Involvement of metabolic pathway in the sperm spontaneous acrosome reaction.
In order to fertilize the egg, spermatozoa must undergo a series of biochemical processes in the female reproductive tract collectively called capacitation. Only capacitated sperm can interact with the egg resulting in the acrosome reaction (AR), allowing egg penetration and fertilization. Sperm can undergo spontaneous AR (sAR) before reaching the egg, preventing successful fertilization. Here we investigated the metabolic pathways involved in sperm capacitation and sAR. Inhibition of glycolysis or oxidative phosphorylation did not affect capacitation or sAR levels; however, when both systems were inhibited, no capacitation occurred, and there was a significant increase in sAR. Under such ATP-starvation, the increase in sAR is triggered by Ca2+ influx into the sperm via the CatSper cation channel. Protein kinase A (PKA) is an essential key enzyme in sperm capacitation; there was no change in its activity when a single metabolic system was inhibited, while complete inhibition of was observed when the two systems were inhibited. Protein tyrosine phosphorylation (PTP), also known to occur in sperm capacitation, was partially reduced by inhibition of one metabolic system, and completely blocked when the two metabolic systems were inhibited. We conclude that ATP, PKA and PTP are involved in the mechanisms protecting sperm from sAR.


Identification of up-regulated genes in human uterine leiomyoma by suppression subtractive hybridization.
In searching for differentially expressed genes in human uterine leiomyomas (ULs), suppression subtractive hybridization was used to construct an UL up-regulated library, which turned out to represent 88 genes. After two rounds of screening by reverse Northern analysis, twenty genes were proved to be up-regulated, including seventeen known genes and three genes with unknown function. All these genes were firstly associated with UL. Three genes with notable difference were selected for Northern confirmation. Our results proved the authenticity of the twenty genes. One gene named Phospholipase A2 (PLA2) showed up-regulation in 4/6 of the patients and investigation of tissue distribution indicated that it had obvious expression in prostate, testis, liver, heart and skeletal muscle.


Functional adaptation of crustacean exoskeletal elements through structural and compositional diversity: a combined experimental and theoretical study.
The crustacean cuticle is a composite material that covers the whole animal and forms the continuous exoskeleton. Nano-fibers composed of chitin and protein molecules form most of the organic matrix of the cuticle that, at the macroscale, is organized in up to eight hierarchical levels. At least two of them, the exo- and endocuticle, contain a mineral phase of mainly Mg-calcite, amorphous calcium carbonate and phosphate. The high number of hierarchical levels and the compositional diversity provide a high degree of freedom for varying the physical, in particular mechanical, properties of the material. This makes the cuticle a versatile material ideally suited to form a variety of skeletal elements that are adapted to different functions and the eco-physiological strains of individual species. This review presents our recent analytical, experimental and theoretical studies on the cuticle, summarising at which hierarchical levels structure and composition are modified to achieve the required physical properties. We describe our multi-scale hierarchical modeling approach based on the results from these studies, aiming at systematically predicting the structure-composition-property relations of cuticle composites from the molecular level to the macro-scale. This modeling approach provides a tool to facilitate the development of optimized biomimetic materials within a knowledge-based design approach.


Predictive role of the monocyte-to-lymphocyte ratio in advanced hepatocellular carcinoma patients receiving anti-PD-1 therapy.
The immune checkpoint inhibitor (ICIs) therapy has been proven effective in a range of solid tumors including hepatocellular carcinoma (HCC), non-small cell lung carcinoma and metastatic melanoma. However, only a subset of approximately 20% of patients shows an objective response to anti-PD-1 therapy in HCC. Furthermore, the response to anti-PD-1 therapy is not correlated with programmed cell death 1 ligand expression in tumor tissue. Therefore, it is urgent to identify a biomarker to predict the response of anti-PD-1 therapy. This retrospective study was conducted at the Fudan University Shanghai Cancer Center from December 2019 to June 2021. The monocyte-to-lymphocyte ratio (MLR) was analyzed using a receiver operating characteristic (ROC) curve. A Cox regression model and the log-rank test were used to analyze the relationship between the MLR value and the time to progression (TTP). A total of 34 advanced HCC patients were enrolled in this study. The cut-off point for the MLR at baseline was 0.35. Univariate and multivariate Cox regression models showed that the MLR at baseline was significantly correlated with the TTP (P<0.05). Consistent results were found for disease progression. The log-rank test showed that patients in the low MLR group had a longer TTP (P=0.0027). At the time of disease progression, the median TTP in the low and high MLR groups were 33 and 18 weeks, respectively (P=0.0047). The MLR can predict the response to anti-PD-1 therapy, and a high MLR is correlated with a short TTP in anti-PD-1-treated HCC patients.


HOXA5 inhibits the proliferation and induces the apoptosis of cervical cancer cells via regulation of protein kinase B and p27.
Homeobox A5 (HOXA5) is a member of the homeobox gene (HOX) family, which plays an important role in the development of various malignant tumors. Here, we speculated that HOXA5 has an effect on cervical cancer development. In our study, we aimed to explore the role and molecular mechanism of HOXA5 in regards to the cell proliferation and apoptosis in cervical cancer. We found that expression levels of HOXA5 measured by RT‑qPCR and western blot assays in cervical cancer cell lines and tissues were both significantly downregulated. We performed a gain‑of‑function experiment by the transfection with pcDNA.3.1‑HOXA5 in ME‑180 and HT‑3 cells to overexpress HOXA5, and the caspase‑3 activity measured by caspase‑3 activity assay kit and cell apoptosis detected by flow cytometry were obviously promoted. Meanwhile, cell proliferation tested by BrdU assay, invasion determined by Transwell and cell viability tested by MTT were inhibited. Moreover, protein kinase B (AKT) was activated by incubation with SC79 (AKT activator; 1 µg/ml) after HOXA5 overexpression, and reversed the effect of HOXA5 overexpression on p27 expression. Additionally, significant elevation of AKT activation measured by western blot analysis abrogated the effect of HOXA5 on caspase‑3 activity, cell apoptosis, proliferation, invasion and cell viability. Taken together, this study revealed that HOXA5 inhibits cervical cancer progression by regulating AKT/p27, proposing the potential role of HOXA5 in the prevention and treatment of cervical cancer.


Immunological consequences of methamphetamine protein glycation.
The drug of abuse methamphetamine has been found to participate in the aberrant glycation of proteins. The importance of this chemical process has been shown wherein mouse albumin was readily modified with methamphetamine, and injection of this protein into mice yields a significant immune response, even in the absence of adjuvants. Competition experiments revealed that although methamphetamine binds weakly to the elicited antibodies, the primary epitope is composed of both the methamphetamine moiety and glucose-derived cross-linking region. Implications of this phenonomenon in the context of drug addiction are discussed.


LncRNA-TCONS_00034812 is upregulated in atherosclerosis and upregulates miR-21 through methylation in vascular smooth muscle cells.
LncRNA-TCONS_00034812 is a critical player in the proliferation of aortic smooth muscle cells. It is known that artery injury plays an important role in atherosclerosis. However, the potential implication of LncRNA-TCONS_00034812 in atherosclerosis remains unclear. In this study, we collected artery specimens from patients with atherosclerosis and healthy controls to investigate the involvement of LncRNA-TCONS_00034812 in atherosclerosis. Sixty patients with atherosclerosis and 60 controls, admitted at The First Hospital of Changsha (Changsha, China), between March 2017 and March 2019, were included. An artery biopsy was performed on all participants to obtain the artery specimens. Real-time quantitative PCR were performed to quantify the relative expression level of LncRNA-TCONS_00034812. Its role in atherosclerotic lesion was evaluated in (high fat diet) HFD-induced ApoE-/- mice. Moreover, human aortic smooth muscle cells (HAOSMCs) was employed to study functional role of LncRNA-TCONS_00034812 overexpression and knockdown by methylation-specific PCR and cell proliferation assay. Overexpression of TCONS_00034812 resulted in miR-21 upregulation and a decrease of miR-21 gene methylation. In contrast, silencing of TCONS_00034812 caused miR-21 downregulation and an increase of miR-21 gene methylation. Cell proliferation analysis indicated that the overexpression of TCONS_00034812 and miR-21 promoted cell proliferation, while silencing of TCONS_00034812 played an opposite role. Moreover, miR-21 overexpression weakened the effects of silencing TCONS_00034812 on cell proliferation. In summary, LncRNA-TCONS_00034812 is upregulated in atherosclerotic samples, and its overexpression upregulates miR-21 through methylation in human aortic smooth muscle cells (HAOSMCs). Our study indicates that LncRNA-TCONS_00034812 could serve as a potential biomarker for diagnosis of atherosclerosis.


New proteins related to the Ser-Arg family of splicing factors.
A family of six highly conserved proteins that contain domains rich in alternating serine/arginine residues (SR proteins) function in the regulation of splice site selection and are required for splicing. Using a selective precipitation method, more than 35 proteins were detected in nuclear extracts of HeLa cells that co-fractionate with the defined SR family. Many of these proteins were recognized by three monoclonal antibodies that bind to distinct phosphoepitopes on SR proteins. Two of these SR-related proteins were identified as the nuclear matrix antigens B1C8 and B4A11, which previously have been implicated in splicing. A subset of SR proteins, in their phosphorylated state, are associated with spliceosome complexes through both steps of the splicing reaction, remaining preferentially bound to complexes containing the exon-product. In contrast, other SR-related proteins appear to remain specifically associated with the intron-Iariat complex. The results indicate the existence of a potentially large group of SR-related proteins, and also suggest possible additional functions of SR proteins at a post-splicing level.


Primary pulmonary undifferentiated pleomorphic sarcoma (PPUPS).
Undifferentiated pleomorphic sarcoma (UPS) is a high-grade pleomorphic neoplasm with no identifiable line(s) of differentiation using currently available diagnostic techniques. Therefore, it is essentially a diagnosis of exclusion, which requires generous tissue sampling, adequate contextually interpreted immunohistochemistry, and relevant molecular studies. UPS is a common soft tissue sarcoma (historically one of the entities referred to as malignant fibrous histiocytoma (MFH)), which can develop in various organs, but lung involvement is usually due to metastasis. Primary Pulmonary UPS (PPUPS) is exceptionally rare and here we present a 66-year-old man who presented with anemia and weight loss, found to have a 17 cm right lung mass with invasion to the chest wall and diaphragm. Extensive sampling and immunohistochemistry studies failed to reveal any line of differentiation. Upon exclusion of a possible extrapulmonary origin, a diagnosis of PPUPS was rendered. In addition, we reviewed all 84 previously reported cases of PPUPS/PPMFH in the literature since 1979 and summarized the clinical information.


