vaccinations. Unfortunately, the booster uptake has been slow and vaccine response has shown to wane over time. Therefore, it's critical to evaluate the role of vaccinations on outcomes with newer sub-lineages of omicron. Utilising a Hospital Information System established in Chiang Mai, Thailand, we conducted a cohort study by linking patient-level data of laboratory-confirmed COVID-19 cases to the national immunization records, during BA.2 and BA.4/BA.5 predominance. In adjusted cox-proportional hazard models, BA.4/BA.5 was not associated with more severe COVID-19 outcomes or deaths as compared to BA.2. Risk of severe outcomes and deaths were significantly reduced with third (87% and 95%) and fourth (88% and 95%) dose vaccination, while events were not observed with a fifth dose. Across the regimens, vaccination within 14-90 days prior showed the highest level of protection. All the vaccine types used for boosting in Thailand offered similar protection against severe COVID-19. Boosters provide high level of protection against severe COVID-19 outcomes and deaths with newer omicron sub-lineages. Booster campaigns should focus on improving coverage utilising all available vaccines to ensure optimal protection.


Brucellosis complicating chronic non-infectious disorders: diagnostic and therapeutic dilemmas.
There is little information in the literature on the clinical progress of brucellosis in patients affected by other non-infectious diseases; however, the infection can often trigger an exacerbation of existing underlying conditions in certain target organs. In this report we present four cases of brucellosis complicating previous diseases, and the difficulties in relation to their diagnosis and treatment. The study involved four patients with the following disorders: polycythaemia vera, pulmonary fibrosis, cirrhosis of the liver and arthritis of the knee. Brucellosis was diagnosed by classical serological and bacteriological methods. The strains involved could be isolated only in three of the four patients: two strains were Brucella abortus biovar 1 and one was Brucella suis biovar 1. Two patients relapsed 10 and 7 months after admission, another presented chronic brucellosis and received various therapy schemes, and one died. Since the best selection of antibiotics and the optimal duration of therapy remain unknown for patients having brucellosis complicated by previous pathologies, these remain at the discretion of the attending physician. Management of our patients was controversial in terms of the selection of antibiotics, duration of treatment and decision regarding surgery.


Evaluation of two types of 'medically significant error limits' and two quality control procedures on a multichannel analyzer.
Analytical error limits based on two different concepts of "medical significance" are derived. The ability of the system to detect medically significant errors using standard (1(3s] and enhanced (mean and range) quality control procedures is investigated. Error detection is found to be adequate for most analytes when error limits are based on physicians' opinions and a mean and range quality control system is used. Error detection is unsatisfactory when error limits based on intraindividual biological variation are used and this cannot be improved by using better quality control systems. The limiting factor is analytical precision. The use of error limits based on biological variation as proposed by major clinical chemistry professional bodies cannot be achieved using most current types of analytical technology.


Fatal COVID-19 in a neonate after probable late intrauterine transmission.
We report the case of a 35-week gestation infant girl born by emergent cesarean section for fetal distress in a woman with recent coronavirus disease 2019 (COVID-19). Tests for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) using polymerase chain reaction (PCR) on the infant at 24 and 48 hours of life were negative. However, at 72 hours of life, the infant's respiratory status worsened, and a repeat SARS-CoV-2 PCR was positive. The infant developed leukopenia, thrombocytopenia, and progressive respiratory failure, and died on the ninth day of life. Pathologic examination of the placenta revealed findings consistent with COVID-19 placentitis, and SARS-CoV-2 RNA staining was positive, suggesting intrauterine transmission of the infection.


Detection of hepatitis C virus by PCR in second-generation enzyme immunoassay-seropositive blood donors by using matched pairs of fresh frozen plasma and pilot tube sera.
Between April 1993 and March 1995, 429 of 334,454 (0.13%) blood donations at the Toronto Centre of the Canadian Red Cross were reactive for hepatitis C virus (HCV) by second-generation enzyme immunoassay (EIA-2). Of the 429 EIA-2-positive donations, 189 (44%), 138 (32%), and 102 (24%) were positive, indeterminate, and negative by Second-Generation Recombinant Immunoblot Assay (RIBA-2). To assess HCV viremia and minimize the risk that specimen handling affected PCR-based detection, the qualitative AMPLICOR HCV test was performed on both pilot tube sera (PTS) and the corresponding fresh frozen plasma (FFP) from 294 EIA-2-reactive donations. AMPLICOR PCR results for PTS and FFP were 100% concordant and were confirmed by nested HCV PCR for 27 of 294 donations. The AMPLICOR HCV test was positive for 127 of 140 (91%) of RIBA-2-positive donations (81, 91, and 96% of donations with two, three, and four reactive bands, respectively), 5 of 88 (5.7%) indeterminate donations, and 0 of 66 (0%) RIBA-2-negative donations. The Third-Generation Recombinant Immunoblot Assay (RIBA-3) was performed on RIBA-2-negative, -indeterminate, and -positive, PCR-negative donations. RIBA-3 demonstrated enhanced specificity and resolved 18 of 88 (20%) of RIBA-2-indeterminate samples as HCV antibody positive. The study demonstrates that PTS are as suitable as FFP for PCR-based detection of HCV and can be used to determine if EIA-2-reactive blood donors are viremic at the time of donation.


Genotypic approaches to the diagnosis of bacterial infections: plasmid analyses and gene probes.
Practical genetic approaches have been helpful in the diagnosis, epidemiology, and taxonomy of bacterial pathogens encountered in our laboratory at the Centers for Disease Control. There are many examples in which plasmid profiles have been used to define epidemic strains of enteric bacteria, staphylococci, pseudomonads, vibrios, and other pathogenic bacteria. Current methodologies should allow the microbiology laboratory to use plasmid profiles routinely and to identify plasmids associated with bacterial pathogenesis. Simplified DNA-DNA hybridization procedures have been used in our laboratory to survey or "probe' thousands of Escherichia coli colonies for the presence of enterotoxin genes, eliminating traditional tissue culture or animal assays. Research scientists continue to develop gene probes for a number of bacterial toxins and hemolysins and for the identification of pathogens such as legionellae and salmonellae. These and other probes as well as hybridization "kits' may be commercially available to diagnostic laboratories within the next few years.


