Salt-sensitive hypertension in GR mutant rats is associated with altered plasma polyunsaturated fatty acid levels and aortic vascular reactivity.

In humans, glucocorticoid resistance is attributed to mutations in the glucocorticoid receptor (GR). Most of these mutations result in decreased ligand binding, transactivation, and/or translocation, albeit with normal protein abundances. However, there is no clear genotype‒phenotype relationship between the severity or age at disease presentation and the degree of functional loss of the receptor. Previously, we documented that a GR+/- rat line developed clinical features of glucocorticoid resistance, namely, hypercortisolemia, adrenal hyperplasia, and salt-sensitive hypertension. In this study, we analyzed the GR+/em4 rat model heterozygously mutant for the deletion of exon 3, which encompasses the second zinc finger, including the domains of DNA binding, dimerization, and nuclear localization signals. On a standard diet, mutant rats exhibited a trend toward increased corticosterone levels and a normal systolic blood pressure and heart rate but presented with adrenal hyperplasia. They exhibited increased adrenal soluble epoxide hydroxylase (sEH), favoring an increase in less active polyunsaturated fatty acids. Indeed, a significant increase in nonactive omega-3 and omega-6 polyunsaturated fatty acids, such as 5(6)-DiHETrE or 9(10)-DiHOME, was observed with advanced age (10 versus 5 weeks old) and following a switch to a high-salt diet accompanied by salt-sensitive hypertension. In thoracic aortas, a reduced soluble epoxide hydrolase (sEH) protein abundance resulted in altered vascular reactivity upon a standard diet, which was blunted upon a high-salt diet. In conclusion, mutations in the GR affecting the ligand-binding domain as well as the dimerization domain resulted in deregulated GR signaling, favoring salt-sensitive hypertension in the absence of obvious mineralocorticoid excess.

Effect of sex-sorting and cryopreservation on the post-thaw sperm quality of Iberian red deer spermatozoa.

This study investigated the effect of sex-sorting and cryopreservation on post-thaw characteristics and fertility of red deer (Cervus elaphus) sperm for the first time. Semen was collected by electroejaculation from 10 mature stags during the breeding season, and each ejaculate split into four experimental groups: Bulk sorted spermatozoa, sorted but not sexed (BSS); sorted high purity X-spermatozoa (XSS); sorted high purity Y-spermatozoa (YSS); and, control non-sorted spermatozoa (NS). Following, all samples were frozen over liquid nitrogen. Two straws per stag and sample type were analyzed immediately post-thaw and following a 2-h incubation period at 37 °C. Post-thaw total motility (TM) as assessed by CASA was not different (P < 0.05) among NS, BSS and YSS sperm. For XSS, post-thaw TM was lower (39%, P < 0.05) than that for NS (54%) or BSS (50%), but similar (P > 0.05) to that of YSS (47%) sperm. The percentage of apoptotic spermatozoa as assessed by PI/YO-PRO-1 and flow cytometry analysis, was higher (17%, P ≤ 0.05) for XSS sperm than NS (12%), BSS (13%) and YSS (14%) sperm. Following incubation there were no differences (P > 0.05) in TM or percent apoptosis among treatments. Post-thaw chromatin stability calculated as the DNA fragmentation index (%DFI) was similar among treatments; following incubation %DFI increased in all except YSS, which displayed the lowest value (P < 0.05). Artificial insemination of synchronized hinds yielded 44, 52 and 62% delivery rates for YSS, NS and standard frozen-thawed sperm, respectively (P < 0.05). Notably, 93 and 55% of fawns born were males for the YSS and NS spermatozoa, respectively (P < 0.05). In summary, Y-sorted sperm displayed acceptable post-thaw sperm evaluation parameters and the expected offspring sex ratio. More studies are needed to understand the source of sperm damage that may compromise the fertility of Y-sorted red deer sperm.

The Effect of Oxidative Stress on Thawed Bulk-Sorted Red Deer Sperm.