Targeting STAT proteins via computational analysis in colorectal cancer.
Colorectal cancer (CRC) is the third most common cancer diagnosed worldwide making it a serious global challenge. CRC progression results from dysregulated cytoplasmic transcription factors, including signal transducer and activator of transcription (STAT) proteins that are involved in JAK-STAT pathway. The STAT proteins contain a conserved SH2 domain that facilitates the initiation of STAT activation via binding to tyrosine motifs followed by STAT dimerization. The STAT proteins include STAT1, STAT2 and STAT3 which all facilitate therapeutic targets for many drugs, since they are associated with pathogenesis in various cancers such as CRC. Genistein is an efficient chemopreventive phytochemical drug against CRC. The current investigation presents a computational study performed to investigate the molecular interaction between STAT1, STAT2 and STAT3 proteins with genistein. The molecular dynamic simulation was conducted for STAT2 protein. The studies from molecular docking revealed that the interaction of STAT proteins and genistein is predicted to be effective with better binding energies. Furthermore, targeting STAT3 could be an efficient therapeutic target and understanding the interaction between STAT3 and genistein can help to contribute to a better inhibition process for CRC progression. Treatment with genistein led to significant suppression of cell proliferation and STAT3 protein expression in both CRC (HCT 116 and HT-29) cell lines. This further provides development of efficient STAT inhibitors with better potency and bioavailability.


Adrenergic innervation of bowel in Hirschsprung's disease.
The adrenergic innervation of normal and aganglionic regions of bowel from patients with Hirschsprung's disease was investigated by a fluorescent histochemical technique. In normal bowel the adrenergic nerves end about intramural ganglion cells. In aganglionic bowel the adrenergic nerves form a dense varicose plexus in both muscularis externa and muscularis mucosae. It is suggested that the cause of megacolon in Hirschsprung's disease is due to a lack of nervous pathways controlling the intrinsic reflexes, which is probably congenital in origin.


Partial purification and characterization of the active entity responsible for inducing interleukin-6 production by human gingival fibroblasts from Mycoplasma salivarium cells.
The active entity responsible for inducing interleukin-6 production by human gingival fibroblasts was partially purified by ion-exchange chromatography from the water-soluble fraction of Mycoplasma salivarium cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the final preparation revealed one densely stained band with a molecular weight of 20.6 kilodaltons and two faint bands with molecular weights of 40.5 and 82.5 kilodaltons. The specific activity of the final preparation was 34-fold higher than that of the starting water-soluble fraction. The interleukin-6-inducing activity was destroyed by proteinase K and reduced 70% by lipoprotein lipase and heat treatment, but was not affected by deoxyribonuclease I or endoglucosidase D. The final preparation induced small amounts of tumor necrosis factor-alpha and interleukin-lbeta in a myelomonocytic cell line, THP-1 cells, but did not induce interleukin-6. The ability of Escherichia coli lipopolysaccharide to stimulate human gingival fibroblasts to release interleukin-6 was dependent upon the presence of serum in the assay medium, but that of the final preparation from M. salivarium was not. Thus, we partially purified the protein(s) from M. salivarium which were capable of stimulating human gingival fibroblasts to release interleukin-6 by a mechanism different from that of E. coli lipopolysaccharide.


Resveratrol inhibits TNF-alpha-induced proliferation and matrix metalloproteinase expression in human vascular smooth muscle cells.
Resveratrol (RV), a polyphenolic substance found in grape skin, was suggested to play a role in preventing the development of atherosclerotic disease. Although RV has antiatherogenic effects on vascular smooth muscle cells (VSMC), the molecular mechanisms associated with tumor necrosis factor (TNF)-alpha-induced VSMC are unclear. The goal of this study was to determine the effect of RV on the modulation of cell proliferation, cell-cycle regulation, and matrix metalloproteinase (MMP)-9 expression in TNF-alpha-induced human VSMC. RV treatment inhibited DNA synthesis in cultured VSMC in the presence of TNF-alpha. These inhibitory effects were associated with reduced levels of extracellular signal-regulated kinase (ERK) 1/2 activity and G(1) cell-cycle arrest. Treatment with RV, which blocks the cell cycle in the G(1) phase, downregulated the expression of cyclins and cyclin-dependent kinases (CDKs) and upregulated the expression of p21/WAF1, a CDK inhibitor. RV did not upregulate p27. Moreover, RV increased the promoter activity of the p21/WAF1 gene. Immunoblot and deletion analysis of the p21/WAF1 promoter showed that RV induced the expression of p21/WAF1 and that this expression was independent of the p53 pathway. Furthermore, zymographic and immunoblot analyses showed that RV dose dependently suppressed the TNF-alpha-induced expression of MMP-9. This inhibition was characterized by the downregulation of MMP-9, which was transcriptionally regulated at the activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB) sites in the MMP-9 promoter. Collectively, these results suggest that RV inhibits cell proliferation, G(1) to S phase cell-cycle progress, and MMP-9 expression through the transcription factors NF-kappaB and AP-1 in TNF-alpha-induced VSMC.


Native Bacterial Community Convergence in Augmented and Stimulated Ureolytic MICP Biocementation.
Microbially induced calcite precipitation is a biomineralization process with numerous civil engineering and ground improvement applications. In replicate soil columns, the efficacy and microbial composition of soil bioaugmented with the ureolytic bacterium Sporosarcina pasteurii were compared to a biostimulation method that enriches native ureolytic soil bacteria in situ under conditions analogous to field implementation. The selective enrichment resulting from sequential stimulation treatments strongly selected for Firmicutes (>97%), with Sporosarcina and Lysinibacillus comprising 60 to 94% of high-throughput 16S rDNA sequences in each suspended community sample. Seven species of the former and two of the latter were present in greater than 10% abundance at different times, demonstrating unexpected within-genus diversity and robustness in the suspended phase of this highly selective environment. Based on longer 16S sequences, it was inferred that augmented S. pasteurii competed poorly with natural bacteria, decreasing to below detection after nine treatments, while the native microbial community was enriched to approximately that present in the stimulated columns. These analyses were corroborated by the observed convergence in bulk ureolytic rates and calcite contents between techniques. However, a 10-fold discrepancy between the observed cell density and an activity-based estimate indicates the attached community, uncharacterized despite efforts, substantially contributes to bulk behavior.


Comparison of clinician-rated and self-report insight in Korean patients with schizophrenia using VAGUS insight scale.
This study was aimed to explore self-report auditory verbal hallucinations to provide unique and valuable information in addition to clinician-rated assessment in patients with schizophrenia. The VAGUS (http://www.vagusonline.com) is a recently developed insight scale that includes both clinician-rated (CR) and self-report (SR) versions. Insight measures obtained by the two versions of the VAGUS from the clinicians and the patients, respectively, in forty-one patients diagnosed with schizophrenia by DSM-IV-TR criteria were compared. Correlation coefficients for inter-scale convergence and 3-D biplots for multivariate relationship were derived from the subscales of the VAGUS. For external validation, correlation analyses with abridged version of Scale to Assess Unawareness in Mental Disorder (SUMD-A) and PANSS G12 item were conducted. Total scores of VAGUS-CR and -SR were 5.2 ± 2.6 and 4.9 ± 2.2, respectively. There was a strong correlation between them along with moderate pairwise correlations among the subscales. The 3-D biplots demonstrated that most subscales were clustered as a single factor apart from self-report Symptom Attribution separated as an independent factor. The VAGUS-CR, not -SR correlated significantly with the SUMD-A and PANSS G12. The utility of the VAGUS in reaching more overall understanding of the elusive phenomenon of insight in patients with schizophrenia is discussed.


MetDAT: a modular and workflow-based free online pipeline for mass spectrometry data processing, analysis and interpretation.
Analysis of high throughput metabolomics experiments is a resource-intensive process that includes pre-processing, pre-treatment and post-processing at each level of experimental hierarchy. We developed an interactive user-friendly online software called Metabolite Data Analysis Tool (MetDAT) for mass spectrometry data. It offers a pipeline of tools for file handling, data pre-processing, univariate and multivariate statistical analyses, database searching and pathway mapping. Outputs are produced in the form of text and high-quality images in real-time. MetDAT allows users to combine data management and experiment-centric workflows for optimization of metabolomics methods and metabolite analysis. MetDAT is available free for academic use from http://smbl.nus.edu.sg/METDAT2/. email@example.com


Is modulation of the rate of proton pumping a key event in osmoregulation?
The net uptake of 3-O-methylglucose into leaf segments obtained from Senecio mikanioides Otto, and net proton efflux from the segments, were both promoted when the osmotic potential of the medium was decreased by addition of mannitol, sorbitol, or polyethylene glycol (optimal osmolarity, 0.3 Osmolar for mannitol and sorbitol). The effect was not due to promotion of ;aging', since the antibiotic cerulenin suppressed aging without reducing the size of the mannitol stimulation; further, mannitol did not accelerate aging. Neither was the effect ascribable to diminished efflux (i.e. reduced ;leak' because: first, visualization of the unidirectional sugar fluxes by double labeling indicated that the effect of added osmoticum was to promote influx rather than to reduce efflux; second, compartment analysis did not suggest any effect of mannitol on the rate constants for efflux from either the slowly equilibrating or more rapidly equilibrating compartment. The effect was not specific to poly-ols since it was also obtained with betaine and choline chloride. Since methyl glucose is not taken up into the phloem it could not be ascribed to a turgor effect on phloem loading. We conclude that the effect may reflect osmoregulation. As the sugar flux is probably driven by protonmotive force, it is likely that the effects on proton flux and on sugar flux are related. We suggest that the plasmalemma-sited proton pump is sensitive to the hydrostatic pressure gradient across the plasmalemma-cell wall complex, and functions both as detector and as effector in osmoregulation.


Transitional health care for offenders being released from United States prisons.
Ex-offender managed health care can enhance post-release continuity of care by increasing access, decreasing acute-care episodes, controlling the spread of communicable diseases, and reducing the financial impact on public health-care systems. This study describes transitional health care for inmates with AIDS, tuberculosis (TB), hepatitis, mental illness, and substance abuse. The relationship between size of prison system and coordination of care was also investigated. A mail survey was completed by 33 chief medical officers of prison systems in the United States. Transitional health-care programs for ex-offenders vary widely and no significant relationship was found between number of inmates released per state annually and state coordination of transitional health care for supervised ex-offenders. All respondents reported some type of transitional health-care planning, usually either 1 month or 6 months prior to release. This included provision of post-release medication, referral to community health agencies, scheduling of appointments, and instruction in prevention of transmission. The majority of respondents reported that transitional health-care planning was coordinated by registered nurses. Specific measures for inmates with HIV/AIDS,TB, mental illness, and substance abuse were reported. Information about existing transitional health-care programs can help nurses and other health-care providers identify trends in transitional health-care planning and ensure continuity of care for released offenders.


