Miracle-dz/UniFAR-FTUs huggingface dataset:
Impact of adrenalectomy and dexamethasone treatment on testicular morphology and sperm parameters in rats: insights into the adrenal control of male reproduction

Here we investigated the hypothesis that normal levels of glucocorticoids, a class of adrenal steroid hormones, are required for normal testicular and epididymal functions. We examined the effects of the manipulation of glucocorticoid plasma levels by bilateral adrenalectomy (1, 2, 7 and 15 days) alone or in combination with daily treatment with the synthetic glucocorticoid dexamethasone (DEX; 5 μg/kg, i.p., 6 days) on the morphology of the testis and sperm parameters in rats. We showed that adrenalectomy led to a reduction in testicular sperm count and daily sperm production starting 2 days after surgery and a differential decrease in sperm count in the epididymis, according to the region and time post‐adrenalectomy analysed. In parallel, testes from 7‐day adrenalectomized (ADX) rats displayed a higher frequency of damaged seminiferous tubules and the presence of elongated spermatids retained in the basal epithelial compartment in stages IX–XVII, which is indicative of defective spermiation. The alkaline comet assay revealed a late effect of adrenalectomy on epididymal sperm DNA fragmentation, which was increased only 15 days after surgery. DEX treatment prevented the changes in testicular and epididymal sperm count observed in 7‐day ADX rats, but failed to protect the testis from ADX‐induced morphological abnormalities. Thus, our results indicated that glucocorticoids may be involved in events related to the maintenance of spermatogenesis and sperm maturation during adulthood. These findings provide new insights into the importance of adrenal steroids to male fertility.
COMBINATORIAL PROBLEMS IN THE THEORY OF GRAPHS. I.

1953. The results referred to are contained in Theorems 10, 12, and 13. Theorem 12 it shown that congruence (3) of the present paper is if m = 2, P odd, (aroy, P) = 1. This result is also an immediate corollary of 2. in case m = 2.
Glycosaminoglycan Content of the Lateral Compartment Cartilage in Knees Conforming to the Indications for Oxford Medial Unicompartmental Knee Arthroplasty

Background: The quality of the lateral compartment cartilage is important to preoperative evaluation and prognostic prediction of unicompartmental knee arthroplasty (UKA). Delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) enables noninvasive assessment of glycosaminoglycan (GAG) content in cartilage. This study aimed to determine the GAG content of the lateral compartment cartilage in knees scheduled to undergo Oxford medial UKA. Methods: From December 2016 to May 2017, twenty patients (20 osteoarthritic knees) conforming to the indications for Oxford medial UKA were included as the osteoarthritis (OA) group, and 20 healthy volunteers (20 knees) paired by sex, knee side, age (±3 years), and body mass index (BMI) (±3 kg/m2) were included as the control group. The GAG contents of the weight-bearing femoral cartilage (wbFC), the posterior non-weight-bearing femoral cartilage (pFC), the lateral femoral cartilage (FC), and tibial cartilage (TC) were detected using dGEMRIC. The dGEMRIC indices (T1Gd) were calculated in the middle three consecutive slices of the lateral compartment. Paired t-tests were used to compare the T1Gd in each region of interest between the OA group and control group. Results: The average age and BMI in the two groups were similar. In the OA group, T1Gd of FC and TC was 386.7 ± 50.7 ms and 429.6 ± 59.9 ms, respectively. In the control group, T1Gd of FC and TC was 397.5 ± 52.3 ms and 448.6 ± 62.5 ms, respectively. The respective T1Gd of wbFC and pFC was 380.0 ± 47.8 ms and 391.0 ± 66.3 ms in the OA group and 400.3 ± 51.5 ms and 393.6 ± 57.9 ms in the control group. Although the T1Gd of wbFC and TC tended to be lower in the OA group than the control group, there was no significant difference between groups in the T1Gd in any of the analyzed cartilage regions (P value of wbFC, pFC, FC, and TC was 0.236, 0.857, 0.465, and 0.324, respectively). Conclusions: The GAG content of the lateral compartment cartilage in knees conforming to indications for Oxford medial UKA is similar with those of age- and BMI-matched participants without OA.
Multiple Genes in a Single Host: Cost-Effective Production of Bacterial Laccase (cotA), Pectate Lyase (pel), and Endoxylanase (xyl) by Simultaneous Expression and Cloning in Single Vector in E. coli