Cannabinoid-based drugs as anti-inflammatory therapeutics.
In the nineteenth century, marijuana was prescribed by physicians for maladies ranging from eating disorders to rabies. However, as newer, more effective drugs were discovered and as the potential for abuse of marijuana was recognized, its use as a therapeutic became restricted, and only recently has its therapeutic potential been re-evaluated. Recent studies in animal models and in humans have produced promising results for the treatment of various disorders - such as obesity, cancer, and spasticity and tremor due to neuropathology - with drugs based on marijuana-derived cannabinoids. Moreover, as I discuss here, a wealth of information also indicates that these drugs have immunosuppressive and anti-inflammatory properties; therefore, on the basis of this mode of action, the therapeutic usefulness of these drugs in chronic inflammatory diseases is now being reassessed.


Effects of carbon dioxide and nitrogen on adhesive growth and expressions of E-cadherin and VEGF of human colon cancer cell CCL-228.
To study the effects of carbon dioxide on the metastatic capability of cancer cells, and to compare them with that of nitrogen. The colon cancer cell CCL-228 was treated with 100 % carbon dioxide or nitrogen at different time points and then cultured under normal condition. Twelve hours after the treatment, the survival rates of suspension cells and the expressions of e-cadherin and VEGF were examined. After 60 min of carbon dioxide and longer time of nitrogen treatment, the suspended cells increased and the expression of e-cadherin decreased while the expression of VEGF was enhanced significantly. And the effects of nitrogen were similar to, but weaker than, those of carbon dioxide. Carbon dioxide may improve the metastatic capability of cancer cells and its effects are significantly stronger than that of nitrogen. A sequential use of carbon dioxide and nitrogen in pneumoperitoneum may take the advantage of both gases.


Isolated traumatic aortic valvular insufficiency with rapid pulmonary deterioration. Report of two cases.
Blunt trauma followed by aortic valvular insufficiency occurs rarely. Valve replacement or repair has most often taken place several months or years after injury; only a few cases have been reported of acute operative intervention performed within a few days after injury. However, we herein report two cases of isolated aortic valvular trauma in young men, in whom fulminant pulmonary edema ensued so rapidly that urgent aortic valve replacement was necessitated within hours after injury.


Modified denatured lysozyme effectively solubilizes fullerene c60 nanoparticles in water.
Fullerenes, allotropic forms of carbon, have very interesting pharmacological effects and engineering applications. However, a very low solubility both in organic solvents and water hinders their use. Fullerene C60, the most studied among fullerenes, can be dissolved in water only in the form of nanoparticles of variable dimensions and limited stability. Here the effect on the production of C60 nanoparticles by a native and denatured hen egg white lysozyme, a highly basic protein, has been systematically studied. In order to obtain a denatured, yet soluble, lysozyme derivative, the four disulfides of the native protein were reduced and exposed cysteines were alkylated by 3-bromopropylamine, thus introducing eight additional positive charges. The C60 solubilizing properties of the modified denatured lysozyme proved to be superior to those of the native protein, allowing the preparation of biocompatible highly homogeneous and stable C60 nanoparticles using lower amounts of protein, as demonstrated by dynamic light scattering, transmission electron microscopy and atomic force microscopy studies. This lysozyme derivative could represent an effective tool for the solubilization of other carbon allotropes.


Naturalistic, prospective study of glaucoma and ocular hypertension treatment in France: strategies, clinical outcomes, and costs at 1 year.
To prospectively observe second-line treatment strategies, their clinical outcomes, and treatment costs in patients with glaucoma or ocular hypertension (OH) in France. Patients were recruited between 1998 and 2000 in 37 centers and were followed for up to 2 years. Outcomes were numbers of and reasons for treatment changes, changes in clinical parameters (intraocular pressure [IOP] levels, visual field defects, and optic nerve excavation), and direct medical costs associated with glaucoma management in patients receiving monotherapy or combination therapy. This article reports results of an interim analysis of 1-year follow-up data for patients having at least two contacts with a study ophthalmologist. Data were analyzed for 283 patients and 549 treated eyes. Ocular hypotensive monotherapy was used as first-line therapy in 92.0% of eyes. Second-line treatment was initiated an average of 3.4 +/- 0.5 years after diagnosis, primarily due to insufficient IOP control (62.8%). Mean IOP reductions after 1 year of second-line therapy were 3.0 mmHg in eyes treated with latanoprost monotherapy versus 2. 1 mmHg in those receiving beta-blocker monotherapy (p = 0.02) and 5.4 mmHg in eyes treated with the latanoprost + timolol combination versus 4.1 mmHg in those receiving combination therapies that did not include latanoprost (p = 0.01). Although second-line treatment with latanoprost was more costly than treatment with beta blockers, the average daily cost for latanoprost monotherapy was similar to that for patients who failed beta-blocker monotherapy, and latanoprost + timolol was less costly than therapeutic combinations without latanoprost. Insufficient IOP control is the main reason for changing first-line treatment in patients with glaucoma or OH. After 1 year, second-line treatment with latanoprost, as monotherapy or combined with timolol, provides superior IOP control at an acceptable cost.


Group differences in physician responses to handheld presentation of clinical evidence: a verbal protocol analysis.
To identify individual differences in physicians' needs for the presentation of evidence resources and preferences for mobile devices. Within-groups analysis of responses to semi-structured interviews. Interviews consisted of using prototypes in response to task-based scenarios. The prototypes were implemented on two different form factors: a tablet style PC and a pocketPC. Participants were from three user groups: general internists, family physicians and medicine residents, and from two different settings: urban and semi-urban. Verbal protocol analysis, which consists of coding utterances, was conducted on the transcripts of the testing sessions. Statistical relationships were investigated between staff physicians' and residents' background variables, self-reported experiences with the interfaces, and verbal code frequencies. 47 physicians were recruited from general internal medicine, family practice clinics and a residency training program. The mean age of participants was 42.6 years. Physician specialty had a greater effect on device and information-presentation preferences than gender, age, setting or previous technical experience. Family physicians preferred the screen size of the tablet computer and were less concerned about its portability. Residents liked the screen size of the tablet, but preferred the portability of the pocketPC. Internists liked the portability of the pocketPC, but saw less advantage to the large screen of the tablet computer (F[2,44] = 4.94, p = .012). Different types of physicians have different needs and preferences for evidence-based resources and handheld devices. This study shows how user testing can be incorporated into the process of design to inform group-based customization.