Accuracy of Carbohydrate Counting in Adults.
In Brief This study investigates carbohydrate counting accuracy in patients using insulin through a multiple daily injection regimen or continuous subcutaneous insulin infusion. The average accuracy test score for all patients was 59%. The carbohydrate test in this study can be used to emphasize the importance of carbohydrate counting to patients and to provide ongoing education.


Association of BMI with total mortality and recurrent stroke among stroke patients: A meta-analysis of cohort studies.
Studies of the association between obesity and total mortality and recurrent stroke events have shown contradictory results. Therefore, we aimed to conduct a meta-analysis to examine the association of body mass index (BMI) with total mortality and recurrent stroke events among patients after stroke onset. We performed an electronic search of PubMed, EMBASE and the Cochrane Library Database, as well as a bibliography review to identify relevant cohort studies published prior to 15th December 2015. Estimates of relative risks (RRs) and corresponding 95% confidence intervals (CIs) comparing underweight, overweight and obese groups with normal weight were pooled using random effects models. In total, 15 studies with 122,472 stroke patients were eligible for inclusion in the meta-analysis. Compared with the normal weight group, obese stroke patients had a significant decreased risk for total mortality (RR = 0.83, 95% CI, 0.73-0.93, p = 0.002), while underweight patients had a significant increased risk for total mortality (RR = 1.54, 95% CI, 1.31-1.82, p = 3.66 × 10-7). A similar, but not significant, association of BMI categories with recurrent stroke events was also observed. Furthermore, the dose-response meta-analysis identified a nonlinear trend for total mortality and a linear trend for recurrent stroke events, associated with BMI. Our results suggested that obesity may have a protective effect on total mortality and recurrent stroke events among patients with established stroke.


Cardiac oxytocin receptor blockade stimulates adverse cardiac remodeling in ovariectomized spontaneously hypertensive rats.
An increasing amount of evidence demonstrates the beneficial role of oxytocin (OT) in the cardiovascular system. Similar actions are attributed to genistein, an isoflavonic phytoestrogen. The treatment with genistein activates the OT system in the aorta of ovariectomized (OVX) Sprague-Dawley (SD) rats. The objective of this study was to determine the effects of low doses of genistein on the OT-induced effects in rat hypertension. The hypothesis tested was that treatment of OVX spontaneously hypertensive rats (SHRs) with genistein improves heart structure and heart work through a mechanism involving the specific OT receptor (OTR). OVX SHRs or SD rats were treated with genistein (in microg/g body wt sc, 10 days) in the presence or absence of an OT antagonist (OTA) [d(CH(2))(5), Tyr(Me)(2), Orn(8)]-vasotocin or a nonspecific estrogen receptor antagonist (ICI-182780). Vehicle-treated OVX rats served as controls. RT-PCR and Western blot analysis demonstrated that left ventricular (LV) OTR, downregulated by ovariectomy, increased in response to genistein. In SHRs or SD rats, this effect was blocked by OTA or ICI-182780 administration. The OTR was mainly localized in microvessels expressing the CD31 marker and colocalized with endothelial nitric oxide synthase. In SHRs, the genistein-stimulated OTR increases were associated with improved fractional shortening, decreased blood pressure (12 mmHg), decreased heart weight-to-body weight ratio, decreased fibrosis, and lowered brain natriuretic peptide in the LV. The prominent finding of the study is the detrimental effect of OTA treatment on the LV of SHRs. OTA treatment of OVX SHRs resulted in a dramatic worsening of ejection fractions and an augmented fibrosis. In conclusion, these results demonstrate that cardiac OTRs are involved in the regulation of cardiac function of OVX SHRs. The decreases of OTRs may contribute to cardiac pathology following menopause.


The type I human T-cell leukemia virus (HTLV-I) Rex trans-activator binds directly to the HTLV-I Rex and the type 1 human immunodeficiency virus Rev RNA response elements.
The Rex protein of the type I human T-cell leukemia virus (HTLV-I) is essential for the replication of this pathogenic retrovirus and, surprisingly, can also replace the function of the structurally distinct Rev protein of the type 1 human immunodeficiency virus (HIV-1). Rex action requires a 255-nucleotide viral RNA stem-loop structure termed the Rex RNA response element (RexRE) located in the 3' retroviral long terminal repeat. Rex function leads to the induced cytoplasmic expression of the incompletely spliced family of viral mRNAs that uniquely encode the HTLV-I structural and enzymatic proteins (Gag, Pol, and Env). Our studies now demonstrate that Rex acts by binding directly to the RexRE in a sequence-specific manner. These effects of Rex require the presence of a 10-nucleotide subregion of the RexRE that is essential for Rex function in vivo. Dominant-negative mutants of Rex also bind to the RexRE with high affinity, while a recessive-negative Rex mutant altered within its arginine-rich, positively charged domain fails to engage the RexRE. Analogously, both the wild-type and dominant-negative Rex proteins specifically bind to the structurally distinct HIV-1 Rev response element, a finding that likely underlies the respective stimulatory and inhibitory effects of these HTLV-I proteins in the heterologous HIV-1 system. However, consistent with their lack of amino acid homology, the binding sites for Rex and Rev within the HIV-1 Rev response element are distinct.


Can Pentoxifylline be used as Adjunct Therapy to ACE Inhibitors and ARBs in Preserving Kidney Function?
To determine if there is sufficient evidence to recommend the addition of pentoxifylline to standard ACE inhibitor and ARB therapy in chronic kidney disease patients to reduce proteinuria and preserve kidney function. A search of the literature was conducted using the PubMed.gov and ClinicalTrials.gov search engines and the search terms "pentoxifylline renoprotection", "pentoxifylline CKD", and "pentoxifylline nephropathy". RESULTS were limited to studies in human subjects and published in the English language. No date range was specified. Studies focused on the effects of pentoxifylline on drug induced nephropathy were excluded. Nine relevant articles were retrieved and evaluated. The two main populations studied were patients with chronic kidney disease (CKD) and patients with CKD and comorbid type 2 diabetes. Six of the nine studies reported a significant reduction in proteinuria in pentoxifylline treated patients. Four studies reported a significant change in estimated glomerular filtration rate (eGFR). Addition of pentoxifylline to ACE inhibitor and ARB therapy may improve proteinuria in CKD patients. There is conflicting evidence as to whether pentoxifylline will improve kidney function as measured by eGFR. pentoxifylline renoprotection, pentoxifylline CKD , pentoxifylline nephropathy.This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on the issue's contents page.


The Current Directions of Searching for Antiparasitic Drugs.
Parasitic diseases are still a huge problem for mankind. They are becoming the main cause of chronic diseases in the world. Migration of the population, pollution of the natural environment, and climate changes cause the rapid spread of diseases. Additionally, a growing resistance of parasites to drugs is observed. Many research groups are looking for effective antiparasitic drugs with low side effects. In this work, we present the current trends in the search for antiparasitic drugs. We report known drugs used in other disease entities with proven antiparasitic activity and research on new chemical structures that may be potential drugs in parasitic diseases. The described investigations of antiparasitic compounds can be helpful for further drug development.


Neurocytopathic effects of beta-amyloid-stimulated monocytes: a potential mechanism for central nervous system damage in Alzheimer disease.
Growing evidence indicates that cells of the mononuclear phagocyte lineage, which includes peripheral blood monocytes (PBM) and tissue macrophages, participate in a variety of neurodestructive events and may play a pivotal role in neurodegenerative conditions such as Alzheimer disease. The present study sought to determine whether exposure of PBM to beta-amyloid peptide (A beta), the major protein of the amyloid fibrils that accumulate in the brain in Alzheimer disease, could induce cytopathic activity in these cells upon their subsequent incubation with neural tissue. PBM were incubated with A beta for 3 days, centrifuged and washed to remove traces of cell-free A beta, and then applied to organotypic cultures of rat brain for varying periods of time. By using a cell-viability assay to quantitate neurocytopathic effect, an increase in the ratio of dead to live cells was detected in cultures containing A beta-stimulated PBM versus control PBM (stimulated with either bovine serum albumin or reverse A beta peptide) as early as 3 days after coculture. The ratio of dead to live cells increased further by 10 days of coculture. By 30 days of coculture, the dead to live cell ratio remained elevated, and the intensity of neurocytopathic effect was such that large areas of brain mass dissociated from the cultures. These results indicate that stimulation of PBM with A beta significantly heightens their neurocytopathic activity and highlight the possibility that inflammatory reactions in the brain play a role in the neurodegeneration that accompanies Alzheimer disease.


Successful treatment of primary granulocytic sarcoma by non-myeloablative stem cell transplant.
Pre-leukemic granulocytic sarcoma (GS) may pose an initial diagnostic problem and its therapeutic approach has never been formally established. To our knowledge, non-myeloablative stem cell transplantation has been reported in cases of leukemic GS, but not in primary GS. We report a case of primary GS with extensive and aggressive presenting features and successfully treated with intensive chemotherapy followed by non-myeloablative allogeneic stem cell transplant. This resulted in complete remission with minimal complications. Our case demonstrates the potential of graft-vs.-tumour effect in the treatment of GS and suggests that non-myeloablative allogeneic stem cell transplant may be a feasible therapeutic approach for primary GS.


Isolation, characterization and application of bacteriophage PSDA-2 against Salmonella Typhimurium in chilled mutton.
Salmonella is a common foodborne pathogen, especially in meat and meat products. Lytic phages are promising alternatives to conventional methods for Salmonella biocontrol in food and food processing. In this study, a virulent bacteriophage (PSDA-2) against Salmonella enterica serovar Typhimurium was isolated from the sewage and it was found that PSDA-2 belongs to Cornellvirus genus of Siphoviridae family by morphological and phylogenetic analysis. Based on the one-step growth curve, PSDA-2 has a short latent period (10 min) and a high burst size (120 PFU/cell). The stability test in vitro reveals that PSDA-2 is stable at 30-70°C and pH 3-10. Bioinformatics analysis show that PSDA-2 genome consists of 40,062 bp with a GC content of 50.21% and encodes 63 open reading frames (ORFs); no tRNA genes, lysogenic genes, drug resistance genes and virulence genes were identified in the genome. Moreover, the capacity for PSDA-2 to control Salmonella Typhimurium in chilled mutton was investigated. The results show that incubation of PSDA-2 at 4°C reduced recoverable Salmonella by 1.7 log CFU/mL and 2.1 log CFU/mL at multiplicity of infection (MOI) of 100 and 10,000 respectively, as relative to the phage-excluded control. The features of phage PSDA-2 suggest that it has the potential to be an agent to control Salmonella.