This study attempted to reduce the enzyme production cost for exploiting lignocellulosic materials by expression of multiple genes in a single host. Genes for bacterial laccase (CotA), pectate lyase (Pel) and endoxylanase (Xyl), which hold significance in lignocellulose degradation, were cloned in pETDuet-1 vector containing two independent cloning sites (MCS). CotA and xyl genes were cloned in MCS1 and MCS 2, respectively. Pel gene was cloned by inserting complete cassette (T7 promoter, ribosome binding site, pel gene, His tag and complete gene ORF) preceded by cotA open reading frame in the MCS1. IPTG induction of CPXpDuet-1 construct in E. coli BL21(DE3) resulted in expression of all three heterologous proteins of ~65 kDa (CotA), ~45 kDa (Pel) and ~25 kDa (Xyl), confirmed by SDS-PAGE and western blotting. Significant portions of the enzymes were also found in culture supernatant (~16, ~720 and ~370 IU/ml activities of CotA, Pel and Xyl, respectively). Culture media optimization resulted in 2, 3 and 7 fold increased secretion of recombinant CotA, Pel and Xyl, respectively. Bioreactor level optimization of the recombinant cocktail expression resulted in production of 19 g/L dry cell biomass at OD600nm 74 from 1 L induced culture after 15 h of cultivation, from which 9, 627 and 1090 IU/ml secretory enzyme activities of CotA, Xyl and Pel were obtained, respectively. The cocktail was also found to increase the saccharification of orange peel in comparison to the xylanase alone. Thus, simultaneous expression as well as extra cellular secretion of these enzymes as cocktail can reduce the enzyme production cost which increases their applicability specially for exploiting lignocellulosic materials for their conversion to value added products like alcohol and animal feed.
Developmental regulation of the composite CAG promoter activity in the murine T lymphocyte cell lineage

Promoter selection is of utmost importance for the study of in vivo gene function using transgenic models. In the present study, we have analyzed the expression of the GFP marker under the control of the composite CAG promoter in the lymphoid compartment of several transgenic mouse strains. Despite the ability of the CAG promoter to drive gene expression in almost all tissues examined to date, its activity appears to be developmentally regulated within the T lymphocyte cell lineage. In particular, CD4 and CD8‐expressing, thymic immature T cells displayed lower levels of the GFP marker when compared with both bone marrow precursors and mature circulating T cells, suggesting a transient downregulation of CAG activity during T cell development. Alternative promoters may therefore be preferred for the study of T cell development in vivo using a transgenic approach. genesis 47:799–804, 2009. © 2009 Wiley‐Liss, Inc.
N-Myristoylation and Ca2+ Binding of Calcineurin B Homologous Protein CHP3 Are Required to Enhance Na+/H+ Exchanger NHE1 Half-life and Activity at the Plasma Membrane*

Background: CHP3 is an N-myristoylated Ca2+-binding protein that up-regulates the cell surface expression and stability of the Na+/H+ exchanger NHE1 isoform. Results: N-Myristoylation or the Ca2+-binding site of CHP3 regulates the half-life and activity of NHE1 at the cell surface. Conclusion: CHP3 possesses a Ca2+-myristoyl switch mechanism to promote optimal NHE1 activity at the cell surface. Significance: These findings provide fundamental insight into the molecular mechanisms that regulate NHE1. Calcineurin B homologous proteins (CHP) are N-myristoylated, EF-hand Ca2+-binding proteins that regulate multiple cellular processes, including intracellular pH homeostasis. Previous work has shown that the heart-enriched isoform, CHP3, regulates the plasmalemmal Na+/H+ exchanger NHE1 isoform by enhancing its rate of oligosaccharide maturation and exocytosis as well as its half-life and transport activity at the cell surface (Zaun, H. C., Shrier, A., and Orlowski, J. (2008) J. Biol. Chem. 283, 12456–12467). However, the molecular basis for this effect is not well understood. In this report, we investigated whether the N-myristoylation and Ca2+-binding domains of CHP3 are important elements for regulating NHE1. Mutation of residues essential for either N-myristoylation (G2A) or calcium binding (D123A) did not prevent the interaction of CHP3 with NHE1, although the D123A mutant no longer showed elevated binding to NHE1 in the presence of Ca2+ when assessed using in vitro binding assays. Disruption of either site also did not impair the ability of CHP3 to stimulate the biosynthetic processing and trafficking of NHE1 to the plasma membrane nor did it affect the H+ sensitivity of the exchanger. However, they did significantly reduce the cell surface half-life and near maximal transport velocity of NHE1 to a similar extent. Simultaneous mutation of both sites (G2A/D123A) gave results identical to the individual substitutions. This finding suggests that both domains in CHP3 are interdependent and may function cooperatively as a Ca2+-myristoyl switch mechanism to selectively stabilize the NHE1·CHP3 complex at the cell surface in a conformation that promotes optimal transport activity.
Late Onset Neuropathy with Spontaneous Clinical Remission in Mice Lacking the POZ Domain of the Transcription Factor Myc-interacting Zinc Finger Protein 1 (Miz1) in Schwann Cells*