The effect of ball characteristics on head acceleration during purposeful heading in male and female youth football players.
Objective: The objective of this cross-sectional study was to explore the effects of different ball types and characteristics on head acceleration during purposeful heading in youth football players. Methods: Experienced male and female players (n = 61) aged between 12-17 years completed heading trials with 4 different balls (Ball 1 mass 192 grams (g), pressure 5.0 pounds per square inch (psi); Ball 2 432 g, 5.0 psi; Ball 3 255 g, 5.0 psi; Ball 4 430 g, 10.5 psi) whilst wearing a head-mounted accelerometer and gyroscope. Balls 1, 2 and 4 were size 5 balls; Ball 3 was a size 4 ball.Results: Multivariate analysis of variance and post-hoc univariate analyses revealed a statistically significant difference between ball type and head acceleration during heading for both linear acceleration (adjusted R2 = 0.68; F = 140.90; p = <0.001) and angular velocity (adjusted R2 = 0.28; F = 26.52; p = <0.001). Ball 1 (lightest size 5 ball) and Ball 3 (size 4 ball) demonstrated linear head accelerations up to 59% lower (p = <0.01) when compared with Ball 4 (size 5 regulated match ball).Discussion: Head acceleration during purposeful heading is influenced by changes to ball pressure, ball size and/or ball mass. Changing ball characteristics, particularly in youth football training when heading is being taught, should be an easy strategy to implement.


Comparison of methods to measure heart size using noncontrast-enhanced computed tomography: correlation with left ventricular mass.
Left ventricular (LV) mass is a useful independent predictor of cardiovascular events. We sought to develop a new correlate of LV mass using noncontrast-enhanced cardiac computed tomography (NCE-CCT). We assessed 22 different ventricular measurements made with NCE-CCT in 60 participants in the Multi-Ethnic Study of Atherosclerosis. The primary outcome was the correlation between the NCE-CCT measurements and magnetic resonance imaging (MRI)-derived LV mass. Correlation coefficients (r) for the 22 NCE-CCT techniques in comparison to MRI-derived LV mass ranged from 0.12 to 0.80, with 14 of the 22 techniques having r > 0.7. The highest correlation was achieved using the modified Simpson Rule method to determine the biventricular volume (r = 0.80; P < 0.001). Interrater reliability was good, with intraclass correlation coefficients of 0.84 to 0.90 for the best (r > 0.75) NCE-CCT methods. Noncontrast-enhanced cardiac computed tomography measurements of both biventricular volume and LV volume correlated well with MRI-derived LV mass in a population free of clinical cardiovascular disease.


Fetal macrosomia--a continuing obstetric challenge.
Macrosomic fetuses represent a continuing challenge in obstetrics. We studied maternal risk factors of fetal macrosomia and maternal and infant outcome in such cases. A retrospective cohort study was carried out with a total of 26,961 singleton pregnancies between 1989 and 2001. Records of 886 mothers who gave birth to live born infants weighing > or =4,500 g were compared to those of 26,075 mothers with normal weight (<4,500 g) infants. Multiple regression analysis was used to identify independent reproductive risk factors. Perinatal complications were also assessed. The incidence of fetal macrosomia was 3.4%. Diabetes, previous macrosomic birth, postdatism (>42 weeks of gestation), obesity (BMI > 25 before pregnancy), male infant, gestational diabetes mellitus, and non-smoking were independent risk factors of fetal macrosomia, with adjusted risks of 4.6, 3.1, 3.1, 2.0, 1.9, 1.6, 1.4, respectively. In the macrosomic group, birth and maternal traumas occurred significantly more often than in the control group. However, records of subsequent pregnancies (n = 250) after the study period showed that a previous uncomplicated birth appeared to decrease complication risks. Most cases of fetal macrosomia occur in low-risk pregnancies and evaluation of maternal risks cannot accurately predict which women will eventually give birth to an overweight newborn. After an uncomplicated birth of a macrosomic infant, vaginal delivery may be a safe option for the infant and mother.


Delirium in the ICU: time to probe the hard questions.
Prevalent in critically ill patients, delirium remains poorly understood and difficult to treat. In a cross-sectional study conducted in 12 countries, delirium was identified in close to one third of patients and was independently associated with increased mortality. While such epidemiological accounts represent an important cornerstone for research, scientific efforts are needed to elucidate the causes of delirium and the mechanisms underlying its association with poor outcomes.


Identification of gap junctional connexin-32 mRNA and protein in gonadotropin-releasing hormone neurons of the female rat.
Pulsatile gonadotropin-releasing hormone (GnRH) release from the median eminence is critical for the appropriate function of the pituitary gonadotropes and for the generation of a preovulatory gonadotropin surge. The mechanisms by which many GnRH axon terminals are synchronized to release GnRH in a coordinated fashion into the capillaries of the primary plexus are unknown as are the anatomical sites at which the regulation of GnRH neurons takes place. While many neurotransmitters have been shown to influence GnRH release, it is not clear if such neurotransmitters regulate GnRH neurons directly through synaptic interactions or through intermediate neurons. An alternative mechanism of interneuronal communication is provided by gap junctions which allow a rapid, bidirectional exchange of signals. In order to explore if GnRH neurons synthesize the appropriate proteins to form gap junctions with adjacent cells we used double immunohistochemistry for GnRH and connexins-26, -32 or -43 as well as dual in situ hybridization to identify GnRH mRNA and connexin-32 mRNA. The results show that all GnRH neurons contain connexin-32 immunoreactive puncta at their perikarya and, occasionally, at their axon terminals in the median eminence while connexin-26 and -43 immunoreactivity was absent in GnRH neurons. In addition, connexin-32 mRNA was detected in GnRH mRNA containing neurons. However, gap junctional connections between adjacent GnRH neurons were not observed. The data suggest that gap junctional coupling of GnRH neurons with neighboring non-GnRH containing cells may occur and may represent a mechanism by which GnRH neurons can be synchronized or by which hormonal or neurotransmitter signals can be conveyed to the GnRH neurons.


