Effects of paclitaxel before vitrification on the nuclear maturation and development of immature porcine oocytes.

BACKGROUND: Cryopreservation of porcine oocytes is difficult compared with other species and immature oocytes particularly so compared to the meiotic stage. OBJECTIVE: To evaluate the efficacy of a pretreatment with 1 micromole per L paclitaxel (PTX, 30 min exposure) before vitrification to promote the maturation of porcine immature oocytes. MATERIALS AND METHODS: Cumulus cell-enclosed oocytes (COs) aspirated from porcine ovaries were divided into three groups: i) non-pretreated with PTX and non-vitrified group (control group); ii) pretreated with PTX and vitrified group (PTX-V group); and iii) non-pretreated with PTX and vitrified group (nPTX-V group). RESULTS: The nuclear maturation rate up to the preovulatory stage was significantly lower (P < 0.05) in the nPTX-V group than in the control group, but was similar in the PTX-V and control groups. No significant differences were observed in viability assessed by a normal CO morphology and the embryonic development of oocytes activated by the parthenogenetic stimulation between the PTX-V and control groups, but not the non-PTX-V group. CONCLUSION: PTX may promote the maturation of vitrified porcine immature oocytes. Doi.org/10.54680/fr23510110812.

Demonstration of nuclear gamma-ray polarimetry based on a multi-layer CdTe Compton camera.

To detect and track structural changes in atomic nuclei, the systematic study of nuclear levels with firm spin-parity assignments is important. While linear polarization measurements have been applied to determine the electromagnetic character of gamma-ray transitions, the applicable range is strongly limited due to the low efficiency of the detection system. The multi-layer Cadmium-Telluride (CdTe) Compton camera can be a state-of-the-art gamma-ray polarimeter for nuclear spectroscopy with the high position sensitivity and the detection efficiency. We demonstrated the capability to operate this detector as a reliable gamma-ray polarimeter by using polarized 847-keV gamma rays produced by the [Formula: see text]([Formula: see text]) reaction. By combining the experimental data and simulated calculations, the modulation curve for the gamma ray was successfully obtained. A remarkably high polarization sensitivity was achieved, compatible with a reasonable detection efficiency. Based on the obtained results, a possible future gamma-ray polarimetery is discussed.

Trans fatty acids exacerbate dextran sodium sulphate-induced colitis by promoting the up-regulation of macrophage-derived proinflammatory cytokines involved in T helper 17 cell polarization.

Numerous reports have shown that a diet containing large amounts of trans fatty acids (TFAs) is a major risk factor for metabolic disorders. Although recent studies have shown that TFAs promote intestinal inflammation, the underlying mechanisms are unknown. In this study, we examined the effects of dietary fat containing TFAs on dextran sodium sulphate (DSS)-induced colitis. C57 BL/6 mice were fed a diet containing 1·3% TFAs (mainly C16:1, C18:1, C18:2, C20:1, C20:2 and C22:1), and then colitis was induced with 1·5% DSS. Colonic damage was assessed, and the mRNA levels of proinflammatory cytokines and major regulators of T cell differentiation were measured. The TFA diet reduced survival and exacerbated histological damage in mice administered DSS compared with those fed a TFA-free diet. The TFA diet significantly elevated interleukin (IL)-6, IL-12p40, IL-23p19 and retinoic acid-related orphan receptor (ROR)γt mRNA levels in the colons of DSS-treated animals. Moreover, IL-17A mRNA levels were elevated significantly by the TFA diet, with or without DSS treatment. We also examined the expression of proinflammatory cytokines in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and peritoneal macrophages. These cells were exposed to TFAs (linoelaidic acid or elaidic acid) with or without LPS and the mRNA levels of various cytokines were measured. IL-23p19 mRNA levels were increased significantly by TFAs in the absence of LPS. Cytokine expression was also higher in LPS-stimulated cells exposed to TFAs than in unexposed LPS-stimulated cells. Collectively, our results suggest that TFAs exacerbate colonic inflammation by promoting Th17 polarization and by up-regulating the expression of proinflammatory cytokines in the inflamed colonic mucosa.

In vivo measurement of gene expression, angiogenesis and physiological function in tumors using multiphoton laser scanning microscopy.

