Sperm yield after single layer centrifugation with Androcoll-E is related to the potential fertility of the original ejaculate.

Many attempts have been made to identify laboratory tests that are predictive of sperm fertility, both to improve the quality of stallion semen doses for artificial insemination (AI) and to identify potential breeding sires if no fertility data are available. Sperm quality at the stud is mostly evaluated by assessing subjective motility, although this parameter can be poorly indicative of fertility. Sperm morphology and chromatin integrity in Swedish stallions are correlated to pregnancy rate after AI. Because single layer centrifugation (SLC) selects for spermatozoa with normal morphology and good chromatin, retrospective analysis was carried out to investigate whether sperm yield after SLC is linked to potential fertility. Commercial semen doses for AI from 24 stallions (five stallions with four ejaculates each, 19 stallions with three ejaculates each; n = 77) obtained during the breeding season were cooled, and sent overnight to the Swedish University of Agricultural Sciences in an insulated box for evaluation, with other doses being sent to studs for commercial AI. On arrival at Swedish University of Agricultural Sciences, the semen was used for SLC and also for evaluation of sperm motility, membrane integrity, chromatin integrity, and morphology. The seasonal pregnancy rates for each stallion were available. The yield of progressively motile spermatozoa after SLC (calculated as a proportion of the initial load) was found to be highly correlated with pregnancy rate (r = 0.75; P < 0.001). Chromatin damage was highly negatively correlated with pregnancy rate (r = -0.69; P < 0.001). Pregnancy rate was also correlated with membrane integrity (r = 0.58; P < 0.01), progressive motility (r = 0.63; P < 0.01), and normal morphology (r = 0.45; P < 0.05). In conclusion, these preliminary results show that sperm yield after SLC is related to the potential fertility of the original ejaculate, and could be an alternative indicator of stallion fertility if breeding data are not available. Single layer centrifugation is fast (30 minutes) and does not require expensive equipment, whereas other assays require a flow cytometer and/or specialist skills. An additional option could be to transport semen doses to a laboratory for SLC if the stud personnel do not want to perform the procedure themselves.

Pregnancies following artificial insemination with spermatozoa from problem stallion ejaculates processed by single layer centrifugation with Androcoll-E.

Some stallions produce ejaculates of low quality and/or low fertility when used for artificial insemination (AI). The purpose of these five case studies was to use Single Layer Centrifugation (SLC) to select the best spermatozoa from 'problem' ejaculates for subsequent use in AI. Sperm quality, in terms of motility, morphology and chromatin integrity, was improved in the SLC-selected samples compared to the corresponding uncentrifuged samples, with the exception of one stallion thought to have ampullary stasis. In this stallion, neither the incidence of spermatozoa with detached heads nor the proportion of damaged chromatin was decreased by SLC, in contrast to previous results. Pregnancies were obtained after using SLC-selected spermatozoa from the five stallions for AI, indicating that the spermatozoa were functional after SLC. Overall, the results suggest that SLC may be useful when preparing AI doses from some 'problem' ejaculates.

Metabolic effects in neonates receiving intravenous medium-chain triglycerides.

UNLABELLED: The effects of two lipid emulsions, one with 50% each of medium-chain and long-chain triglycerides, and a long-chain triglycerides lipid emulsion as a control, were evaluated for lipid and carnitine metabolism and respiratory quotient when given to neonates after major surgery during a short period of total parenteral nutrition. Each group included 10 neonates, and all tolerated the total parenteral nutrition well. The relative contents of linoleic acid and alpha-linolenic acid increased in all lipid esters in plasma and adipose tissue in both groups, indicating that the content of these fatty acids is sufficient even in the medium-chain triglycerides emulsion. The serum concentration of ketones was within normal limits. Free fatty acids in plasma did not increase in either group. The total plasma carnitine concentration decreased in both groups but the distribution of free carnitine and acylcarnitine did not change. The total muscle carnitine did not change significantly but the ratio of acylcarnitine to free carnitine tended to increase in muscle in the treatment group, probably an effect of the medium-chain triglyceride supplementation. CONCLUSIONS: The two groups displayed the same fatty acid pattern in plasma and adipose tissue and the same respiratory quotient during the treatment period. Regarding carnitine status, essentially the same changes were seen in the two groups. However, discrete changes were seen in muscle tissue in the treatment group.