Macrophage Inhibitory Factor-1 (MIF-1) controls the plasticity of multiple myeloma tumor cells.
Multiple Myeloma (MM) is the second most common hematological malignancy with a median survival of 5-10 years. While current treatments initially cause remission, relapse almost always occurs, leading to the hypothesis that a chemotherapy-resistant cancer stem cell (CSC) remains dormant, and undergoes self-renewal and differentiation to reestablish disease. Our finding is that the mature cancer cell (CD138+, rapidly proliferating and chemosensitive) has developmental plasticity; namely, the ability to dedifferentiate back into its own chemoresistant CSC progenitor, the CD138-, quiescent pre-plasma cell. We observe multiple cycles of differentiation and dedifferentiation in the absence of niche or supportive accessory cells, suggesting that soluble cytokines secreted by the MM cells themselves are responsible for this bidirectional interconversion and that stemness and chemoresistance are dynamic characteristics that can be acquired or lost and thus may be targetable. By examining cytokine secretion of CD138- and CD138+ RPMI-8226 cells, we identified that concomitant with interconversion, Macrophage Migration Inhibitory Factor (MIF-1) is secreted. The addition of a small molecule MIF-1 inhibitor (4-IPP) or MIF-1 neutralizing antibodies to CD138+ cells accelerated dedifferentiation back into the CD138- progenitor, while addition of recombinant MIF-1 drove cells towards CD138+ differentiation. A similar increase in the CD138- population is seen when MM tumor cells isolated from primary bone marrow aspirates are cultured in the presence of 4-IPP. As the CD138+ MM cell is chemosensitive, targeting MIF-1 and/or the pathways that it regulates could be a viable way to modulate stemness and chemosensitivity, which could in turn transform the treatment of MM.


Assessing the reliability of the five minute speech sample against the Camberwell family interview in a chronic fatigue syndrome sample.
The current study aimed to examine the reliability of the Five Minute Speech Sample (FMSS) for assessing relative Expressed Emotion (EE) compared with the Camberwell Family Interview (CFI) in a sample of relatives of adult patients with Chronic Fatigue Syndrome (CFS). 21 relatives were recruited and completed both assessments. The CFI was conducted first for all participants, with the FMSS conducted approximately one month later. Trained raters independently coded both EE measures; high levels of rating reliability were established for both measures. Comparisons were conducted for overall EE status, emotional over-involvement (EOI) and criticism. The distribution of high and low-EE was equivalent across the two measures, with the FMSS correctly classifying EE is 71% of cases (n=15). The correspondence between the FMSS and CFI ratings was found to be non-significant for all categorical variables. However, the number of critical comments made by relatives during the FMSS significantly correlated with the number of critical comments made during the CFI. The poorest correspondence between the measures was observed for the EOI dimension. The findings suggest that the FMSS may be a useful screening tool for identifying high-EE, particularly criticism, within a sample of relatives of patients with CFS. However, the two measures should not be assumed equivalent, and the CFI should be used where possible, particularly with respect to understanding EOI.


Caffeic acid derivatives from Bupleurum chinense.
In this study, caffeic acid (CA) and its three derivatives including 3-caffeoylquinic acid (3-CQA, neochlorogenic acid), 4-caffeoylquinic acid (4-CQA, cryptochlorogenic acid), and 5-caffeoylquinic acid (5-CQA, chlorogenic acid) were identified in Bupleurum chinense aerial parts using reverse-phase high-performance liquid chromatography (RP-HPLC) with photodiode array (PDA) detector, reference compounds and chemical reactions. Separation was performed on a C18 column using gradient elution with 4% (v/v) aqueous acetic acid and acetonitrile as mobile phase at ambient temperature. In addition, the flavonoid aglycones were characterized and quantified after acid hydrolysis of the plant material. The flavonols profile showed quercetin (0.36 g per 100 g), kaempferol (1.11 g per 100 g) and isorhamnetin (0.16 g per 100 g). Total phenolic and total flavonoid contents ranged from 7.3 to 18.7% and 0.58 to 2.72% in dry plant material, respectively.


Three-Dimensional Printing of Cellulose/Covalent Organic Frameworks (CelloCOFs) for CO2 Adsorption and Water Treatment.
The development of porous organic polymers, specifically covalent organic frameworks (COFs), has facilitated the advancement of numerous applications. Nevertheless, the limited availability of COFs solely in powder form imposes constraints on their potential applications. Furthermore, it is worth noting that COFs tend to undergo aggregation, leading to a decrease in the number of active sites available within the material. This work presents a comprehensive methodology for the transformation of a COF into three-dimensional (3D) scaffolds using the technique of 3D printing. As part of the 3D printing process, a composite material called CelloCOF was created by combining cellulose nanofibrils (CNF), sodium alginate, and COF materials (i.e., COF-1 and COF-2). The intervention successfully mitigated the agglomeration of the COF nanoparticles, resulting in the creation of abundant active sites that can be effectively utilized for adsorption purposes. The method of 3D printing can be described as a simple and basic procedure that can be adapted to accommodate hierarchical porous materials with distinct micro- and macropore regimes. This technology demonstrates versatility in its use across a range of COF materials. The adsorption capacities of 3D CelloCOF materials were evaluated for three different adsorbates: carbon dioxide (CO2), heavy metal ions, and perfluorooctanesulfonic acid (PFOS). The results showed that the materials exhibited adsorption capabilities of 19.9, 7.4-34, and 118.5-410.8 mg/g for CO2, PFOS, and heavy metals, respectively. The adsorption properties of the material were found to be outstanding, exhibiting a high degree of recyclability and exceptional selectivity. Based on our research findings, it is conceivable that the utilization of custom-designed composites based on COFs could present new opportunities in the realm of water and air purification.