Intravital microscopy coupled with chronic animal window models has provided stunning insight into tumor pathophysiology, including gene expression, angiogenesis, cell adhesion and migration, vascular, interstitial and lymphatic transport, metabolic microenvironment and drug delivery. However, the findings to date have been limited to the tumor surface (< 150 microm). Here, we show that the multiphoton laser-scanning microscope can provide high three-dimensional resolution of gene expression and function in deeper regions of tumors. These insights could be critical to the development of novel therapeutics that target not only the tumor surface, but also internal regions.

Municipal wastewater characteristics in Thailand and effects of soft intervention measures in households on pollutant discharge reduction.

In developing countries with large Millennium Development Goals (MDGs) sanitation indicator, pollutant discharge reduction function of wastewater treatment systems should be considered. In this paper, pollutant generations per capita (PGCs) and pollutant discharges per capita (PDCs) are estimated as a base dataset for wastewater management in Thailand. PDCs of black water, i.e. toilet wastewater, are found to be much smaller than PGCs of black water. However, PDCs of gray water, i.e. municipal wastewater other than toilet wastewater are large. Gray water is often discharged without treatment and contributes much to ambient water deterioration. Moreover, possible 5-day biological oxygen demand (BOD5) discharge reductions with "soft interventions", i.e. measurements in households to reduce wastewater pollutant discharge such as using a paper filter or a plastic net in kitchen sinks and so on, are estimated as 39, 21 and 34% for BOD5, total Kjeldahl nitrogen (TKN) and phosphate (PO4-P), respectively. For the estimation, environmental accounting housekeeping (EAH) books of domestic wastewater, spreadsheets with pollutant discharges by water usages and possible effects of "soft interventions" are applied. The framework of this study with "soft intervention" effects on pollutant discharge reductions should enhance wastewater management especially in the areas under development of wastewater treatment systems.

Omega-3 polyunsaturated fatty acids ameliorate the severity of ileitis in the senescence accelerated mice (SAM)P1/Yit mice model.

Clinical studies using omega-3 polyunsaturated fatty acids (omega3-PUFA) to Crohn's disease (CD) are conflicting. Beneficial effects of dietary omega3-PUFA intake in various experimental inflammatory bowel disease (IBD) models have been reported. However, animal models of large intestinal inflammation have been used in all previous studies, and the effect of omega3 fat in an animal model of small intestinal inflammation has not been reported. We hypothesized that the effects of omega3 fat are different between large and small intestine. The aim of this study was to determine whether the direct effect of omega3 fat is beneficial for small intestinal inflammation. Senescence accelerated mice (SAM)P1/Yit mice showed remarkable inflammation of the terminal ileum spontaneously. The numbers of F4/80-positive monocyte-macrophage cells as well as beta7-integrin-positive lymphocytes in the intestinal mucosa were increased significantly compared with those in the control mice (AKR-J mice). The area of mucosal addressin cell adhesion molecule-1 (MAdCAM-1)-positive vessels was also increased. The degree of expression levels of monocyte chemoattractant protein-1 (MCP-1), interleukin (IL)-6 and interferon (IFN)-gamma mRNA were increased significantly compared with those in the control mice. The feeding of two different kinds of omega3 fat (fish-oil-rich and perilla-oil-rich diets) for 16 weeks to SAMP1/Yit mice ameliorated inflammation of the terminal ileum significantly. In both the omega3-fat-rich diet groups, enhanced infiltration of F4/80-positive monocytes/macrophages in intestinal mucosa of SAMP1/Yit mice cells and the increased levels of MCP-1, IL-6 and IFN-gamma mRNA expression were ameliorated significantly compared with those in the control diet group. The results suggest that omega3 fat is beneficial for small intestinal inflammation by inhibition of monocyte recruitment to inflamed intestinal mucosa.

Vascular endothelial growth factor (VEGF) modulation by targeting hypoxia-inducible factor-1alpha--> hypoxia response element--> VEGF cascade differentially regulates vascular response and growth rate in tumors.