Background: The Myc-binding transcription factor Miz1 has multiple functions in different cells and tissues. Results: Ablation of the Miz1 POZ domain in Schwann cells causes a late onset peripheral neuropathy with spontaneous remission. Conclusion: Miz1 plays an essential role in myelin homeostasis of peripheral nerves. Significance: Ablation of Miz1 function in Schwann cells leads to a new model for remitting peripheral neuropathies. The transcription factor Miz1 (Myc-interacting zinc finger 1) is a known regulator of the cell cycle but also has cell cycle-independent functions. Here we analyzed the role of Miz1 in the peripheral nervous system, using an early embryonic conditional knock-out model in which the Miz1 POZ domain is ablated in Schwann cells. Although the development of myelinated nerve fibers was not impaired, Miz1ΔPOZ mice acquired behavioral signs of a peripheral neuropathy at the age of 3 months. At this time, ultrastructural analysis of the sciatic nerve showed de- and dysmyelination of fibers, with massive outfoldings and a focal infiltration of macrophages. Although the expression of genes encoding structural myelin proteins, such as periaxin, myelin basic protein, and myelin protein zero, was decreased, genes associated with a negative regulation of myelination, including c-Jun, Sox2, and Id2, were up-regulated in Miz1ΔPOZ mice compared with controls. In animals older than 4 months, the motor disabilities vanished, and the ultrastructure of the sciatic nerve exhibited numerous tomacula and remyelinated fibers, as indicated by thinner myelin. No second acute attack was observed up to the age of 1 year. Thus, the deletion of the Miz1 POZ domain in Schwann cells induces an acute neuropathy with a subsequent regeneration in which there is ongoing balancing between de- and remyelination. Miz1ΔPOZ mice are impaired in the maintenance of myelinated fibers and are a promising model for studying remyelination in adult peripheral nerves.
Mechanistic and physiological consequences of HPr(ser) phosphorylation on the activities of the phosphoenolpyruvate:sugar phosphotransferase system in gram‐positive bacteria: studies with site‐specific mutants of HPr.

The bacterial phosphotransferase system (PTS) catalyzes the transport and phosphorylation of its sugar substrates. The protein‐kinase‐catalyzed phosphorylation of serine 46 in the phosphocarrier protein, HPr, inhibits PTS activity, but neither the mechanism of this inhibition nor its physiological significance is known. Site‐specific HPr mutants were constructed in which serine 46 was replaced by alanine (S46A), threonine (S46T), tyrosine (S46Y) or aspartate (S46D). The purified S46D protein exhibited markedly lower Vmax and higher Km values than the wild‐type, S46T or S46A protein for the phosphoryl transfer reactions involving HPr(His approximately P). Interactions of HPr with the enzymes catalyzing phosphoryl transfer to and from HPr regulated the kinase‐catalyzed reaction. These results establish the inhibitory effect of a negative charge at position 46 on PTS‐mediated phosphoryl transfer and suggest that HPr is phosphorylated on both histidyl and seryl residues by enzymes that recognize its tertiary rather than its primary structure. In vivo studies showed that a negative charge on residue 46 of HPr strongly inhibits PTS‐mediated sugar uptake, but that competition of two PTS permeases for HPr(His approximately P) is quantitatively more important to the regulation of PTS function than serine 46 phosphorylation.
Metabolic syndrome components worsen lower urinary tract symptoms in women with type 2 diabetes.