Mouse oocytes enable LH-induced maturation of the cumulus-oocyte complex via promoting EGF receptor-dependent signaling.
LH triggers the maturation of the cumulus-oocyte complex (COC), which is followed by ovulation. These ovarian follicular responses to LH are mediated by epidermal growth factor (EGF)-like growth factors produced by granulosa cells and require the participation of oocyte-derived paracrine factors. However, it is not clear how oocytes coordinate with the EGF receptor (EGFR) signaling to achieve COC maturation. The aim of the present study was to test the hypothesis that oocytes promote the expression of EGFR by cumulus cells, thus enabling them to respond to the LH-induced EGF-like peptides. Egfr mRNA and protein expression were dramatically reduced in cumulus cells of mutant mice deficient in the production of the oocyte-derived paracrine factors growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15). Moreover, microsurgical removal of oocytes from wild-type COCs dramatically reduced expression of Egfr mRNA and protein, and these levels were restored by either coculture with oocytes or treatment with recombinant GDF9 or GDF9 plus recombinant BMP15. Blocking Sma- and Mad-related protein (SMAD)2/3 phosphorylation in vitro inhibited Egfr expression in wild-type COCs and in GDF9-treated wild-type cumulus cells, and conditional deletion of Smad2 and Smad3 genes in granulosa cells in vivo resulted in the reduction of Egfr mRNA in cumulus cells. These results indicate that oocytes promote expression of Egfr in cumulus cells, and a SMAD2/3-dependent pathway is involved in this process. At least two oocyte-derived growth factors, GDF9 and BMP15, are required for EGFR expression by cumulus cells.


Grey matter volume in a large cohort of MS patients: relation to MRI parameters and disability.
Although grey matter damage in multiple sclerosis is currently recognized, determinants of grey matter volume and its relationship with disability are not yet clear. The objectives of the study were to measure grey and white matter volumes across different disease phenotypes; identify MRI parameters associated with grey matter volume; and study grey and white matter volume as explanatory variables for clinical impairment. This is a cross-sectional study in which MRI data of 95 clinically isolated syndrome, 657 relapsing-remitting, 125 secondary-progressive and 50 primary-progressive multiple sclerosis patients from three centres were acquired. Grey and white matter volumes were determined, together with T2 and T1 lesion volumes. Physical disability was assessed with the Expanded Disability Status Scale, cognitive impairment with the Paced Auditory Serial Addition Task. Data were analysed using multiple regression. Grey matter volume was lower in relapsing-remitting patients (mean [SD]: 0.80 [0.05] L) than in clinically isolated syndrome patients (0.82 [0.05] L), and even greater relative atrophy was found in secondary-progressive patients (0.77 [0.05] L). In contrast, white matter volume in secondary-progressive patients was comparable to that in relapsing-remitting patients. Grey matter volume was the strongest independent predictor of physical disability and cognitive impairment, and was associated with both T2 and T1 lesion volume. Our findings show that grey matter volume is lower in secondary-progressive than in relapsing-remitting disease. Grey matter volume explained physical and cognitive impairment better than white matter volume, and is itself associated with T2 and T1 lesion volume.


Neuroadapted yellow fever virus 17D: determinants in the envelope protein govern neuroinvasiveness for SCID mice.
A molecular clone of mouse-neuroadapted yellow fever 17D virus (SPYF-MN) was used to identify critical determinants of viral neuroinvasiveness in a SCID mouse model. Virus derived from this clone differs from nonneuroinvasive YF5.2iv virus at 29 nucleotide positions, encoding 13 predicted amino acid substitutions and 2 substitutions in the 3' untranslated region (UTR). The virulence determinants of SPYF-MN for SCID mice were identified by constructing and characterizing intratypic viruses in which the E protein of SPYF-MN was expressed in the YF5.2iv background (SPYF-E) or the E protein of YF5.2iv was expressed in the SPYF-MN background (YF5.2-E). SPYF-E caused lethal encephalitis in young adult SCID mice after intraperitoneal inoculation, with average survival times and tissue virus burdens resembling those of mice inoculated with the parental SPYF-MN virus. To define which domains of the E protein are involved in neuroinvasiveness, two viruses were tested in which the amino acid substitutions in domains I-II and III were segregated. This revealed that substitutions in domain III (residues 305, 326, and 380) were critical for the neuroinvasive phenotype, based on average survival times and tissue burdens of infectious virus. Comparison of growth properties of the various intratypic viruses in cell culture indicated that no inherent defects in replication efficiency were likely to account for the biological differences observed in these experiments. These findings demonstrate that the E protein is a critical factor for yellow fever virus neuropathogenesis in the SCID mouse model and that the neuroinvasive properties depend principally on functions contributed by domain III of this protein. To assess whether critical determinants for neuroinvasion of normal ICR mice by SPYF virus were also in the E protein, sequences of viruses recovered from brains of ICR mice succumbing to encephalitis with the parental SPYF virus were derived. No differences were found in the E protein; however, two substitutions were present in the 3' UTR compared to that of SPYF-MN, one of which is predicted to alter RNA secondary structure in this region. These findings suggest that the 3' UTR may also affect neuroinvasiveness of SPYF virus in the mouse model.


Dynamic Network of Interactions in the Wildlife-Livestock Interface in Mediterranean Spain: An Epidemiological Point of View.
The correct management of diseases that are transmitted between wildlife and livestock requires a reliable estimate of the pathogen transmission rate. The calculation of this parameter is a challenge for epidemiologists, since transmission can occur through multiple pathways. The social network analysis is a widely used tool in epidemiology due to its capacity to identify individuals and communities with relevant roles for pathogen transmission. In the present work, we studied the dynamic network of interactions in a complex epidemiological scenario using information from different methodologies. In 2015, nine red deer, seven fallow deer, six wild boar and nine cattle were simultaneously monitored using GPS-GSM-Proximity collars in Doñana National Park. In addition, 16 proximity loggers were set in aggregation points. Using the social network analysis, we studied the dynamic network of interactions, including direct and indirect interactions, between individuals of different species and the potential transmission of pathogens within this network. The results show a high connection between species through indirect interactions, with a marked seasonality in the conformation of new interactions. Within the network, we differentiated four communities that included individuals of all the species. Regarding the transmission of pathogens, we observed the important role that fallow deer could be playing in the maintenance and transmission of pathogens to livestock. The present work shows the need to consider different types of methodologies in order to understand the complete functioning of the network of interactions at the wildlife/livestock interface. It also provides a methodological approach applicable to the management of shared diseases.