The aims of this study were to assess the effects of the sex-sorting process on post-thaw sperm quality as well as on induced oxidative stress damage (H2 O2 0 mm = H000; H2 O2 50 mm = H050; H2 O2 100 mm = H100) and the protective action of reduced glutathione (GSH) and Trolox, when comparing sorted (BSS) and non-sorted (NS) red deer spermatozoa incubated at 37°C. Sperm samples from three stags were collected by electroejaculation and frozen. Immediately after thawing, sperm motility was higher (p < 0.05) for NS (59% ± 3.3) than BSS (36.9% ± 5.8) sperm. Furthermore, the percentage of apoptotic sperm was higher (p < 0.05) for BSS (21.6% ± 5.0) than NS sperm (14.6% ± 1.2). The presence of H2 O2 increased DNA damage in NS (H000 = 4.1% ± 0.9; H050 = 9.3% ± 0.7; and H100 = 10.9% ± 2.3), but not in BSS sperm. However, in the presence of oxidant, GSH addition improved (p < 0.05) sperm motility in both groups of sperm samples as compared to their controls (NS: 44.5 ± 4.8 vs 21.1 ± 3.9 and BSS: 33.3 ± 8.1 vs 8.9 ± 1.8). These results demonstrate that the sperm-sorting process induces sublethal effects, albeit selecting a sperm population with a chromatin more resistant to oxidative stress than that in non-sorted sperm. Moreover, addition of GSH at 1 mm may be a good choice for maintaining the quality of stressed sperm samples, unlike Trolox, which inhibited sperm motility.

The effects of hoechst 33342 staining and the male sample donor on the sorting efficiency of canine spermatozoa.

The aim of this study was to evaluate the influence of Hoechst 33342 (H-42) concentration and of the male donor on the efficiency of sex-sorting procedure in canine spermatozoa. Semen samples from six dogs (three ejaculates/dog) were diluted to 100 × 10(6) sperm/ml, split into four aliquots, stained with increasing H-42 concentrations (5, 7.5, 10 and 12.5 µl, respectively) and sorted by flow cytometry. The rates of non-viable (FDA+), oriented (OS) and selected spermatozoa (SS), as well as the average sorting rates (SR, sorted spermatozoa/s), were used to determine the sorting efficiency. The effects of the sorting procedure on the quality of sorted spermatozoa were evaluated in terms of total motility (TM), percentage of viable spermatozoa (spermatozoa with membrane and acrosomal integrity) and percentage of spermatozoa with reacted/damaged acrosomes. X- and Y-chromosome-bearing sperm populations were identified in all of the samples stained with 7.5, 10 and 12.5 µl of H-42, while these two populations were only identified in 77.5% of samples stained with 5 µl. The values of OS, SS and SR were influenced by the male donor (p < 0.01) but not by the H-42 concentration used. The quality of sorted sperm samples immediately after sorting was similar to that of fresh samples, while centrifugation resulted in significant reduction (p < 0.05) in TM and in the percentage of viable spermatozoa and a significant increase (p < 0.01) in the percentage of spermatozoa with damage/reacted acrosomes. In conclusion, the sex-sorting of canine spermatozoa by flow cytometry can be performed successfully using H-42 concentrations between 7.5 and 12.5 µl. The efficiency of the sorting procedure varies based on the dog from which the sperm sample derives.

Handling of boar spermatozoa during and after flow cytometric sex-sorting process to improve their in vitro fertilizing ability.

The objective of this study was to develop an adequate sperm handling protocol in order to obtain a sex-sorted sperm population with an optimal fertilizing ability. For this purpose, different aspects of the sorting procedure were examined. The effects of the high dilution rates (experiment 1), type of collection medium used (experiment 2), and sheath fluid composition (experiment 3) on sorted boar sperm quality and function were evaluated. Sperm quality was assessed by motility and viability tests, whereas sperm function was evaluated by an in vitro fertilization assay which determined the penetration and polyspermy rates as well as the mean number of sperm penetrating each oocyte. In experiment 1, the results obtained indicated that the high dilution rates did not cause a decrease either in the sperm quality parameters evaluated or the in vitro fertilization ability of spermatozoa. In experiment 2, although sperm quality was not affected, fertilizing ability was compromised after sorting, regardless of the collection medium that was used. In the experiment 3, all groups displayed adequate sperm quality values, but higher in vitro fertility parameters were obtained for spermatozoa sorted in presence of EDTA in the sheath fluid and egg yolk (EY) in the collection media when compared with those sorted in absence of these protective agents. No differences in penetration rates between unsorted highly diluted (control) and sorted sperm in the presence of EDTA and EY were observed. In conclusion, fertilizing ability was compromised in sex-sorted sperm. The addition of EDTA to sheath fluid and EY to collection medium improved boar sperm fertilizing ability, and both agents should be included as essential media components in future studies.