Bacterial translocation from defunctionalized rat small bowel.

Bacterial translocation from the intestine may cause severe infectious complications in a number of clinical situations, including the short bowel syndrome and after small bowel transplantation. The aim of the present study was to develop a simplified model for the study of bacterial translocation from a defunctionalized intestine. An ileal segment from untreated or cyclosporine-treated rats was exteriorized as a Thiry-Vella loop. After 1, 3. or 7 days, bacterial translocation and distribution of immunocompetent cells were assessed. The data obtained were compared with data from animals subjected to intestinal transplantation. Translocation to the mesenteric lymph nodes was detected in 60% of the Thiry-Vella loop animals on day 1. in 100% on day 3, and in 83% on day 7: concomitantly, the number of macrophages and T-cells in the mesenteric lymph nodes increased from day I until day 7. The degree of bacterial translocation on days 3 and 7 in animals with a Thiry-Vella loop was comparable with that observed 7 days after intestinal transplantation. Furthermore, treatment with cyclosporine A enhanced the number of translocating bacteria. In the model presented here bacterial translocation occurs from the small bowel to the mesenteric lymph nodes. The model offers possibilities to study the mechanisms and immunological phenomena associated with microbial translocation.

Resorbable bioscaffold for esophageal repair in a dog model.

PURPOSE: Porcine-derived, xenogeneic extracellular matrix (ECM) derived from either the small intestinal submucosa (SIS) or urinary bladder submucosa (UBS) was used as a tissue scaffold for esophageal repair in a dog model. METHODS: Patch defects measuring approximately 5 cm in length and encompassing 40% to 50% of the circumference of the esophagus or complete circumferential segmental defects measuring 5 cm in length were created by surgical resection in healthy adult female dogs. The defects were repaired with ECM scaffolds derived from either SIS or UBS. The animals were kept alive for periods ranging from 4 days to 15 months. RESULTS: The xenogeneic scaffolds used for repair of the patch defects were resorbed completely within 30 to 60 days and showed replacement by skeletal muscle, which was oriented appropriately and contiguous with adjacent normal esophageal skeletal muscle, organized collagenous connective tissue, and a complete and intact squamous epithelium. No signs of clinical esophageal dysfunction were seen in any of the animals with the patch defect repair. The xenogeneic scaffolds configured into tubes for repair of the segmental defects all showed stricture within 45 days of surgery. CONCLUSION: These ECMs show promise as a treatment option for esophageal repair, but stricture remains problematic for complete tube grafts.

[Intestinal transplantation. The first Swedish transplantation of the small intestine in an adult patient with pseudoobstruction]. / Tarmtransplantation. Första svenska tunntarmstransplantationen till en vuxen patient med pseudoobstruktion.

Recent advances, first and foremost the development of new immunosuppressive agents, have markedly improved the outcome of intestinal transplantation, which is a treatment option for patients with serious intestinal diseases who have become dependent on total parenteral nutrition. The first small bowel transplantation in Sweden was performed at Huddinge Hospital in 1997, in the adult patient with intestinal pseudo-obstruction. The article reports the course of this patient and an update of international progress in intestinal transplantation.

Fetal rat hepatocytes: isolation, characterization, and transplantation in the Nagase analbuminemic rats.