Pancreatitis-associated protein (PAP) in patients with pancreatic cancer.
Pancreatitis-associated protein (PAP) is known as a marker for pancreatitis and cystic fibrosis. The aim of our study was to evaluate PAP in patients with pancreatic cancer, to assess its correlation to the extent of the disease and to compare it to CA19-9. This prospective study comprised 75 individuals. Thirty had pancreatic cancer, 30 were healthy controls and 15 had benign lesions of the pancreas. PAP was determined by enzyme-linked immunosorbent assay. Statistical analysis was by Wilcoxon test and Spearman correlation coefficients. As compared to healthy individuals and using a cut-off of 18 micrograms/l corresponding to a sensitivity of 90%, the specificity of PAP for pancreatic cancer was 82.8%. PAP elevation in cancer patients could not be explained by concomitant pancreatitis (p = 0.649). PAP did not show correlation to tumour size (p = 0.14), T-stages (p = 0.706) or tumour grading (p = 0.105), but was significantly correlated to the overall extent of the disease according to the UICC stages (p = 0.002). No correlation between PAP and CA19-9 was seen. Jaundice was not found to influence PAP values (p = 0.4). Elevation of PAP in patients with pancreatic cancer is not merely explainable by concomitant pancreatitis, but seems to be due to increased PAP production by the cancer cells and is also correlated to tumour load as expressed by the UICC stages.


Do hyperechoic thyroid nodules on B-ultrasound represent calcification?
To explore the correlation between hyperechoic thyroid nodules observed on B-ultrasound and histological calcification seen in paraffin-wax sections. Records of patients who underwent surgical removal of thyroid nodules diagnosed on preoperative B-ultrasound were analysed retrospectively. Calcification present on B-ultrasound was compared with calcification seen in postoperative pathology specimens. Of the 1,655 patients included in the study, 518 had malignant and 1,137 had benign thyroid nodules. Calcification on B-ultrasound was seen in 366 patients with malignant, and 414 with benign nodules. Calcification was confirmed on histology in 209 and 127 of these patients, respectively, giving a sensitivity and specificity for B-ultrasound in diagnosing calcification (compared with histology) of 95.87% and 47.67%, respectively, in thyroid cancer and 90.71%, and 71.21% respectively in benign thyroid nodules. Microcalcification was seen in 483 patients on B-ultrasound and in 186 on histology, of whom 294 (60.87%) and 152 (81.72%), respectively, had thyroid cancer. B-ultrasound is a useful and accurate test for detecting calcification in thyroid nodules, with a high sensitivity. There is a close association between calcification (especially microcalcification) and thyroid cancer on both B-ultrasound and pathological examination.


Genomically informed small-molecule drugs overcome resistance to a sustained-release formulation of an engineered death receptor agonist in patient-derived tumor models.
Extrinsic pathway agonists have failed repeatedly in the clinic for three core reasons: Inefficient ligand-induced receptor multimerization, poor pharmacokinetic properties, and tumor intrinsic resistance. Here, we address these factors by (i) using a highly potent death receptor agonist (DRA), (ii) developing an injectable depot for sustained DRA delivery, and (iii) leveraging a CRISPR-Cas9 knockout screen in DRA-resistant colorectal cancer (CRC) cells to identify functional drivers of resistance. Pharmacological blockade of XIAP and BCL-XL by targeted small-molecule drugs strongly enhanced the antitumor activity of DRA in CRC cell lines. Recombinant fusion of the DRA to a thermally responsive elastin-like polypeptide (ELP) creates a gel-like depot upon subcutaneous injection that abolishes tumors in DRA-sensitive Colo205 mouse xenografts. Combination of ELPdepot-DRA with BCL-XL and/or XIAP inhibitors led to tumor growth inhibition and extended survival in DRA-resistant patient-derived xenografts. This strategy provides a precision medicine approach to overcome similar challenges with other protein-based cancer therapies.


Targeted inactivation of CTNNB1 reveals unexpected effects of beta-catenin mutation.
Inactivating mutations of the adenomatous polyposis coli gene (APC) or activating mutations of the beta-catenin gene (CTNNB1) initiate colorectal neoplasia. To address the biochemical and physiologic effects of mutant beta-catenin, we disrupted either the mutant or wild-type CTNNB1 allele in a human colorectal cancer cell line. Cells with only wild-type beta-catenin had decreased colony-forming ability when plated at low density, although their growth was similar to that of parental cells when passaged under routine conditions. Immunohistochemistry and cell-fractionation studies suggested that mutant beta-catenin activity was distinguished primarily by cellular localization and not by protein degradation. Surprisingly, we found mutant beta-catenin bound less well to E-cadherin than did wild-type beta-catenin, and the membranous localization of wild-type and mutant beta-catenin was accordingly distinct. These findings pose several challenges to current models of APC/beta-catenin function.


Air Oxidation of Activated Carbon to Synthesize a Biomimetic Catalyst for Hydrolysis of Cellulose.
Oxygenated carbon catalyzes the hydrolysis of cellulose present in lignocellulosic biomass by utilizing the weakly acidic functional groups on its surface. Here we report the synthesis of a biomimetic carbon catalyst by simple and economical air-oxidation of a commercially available activated carbon. Air- oxidation at 450-500 °C introduced 2000-2400 μmol g(-1) of oxygenated functional groups on the material with minor changes in the textural properties. Selectivity towards the formation of carboxylic groups on the catalyst surface increased with the increase in oxidation temperature. The degree of oxidation on carbon catalyst was found to be proportional to its activity for hydrolysis of cellulose. The hydrolysis of eucalyptus in the presence of carbon oxidized at 475 °C afforded glucose yield of 77 % and xylose yield of 67 %.


Factors associated with malaria in indigenous populations: A retrospective study from 2007 to 2016.
In Brazil malaria is most frequent in the Amazon region, mainly in the Amazonas state, where it is found the most proportion of indigenous people of the whole country. It is remarkable publications about malaria in the Amazon, although information on malaria in indigenous populations is still poorly explored. Identify factors associated with malaria in indigenous populations. Cross-sectional study of positive cases of malaria in the state of Amazonas, Brazil, from 2007 to 2016. Secondary data were obtained from the Epidemiological Surveillance Information System for Malaria and from the Mortality Information System, both from Brazil. To tackle with race missing data, cases with no race fulfilled were classified according to the probable location where infection occurred. This way, was imputed indigenous race for those which the probable infection location was indigenous village (aldeia). Variables tested with race were: sex, age, schooling, microscope surveillance slide type, parasitic infection species, parasitemia level, and timeliness of treatment. Multivariate logistic regression was used. A total of 1,055,852 cases of malaria were notified in the state of Amazonas. Among the factors that associate malaria and indigenous peoples, the most significant were sex, children and high levels of parasitemia. The magnitude of Plasmodium vivax infection is higher than Plasmodium falciparum, although this parasite was more frequent in indigenous than other races. In regards to mortality, 109 deaths were registered, most of them related to P. vivax. The findings underscore the importance of look at indigenous people differently of other races. The associated factors highlight a profile of cases severity, because of highest parasitemia, many cases of P. falciparum although high frequency of P. vivax, and children. Furthermore, the mortality in indigenous, specially in older people is worrying.


Role of human cytomegalovirus UL131A in cell type-specific virus entry and release.
The human cytomegalovirus (HCMV) genes UL128, UL130 and UL131A are essential for endothelial cell infection. Complementation of the defective UL131A gene of the non-endotheliotropic HCMV strain AD169 with wild-type UL131A in cis in an ectopic position restored endothelial cell tropism. The UL131A protein was found in virions in a complex with gH. Coinfection of fibroblasts with UL131A-negative and -positive viruses restored the endothelial cell tropism of UL131A-negative virions by complementing the virions with UL131A protein. Virus entry into endothelial cells, but not into fibroblasts, was blocked by an antipeptide antiserum to pUL131A. AD169, cis-complemented with wild-type UL131A, showed an impaired release of infectious particles from fibroblasts. A comparable defect in virus release was observed when UL131A was expressed ectopically in a virus background already expressing an intact copy of UL131A. In contrast, virus release from infected endothelial cells was not affected by UL131A. These data suggest a dual role for pUL131A in virus entry and virus exit from infected cells.


The neglected role of Faith-based Organizations in prevention and control of COVID-19 in Africa.
The COVID-19 pandemic has exposed health system weaknesses of economically wealthy countries with advanced technologies. COVID-19 is now moving fast across Africa where small outbreaks have been reported so far. There is a concern that with the winter transmission will grow rapidly. Despite efforts of African Governments to promptly establish mitigating measures, rural areas, especially in sub-Saharan Africa, risk being neglected. In those settings, faith-based and other non-governmental organizations, if properly equipped and supported, can play a crucial role in slowing the spread of COVID-19. We describe our experience in two rural health facilities in eSwatini and Ethiopia highlighting the struggle towards preparedness and the urgency of international support to help prevent a major public health disaster.


[How I treat cryptorchidism].
The author describes the technic of septal orchidopexy, which he use for the treatment of cryptorchidism with palpable testis. The question of high level cryptorchidism is raised.


Measures of contextuality and non-contextuality.
We discuss three measures of the degree of contextuality in contextual systems of dichotomous random variables. These measures are developed within the framework of the Contextuality-by-Default (CbD) theory, and apply to inconsistently connected systems (those with 'disturbance' allowed). For one of these measures of contextuality, presented here for the first time, we construct a corresponding measure of the degree of non-contextuality in non-contextual systems. The other two CbD-based measures do not suggest ways in which degree of non-contextuality of a non-contextual system can be quantified. We find the same to be true for the contextual fraction measure developed by Abramsky, Barbosa and Mansfield. This measure of contextuality is confined to consistently connected systems, but CbD allows one to generalize it to arbitrary systems. This article is part of the theme issue 'Contextuality and probability in quantum mechanics and beyond'.