CONTEXT Diabetic women are more susceptible to develop lower urinary tract symptoms (LUTS), especially overactive bladder (OAB). However, data regarding the effect of components of metabolic syndrome (MS) on this association are conflicting. OBJECTIVE The objective of the study was to examine the potential role of MS in the development of LUTS in diabetic women. DESIGN The study was a prevalence study conducted between 2005 and 2007. SETTING The study was conducted in a university hospital. PARTICIPANTS A total of 518 women with type 2 diabetes aged 50-75 yr were included. They were subgrouped as MS (47.5%) and non-MS (52.5%) groups according to whether they fulfilled the criteria of MS. MAIN OUTCOME MEASURE We used American Urological Association Symptom Index (AUA-SI) to evaluate LUTS and Indevus Urgency Severity Scale to evaluate OAB, respectively. RESULTS Women in the MS group had significantly higher storage and total AUA-SI scores as well as a higher prevalence of LUTS and OAB. Most intriguingly, the number of MS components was strongly associated with the LUTS severity because the AUA-SI scores increased in parallel to the number of components were present. Similar results were found between MS and OAB. Multivariate analysis revealed that peripheral neuropathy, but not MS, significantly predicted LUTS in diabetic women after age adjustment. However, MS remained significantly predictive for LUTS and OAB after additional adjustment for neuropathy. CONCLUSIONS Our results suggest that MS may especially influence LUTS and OAB in diabetic women, probably by compounding the effect of peripheral neuropathy.
Assessment of the biologic markers p53, Ki-67, and apoptotic index as predictive indicators of prostate carcinoma recurrence after surgery.

BACKGROUND This study was designed to evaluate the potential of the molecular and cellular markers p53, Ki-67, and apoptotic index (AI) as adjuncts to the commonly available variables of tumor grade, clinical stage, and serum prostate specific antigen to predict prostate carcinoma recurrence after radical prostatectomy. METHODS Representative punch biopsy specimens of prostate carcinoma from whole mount paraffin blocks were evaluated from 47 men who underwent radical prostatectomy. Two groups were defined: those without evidence of prostate carcinoma recurrence after 5 years of follow-up (N = 30) and those with carcinoma recurrence (N = 17). Gleason grade, clustered p53 immunostaining, Ki-67 immunostaining, and AI were determined by standard techniques. RESULTS All variables tested were associated with disease recurrence by univariate analysis: AI (P = 0.005), clustered p53 immunostaining (P = 0.0070), and Ki-67 immunostaining (P = 0.0390). Using multivariate analyses that included each biomarker with routinely available features, only AI (P = 0.0234) and clustered p53 immunostaining (P = 0.0389) added independent prognostic information (Ki-67 immunostaining, P = 0.1285). In the final logistic regression model that included standard variables with AI and p53, only AI reached statistical significance (P = 0.0332). CONCLUSIONS The continued assessment of additional biomarkers for prostate carcinoma recurrence is important to identify better those patients who may be candidates for early adjuvant therapy and also to further our understanding of the neoplastic potential of a particular malignancy.
The Use of Glycerol-Preserved Corneas in the Developing World

Corneal opacity is the third leading cause of blindness in the developing world and encompasses a wide variety of infectious, inflammatory and degenerative eye diseases. Most caes of corneal blindness are treatable with partial or full-thickness keratoplasty, provided adequate corneal tissue and surgical skill is available. However, access to sightrestoring keratoplasty in developing countries is limited by the lack of developed eye banking networks and a critical shortage of tissue suitable for transplantation. Beyond the developed world, corneal transplantation using fresh corneal tissue (FCT) is further hindered by unreliable storage and transportation facilities, unorganized distribution networks, the cost-prohibitive nature of imported tissue, unreliable compliance with medications and follow-up instructions and inadequate health and education services. Glycerol-preserved corneas overcome many of these limitations inherent to the use of FCT. As surgical innovation in lamellar corneal surgery expands the potential use of acellular corneal tissue, long-term preservation techniques are being revisited as a way to increase availability of corneal tissue to corneal surgeons throughout the developing world. Herein, we discuss the advantages of using and the applications for glycerol-preserved corneal tissue throughout the developing world.
Interactions of spatial and luminance information in the retina of chickens during myopia development.