Lola regulates Drosophila olfactory projection neuron identity and targeting specificity.
Precise connections of neural circuits can be specified by genetic programming. In the Drosophila olfactory system, projection neurons (PNs) send dendrites to single glomeruli in the antenna lobe (AL) based upon lineage and birth order and send axons with stereotyped terminations to higher olfactory centers. These decisions are likely specified by a PN-intrinsic transcriptional code that regulates the expression of cell-surface molecules to instruct wiring specificity. We find that the loss of longitudinals lacking (lola), which encodes a BTB-Zn-finger transcription factor with 20 predicted splice isoforms, results in wiring defects in both axons and dendrites of all lineages of PNs. RNA in situ hybridization and quantitative RT-PCR suggest that most if not all lola isoforms are expressed in all PNs, but different isoforms are expressed at widely varying levels. Overexpression of individual lola isoforms fails to rescue the lola null phenotypes and causes additional phenotypes. Loss of lola also results in ectopic expression of Gal4 drivers in multiple cell types and in the loss of transcription factor gene lim1 expression in ventral PNs. Our results indicate that lola is required for wiring of axons and dendrites of most PN classes, and suggest a need for its molecular diversity. Expression pattern changes of Gal4 drivers in lola-/- clones imply that lola normally represses the expression of these regulatory elements in a subset of the cells surrounding the AL. We propose that Lola functions as a general transcription factor that regulates the expression of multiple genes ultimately controlling PN identity and wiring specificity.


Development of depressive symptoms and depression during organizational change--a two-year follow-up study of civil servants.
On 1 January 2007, Denmark went through a major reorganization, where most of its 275 municipalities and 14 counties merged into larger units. Our study aimed to examine the development of depressive symptoms and incident depression among employees affected by this organizational change. A total of 685 civil servants employed in the administration of 5 municipalities and 2 counties participated in the study. They answered a postal questionnaire, 8 months prior to and 16 months after the reorganization, regarding working conditions, psychosocial work environment factors, and depressive symptoms, based on the Major Depression Inventory (MDI). During the follow-up period of 2006-2008, 295 employees had experienced a merger with other workplaces (hereafter the merger group), 259 had got a new job (the new job group), and 131 who experienced no change in workplace served as the control group. The three groups were compared to each other for (i) mean score of MDI and (ii) incident cases of depression using general linear models and logistic regression analyses, separately by gender. After adjustment of the MDI for age, occupation, supervisor function, and department at baseline in 2006, no significant differences in increase in MDI were found between the groups. The incidence of depression in the merger group was not significantly higher than the control group [women: odds ratio (OR) 1.5 (95% confidence interval [95% CI] 0.6-3.9), men: OR 1.7 (95% CI 0.2-18.7)], after adjustment for confounders. This study showed no significantly increased risk of depression or increase in depressive symptoms among employees exposed to organizational change as a part of a major local government reform.


Brief Report: Human Acute Myeloid Leukemia Reprogramming to Pluripotency Is a Rare Event and Selects for Patient Hematopoietic Cells Devoid of Leukemic Mutations.
Induced pluripotent stem cell reprogramming has provided critical insights into disease processes by modeling the genetics and related clinical pathophysiology. Human cancer represents highly diverse genetics, as well as inter- and intra-patient heterogeneity, where cellular model systems capable of capturing this disease complexity would be invaluable. Acute myeloid leukemia (AML) represents one of most heterogeneous cancers and has been divided into genetic subtypes correlated with unique risk stratification over the decades. Here, we report our efforts to induce pluripotency from the heterogeneous population of human patients that represents this disease in the clinic. Using robust optimized reprogramming methods, we demonstrate that reprogramming of AML cells harboring leukemic genomic aberrations is a rare event with the exception of those with de novo mixed-lineage leukemia (MLL) mutations that can be reprogrammed and model drug responses in vitro. Our findings indicate that unlike hematopoietic cells devoid of genomic aberrations, AML cells harboring driver mutations are refractory to reprogramming. Expression of MLL fusion proteins in AML cells did not contribute to induced reprogramming success, which continued to select for patient derived cells devoid of AML patient-specific aberrations. Our study reveals that unanticipated blockades to achieving pluripotency reside within the majority of transformed AML patient cells. Stem Cells 2017;35:2095-2102.