Influence of Concentration Levels of β-Tricalcium Phosphate on the Physical Properties of a Dental Adhesive.
Our study assessed the influence of integrating 5% and 10% tricalcium phosphate (β-TCP-Ca3(PO4)2.) nanoparticles into a dental adhesive on the adhesive's bonding. To evaluate the filler nanoparticles, scanning electron microscopy (SEM), Energy Dispersive X-Ray (EDX) spectroscopy, Fourier-transform infrared (FTIR) spectroscopy, and micro-Raman spectroscopy techniques were used. Shear Bond strength (SBS) testing, degree of conversion (DC) analysis, investigation of the adhesive-dentin interface, and biofilm experiments were conducted. The SEM micrographs revealed non-uniform agglomerates, while the EDX demonstrated the existence of oxygen 'O' (24.2%), phosphorus 'P' (17.4%) and calcium 'Ca' (60.1%) in the β-TCP nanoparticles. The FTIR and micro-Raman spectra indicated characteristic bands for β-TCP containing materials. The 10 wt.% β-TCP adhesive presented the highest SBS values (NTC-10 wt.% β-TCP: 33.55 ± 3.73 MPa, TC-10 wt.% β-TCP: 30.50 ± 3.25 MPa), followed by the 5 wt.% β-TCP adhesive (NTC-5 wt.% β-TCP: 32.37 ± 3.10 MPa, TC-5 wt.% β-TCP: 27.75 ± 3.15 MPa). Most of the detected failures after bond strength testing were adhesive in nature. The β-TCP adhesives demonstrated suitable dentin interaction by forming a hybrid layer (with few or no gaps) and resin tags. The β-TCP adhesives (10 wt.%) revealed lower DC values compared to control. The incorporation of 5 and 10 wt.% concentrations of β-TCP particles resulted in an increase in SBS values. A linear decline in DC values was witnessed when the nanoparticle concentration was increased. Further research focusing on exploring the influence of higher filler concentrations on adhesive's properties is recommended.


Diabetes Effects on Dental Implant Survival.
The goal of this review is to critically appraise the clinical evidence guiding our application of dental implant therapy relative to glycemic control for patients with diabetes. Our initial searches of the literature identified 129 publications relevant to both dental implants and diabetes. These were reduced to 17 clinical studies for inclusion. Reported implant failure rates in these 17 reports ranged from 0 to 14.3% for patients with diabetes. Unfortunately, the majority of these reports lacked sufficient information relative to glycemic control to allow the application of the findings toward clinical care. However, clinical evidence is emerging from several investigations that diabetes and glycemic control are important considerations that may require modifications to therapeutic protocols, but may not be contraindications to implant therapy in diabetes patients. Also, a potentially important role for implant therapy to support oral function in diabetes dietary management remains to be determined.


Altered phenotype of regulatory T cells associated with lack of human immunodeficiency virus (HIV)-1-specific suppressive function.
Mechanisms by which CD4+ regulatory T cells (T(regs)) mediate suppression of virus-specific responses remain poorly defined. Adenosine, mediated via CD39 and CD73, has been shown to play a role in the action of murine T(regs) . In this study we investigate the phenotype of T(regs) in the context of human immunodeficiency virus (HIV)-1 infection, and the function of these cells in response to HIV-1-Gag and cytomegalovirus (CMV) peptides. Phenotypic data demonstrate a decrease in forkhead box transcription factor 3 (FoxP3+) T(reg) numbers in the peripheral blood of HIV-1+ individuals compared to healthy controls, which is most pronounced in those with high HIV-1 RNA plasma load. Due to aberrant expression of CD27 and CD127 during HIV-1 disease, these markers are unreliable for T(reg) identification. The CD3+ CD4+ CD25(hi) CD45RO+ phenotype correlated well with FoxP3 expression in both the HIV-1+ and seronegative control cohorts. We observed expression of CD39 but not CD73 on T(regs) from HIV-1+ and healthy control cohorts. We demonstrate, through T(reg) depletion, the suppressive potential of T(regs) over anti-CMV responses in the context of HIV-1 infection; however, no recovery of the HIV-1-specific T cell response was observed indicating a preferential loss of HIV-1-specific T(reg) function. We propose that before immunotherapeutic manipulation of T(regs) is considered, the immunoregulatory profile and distribution kinetics of this population in chronic HIV-1 infection must be elucidated fully.


Beamformed nearfield imaging of a simulated coronary artery containing a stenosis.
This paper is concerned with the potential for the detection and location of an artery containing a partial blockage by exploiting the space-time properties of the shear wave field in the surrounding elastic soft tissue. As a demonstration of feasibility, an array of piezoelectric film vibration sensors is placed on the free surface of a urethane mold that contains a surgical tube. Inside the surgical tube is a nylon constriction that inhibits the water flowing through the tube. A turbulent field develops in and downstream from the blockage, creating a randomly fluctuating pressure on the inner wall of the tube. This force produces shear and compressional wave energy in the urethane. After the array is used to sample the dominant shear wave space-time energy field at low frequencies, a nearfield (i.e., focused) beamforming process then images the energy distribution in the three-dimensional solid. Experiments and numerical simulations are included to demonstrate the potential of this noninvasive procedure for the early identification of vascular blockages-the typical precursor of serious arterial disease in the human heart.


Characteristics of uncontrolled hemorrhage in cardiac surgery.
Patients with uncontrolled hemorrhage require massive transfusion therapy and consume a large fraction of blood bank resources. Institutional guidelines have been established for treatment, but early identification and prevention in susceptible patients remains challenging. Uncontrolled hemorrhage was defined as meeting institutional guidelines for recombinant FVIIa administration. Patients who received rFVIIa were compared with patients who did not require the therapy but who were operated on during the same time period. After institutional review board approval, demographic, operative, and transfusion data were analyzed from a prospective database. Patients receiving rFVIIa were more likely to undergo multiple procedures (2.6 +/- 0.8 vs. 1.8 +/- 0.8; p < .001); aortic surgery (59% vs. 11%; p < .005); have a higher Cleveland Clinic Clinical Severity score (7.8 +/- 2.7 vs. 5.5 +/- 4.0; p < .005); require longer bypass (265 +/- 92 min vs. 159 +/- 63 min; p < .001), cross-clamp (182 +/- 68 min vs. 112 +/- 56 min; p < .001), and circulatory arrest (15 +/- 24 min vs. 2 +/- 7 min; p < .05) times; and require more autotransfusion (2580 +/- 1847 mL vs. 690 +/- 380 mL; p < .05). Uncontrolled hemorrhage is associated with more complex surgery requiring longer bypass times and more autotransfusion.


Characterization of the enhancer region for germline transcription of the gamma 3 constant region gene of human immunoglobulin.
A constant region gene (C) of the immunoglobulin heavy chain can be transcribed as germline transcripts from a promoter located upstream of a switch region. We have studied the structure and function of the human C gamma 3 promoter region. When the human IgM-producing cell line SSK41 is stimulated with interleukin 4 (IL-4) in the presence of phorbol myristate acetate (PMA) or Staphylococcus aureus Cowan I, expression of germline C gamma 3 transcripts was specifically augmented within 4 h. Upstream DNA fragments flanking the I gamma 3 exon were fused with a reporter gene and tested for IL-4-induced promoter/enhancer activity by transfection of SSK41 cells. The DNA fragment between 450 and 250 bp upstream of the transcription initiation site of the C gamma 3 gene was shown to be required for transcriptional up-regulation by PMA and IL-4. The upstream 514 bp fragment of the I gamma 3 flanking region was shown to contain an enhancer activity in response to PMA and IL-4.


Follistatin promotes LIN28B-mediated supporting cell reprogramming and hair cell regeneration in the murine cochlea.
Hair cell (HC) loss within the inner ear cochlea is a leading cause for deafness in humans. Before the onset of hearing, immature supporting cells (SCs) in neonatal mice have some limited capacity for HC regeneration. Here, we show that in organoid culture, transient activation of the progenitor-specific RNA binding protein LIN28B and Activin antagonist follistatin (FST) enhances regenerative competence of maturing/mature cochlear SCs by reprogramming them into progenitor-like cells. Transcriptome profiling and mechanistic studies reveal that LIN28B drives SC reprogramming, while FST is required to counterbalance hyperactivation of transforming growth factor-β-type signaling by LIN28B. Last, we show that LIN28B and FST coactivation enhances spontaneous cochlear HC regeneration in neonatal mice and that LIN28B may be part of an endogenous repair mechanism that primes SCs for HC regeneration. These findings indicate that SC dedifferentiation is critical for HC regeneration and identify LIN28B and FST as main regulators.


Epstein-Barr Virus: Diseases Linked to Infection and Transformation.
Epstein-Barr virus (EBV) was first discovered in 1964, and was the first known human tumor virus now shown to be associated with a vast number of human diseases. Numerous studies have been conducted to understand infection, propagation, and transformation in various cell types linked to human diseases. However, a comprehensive lens through which virus infection, reactivation and transformation of infected host cells can be visualized is yet to be formally established and will need much further investigation. Several human cell types infected by EBV have been linked to associated diseases. However, whether these are a direct result of EBV infection or indirectly due to contributions by additional infectious agents will need to be fully investigated. Therefore, a thorough examination of infection, reactivation, and cell transformation induced by EBV will provide a more detailed view of its contributions that drive pathogenesis. This undoubtedly expand our knowledge of the biology of EBV infection and the signaling activities of targeted cellular factors dysregulated on infection. Furthermore, these insights may lead to identification of therapeutic targets and agents for clinical interventions. Here, we review the spectrum of EBV-associated diseases, the role of the encoded latent antigens, and the switch to latency or lytic replication which occurs in EBV infected cells. Furthermore, we describe the cellular processes and critical factors which contribute to cell transformation. We also describe the fate of B-cells and epithelial cells after EBV infection and the expected consequences which contribute to establishment of viral-associated pathologies.


[Smell impairment in COVID-19 patients: mechanisms and clinical significance].
Numerous studies have shown that loss of smell is a serious symptom that requires careful differential diagnosis. There is convincing evidence that a violation of the sense of smell is not so much a sign of the pathology of the nasal cavity and paranasal sinuses, but it can be a manifestation of neurodegenerative diseases. Some patients with the identified SARS-CoV-2 virus have neurological symptoms. Most of them are not specific - headache, dizziness, fatigue, myalgia. A small percentage of patients with COVID-19 infection have convulsions, impaired consciousness, and the presence of 2019-NCoV RNA was found in the cerebrospinal fluid. Data on the development of new symptoms of the disease, in the form of anosmia and dysgeusia, are presented.


A method to infer positive selection from marker dynamics in an asexual population.
The observation of positive selection acting on a mutant indicates that the corresponding mutation has some form of functional relevance. Determining the fitness effects of mutations thus has relevance to many interesting biological questions. One means of identifying beneficial mutations in an asexual population is to observe changes in the frequency of marked subsets of the population. We here describe a method to estimate the establishment times and fitnesses of beneficial mutations from neutral marker frequency data. The method accurately reproduces complex marker frequency trajectories. In simulations for which positive selection is close to 5% per generation, we obtain correlations upwards of 0.91 between correct and inferred haplotype establishment times. Where mutation selection coefficients are exponentially distributed, the inferred distribution of haplotype fitnesses is close to being correct. Applied to data from a bacterial evolution experiment, our method reproduces an observed correlation between evolvability and initial fitness defect.