Non-surgical deep intrauterine transfer of superfine open pulled straw (SOPS)-vitrified porcine embryos: evaluation of critical steps of the procedure.

Previous trials achieved extremely poor results when using the one-step warming method in a syringe in combination with non-surgical deep intrauterine transfer (NET) of superfine open pulled straw (SOPS)-vitrified embryos. This study aimed to assess the effect of the warming procedure on the in vitro and in vivo development of SOPS-vitrified embryos. The effect of the passage of the vitrified-warmed (VW) embryos through the NET catheter was also evaluated. Groups of 4 to 6 morulae and blastocysts, collected from weaned sows, were SOPS-vitrified in 1 µL of vitrification medium, warmed by the one-step warming method in a dish or in a 1-mL syringe and cultured in vitro for 48 h to evaluate the embryo survival (ES) and hatching rates (HR). Warming in syringe had a deleterious effect (P < 0.05) on the in vitro ES (60.5 ± 10.4%) and HR (39.6 ± 9.5%) of VW embryos in comparison with embryos warmed in a dish (85.4 ± 10.6% and 69.0 ± 8.4%, respectively). This decreased embryonic development was due to the increased time required between the removal of the straws from the liquid nitrogen and the contact of the embryos with the warming medium when the warming was performed in a syringe in comparison with that for the warming in a dish. After verifying that the passage of VW embryos through the NET catheter does not have a damaging effect on their further in vitro development, the negative effect of warming in a syringe was also confirmed after NET. Fifteen fresh and SOPS-vitrified embryos warmed in a syringe or in a dish were transferred to each recipient (n = 28) and recovered 24 h later to assess their developmental progression. All embryos from the syringe group were found to have degenerated at recovery. The in vivo ES and HR from the dish group (80.4 ± 3.4% and 14.2 ± 7.2%, respectively) were lower (P < 0.05) than those from the fresh group (94.0 ± 4.1% and 36.8 ± 7.8%, respectively). Combining the warming in a dish and the NET procedure, 35 VW embryos were transferred to each of 10 gilts. Five recipients farrowed an average of 10.4 ± 0.9 piglets. In conclusion, the method of one-step warming in a syringe has a negative effect on the in vitro and in vivo viability of SOPS-vitrified porcine embryos. In addition, NET of SOPS-vitrified embryos warmed by the one-step method in a dish showed promising reproductive performance of recipients. However, despite the great potential of this technology, further developments are required for large-scale commercial applications.

Seminal plasma proteins as modulators of the sperm function and their application in sperm biotechnologies.

Seminal plasma (SP) is known to play an important role in mammalian fertilization. However, the variability found in its composition among species, males and even fractions of the same ejaculate has made difficult to completely understand its effect in sperm function. Proteins are one of the major SP components that modulate sperm functionality. During the last years, intensive work has been performed to characterize the role of these proteins. They have been found to influence sperm capacitation, formation of the oviductal sperm reservoir and sperm-oocyte interaction. Sperm biotechnologies, such as sperm cryopreservation and flow cytometric sex-sorting, that involve a substantial dilution of the SP are detrimental to sperm quality. Attempts to improve the outcome of these biotechnologies include the restoration of SP, which has produced contradictory results. To overcome this variability, different research groups have proposed the application of isolated SP proteins. Herein, we will review the current knowledge in the role of the major SP proteins as modulators of sperm functionality. Furthermore, we will discuss the possible applications of the SP proteins in sperm cryopreservation and flow cytometric sex-sorting.

The effect of glycerol concentrations on the post-thaw in vitro characteristics of cryopreserved sex-sorted boar spermatozoa.