Although tumors can activate vascular endothelial growth factor (VEGF) promoter in host stromal cells, the relative contribution to VEGF production of host versus tumor cells and the resulting vascular response have not been quantitated to date. To this end, we implanted VEGF-/- and wild-type (WT) embryonic stem (ES) cells in transparent dorsal skin windows in severe combined immunodeficient mice. VEGF-/- ES cell-derived tumors produced approximately 50% of VEGF compared with the WT tumors, suggesting significant contribution of host stromal cells. To discern the hypoxia-induced hypoxia inducible factor (HIF)-1alpha --> hypoxia response element (HRE) --> VEGF signaling cascade, we also examined tumors derived from HIF-1alpha-/- and HRE-/- ES cells. As expected, the VEGF protein level in HIF-1alpha-/- ES tumors was intermediate between VEGF-/- and WT ES cell tumors. Surprisingly, HRE-/- ES tumors produced the same level of VEGF as the VEGF-/- ES tumors, suggesting a critical role of HRE in tumor cell VEGF production. Angiogenesis in these tumors was proportional to their VEGF levels (VEGF-/- approximate to HRE-/- < HIF-1alpha-/- < WT). In contrast, vascular permeability, leukocyte-endothelial adhesion, and tumor growth were reduced in VEGF-/- and HRE-/- tumors but were comparable in HIF-1a-/- and WT tumors. This discrepancy suggests that different intracellular signaling pathways may be involved in each of these functions of VEGF. More importantly, these data suggest that host cells are active players in tumor angiogenesis and growth and need to be taken into account in the design of any therapeutic strategy.

Induction of protein kinase Czeta-related protein kinase by growth suppression in carcinogen-initiated epidermal cell-line WYF31 cells.

In primary cultured mouse epidermal cells, protein kinase C isozyme zeta (PKCzeta) consists of multiple forms, for example, low-salt eluted PKCzeta (1-PKCzeta; 79 and 85 kDa) and high-salt eluted PKCzeta (h-PKCzeta; 79 and 85 kDa) on anion-exchange column chromatography. In this study, biochemical and biophysical differences between 1-PKCzeta and h-PKCzeta were examined by using carcinogen-initiated mouse epidermal cell-line WYF31 cells, whose growth is stimulated by tumour promoter phorbol 12-myristate 13-acetate (PMA). The binding efficiency of h-PKCzeta to anti-PKCzeta antibody-affinity column was 10 times higher than that of 1-PKCzeta. T7-tagged rat PKCzeta overexpressed in WYF31 cells was recovered only in the high-salt eluted area on the anion-exchange column. Furthermore, when rat PKCzeta was stably overexpressed in WYF31 cells, the content of h-PKCzeta increased 4 to 5 times compared to that of parental cells, but the content of 1-PKCzeta was not altered. All of these results indicate that h-PKCzeta is the product of the PKCzeta gene (referred to as PKCzeta) and that 1-PKCzeta is closely related but different from PKCzeta (referred to as PKCzeta-related kinase). Interestingly, serum starvation of WYF31 cells caused a marked increase of the content of PKCzeta-related kinase with a concomitant decrease of PKCzeta content. These changes were reversed by stimulating the cell growth with 10% foetal calf serum. Prolonged treatment of starved cells with PMA, which induces the proliferation of WYF31 cells, also caused the downregulation of PKCzeta-related kinase. These results suggest that the expression levels of PKCzeta-related kinase and PKCzeta are differently regulated, and that the increased expression of PKCzeta-related kinase might play a significant role in the growth-suppression processes of WYF31 cells.

Familial aggregation and coaggregation of history of hypertension and stroke.

We attempted to evaluate familial aggregation and coaggregation of history of hypertension and stroke. Past and family history of hypertension and stroke for 83 089 probands and their relatives were obtained from a data set for the Japan Collaborative Cohort Study for Evaluation of Cancer Risk sponsored by the Ministry of Education (JACC Study), which was initiated from 1988 to 1990. First, evaluation was performed for familial aggregation of each of two disorders using ordinal logistic regression of the generalized estimation equations (GEE) to account for dependence of observations within families. Secondly, in order to evaluate the familial congregation of the history of hypertension and stroke, a GEE-based multivariate probed predictive model was applied. After adjusting for the proband's age, level of obesity, smoking status, drinking status, habitation area, and the gender and type of the relatives, the estimated odds ratios for the intraindividual clustering and familial aggregation of the disease history showed statistically significant relationships. In addition, the history of the two disorders showed a significant relationship in terms of familial coaggregation independently of the aggregation of each disorder itself. Our results confirmed that hypertension and stroke coaggregate strongly within families through possible effects of genetic factors, which, alone or in conjunction with environmental factors, influence susceptibility to both hypertension and stroke.

Inhibition of nitric oxide synthase attenuates the cerebral blood flow response to stimulation of postganglionic parasympathetic nerves in the rat.