BACKGROUND: In contrast to adult hepatocytes, fetal hepatocytes (FH) are thought to be highly proliferative, less immunogenic, and resistant to cryopreservation and ischemic injury. These qualities could enhance FH engraftment, proliferation, and gene transfer requiring active DNA synthesis. METHODS: Rat FH were obtained using the nonperfusion collagenase/DNase digestion method. Free and cultured cells were studied using electron microscopy, fluorescence-activated cell sorting, and Northern analysis using alpha-fetoprotein and albumin as markers of hepatocyte lineage. DNA synthetic activity was measured in quiescent and mitogen-stimulated fetal and adult hepatocytes by [3H]thymidine incorporation. Susceptibility of cultured FH to retrovirally mediated gene transfer was studied using an amphotropic retroviral vector carrying the Escherichia coli lac-Z gene. Nagase analbuminemic rats were used as recipients to study the effects of intraportal FH transplantation. Analysis of serum albumin was carried out by enzyme-linked immunosorbent assay. RESULTS: In fetal liver, 87+/-2% of the cells showed morphological and molecular features of hepatocytes. DNA synthetic activity in nonstimulated cultured FH was 10 times greater than the maximal hepatocyte growth factor-driven response in adult rat hepatocytes. A total of 5-15% FH stained positive for X-gal; results of transduction in adult hepatocyte cultures were negative. In Nagase analbuminemic rat recipients, FH produced significant amounts of albumin only when a hepatic regenerative stimulus was applied. Immunohistochemistry confirmed presence of albumin-positive hepatocytes. CONCLUSIONS: Fetal rat liver from the late gestation period is highly enriched with hepatocyte progenitors. They are highly proliferative and susceptible to retroviral transduction and can engraft and function in the adult rat liver if transplanted under a hepatic regenerative stimulus.

Plasma and tissue levels of lipids, fatty acids and plasma carnitine in neonates receiving a new fat emulsion.

This study was undertaken to compare Intralipid with a new fat emulsion containing gamma-linolenic acid and carnitine, named Pediatric Fat Emulsion 4501, in neonates with regard to lipid and carnitine metabolism over a short period of total parenteral nutrition. There were 10 neonates in each group and they tolerated the total parenteral nutrition well. In spite of the gamma-linolenic acid supplementation in the new emulsion, arachidonic acid decreased significantly in plasma lipid esters and adipose tissue in both groups after 5 d of treatment. Also, there was a decrease in plasma docosahexaenoic acid which was more pronounced in the treatment group. The relative percentage values of linoleic and linolenic acids in adipose tissue were increased, indicating that newborns have a rapid accretion of fatty acids. Plasma-triglycerides were effectively cleared during the periods without fat infusion. In the group that received Pediatric Fat Emulsion 4501 the means of both free and total plasma carnitine concentrations increased significantly, whereas they tended to decrease in the Intralipid group.

Interactions of vitamins A and E and retinol-binding protein in healthy Swedish children--evidence of thresholds of essentiality and toxicity.

The serum levels of vitamins A and E and retinol-binding protein (RBP) were determined and their relationships investigated in 144 healthy Swedish children between the ages of 1 month and 16 years. The median level for vitamin A was 540 micrograms l-1 (range 181-1720 micrograms l-1); for vitamin E 6.8 mg l-1 (range 1.0-14.0 mg l-1) and for RBP 0.030 g l-1 (0.018-0.045 g l-1). Retinol-binding protein levels rose steeply in relation to vitamin A levels of up to approximately 800 micrograms l-1. However, RBP levels remained constant when vitamin A levels were between 800 and 1600 micrograms l-1 but displayed a further steep rise for vitamin A levels above this range. Fourteen individuals had levels that did not follow this pattern but fell rather neatly below the correlation line. These individuals were also found to have significantly higher mean levels of acute phase reactants than the rest of the study group, suggesting the possibility of subclinical infection or inflammation in these subjects. Vitamins A and E showed an inverse curvilinear relationship. This is the first report of an interaction between vitamins A and E and RBP in healthy Swedish children. The study demonstrates that a linear one-to-one relationship between RBP and vitamin A may not be of regular occurrence as previously assumed. The pattern described suggests the existence of thresholds of essentiality and toxicity in the interaction between the two vitamins and RBP.(ABSTRACT TRUNCATED AT 250 WORDS)

Plasma levels of fat-soluble vitamins A and E in neonates, after administration of two different vitamin solutions.