Degrading the retinal image by frosted eye occluders produces elongated eyes and 'deprivation myopia' in a variety of animal models. The postulated retinal 'deprivation detector' is quite sensitive to even small changes in image contrast or spatial frequency composition. Because psychophysical experiments have shown that a decline in luminance shifts the contrast sensitivity function to lower spatial frequencies, it is likely that only a reduced spatial frequency range is available for image analysis to control eye growth. It is even possible that the compression might be sufficient to promote deprivation myopia. We have tested this hypothesis, using the animal model of the chicken. (1) At an ambient illumination of 550 lux (about 76 cd m-2), neutral density (ND) filters placed in front of the eye with 0.0, 0.5 or 1.0 log unit attenuation did not change refractive development. However, monocularly or binocularly attached filters with 2 log units attenuation produced 5-7 D of myopia relative to normal eyes. Black occluders were not more effective. Frosted eye occluders with little effect on image brightness (about 0.5 log units attenuation) produced much more myopia (about 16 D compared with the controls). (2) The effects of the ND filters on refractive development could not be reproduced if the ambient illumination was reduced by 2 log units. Probably, minor effects on image quality were introduced by optical imperfections of the ND filters which were more critical at low retinal image brightness. (3) In an optomotor experiment (spatial frequency 0.2 cyc deg-1, stripe speed 57 deg sec-1), it was shown that the chickens' contrast sensitivity was severely reduced when the eyes were covered by 2.0 ND filters. (4) Since there is evidence that changes in dopamine release from the retina may be one of the factors affecting the development of myopia, we have tested how selective these changes were for spatial information. It was found that dopamine release was controlled by both spatial and luminance information and that the inputs of both could be scarcely separated. (5) Because the experiments show that the eye becomes more sensitive to image degradation at low light, the human eye may also be more prone to develop myopia if the light levels are low during extended periods of near work.
Activation of Mast Cells by Trimeric G Protein Gi3; Coupling to the A3 Adenosine Receptor Directly and upon T Cell Contact

Mast cells are key players in mediating and amplifying allergic and inflammatory reactions. Previously, we identified the G-protein, Gi3, as the cellular target of receptor mimetic basic secretagogues that activate mast cell independently of IgE. In this study, we demonstrate that Gi3 is the cellular target of the adenosine A3 receptor (A3R), a G-protein coupled receptor involved in inflammation and the pathophysiology of asthma. By using a cell permeable peptide comprising the C-terminal end of Gαi3 fused to an importation sequence (ALL1) as a selective inhibitor of Gi3 signaling, we show that by coupling to Gi3, the A3R stimulates multiple signaling pathways in human mast cells, leading to upregulation of cytokines, chemokines, and growth factors. We further show that after contact with activated T cell membranes, endogenous adenosine binds to and activates the A3R, resulting in Gi3-mediated signaling. Specifically, the majority of ERK1/2 signaling initiated by contact with activated T cell membranes, is mediated by Gi3, giving rise to ALL1-inhibitable cellular responses. These results unveil the physiological G-protein coupled receptor that couples to Gi3 and establish the important role played by this G-protein in inflammatory conditions that involve adenosine-activated mast cells.
Male fertility and varicocoele: role of immune factors

The role of antisperm antibodies (ASA) in the aetiopathogenesis of varicocoele‐related male infertility remains unclear. The objective of this study was to determine whether varicocoele is associated with antisperm immune response and whether this factor provides additional affect on male fertility. We performed a multicentral, prospective study that included the clinical examination of 1639 male subjects from infertile couples and 90 fertile men, the evaluation of the absolute and relative risks of immune infertility associated with varicocoele and the impact of the autoimmune response on the semen quality. The methods used were as follows: standard examination of seminal fluid according to WHO criteria; ASA detection in seminal fluid using mixed antiglobulin reaction (MAR) and direct flow cytometry; measurement of spontaneous and ionophore‐induced acrosome reactions; oxidative stress evaluation with luminal‐dependent chemiluminescence method and evaluation of DNA fragmentation by sperm chromatin dispersion. The prevalence of varicocoele‐related immune infertility is about 15% and does not depend on the grade of vein dilatation both in primary and secondary fertility disorders. Varicocoele is not an immediate cause of autoimmune reactions against spermatozoa, but is a cofactor increasing ASA risk; the OR of immune infertility after a testicular trauma in varicocoele patients increases twofold. In varicocoele patients, the autoimmune antisperm reaction is accompanied by a more significant decrease in the semen quality (concentration and number of progressively motile and morphologically normal spermatozoa in the ejaculate), acrosome reaction disorders (presence of pre‐term spontaneous and lack of induced reactions) and an increase in the proportion of spermatozoa with DNA fragmentation. These disorders correlate with the level of sperm oxidative stress; reactive oxygen species (ROS) production in ASA‐positive varicocoele patients is 2.8 and 3.5 times higher than in ASA‐negative varicocoele patients and fertile men respectively. We did not find correlation between the grade of spermatic cord vein dilatation and ROS production.