The objective of this study was to optimize protocols for the cryopreservation of sex-sorted boar spermatozoa. In the experiment 1, we evaluated the effects of a standard boar sperm cryopreservation procedure (3% final glycerol concentration) on the in vitro characteristics of sex-sorted sperm frozen at low sperm concentrations (20 × 10(6) sperm/ml; S20 group). Non-sorted spermatozoa frozen at 1000 × 10(6) (C1000 group) and 20 × 10(6) (C20 group) sperm/ml were used as the freezing control groups. In experiment 2, the effects of different final glycerol concentrations (0.16%, 0.5%, 1.0%, 2.0% and 3.0%) on post-thaw quality of the S20 and C20 groups were evaluated. In both experiments, the samples were evaluated prior to freezing (5°C) and at 30, 90 and 150 min after thawing. Experiment 1 indicated that freezing sperm at low concentrations decreased (p < 0.05) the total motility (TM) and progressive motility (PM) at 90 and 150 min after thawing regardless of whether the sperm were sorted or not. However, the sperm membrane integrity was not affected at any evaluation step. Inexperiment 2, significant effects on the TM and PM because of increased glycerol concentrations in the S20 and C20 groups were observed only at 90 and 150 min after thawing. The samples frozen in 3% glycerol showed lower (p < 0.05) TM and PM values when compared to those frozen in the presence of 0.5% and 1% glycerol. In both experiments, non-sorted control samples displayed higher percentages of spermatozoa with damaged DNA than sorted spermatozoa. In conclusion, the optimization of cryopreservation conditions by decreasing the glycerol concentrations can improve post-thaw motility of sex-sorted spermatozoa frozen at low concentrations.

[Fiber in enteral nutrition: systematic review of the literature]. / La fibra en nutrición enteral: revisión sistemática de la literatura.

UNLABELLED: It has been suggested that adding fibre to enteral nutrition (EN) formulas may achieve a normalization of the gastrointestinal tract's functions by reducing the incidence of diarrhoea and constipation. However, this supposition is based on the demonstrated effect of fibre in natural feeding. There have so far been no firm recommendations on the use, amount or type of fibre that is most advisable in EN. GOAL: To establish the scientific evidence existing on the benefits gained by adding fibre to EN formulas compared with the use of fibre-free formulas. MATERIAL AND METHODS: Bibliographical search on PubMed, on the register of Randomized Prospective Trials (RPT) of the Cochrane Library and manual search. INCLUSION CRITERIA: RPT comparing fibre-free formulas with isocaloric ad isonitrogen formulas containing fibre in total EN. Four independent reviewers revised the references selected. The data were analyzed using the RevMan 4.1 programme from the Cochrane Library. RESULTS: Of 286 references, 25 fulfilled the inclusion criteria. The data were grouped by population and outcome variable. The addition of fermentable fibre to the EN formulas tends to reduce the incidence of diarrhoea in critically-ill and post-surgery patients (OR = 0.66; 95% CI 0.46-0.95; p = 0.19). The use of formulas with insoluble fibre seems to increase the frequency of depositions and reduce the need for laxatives in immobilized patients receiving long-term EN but the data are insufficient. In people with normal gastrointestinal functions, the frequency of depositions is similar when using formulas with and without fibre (DPM = -0.97; 95% CI -0.12-[-0.02]; p < 0.0001). CONCLUSION: There is currently insufficient evidence to recommend the systematic use of formulas containing fibre in total EN. However, in critically-ill and post-surgery patients, the use of formulas with fermentable fibre tends to reduce the incidence of diarrhoea. Although information is still lacking. The formulas with insoluble fibre seem to diminish constipation in chronic EN, whereas in other situations where the digestive system is working appropriately, no benefit has been shown. It is necessary to increase the number and quality of these trials in order to be able to establish firm recommendations on the use of EN formulas containing fibre.

[Acute water intoxication as complication of intravenous urography]. / Intoxicación aguda acuosa como complicación de una urografía intravenosa.

Physiologically, two complementary mechanisms regulate plasma osmolality: antduretic hormone ADH) and thirst. ADH release s supressed, thirst s inhbted and renal water loss occurs when plasma osmolality below a threshold level. The rise in plasma osmolalty causes ADH release, stimulation of thirst and water intake. Acute water intoxication is exceptional in patients without a chronic psychiatric disease. Herein, we describe a case of acute water intoxication in a previously healthy patient, after making an intravenous urography. The excessive water intake and the impossibility of renal water loss because of streee-induced ADH release originated t. Only nine cases have been previously described; almost they all were women preparing for diagnostic procedures.

IGF-1 and its binding proteins IGFBP-1 and 3 as nutritional markers in prepubertal children.