The aim of this study was to compare the effects of 2 different vitamin solutions on plasma levels of vitamin A and E during and after short term total parenteral nutrition (TPN), after neonatal surgery. 2 compounds were compared, one with a higher vitamin A content (100 mu g/ml) and no vitamin E and one with vitamin E (0.64 mg/ml) and a lower vitamin A content (69 mu g/ml). 2 randomly chosen groups of 10 neonates were studied each with gastrointestinal malformations. The groups were comparable regarding gestational age (GA), weight and length. Diagnoses within the groups were similar. TPN was given after corrective surgery for a median time of 6.5 days to both groups. Vitamin A and retinol binding protein (RBP) levels increased significantly (p < 0.05) above the pre-treatment level in patients receiving supplementation with a higher dosage of vitamin A (100 mu g/ml). In patients receiving the lower dose of vitamin A (69 mu g/ml) plasma levels of this vitamin were unchanged but RBP levels fell significantly (p < 0.05). There was no significant difference in plasma vitamin E levels in the group of patients receiving vitamin E supplementation as compared with the unsupplemented group. In both groups vitamin E levels increased significantly (p < 0.005) after introduction of oral feeding. The results indicate that even during short term TPN endogenous stores of vitamin A might become depleted. In spite of the fact that no vitamin A deficiency was noted it seems obvious that a higher vitamin A intake is advisable which is in accordance with the recommendations from the Subcommittee on Paediatric Parenteral Nutrient Requirements from the Committee on Clinical Practice Issues of The American Society for Clinical Nutrition.

Plasma concentrations of fat-soluble vitamins A and E in neonates with myelomeningocele.

A group of newborn infants with myelomeningocele was compared with a group of neonates suffering from gastrointestinal malformations and a group of healthy children less than one year of age. The parameters studied were prealbumin, retinol-binding protein and the fat-soluble vitamins A and E. There were no significant differences between the groups in the plasma levels of the proteins or vitamin E. Vitamin A levels in newborn infants with myelomeningocele were found to be significantly lower compared to both the other groups.

Clinical adsorption and photodegradation of the fatsoluble vitamins A and E.

The adsorption of the fat-soluble vitamins A and E to administration sets and their photodegradation was studied under varying conditions. Infusion bottles with or without photoprotectors (red plastic bags) were studied, as were systems exposed to the blue light, used in the treatment of hyperbilirubinaemia, which has a maximum emission spectrum at 450 nm. An IV bag containing lipids, carbohydrates, amino-acids, minerals and vitamins was also studied. Vitamin levels in all administration sets decreased by approximately 30% during the first 3h, except for those in the IV bag. Vitamins were best preserved in the IV bag, where 85-90% of both vitamins remained after 20h. Vitamin E appeared to suffer from both adsorption and photodegradation but to a lesser extent than vitamin A. Vitamin A displayed a greater tendency for adsorption. In order to avoid loss of vitamins during parenteral administration, we recommend that they should be administered as a bolus or during a limited period of 1 h, and that the system be protected from light.

Megacystis-microcolon-intestinal hypoperistalsis syndrome (MMIHS), an autosomal recessive disorder: clinical reports and review of the literature.

We present 2 new patients with the megacystis-microcolon-intestinal hypoperistalsis syndrome (MMIHS), review the literature, and discuss the prenatal diagnosis and treatment. MMIHS, as reported in 43 cases, is usually lethal. Most children die during the first year of life, and only 3 children survived their first year. We report the 6th pair of sibs with the disease. Overall, 17 patients reported have had sibs with MMIHS or the parents were consanguineous; 4 times the parents were first, cousins, confirming that this is an autosomal recessive disorder. The present 2 children, whose parents also were first cousins, were of different sex. They had typical MMIHS with abdominal distension due to pronounced megacystis, hydronephrosis, microcolon, and microileum, involving the distal part of the ileum, malrotation of the gut, and intestinal hypoperistalsis. Neither surgery nor medical treatment was successful and the children died at the age of 19 days and 2 1/2 months, respectively. There is no cure for the disease. However, a new protkinetic drug, Cisapride might be worth trying in these cases. Prenatal ultrasound diagnosis of MMIHS might be possible.