Tracking chromosomal positions of oligomers - a case study with Illumina's BovineSNP50 beadchip.
High density genotyping arrays have become established as a valuable research tool in human genetics. Currently, more than 300 genome wide association studies were published for human reporting about 1,000 SNPs that are associated with a phenotype. Also in animal sciences high density genotyping arrays are harnessed to analyse genetic variation. To exploit the full potential of this technology single nucleotide polymorphisms (SNPs) on the chips should be well characterized and their chromosomal position should be precisely known. This, however, is a challenge if the genome sequence is still subject to changes. We have developed a mapping strategy and a suite of software scripts to update the chromosomal positions of oligomer sequences used for SNP genotyping on high density arrays. We describe the mapping procedure in detail so that scientists with moderate bioinformatics skills can reproduce it. We furthermore present a case study in which we re-mapped 54,001 oligomer sequences from Ilumina's BovineSNP50 beadchip to the bovine genome sequence. We found in 992 cases substantial discrepancies between the manufacturer's annotations and our results. The software scripts in the Perl and R programming languages are provided as supplements. The positions of oligomer sequences in the genome are volatile even within one build of the genome. To facilitate the analysis of data from a GWAS or from an expression study, especially with species whose genome assembly is still unstable, it is recommended to update the oligomer positions before data analysis.


Endomyocardial biopsies after heart transplantation. The presence of markers indicative of activation.
A series of 104 endomyocardial biopsies (EMB) from patients after heart transplantation was evaluated for the presence of immunological markers on graft component and infiltrating cells. This included markers for cells expressing alpha beta-T-cell receptors and gamma delta-T-cell receptors, and cytotoxic T cells with granules bearing the serine esterase Granzyme B; the presence of activation markers identified by CD25 (interleukin 2 receptor), CD30, CD69 (activation inducer molecule), CDw70; macrophages using antibody CD14 (WT14), and cells with Fc gamma-receptors type III (CD16). Almost all cells in T-cell infiltrates expressed the alpha beta-T cell receptor. Cells bearing the gamma delta-T cell receptor were scarcely found. The analysis with respect to the histopathologic diagnosis for rejection showed an absence of significance for T cell subsets, Granzyme B-positive cells, and activation markers except CD25. The numbers of macrophages labeled by CD14 and cells expressing Fc gamma RIII showed a significant relation to histopathology of rejection. Apart from leukocytes, also endothelium in EMB with rejection was labeled by the two anti-Fc gamma RIII antibodies used. In addition, in a small series of biopsies investigated, Fc gamma RI- and Fc gamma RII-positive cells were increased in EMB with rejection, and endothelium was labeled by Fc gamma RII antibodies. A cluster analysis on the basis of scores for CD25, CD14, and anti-Fc gamma RIII revealed three main clusters, one cluster comprising biopsies without abnormalities, one cluster containing EMB with the histopathology of rejection and high scores in immunophenotyping for lymphocytes and macrophages, and one cluster in between. The present data emphasize the importance of macrophage assessment in evaluating pathologic processes during rejection of heart allografts and diagnosing rejection.


Urdu translation of Shoulder Pain and Disability Index (SPADI) and its validity and reliability on adhesive capsulitis patients.
Excessive scar tissues around the shoulder are the results of shoulder pathologies that lead to pain and disability. The Shoulder pain and disability index (SPADI) is used to measure the level of pain and disability in patients with shoulder pathology. SPADI is translated into Urdu and its validity and reliability are measured on patients with adhesive capsulitis. The study was aimed to translate the SPADI in Urdu and to evaluate its reliability and validity in patients with shoulder adhesive capsulitis. Translation of SPADI in Urdu was conducted by applying the standardized process. Two forward translations in Urdu were made T1 and T2 by bilingual translators. Urdu version of SPADI was drafted after experts' opinion. Two Backward English translations of Urdu SPADI were made BT1 and BT2 and the back translation was finalized by the consensus of all experts. After this process of reviewing by the professional experts, 3rd version of Urdu SPADI was drafted. The Final version was drafted after its application on 10 patients. Its reliability and validity were tested on 150 patients with shoulder adhesive capsulitis. Content Validity Index was good with values of each item > 0.85. For Test-retest reliability, the Intraclass correlation coefficient (ICC2,1) was measured with a value of 0.89 which showed good Test-retest reliability. The internal consistency and reliability of SPADI were calculated by Cronbach's alpha for a total score with a value of 0.94. Construct validity and Concurrent validity were determined. In Construct validity, factor analysis of Urdu SPADI showed two factors and a cumulative variance of 75.443%. It was concluded that the Urdu version of SPADI is a valuable translation that is a valid assessment tool for patients with shoulder adhesive capsulitis. It has good validity and test-retest reliability.


The Mediating Role of Parental Influence on the Relationship Between Adult Children's Filial Piety Beliefs and Mate Preferences.
In the present study, we aim to examine the mediating roles of the two types of perceived parental influence in the relation between the dual filial piety model and adult children's long-term mate preferences. A survey was administered to 499 Chinese adult children on their filial piety beliefs, perceptions about parental influence, and long-term mate preferences. Structural equation modeling revealed the following findings. (a) For both genders, reciprocal filial piety was positively correlated with good father/mother traits, and authoritarian filial piety was positively correlated with good gene traits. (b) For both genders, sensitivity from the perceived parental influence mediated the link between authoritarian filial piety and good provider traits. (c) For females, sensitivity mediated the link between authoritarian filial piety and good gene traits. (d) For males, parental monitoring from perceived parental influence mediated the link between authoritarian filial piety and good gene and good mother traits. In addition, authoritarian filial piety was positively correlated with good provider and good mother traits. In summary, filial piety can link both directly and indirectly to mate preference through parental sensitivity for males and females and through parental monitoring mainly for males.


Resonant plasmon nanofocusing by closed tapered gaps.
We study radiation nanofocusing by closed tapered gaps, i.e. metal V-grooves, under normal illumination, and discover that the local field inside a groove can be resonantly enhanced due to interference of counter-propagating gap plasmons. Considering V-grooves milled in gold, we analyze this phenomenon theoretically, deriving an analytic expression for the resonance condition and predicting more than 550-fold intensity enhancements at resonance, and observe it experimentally with two-photon photoluminescence microscopy, demonstrating more than 100-fold intensity enhancements.


[Idiopathic retroperitoneal fibrosis diagnosed by CT-guided needle biopsy: a case report].
A 72-year-old man was admitted to another hospital because of general fatigue and dyspnea secondary to renal insufficiency. Abdominal computed tomography (CT) and magnetic resonance imaging showed left atrophic kidney, right hydronephrosis, and an intra-pelvic mass of soft-tissue density located anterior to the sacrum, involving the right ureter at the level between the 5th lumber vertebra and the sacrum. He was referred to our hospital after percutaneous nephrostomy was constructed into right kidney. CT-guided needle biopsy revealed the idiopathic retroperitoneal fibrosis, leading us to give him steroid therapy. Three weeks later, radiographic findings showed a remarkable reduction of the mass and the improvement of the right ureteral stricture.


Evaluation of Antimicrobial Photodynamic Therapy Using Indocyanine Green and Near-Infrared Diode Laser Against Enterococcus faecalis in Infected Human Root Canals.
Photodynamic therapy (PDT) is considered a promising adjunct to the currently available endodontic disinfection techniques leading to more effective reduction of intracanal bacteria. The present ex vivo study aimed to assess the antimicrobial effect of PDT using indocyanine green (ICG) as photosensitizer and a near-infrared (NIR) diode laser in root canals of human teeth infected with Enterococcus faecalis. Ninety single-rooted teeth after chemomechanical preparation and sterilization were contaminated with an E. faecalis strain. The specimens were divided, randomly, into eight experimental groups: (1) PDT with ICG and laser (0.5 W output power-medium-energy fluence), (2) PDT with ICG and laser (1 W output power-high-energy fluence), (3) only laser emission, (4) only ICG, (5) 2.5% sodium hypochlorite (NaOCl) as irrigant, (6) 2.5% NaOCl and PDT with ICG and laser, (7) no treatment (positive control), and (8) no bacterial biofilm growth (negative control). The root canal contents were sampled by flushing and the collected washings were plated on an appropriate culture medium, which was incubated for 48 h at 35°C ± 2.0. The colony-forming units (CFUs) were determined to assess the bactericidal effect of the tested experimental combinations. The microbiological tests revealed that PDT groups, regardless of the overall power, showed significant lower mean log10 CFU levels, than groups 3 and 4 (p < 0.001) and similar reduction of viable counts with group 5. The combination treatment (group 6) achieved adequate reduction of log10 CFU levels in the viable counts. However, no significant difference (p > 0.05) was observed between groups 1, 2, 5, and 6 and significant difference was noticed between groups 3, 4, and 5 (p < 0.001). ICG-mediated PDT activated by an NIR diode laser provided increased disinfection of the root canal system, but the overall benefit in total bacterial elimination should be further investigated.


Natural history of motor symptoms in Parkinson's disease and the long-duration response to levodopa.
The natural pattern of progression of Parkinson's disease is largely unknown because patients are conventionally followed on treatment. As Parkinson's disease progresses, the true magnitude of the long-duration response to levodopa remains unknown, because it can only be estimated indirectly in treated patients. We aimed to describe the natural course of motor symptoms by assessing the natural OFF in consecutive Parkinson's disease patients never exposed to treatment (drug-naïve), and to investigate the effects of daily levodopa on the progression of motor disability in the OFF medication state over a 2-year period. In this prospective naturalistic study in sub-Saharan Africa, 30 Parkinson's disease patients (age at onset 58 ± 14 years, disease duration 7 ± 4 years) began levodopa monotherapy and were prospectively assessed using the Unified Parkinson's disease Rating Scale (UPDRS). Data were collected at baseline, at 1-year and 2-years follow-up. First-ever levodopa intake induced a significant improvement in motor symptoms (natural OFF versus ON state UPDRS-III 41.9 ± 15.9 versus 26.8 ± 15.1, respectively; P < 0.001). At 1-year follow-up, OFF state UPDRS-III score after overnight withdrawal of levodopa was considerably lower than natural OFF (26.5 ± 14.9; P < 0 .001). This effect was not modified by disease duration. At the 2-year follow-up, motor signs after overnight OFF (30.2 ± 14.2) were still 30% milder than natural OFF (P = 0.001). The ON state UPDRS-III at the first-ever levodopa challenge was similar to the overnight OFF score at 1-year follow-up and the two conditions were correlated (r = 0.72, P < 0.001). Compared to the natural progression of motor disability, levodopa treatment resulted in a 31% lower annual decline in UPDRS-III scores in the OFF state (3.33 versus 2.30 points/year) with a lower model's variance explained by disease duration (67% versus 36%). Using the equation regressed on pretreatment data, we predicted the natural OFF at 1-year and 2-year follow-up visits and estimated that the magnitude of the long-duration response to levodopa ranged between 60% and 65% of total motor benefit provided by levodopa, independently of disease duration (P = 0.13). Although levodopa therapy was associated with motor fluctuations, overnight OFF disability during levodopa was invariably less severe than the natural course of the disease, independently of disease duration. The same applies to the yearly decline in UPDRS-III scores in the OFF state. Further research is needed to clarify the mechanisms underlying the long-duration response to levodopa in Parkinson's disease. Understanding the natural course of Parkinson's disease and the long-duration response to levodopa may help to develop therapeutic strategies increasing its magnitude to improve patient quality of life and to better interpret the outcome of randomized clinical trials on disease-modifying therapies that still rely on the overnight OFF to define Parkinson's disease progression.