OBJECTIVE: To assess the validity of the use of IGF-1, IGFBP-1 and IGFBP-3 as biochemical markers of nutritional status in prepubertal healthy children. DESIGN: Cross-sectional survey. SETTING: Healthy children from the Madrid area. PARTICIPANTS: Prepubertal children (aged 7-10 years) with a body mass index (BMI) above the 90th percentile (n = 25) and below the 10th percentile (n = 31) were selected from 2,559 included in the CAENPE study (Food Intake and Nutritional Status in Schoolchildren from Madrid). RESULTS: Overweight children were found to have higher serum levels of IGF-1 (306+/-162.2 vs. 209+/-71.2 ng/ml, p<0.001) and IGFBP-3 (3.3+/-1.0 vs. 2.9+/- 0.5 mg/l, p<0.01) and lower serum levels of IGFBP-1 (4.3+/-3.9 vs. 13.8+/-7.4 microg/l, p<0.01). There was a positive correlation with BMI, for IGF-1 and IGFBP-3, and negative correlation for IGFBP-1. No differences in albumin and transferrin concentrations were observed between both groups of children. CONCLUSION: IGF-1, IGFBP-1 and IGFBP-3 clearly classify over- and underweight prepubertal children, showing a good correlation with BMI. They can be used as biochemical markers of caloric nutritional status in this population.

[Nutritional management of radiation-induced enteritis: apropos of a case]. / Manejo nutricional en la enteritis rádica: a propósito de un caso.

We present a case of a 46-year-old woman who is hospitalized in March of 1997 for a picture of severe mixed malnutrition. We describe the clinical history since the diagnosis of an endometrium carcinoma in 1995, treated with radiation therapy, with the aim of determining the etiology of the severe malnutrition picture, as well as the different complications she presented in the course of her hospitalization that justify the decisions made in the choice of the nutritional support.

[Thyroid disorders associated with interferon treatment in patients with chronic viral hepatitis or multiple sclerosis]. / Alteraciones tiroideas asociadas al tratamiento con interferón en pacientes con hepatitis crónica viral o esclerosis múltiple.

BACKGROUND: Alpha and beta interferon (IFN-alpha and beta) treatment is associated with the synthesis of thyroid autoantibodies and the development of autoimmune thyroid diseases. METHODS AND RESULTS: We have retrospectively evaluated their effect in patients with chronic viral hepatitis (CH) (n = 118) and multiple esclerosis (ME) (n = 10). Thyroid dysfunction has been detected in 7.4% of patients, and seroconversion in 4.7%. CONCLUSIONS: Pre-treatment antithyroid antibodies do not predispose to altered thyroid function, after IFN therapy; their presence should not contraindicate IFN treatment.

[Critical analysis of the evolution of commercial preparations for enteral nutrition during 1988-1996]. / Análisis crítico de la evolución de los preparados comerciales enterales en el período 1988-1996.

Enteral nutrition is a form of nutritional support that is continually growing and expanding, and within this area especially the enteral preparations or formulae. The object of the present is to analyze the evolution of the commercially available nutritionally complete enteral preparations between 1988 and 1997, illustrating the variations that have occurred both from the quantitative and form the qualitative points of view. A progressive increase is seen in the absolute number of available enteral formulae, with the increase in both polymeric formulae with dietary fiber, and that of formulae designed for specific diseases being significant (p < 0.05). The standard polymeric formulae and the oligomeric formulae decrease, but not in a significant manner. There is a special discussion or the characteristic and the usefulness of the polymeric formulae with dietary fiber ad the formulae designed especially for specific diseases. The clinical efficacy of most of these special formulae is controversial, with there not being sufficient objective clinical evidence at present that justifies their routine use.

[Description and evaluation of the adequacy of parenteral nutrition in our hospital]. / Descripción y valoración de la adecuación de la nutrición parenteral en nuestro hospital.