The utility of the gonadotrophin releasing hormone (GnRH) test in the diagnosis of polycystic ovary syndrome (PCOS).
Polycystic ovary syndrome (PCOS) is characterised by increased frequency of hypothalamic GnRH pulses leading to a relative increase in LH synthesis by the pituitary. As GnRH stimulation can reveal a relative LH excess, we have endeavoured to assess whether GnRH test might be useful in the diagnosis of PCOS. The study involved 185 subjects: a PCOS group, n = 151, all with oligo- or amenorrhoea, aged (mean ± SD) 24.8 ± ± 5.4 years, BMI 24.5 ± 6.0 kg/m²; and regularly menstruating controls, n = 34, aged 26.6 ± 5.0 years, BMI 24.6 ± 5.5 kg/m². In 121 subjects with PCOS and in 32 controls, serum LH and FSH were measured before (0 minutes) and 30 and 60 minutes after GnRH stimulation (100 μg i.v.). Insulin resistance was assessed by HOMA and Insulin Resistance Index derived from glucose and insulin concentrations during 75 gram oral glucose tolerance test. Women with PCOS had higher testosterone (p = 0.0002), androstendione (p = 0.0021), 17OH-progesterone (p < 0.0001) and were more insulin resistant. Raised concentrations of at least one androgen were, however, found only in 58.1% of women with PCOS. Baseline and stimulated LH concentrations were higher in PCOS (9.09 ± 5.56 vs 4.83 ± 1.71 IU/L, 35.48 ± 31.4 vs 16.30 ± 6.68 IU/L, 33.86 ± 31.8 vs 13.45 ± 5.2 IU/L, at 0, 30 and 60 mins post GnRH, respectively, p < 0.0001). There was no difference in baseline or stimulated FSH concentrations between groups. Relative increases of LH or FSH in comparison to respective baseline values were similar in both groups. There was, however, a marked increase in LH/FSH ratio in PCOS in comparison to controls (LH0 min/FSH(₀ min) 1.59 ± 0.95 vs 0.76 ± 0.2, LH(₃₀ min) /FSH(₃₀ min) 4.07 ± 3.0 vs 1.89 ± 0.79, LH(₆₀ min)/FSH(₆₀ min) 3.56 ± 2.58 vs 1.55 ± 0.63, p < 0.0001 at all time points). Further analysis revealed that LH30 min/FSH(₃₀ min) > 2.11 or LH(₆₀ min)/FSH(₆₀ min) > 1.72 had 78.3% and 87.5% sensitivity and 81.7% and 81.3% specificity for the diagnosis of PCOS, respectively. Women with PCOS have higher baseline and GnRH-stimulated LH concentrations. GnRH stimulation results in an increase in LH/FSH ratio in women with PCOS. Therefore we postulate that this phenomenon might be potentially useful as an additional tool in the diagnosis of PCOS.


Simultaneous imaging of hyperpolarized [1,4-13 C2 ]fumarate, [1-13 C]pyruvate and 18 F-FDG in a rat model of necrosis in a clinical PET/MR scanner.
A co-polarization scheme for [1,4-13 C2 ]fumarate and [1-13 C]pyruvate is presented to simultaneously assess necrosis and metabolism in rats with hyperpolarized 13 C magnetic resonance (MR). The co-polarization was performed in a SPINlab polarizer. In addition, the feasibility of simultaneous positron emission tomography (PET) and MR of small animals with a clinical PET/MR scanner is demonstrated. The hyperpolarized metabolic MR and PET was demonstrated in a rat model of necrosis. The polarization and T1 of the co-polarized [1,4-13 C2 ]fumarate and [1-13 C]pyruvate substrates were measured in vitro and compared with those obtained when the substrates were polarized individually. A polarization of 36 ± 4% for fumarate and 37 ± 6% for pyruvate was obtained. We found no significant difference in the polarization and T1 values between the dual and single substrate polarization. Rats weighing about 400 g were injected intramuscularly in one of the hind legs with 200 μL of turpentine to induce necrosis. Two hours later, 13 C metabolic maps were obtained with a chemical shift imaging sequence (16 × 16) with a resolution of 3.1 × 5.0 × 25.0 mm3 . The 13 C spectroscopic images were acquired in 12 s, followed by an 8-min 18 F-2-fluoro-2-deoxy-d-glucose (18 F-FDG) PET acquisition with a resolution of 3.5 mm. [1,4-13 C2 ]Malate was observed from the tissue injected with turpentine indicating necrosis. Normal [1-13 C]pyruvate metabolism and 18 F-FDG uptake were observed from the same tissue. The proposed co-polarization scheme provides a means to utilize multiple imaging agents simultaneously, and thus to probe various metabolic pathways in a single examination. Moreover, it demonstrates the feasibility of small animal research on a clinical PET/MR scanner for combined PET and hyperpolarized metabolic MR.


Cell-cycle arrest and p53-independent induction of apoptosis in vitro by the new anticancer drugs 5-FdUrd-P-FdCydOct and dCydPam-P-FdUrd in DU-145 human prostate cancer cells.
Current therapies have limited impact on the progression of metastatic hormone-refractory prostate cancer. Therefore, we investigated the utility of new heterodinucleoside phosphate dimers of 5-fluorodeoxyuridine (5-FdUrd) in p53-mutated and androgen-independent DU-145 human prostate tumour cells. The effects of the dimers were assessed in vitro by a cell proliferation assay for cytotoxicity, flow cytometry for cell cycle distribution, confocal laser scanning microscopy for the detection of apoptotic bodies, poly(ADP-ribose) polymerase cleavage for caspase 3 activity and by a thymidylate synthetase assay. The new dimers N4-palmitoyl-2'-deoxycytidylyl-(3'-->5')-5-fluoro-2'-deoxyuridine (dCydPam-P-FdUrd) and 2'-deoxy-5-fluorouridylyl-(3'-->5')-2'-deoxy-5-fluoro-N4-octade cylcytidine (5-FdUrd-P-FdCydOct) caused marked cytotoxicity with IC50 values of 3-4 microM. 5-FdUrd-P-FdCydOct at 200 microM was capable of eradicating 100% of tumour cells whereas 10% of the cells were resistant to 5-FdUrd. Cytotoxicity was caused by a dramatic S-phase arrest, resulting in an increase of this cell population from 34% to 85% with 5-FdUrd-P-FdCydOct and to 81% with dCydPam-P-FdUrd. S-phase arrest was followed by apoptosis, as shown by 85% of the cells staining positive for Apo 2.7 antibody, a six- to eight-fold increased caspase 3 activity and DNA fragmentation. Thymidylate synthase activity was inhibited by 50% at 0.6-0.7 microM dimer concentration. The dimers were hydrolysed in vitro by phosphodiesterase I and human serum to the corresponding nucleosides and nucleoside monophosphates. The new dimers dCydPam-P-FdUrd and 5-FdUrd-P-FdCydOct are effective prodrugs of 5-FdUrd and have potential value for the treatment of p53-mutated and hormone-independent human prostate carcinomas.