[Analysis of risk factors for reversible posterior leukoencephalopathy syndrome in lupus].
To analyze the different risk factors for reversible posterior leukoencephalopathy syndrome (RPLS) in systemic lupus erythematosus (SLE). The clinical and imaging characteristics of 2 cases were described in details. The literature reports for 33 cases of SLE and RPLS were reviewed systematically. The etiologies of SLE and RPLS were analyzed for all 35 cases. Thirty-five cases of SLE patients were complicated with RPLS, 32 cases suffered hypertension, 3 cases had no hypertension, 1 case was explicitly related with rituximab and 1 case was related with cyclosporine therapy. SLE complicated by RPLS has different causes, such as hypertension, SLE involving central nervous system and drug neurotoxicity.


A descriptive study of the blood vessels of the sciatic nerve in the rat, man and other mammals.
The gross blood supply and internal vascular organization of the sciatic nerve in the rat (and other mammals including man) have been studied by injection, dissection, and microradiographic methods, in thick sections stained for alkaline phosphatase and semithin sections, and by electron microscopy. In the rat, an anastomotic vascular connection from the arteria comitans to the medial femoral circumflex artery is illustrated. In all species studied, the microvascular network of the nerve has an unusually large calibre and wide spacing, particularly when compared with that of skeletal muscle. Arterioles and venules are present within fascicles; the arterioles are thin-walled, with only rudimentary internal elastic laminae. Endoneurial capillaries are large, continuous and nonfenestrated, with prominent (often multiple) basal laminae and an unusually complete pericyte investment. Endothelial junctions appear to vary in type; not all are tight. Pinocytosis is common. The extensively interconnected vascular network of the nerve fascicle may act as an adaptable in situ reservoir for blood in the precise maintenance of the endoneurial milieu.


National health expenditures: short-term outlook and long-term projections.
This paper presents projections of national health expenditures by type of expenditure and source of funds for 1981, 1985, and 1990. Rapid growth in national health expenditures is projected to continue through 1990. National health expenditures increased 400 percent between 1965 and 1979, reaching $212 billion in 1979. As a proportion of the Gross National Product (GNP), health expenditures rose from 6.1 percent to 9.0 percent between 1965 and 1979. They are expected to continue to rise, reaching 10.8 percent by 1990. This study projects that, under current legislation, national health expenditures will research $279 billion in 1981, $462 billion in 1985, and $821 billion in 1990. Sources of payments for these expenditures are shifting. From 1965 to 1979, the percentage of total health expenditures financed by public funds increased 17 percentage points--from 26 to 43 percent. The Federal share of public funds during this same period grew rapidly, from 51 percent in 1965 to 67 percent in 1979. This study projects that in 1985 approximately 45 percent of total health spending will be financed from public funds, of which 68 percent will be paid for by the Federal government. Public funds will account for 46 percent of total national health expenditure by 1990.


Loss of bone resulting from accumulation of aluminum in bone of patients undergoing dialysis.
The effects of accumulation of aluminum on cancellous bone mass and histomorphometric parameters of bone formation and resorption are evaluated in three separate studies: a cross-sectional study in 120 patients receiving long-term maintenance dialysis, including patients with and without stainable bone aluminum; a longitudinal retrospective study in eight patients receiving hemodialysis, who had progressive accumulation of aluminum in bone. These patients had two bone biopsies 11 to 16 months apart; a longitudinal prospective study in 10 patients receiving long-term hemodialysis, who had histologically proved renal osteodystrophy and stainable bone aluminum. These patients were given deferoxamine for 9 to 12 months. Repeat bone biopsies were done thereafter, and decrease or disappearance of stainable bone aluminum was observed. The results of the independent studies demonstrate that aluminum accumulation in bone is associated not only with disturbed mineralization but also with loss of cancellous bone mass, and that removal of aluminum from bone results in gain in bone volume. The mechanisms of this phenomenon are related to a disproportionately greater effect of aluminum on bone formation than on bone resorption. The negative effect on bone formation is caused by decreased number and activity of osteoblasts, which is reversed by removal of aluminum from bone. In addition, the data show that aluminum induces suppression of bone turnover, with uncoupling between bone formation and resorption whereby the effects on bone formation exceed those on bone resorption. Thus, fractures in patients with renal failure and accumulation of aluminum may result not only from osteomalacia but also from osteopenia.


Perineal biofeedback versus pelvic floor training in the treatment of urinary incontinence.
Perineal floor training and perineal biofeedback allow to reach a good improvement of clinical discomfort in the treatment of stress and urge urinary incontinence. The aim of this study is the real evaluation of the benefit due to a 6 week perineal biofeedback and pelvic floor training (PFT) versus PFT alone. 8 female patients have been treated with a 6 week trial with perineal biofeedback plus PFT trial and 22 with a 3 month PFT alone one. PFT alone, permits a good improvement but 10% less than if associated to perineal biofeedback.


Cytotoxicity of eight cigarette smoke condensates in three test systems: comparisons between assays and condensates.
Cytotoxic properties of tobacco smoke are associated with chronic tobacco-related diseases. The cytotoxicity of tobacco smoke can be tested with short-term predictive assays. In this study, we compare eight mainstream cigarette smoke condensates (CSCs) from commercial and experimental cigarettes in three different cytotoxicity assays with unique and overlapping endpoints. The CSCs demonstrated cytotoxicity in all assays. In the multiple cytotoxicity endpoint (MCE) assay with TK-6 cells, the cigarette varieties that had the highest EC50s for reduced cell growth also showed a positive dose-response relationship for necrotic cells. In the IdMOC multiple cell-type co-culture (MCTCC) system, all CSCs reduced the viability of the cells. Low concentrations of some CSCs had a stimulatory effect in lung microvascular endothelial cells and small airway epithelial cells. In the neutral dye assay (NDA), except for a 100% flue-cured tobacco CSC, there was little consistency between CSCs producing morphological evidence of moderate or greater toxicity and the CSCs with the lowest EC50s in the MCE or MCTCC assays. Overall, cigarettes made with flue-cured tobacco were the most cytotoxic across the assays. When results were expressed on a per-mg of nicotine basis, lower tar cigarettes were the most cytotoxic in primary human respiratory cells.


Growth and chemical defense in willow seedlings: trade-offs are transient.
Many studies have failed to detect costs of defense and some have even found a positive correlation between growth and the concentrations of chemical defenses. These studies contradict the theoretical assumption that anti-herbivore defenses are costly-produced at the expense of growth and/or reproduction. Costs, however, may be transient and therefore difficult to detect. Here we tested the hypothesis that costs of defense would be pronounced early in development when root growth is prioritized (high percent root allocation), but not later in development. To test this hypothesis, we grew F(2) hybrid willow seedlings from five different families, and harvested cohorts of even-aged seedlings after 6, 7, 8 and 9 weeks of growth. Seedlings were divided into root and shoot tissue and shoots were analyzed for phenolics (condensed tannins and phenolic glycosides). We found evidence for transient costs of defense. The concentrations of phenolics were negatively correlated with total biomass, shoot biomass, and the proportion of biomass allocated to roots in week 6. After week 6, however, the concentrations of phenolics were positively correlated with shoot biomass and total biomass, while phenolics were uncorrelated with the proportion of biomass allocated to roots. These results, the first ever, to our knowledge, with woody plants, suggest that costs of defense were transient; specifically, costs were found in early development, when root establishment was a priority. Our findings suggest that studies should focus more on trade-offs early in plant development.


Macrophage migration inhibitory factor induces ICAM-1and thrombomobulin expression in vitro.
Macrophage migration inhibitory factor (MIF) is an important cytokine in the modulation of inflammatory and immune responses, but its role in coagulation remains to be elucidated. In this study, we investigated the potential role of MIF in coagulation through its influence on two factors, thrombomodulin (TM) and intercellular adhesion molecule-1 (ICAM-1). Recombinant human MIF was added to human microvascular endothelial cell line (HMEC-1) to investigate its influence on the expression of TM and ICAM-1. The results showed that both TM and ICAM-1 were induced with MIF addition in a dose-dependent and time-dependent manner. The expression of ICAM-1 and TM was increased as MIF doses were increased, with the highest expression seen at 12 hr after 400 ng/ml of MIF treatment. Besides, anti-MIF antibody treatment reduced the TM expression in HMEC-1 cells. In conclusion, our data support a role of MIF as an important factor in the regulation of coagulation.


Pentastomiasis.
Pentastomiasis is a parasitic zoonosis caused by pentastomes, members of an unusual phylum--Pentastomida--with characteristics of both arthropods and annelids. Adult pentastomes parasitize the respiratory tracts of reptiles or carnivorous mammals. The infection is generally limited to the tropics and subtropics, but ocular involvement has been reported in the southern United States. The majority of human pentastomiasis is caused by two species. The first, Armillifer armillatus, infects humans as secondary hosts. Infection is usually asymptomatic but has characteristic postmortem and radiologic features. The second, Linguatula serrata, can infect humans as does Armillifer or can cause a self-limited nasopharyngitis--the halzoun or marrara syndrome--with the human acting as a temporary definitive host. This article discusses the biology and parasitology of these metazoans as well as the clinical manifestations, pathology, diagnosis, and epidemiology of pentastomiasis.


The impact of age-related cataract on measures of frailty in an aging global population.
To review the associations among age-related cataract, frailty, and frailty outcomes (e.g., disabilities). It is predicted that the proportion of the population aged 65 and older, in developed and developing nations alike, will rise until at least 2050. The proportion of patients suffering from cataracts and frailty is expected to increase, as are age-related diseases. Although there are many papers reporting on the association between frailty outcomes, cataract, and visual impairment, there is a relative paucity of papers describing associations between frailty markers, cataract, and visual impairment. Reports regarding the relationship between frailty, visual impairment, cataract, and cataract surgery are limited, but gradually increasing. Further research is expected to clarify the mechanism of visual function or the impact of restored vision on frailty. Evidence for the effect of cataract on frailty and frailty outcomes after restoring vision by cataract surgery remains limited.