INTRODUCTION: Parenteral nutrition (PN) is a form of nutrition that can be very beneficial, but it does have some indications and complications that should be taken into consideration. With this work we would like to describe the characteristics of our patients with PN, as well as their follow up, and to evaluate the appropriateness of its prescription. MATERIAL AND METHODS: Retrospective study carried out at a hospital with 415 beds. All patients who received PN during the first six months of 1996 were reviewed. Inadequacy criteria are established and a nutritional assessment was carried out by means of clinical and analytical parameters. Catheter infections were recorded according to specific criteria and the clinical and analytical evolution of the patients was followed. RESULTS: A total of 83 patients were given parenteral nutrition. The most frequent indication was prolonged fasting after a surgical resection of the digestive apparatus. The most common nutritional status when beginning the PN was severe protein malnutrition (23%). There were 33 cases (39.7%) of PN that was not indicated and/or potentially avoidable. A catheter infection was reported in 7 cases (8.4%).

[Nutritional status of patients with HIV infection. Spontaneous evolution during the hospital stay]. / Estado nutricional de pacientes con infección por VIH. Evolución espontánea durante un ingreso hospitalario.

UNLABELLED: BASICS: Evaluate the nutritional status of HIV positive patients who are admitted to hospital for some acute process, and their evaluation, without nutritional support, during the admission. METHODS: Prospective study in HIV positive patients. Nutritional evaluation (on admission and on release); (1) anthropometric (weight, size, triceps, fold, circumference of the arm, and muscular circumference of the arm, and (2) biochemical (albumin, cholesterol, triglycerides, lymphocytes, pre-albumin, and transferrin). Statistical study: comparison of the paired means and chi squared test. RESULTS: 60 patients. Mean age 32 +/- 4.8 years, 76% men and 23% women. Staging: AIDS 84.8%, non-AIDS 15.2%. Main reason for admission: Infection (80%). Mean stay: 14 days +/- 9.5. Initial nutritional evaluation: normal: 1.7%, protein malnutrition: 5.3%, caloric malnutrition: 38.5%, mixed malnutrition: 54.3%, 85% of the patients refer weight loss. 21 patients (35%) were followed up. There were no significant differences in the anthropometric nor in the biochemical parameters, except in the levels of pre-albumin and transferrin, which improved (p < 0.001). Nutritional evaluation on release: normal: 9.5%, caloric malnutrition: 66.5%, mixed malnutrition: 23.7%. There were no cases of protein malnutrition. CONCLUSIONS: The vast majority of the HIV+ patients who are admitted, are malnourished, and after curing the acute process, 90.5% of them remain malnourished. The anthropometric measurements, albumin, cholesterol, and triglycerides do not vary during the hospital admission, despite the treatment and the clinical improvement. The increase of proteins with a short half life is due to controlling the infection, which is why these are not good parameters for the nutritional evaluation in these patients.

[Consumption of sweets and snacks by a population of school children in the Autonomous Community of Madrid. The CAENPE Group]. / Consumo de alimentos del grupo "dulces y golosinas" en la población infantil escolarizada de la Comunidad Autónoma de Madrid. Grupo CAENPE.

BACKGROUND: In our society advertising compaigns promote consumption of superfluous foods, such as sweets and snacks, displacing the intake of other basic nutrients. The aim of this study is to assess the intake of these foods by children, their relative contribution to overall macronutrient intake and their effects on health. SUBJECTS AND METHODS: A dietary assessment was performed on a representative sample of scholar children from the Autonomous Community of Madrid. It reflects food consumption for a 4-days period (3 week-days and 1 weekend day) in 2,698 boys and girls aged 6 to 15 years. The intake is expressed as grams per 1,000 kcal to standardize nutrient consumption in boys and girls. Lipid profile and several markers of nutritional status are also determined. RESULTS: The consumption of sweets and snacks items per 1,000 kcal is higher in boys than in girls (35.8 +/- 20.0 and 34.2 +/- 19.6 g/1,000 kcal, respectively; p = 0.01) and increases during adolescence (12 and 13 years) in both sexes. Sweets and snacks provide 16.1% of dietary total caloric intake, 7.1% of saturated fatty acids, 10.7% of monounsaturated fatty acids, 10.4% of polyunsaturated fatty acids and 11.3% of cholesterol. Children of ages 6, 7, 8 and 9 years with high calculated LDL-cholesterol levels (> or = 120 mg/dl) consume significative higher amounts of sweet foods than children with low calculated LDL-cholesterol levels (< or = 90 mg/dl). CONCLUSIONS: Children from the Autonomous Community of Madrid, Spain consume excessive sweets, which could be responsible for a higher prevalence of diseases related to this intake, such as caries, obesity and obesity-related associate diseases.