Priority areas for research on the intake, composition, and health effects of tree nuts and peanuts.
This article summarizes the main conclusions drawn from a conference on the health effects of nut consumption and identifies priority areas for future research. Individuals with higher intakes of nuts generally have higher intakes of many beneficial dietary constituents. More information is needed on nut composition, the bioavailability of nutrients, and other bioactive constituents. Better methods are needed to assess usual nut intake, including biomarkers, and the types, physical form, and amounts of nuts that are consumed. The feasibility of including nuts and seeds as a separate food group in the Dietary Guidelines should be tested, as should ways to increase nut intake. A moderate intake of nuts can be included in a weight loss regimen and further information is needed on whether nuts improve satiety as well as adherence to and efficacy of diets designed for weight reduction. There is substantial evidence that nut consumption reduces risk of cardiovascular disease. Future research should investigate their benefits for prevention of congestive heart failure, including clinical studies in patients with this condition, to evaluate the effects of nuts on markers of heart disease risk. Higher nut consumption is associated with lower risk of diabetes and associated cardiovascular disease. More remains to be learned about the effects of nuts on postprandial glycemic and insulin response, glycemic control, and improvement of disease risk factors in subjects with prediabetes and diabetes. Information is needed on nut-induced allergic reactions, including their prevalence and consequences, causes of sensitization, biomarkers of severe reactions, and cross-reactivity to different types of nuts.


Cyclosporin inhibits phosphate transport and stimulates alkaline phosphatase activity in renal BBMV.
The activity of various enzymes and transport systems was studied in renal brush-border membrane vesicles (BBMV) isolated from rats injected daily with cyclosporin. Alkaline phosphatase (AP) was strongly stimulated: 55 and 113% increases were obtained in BBMV isolated from rats injected with 10 mg cyclosporin/kg for 5 and 10 days. The affinity of the enzyme remained unaltered, but maximal activity (Vmax) showed a strong increase of 2.4-fold between control and treated animals. In addition to the phosphatase activity, phosphate binding to AP also showed a dose-dependent stimulation by cyclosporin treatment: 44 and 70% increases in animals treated for 5 days with 5 and 10 mg cyclosporin/kg. However, the activity of aminopeptidase M was not affected by these treatments, and polyacrylamide gel electrophoresis of BBMV revealed no alterations in the profile of membrane proteins, suggesting the specificity of cyclosporin interaction with alkaline phosphatase. Na(+)-dependent amino acid and D-glucose transport systems remained unaffected by cyclosporin treatment. The Na(+)-independent transport system for lysine and the Na(+)-H+ antiporter activity were also unaltered. In contrast, the initial rate of phosphate uptake decreased by 28% after administration of cyclosporin (10 mg/kg) for 5 days: the Michaelis constant (Km) and Vmax decreased from 137 to 85 microM and from 1.49 to 1.07 pmol.micrograms-1.5 s-1, respectively. "In vitro" studies with membranes isolated from untreated rats were also undertaken by preincubating membranes with cyclosporin. Neither alkaline phosphatase nor the transport systems were affected under these conditions.(ABSTRACT TRUNCATED AT 250 WORDS)


Estimation of the Cost of Smoking-Attributable Diseases (Five Selected Diseases): A Case in Kerman City, Iran, 2014.
Cigarette seems to be the least valuable of addictive drugs. It is easily accessible to the public, and its harmful personal and social effects have attracted less attention. Therefore, the present research was carried out with the aim of presenting cost estimations of smoking-related diseases in smokers who aged 35 or higher in Kerman City, Iran, in 2014. Using the prevalence approach, the direct and indirect costs of smoking-attributable diseases including lung cancer, gastric cancer, myocardial infarction, stroke, and chronic obstructive pulmonary disease (COPD) were estimated. The initial data were obtained from the information in documents of medical document units as well as the 2014 income statements of teaching hospitals of Kerman University of Medical Sciences, Kerman City. In this research, total economic costs of diseases attributed to smoking were estimated to be 50 million dollars in 2014 in Kerman City, and calculations suggest that this figure accounts for 0.02% of Iran's gross domestic product (GDP). Total direct cost of diseases caused by smoking in Kerman City adds up to 17 million dollars, whereas the estimated indirect cost of diseases caused by smoking is 33 million dollars. The yearly per capita cost of any of the selected five diseases is 270 dollars. Smoking places a high economic burden on health system and society as a whole. Therefore, stronger intervention measures against smoking should be taken without delay to reduce the health and financial losses caused by smoking.


Perceived Interpersonal Discrimination and Older Women's Mental Health: Accumulation Across Domains, Attributions, and Time.
Experiencing discrimination is associated with poor mental health, but how cumulative experiences of perceived interpersonal discrimination across attributes, domains, and time are associated with mental disorders is still unknown. Using data from the Study of Women's Health Across the Nation (1996-2008), we applied latent class analysis and generalized linear models to estimate the association between cumulative exposure to perceived interpersonal discrimination and older women's mental health. We found 4 classes of perceived interpersonal discrimination, ranging from cumulative exposure to discrimination over attributes, domains, and time to none or minimal reports of discrimination. Women who experienced cumulative perceived interpersonal discrimination over time and across attributes and domains had the highest risk of depression (Center for Epidemiologic Studies Depression Scale score ≥16) compared with women in all other classes. This was true for all women regardless of race/ethnicity, although the type and severity of perceived discrimination differed across racial/ethnic groups. Cumulative exposure to perceived interpersonal discrimination across attributes, domains, and time has an incremental negative long-term association with mental health. Studies that examine exposure to perceived discrimination due to a single attribute in 1 domain or at 1 point in time underestimate the magnitude and complexity of discrimination and its association with health.


Unravelling biological macromolecules with cryo-electron microscopy.
Knowledge of the three-dimensional structures of proteins and other biological macromolecules often aids understanding of how they perform complicated tasks in the cell. Because many such tasks involve the cleavage or formation of chemical bonds, structural characterization at the atomic level is most useful. Developments in the electron microscopy of frozen hydrated samples (cryo-electron microscopy) are providing unprecedented opportunities for the structural characterization of biological macromolecules. This is resulting in a wave of information about processes in the cell that were impossible to characterize with existing techniques in structural biology.