Simulated temporary hypoxia triggers the release of CD31+/Annexin+ endothelial microparticles: A prospective pilot study in humans.
Endothelial microparticles (EMP) are small membrane vesicles that originate from activated or apoptotic endothelial cells. Although the exact mechanism of EMP function is still relatively unknown, it has been shown that they modulate inflammatory processes, coagulation and vascular function. In this study we hypothesized that transient hypoxia may act as a trigger for the release of EMP into circulation. Fourteen healthy volunteers were subjected to transient normobaric hypoxia in an air-conditioned chamber simulating an oxygen concentration of a height of up to 5500 meters. Blood samples were evaluated for EMP using flow cytometry. During the experiment oxygen concentration was adjusted to a value equivalent to a height of 5500 meters to achieve hypoxic conditions. Oxygen saturation decreased to 78% . At the final height a significant increase of CD31+/Annexin+ EMP levels was evident (increase from 0.03% ± 0.01% SEM to 0.12% ± 0.04% SEM, p = 0.0188). These experimental results show that temporary hypoxic conditions can trigger the release of CD31+/ Annexin+ EMP also in healthy volunteers. In our previous studies we have shown that apoptotic bodies can confer pro-survival signals to cardiomyocytes during myocardial ischemia. Based on the experimental results of this current study we believe that the release of CD31+/Annexin+ EMP during hypoxia might act as an endogenous survival signal.


Device for microparticle array preparation.
A simple device is described for preparing arrays of microballoons to be characterized by radiography for use as laser fusion fuel containers.


Noncovalent interactions of Zn+ with N-donor ligands (pyridine, 4,4'-dipyridyl, 2,2'-dipyridyl, and 1,10-phenanthroline): collision-induced dissociation and theoretical studies.
The binding interactions in complexes of Zn(+) with nitrogen donor ligands, (N-L) = pyridine (x = 1-4), 4,4'-dipyridyl (x = 1-3), 2,2'-dipyridyl (x = 1-2), and 1,10-phenanthroline (x = 1-2), are examined in detail. The bond dissociation energies (BDEs) for loss of an intact ligand from the Zn(+)(N-L)(x) complexes are reported. Experimental BDEs are obtained from thermochemical analyses of the threshold regions of the collision-induced dissociation cross sections of Zn(+)(N-L)(x) complexes. Density functional theory calculations at the B3LYP/6-31G* level of theory are performed to determine stable structures of these species and to provide molecular parameters needed for the thermochemical analysis of experimental data. Relative stabilities of the various conformations of these N-donor ligands and their complexes to Zn(+) as well as theoretical BDEs are determined from single point energy calculations at the B3LYP/6-311+G(2d,2p) and M06/6-311+G(2d,2p) levels of theory using the B3LYP/6-31G* optimized geometries. The experimental BDEs for the Zn(+)(N-L)(x) complexes are in reasonably good agreement with values derived from density functional theory calculations. BDEs derived from M06 calculations provide better agreement with the measured values than those based on B3LYP calculations. Trends in the sequential BDEs are explained in terms of sp polarization of Zn(+) and repulsive ligand-ligand interactions. Comparisons are made to the analogous Cu(+)(N-L)(x) and Ni(+)(N-L)(x) complexes previously studied.


Heat Shock Proteins in Tobacco Cell Suspension during Growth Cycle.
Tobacco (Nicotiana tabacum L. cv Wisconsin 38) cells grown in suspension culture at 26 degrees C produce heat shock proteins (HSPs) when exposed to elevated temperature of 34 to 42 degrees C. At 34 and 38 degrees C, synthesis of normal proteins is maintained while HSPs are expressed within 30 minutes after initiation of the shock. At 42 degrees C, HSPs are still expressed but normal proteins are made at a reduced rate or not at all. Exposure of cells to 38 degrees C allows for a full expression of HSPs without inhibition of the synthesis of normal proteins. Induced synthesis of HSPs at 38 degrees C is maximal 1 to 2 hours after elevation of temperature and diminishes thereafter through at least 6 hours. Cells growing asynchronously in the logarithmic phase of growth produce HSPs at a much higher rate than those in the stationary phase. The ability to synthesize HSPs disappears about one generation time before the cells reach a growth plateau.


Cytokine induction by 41.8 degrees C whole body hyperthermia.
The potential for 41.8 degrees C whole body hyperthermia (WBH) to enhance ionizing irradiation and cytotoxic chemotherapy without a commensurate increase in normal tissue toxicity is currently receiving renewed clinical interest. Additionally, WBH may have other biological sequela which may be clinically exploited. In this paper, data are summarized revealing the ability of WBH to induce elevated plasma levels of granulocyte-colony stimulating factor (G-CSF), interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), and tumor necrosis factor-alpha (TNF-alpha) within hours of WBH. Data regarding TNF-alpha shows induction in only a proportion of patients. No induction of C-reactive protein (CRP) or the following cytokines was observed: granulocyte macrophage-colony stimulating factor (GM-CSF), interferon-gamma (IFN-gamma), interleukin-1 alpha (IL-1 alpha), interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-7 (IL-7), interleukin-11 (IL-11), interleukin-12 (IL-12), macrophage-colony stimulating factor (M-CSF), and macrophage inflammatory protein-1 alpha (MIP-1 alpha). Data regarding interleukin-3 (IL-3) and transforming growth factor-beta 1 (TGF-beta 1) were variable and inconclusive. The implications of these results to past and future clinical trials are discussed.


Regulation of Glutamine Synthetase by Light and during Nitrogen Deficiency in Synchronous Chlorella sorokiniana.
A method is described to achieve density labeling of proteins in unicellular algae by using (13)CO(2). This is a satisfactory procedure especially for work on nitrogen metabolism. The increase in activity of glutamine synthetase (EC 184.108.40.206.) and glutamate synthase (EC 220.127.116.11.) in Chlorella sorokiniana mediated by a dark/light shift and by nitrogen starvation were investigated. Using the method of density labeling and isopycnic centrifugation, we demonstrated that the increase in enzyme activity after a dark/light shift is based on activation rather than de novo synthesis. The increase in enzyme activity after transfer to nitrogen-deficient medium is based both on activation and de novo synthesis.


A novel surgical procedure for papilla reconstruction using platelet rich fibrin.
Loss of interdental papilla predisposes to phonetic, functional, and esthetic problem. Surgical techniques are manifold, but are challenging and very unpredictable. The purpose of this case report is to present the use of platelet rich fibrin (PRF) in the reconstruction of papilla in the maxiallry anterior region of a forty year old woman who presented with loss of interdental papillae in 11, 12 and 11, 21 region. PRF was tucked in to the pouch created with a semilunar pedicle flap and the entire gingivopapillary unit was displaced coronally. Optimal fill was noted at 3 and 6 months postoperatively. Use of PRF may thus be the panacea for interdental papilla augmentation.


Enantiomeric separation of FMOC-amino acids by nano-LC and CEC using a new chiral stationary phase, cellulose tris(3-chloro-4-methylphenylcarbamate).
A novel polysaccharide-based chiral stationary phase (CSP), cellulose tris(3-chloro-4-methylphenylcarbamate), also known as Sepapak-2 or Lux Cellulose-2, has been evaluated for the enantiomeric separation of FMOC derivatives of amino acids. After mobile-phase optimization in nano liquid chromatography (nano-LC) the column enabled the enantiomeric separation of 19 out of 23 amino acids tested, indicating the high chiral recognition power of this new CSP. Subsequently, a comparison of the driving force employed (pressure or voltage) was carried out comparing nano-LC and CEC under the same conditions. Better peak efficiencies and resolution were observed by using CEC experiments, which enabled the chiral discrimination of 20 out of 23 amino acids tested. Finally, in order to show the potential of this new CSP, the determination of the content and the enantiomeric purity of the non-protein amino acid citrulline in food supplements was performed. For that purpose, the method was optimized, evaluated and applied to different commercial samples.


The UCLA MEDLARS computer system.
Under a subcontract with UCLA the Planning Research Corporation has changed the MEDLARS system to make it possible to use the IBM 7094/7040 direct-couple computer instead of the Honeywell 800 for demand searches. The major tasks were the rewriting of the programs in COBOL and copying of the stored information on the narrower tapes that IBM computers require. (In the future NLM will copy the tapes for IBM computer users.) The differences in the software required by the two computers are noted. Major and costly revisions would be needed to adapt the large MEDLARS system to the smaller IBM 1401 and 1410 computers. In general, MEDLARS is transferrable to other computers of the IBM 7000 class, the new IBM 360, and those of like size, such as the CDC 1604 or UNIVAC 1108, although additional changes are necessary. Potential future improvements are suggested.


Mechanical Stretching-Induced Traumatic Brain Injury Is Mediated by the Formation of GSK-3β-Tau Complex to Impair Insulin Signaling Transduction.
Traumatic brain injury confers a significant and growing public health burden. It is a major environmental risk factor for dementia. Nonetheless, the mechanism by which primary mechanical injury leads to neurodegeneration and an increased risk of dementia-related diseases is unclear. Thus, we aimed to investigate the effect of stretching on SH-SY5Y neuroblastoma cells that proliferate in vitro. These cells retain the dopamine-β-hydroxylase activity, thus being suitable for neuromechanistic studies. SH-SY5Y cells were cultured on stretchable membranes. The culture conditions contained two groups, namely non-stretched (control) and stretched. They were subjected to cyclic stretching (6 and 24 h) and 25% elongation at 1 Hz. Following stretching at 25% and 1 Hz for 6 h, the mechanical injury changed the mitochondrial membrane potential and triggered oxidative DNA damage at 24 h. Stretching decreased the level of brain-derived neurotrophic factors and increased amyloid-β, thus indicating neuronal stress. Moreover, the mechanical injury downregulated the insulin pathway and upregulated glycogen synthase kinase 3β (GSK-3β)S9/p-Tau protein levels, which caused a neuronal injury. Following 6 and 24 h of stretching, GSK-3βS9 was directly bound to p-TauS396. In contrast, the neuronal injury was improved using GSK-3β inhibitor TWS119, which downregulated amyloid-β/p-Taus396 phosphorylation by enhancing ERK1/2T202/Y204 and AktS473 phosphorylation. Our findings imply that the neurons were under stress and that the inactivation of the GSK3β could alleviate this defect.