Stimulation of cerebrovascular parasympathetic nerves markedly increases cortical blood flow. Nitric oxide (NO) or a NO-containing compound is present in these nerves and may therefore, upon release, be partly responsible for the flow increase. In addition, transmitters released from the nerves may cause synthesis and release of this compound from the endothelium. The contribution of NO synthesis to the cortical blood flow (CoBF) increase during parasympathetic stimulation was elucidated in rat by laser-Doppler flowmetry. Thirty-minute exposure to circulating N omega-nitro-L-arginine methyl ester (L-NAME) 50 mg kg-1 eliminated most of the response (from 104 to 8% increase), whereas 10-min exposure to this dose or 30-min exposure to 5 mg kg-1 caused a less marked reduction. The reducing effect was particularly evident after elimination of the systemic blood pressure increase caused by L-NAME (only 3% increase after the high dose). Infusion of L-arginine restored the flow response. Resting CoBF was not substantially affected by blockade of NO formation. Thus, release of an NO-containing compound constitutes a major component of the increase in CoBF caused by parasympathetic nerve stimulation but does not seem to contribute to cortical flow regulation during resting conditions.

Presence of protein phosphatase type 1 and its involvement in temperature-dependent flagellar movement of fowl spermatozoa.

Even in the presence of ATP, the motility of demembranated fowl spermatozoa was negligible at the avian body temperature of 40 degrees C. Motility could be restored by the addition of calyculin A, okadaic acid, specific inhibitors of phosphatase type 1 (PP1) and PP-2A, and inhibitor 1 or inhibitor 2, which are specific inhibitors of protein phosphatase type 1 (PP1). Demembranated spermatozoa, stimulated by calyculin A or okadaic acid, lost their motility following the addition of 1 mM CaCl2, but this was restored gradually by the stepwise addition of EGTA. Immunoblotting of sperm extract using an antibody to PP1 revealed a major cross-reacting protein of 36-37 kDa, which corresponded to the molecular weight of the known catalytic subunit of PP1. These results suggest that PP1 present in the fowl sperm axoneme may be involved in the inhibition of fowl sperm motility at 40 degrees C via Ca(2+)-dependent regulatory systems.

Simple method for cryopreservation of an anaerobic rumen fungus using ethylene glycol and rumen fluid.

The cryopreservation of an anaerobic rumen fungus, Piromyces communis OTS1, was examined at -84 degrees C using dimethyl sulfoxide, propylene glycol or ethylene glycol as cryoprotectants. Ethylene glycol was the most effective agent, combining high survival and low toxicity, followed by dimethyl sulfoxide and propylene glycol. Cell-free rumen fluid in the cryopreservation medium decreased the toxicity of the cryoprotectant agents and also had a protective action per se. A survival of 80% after 1 year storage was obtained when samples with an initial zoospore density of 5 x 10(4) zoospores/ml were equilibrated for 15 min in medium containing 0.64 M ethylene glycol and 5% cell-free rumen fluid, then frozen with dry ice and stored at -84 degrees C.

Controlled ultraviolet irradiation generates endothelial damage without affecting the nerve terminals of the cerebral artery in cats.

The vesicles of adventitial autonomic nerve terminals were examined quantitatively under an electron microscope in controlled ultraviolet ray (UV)-irradiated cerebral vessels. Five cats whose basilar arteries were irradiated with UV (UV group) and 5 cats whose basilar arteries were irradiated with visible rays (control group) were compared. Endothelial vacuolation was observed only in the UV group. There was no statistically significant difference in the diameters of the dense-cored vesicles, related to noradrenaline, and clear vesicles, related to acetylcholine, between the two groups. It is concluded that controlled UV irradiation which generates endothelial damage does not affect the vascular adventitia ultrastructurally.

Moderate hypoglycemia induces ultrastructural changes in perivascular nerve terminals of cat cerebral arteries.

A quantitative morphological analysis of the perivascular nerve terminals of cerebral arteries during moderate hypoglycemia was performed. 5-Hydroxydopamine (5-OHDA) was applied to discriminate dense-cored vesicles, related to noradrenaline, and clear vesicles, related to acetylcholine, under the electron microscope. Five hypoglycemic and 5 normoglycemic cats, all receiving 5-OHDA, were compared. In both the middle cerebral artery and vertebral artery, the dense-cored vesicles were significantly smaller and clear vesicles were significantly larger in hypoglycemia than in normoglycemia. These morphological changes in the vesicles may indicate hyperactivity of the sympathetic system and hypoactivity of the parasympathetic system of the cerebral vessels during hypoglycemia.

Effects of locus ceruleus lesions on the pericapillary nerve terminals in the feline brain.

The effects of bilateral locus ceruleus (LC) lesions on the pericapillary nerve terminals were investigated in the feline brain parenchyma using electron microscopy. LC lesions were induced stereotaxically and the animals were sacrificed after intravenous administration of 5-hydroxydopamine (5-OHDA). The diameter and number of dense-cored vesicles (DCVs) and clear vesicles (CVs) in the pericapillary nerve terminals were measured. The number of DCVs in the nerve terminal was significantly decreased by bilateral LC lesions. The diameters of the DCVs and CVs decreased significantly as compared with those in the non-operated control group. These data suggest that the LC is closely related to the pericapillary nerve terminals in the brain parenchyma and that not only nerve terminals with DCVs but also those with CVs are affected by LC lesions.

Effect of methylmercury exposure on different neurotransmitter systems in rat brain.

The effect of methylmercury (4.0 mg/kg/day, for 50 days) exposure was studied on various neurotransmitter systems in hypothalamus, pons-medulla, brain-stem, and striatum of rats. Although methylmercury exposure resulted in marked increases in dopamine (DA) content of various brain regions and in the brain-stem or striatal norepinephrine (NE) level, the level of metabolite 3,4-dihydroxyphenylacetic acid (DOPAC) was markedly decreased in the same regions. On the other hand, concentrations of 5-hydroxyindoleacetic acid (5-HIAA) in various brain regions were considerably decreased in methylmercury-exposed rats. Similarly, the 5-hydroxytryptamine (5-HT) level was markedly decreased in the hypothalamus, brain-stem, and striatum.

A case of brainstem encephalitis with CT scan abnormality mimicking Fisher syndrome.

A 7-year-old boy with brainstem encephalitis is described. He was drowsy in the acute phase. CSF showed pleocytosis without elevated protein. EEG showed diffuse slow wave activity during wakefulness. CT scan disclosed a low density abnormality in the basal ganglia area, which disappeared as the patient recovered. The clinical signs, CT scan abnormality and EEG findings suggest that this case is not a variant of the Guillain-Barré syndrome but is brainstem encephalitis mimicking Fisher syndrome.

Inhibition of flagellar motility of demembranated fowl spermatozoa by protease substrates.

We investigated the effects of various protease substrates on the motility of demembranated fowl spermatozoa. In the presence of ATP, the motility of demembranated spermatozoa was vigorous at 30 degrees C, but decreased markedly following the addition of protease substrates, such as N alpha-carbobenzoxy-L-lys-thiobenzyl ester (BLT), N-benzoyl-phe-val-arg p-nitroanilide or N alpha-benzoyl-D,L-arg p-nitroanilide (BAPNA) in a dose-dependent manner, within the range 0-1 mM. The subsequent addition of 100 ng/ml trypsin released the inhibitory effect of protease substrates within 10 s. Phosphorylation or dephosphorylation of several proteins of demembranated spermatozoa was observed following the addition of protease substrates, however, no consistent patterns of protein phosphorylation or dephosphorylation were associated with the inhibition of motility. These results suggest that endogenous protease activity is instrumental in the maintenance of fowl sperm motility and that the site of action of this protease is in the axoneme and/or accessory cytoskeletal components. This enzyme may not act directly on the phosphorylation of sperm proteins involved in the regulation of motility.

Malignant proliferating trichilemmal tumor in the skin over the breast: a case report.

A proliferating trichilemmal tumor is relatively uncommon. It is composed of multiple cysts consisting of squamous epithelium with trichilemmal keratinization without granular layer interposition. This lesion usually occurs in the scalp of elderly women. We describe a 67-year-old woman with a malignant proliferating trichilemmal tumor in the skin over the breast. We first misdiagnosed the disease as a primary squamous cell carcinoma of the breast with a metastatic lymph node in the axilla because of the disease site and our unfamiliarity with the disease. The patient underwent radical mastectomy with axillary dissection. Eight months postoperatively, a tumor appeared in her right axilla and progressively enlarged. We subsequently excised the tumor. She is healthy as of 8 months postoperatively. To the best of our knowledge, only one case of a proliferating trichilemmal tumor occurring in the skin over the breast has been reported.