Effects of vitamin A on endotoxaemia in rats.

OBJECTIVES: To find out if survival after peritonitis induced by caecal puncture and injection of endotoxin is correlated with serum endotoxin concentrations in rats; whether this study could be carried out by studying the effects of vitamin A in rats with peritonitis; and if it was possible to differentiate between local and systemic effects of vitamin A on endotoxaemia in rats. DESIGN: Prospective randomised study. MATERIAL: Sixty adult male Sprague-Dawley rats. INTERVENTIONS: Rats were kept on a diet containing no vitamin A for six days; 30 were given vitamin A 1,380 IU/kg/day intramuscularly and 30 were given peanut oil. Peritonitis was then induced in 16 rats by caecal puncture (treated n = 10, untreated n = 6) and the remainder received intraperitoneal injections of either 0.5 mg/kg endotoxin, or 2.0 mg/kg endotoxin. MAIN OUTCOME MEASURES: Plasma concentrations of vitamin A, vitamin E, and serum concentrations of endotoxin, estimation of erythrocyte volume fraction and leucocyte particle concentrations, and survival. RESULTS: Rats that were given 2 mg of endotoxin had higher mortality and did generally less well than others. Serum endotoxin concentrations were lower in those groups that had been pretreated with vitamin A. No animal developed vitamin A deficiency and there were no differences in erythrocyte volume fraction or leucocyte particle concentration between those that did and did not receive vitamin A. CONCLUSION: Vitamin A lowered the concentration of endotoxin in the circulation, it has a central function in the host's defence against endotoxin, and the experimental model seems suitable for the study of the effect of vitamin A or endotoxemia and peritonitis.

Serum trace elements in children receiving long-term parenteral nutrition.

The trace elements iron (Fe), copper (Cu), zinc (Zn), selenium (Se), chromium (Cr), manganese (Mn), nickel (Ni), rubidium (Rb), and bromine (Br) were determined in serum from 18 children, ages 4 to 65 months, who received long-term total parenteral nutrition (TPN). Nine patients (group 1) received TPN as their only source of nutrition; the other nine (group 2) ingested 30% to 70% of their total calorie requirement and received the remainder intravenously. The reference group consisted of 19 healthy children of similar age and sex distribution. Groups 1 and 2 had received parenteral nutrition for 28.4 +/- 13.7 (SD) and 23.1 +/- 17.1 (SD) months, respectively. All patients in group 1 and four in group 2 had short bowel syndrome; four in group 2 had pseudo-obstruction syndrome. Patients in group 1 had normal serum levels of Fe, Cu, and Mn, but increased levels of Zn; they had decreased serum concentrations of Se, Cr, Ni, Rb, and Br. Patients in group 2 had decreased serum levels of Mn, Cr, Ni, Rb, and Br. Those trace elements not supplemented in TPN solutions (Se, Cr, Mn, Ni, Rb, and Br) were lower in the children receiving TPN than in the control children, indicating inadequate intake of these trace elements from food and TPN solutions. However, no obvious clinical signs or symptoms of trace element deficiencies were observed.

Total parenteral nutrition with different ratios of fat/carbohydrate at two energy levels: an animal study.

Different regimens of total parenteral nutrition were evaluated in 64 growing rats, during a 10-day period. Thirty-two rats were kept at a low energy regimen (270 kcal/kg body weight per day), and the remaining rats were infused with a high energy solution (350 kcal/kg/day). At each energy level, four fat/carbohydrate (CHO) nonprotein energy ratios were tested: 0% fat/100% CHO (no fat); 6% fat/94% CHO (low fat); 30% fat/70% CHO (medium fat); 60% fat/40% CHO (high fat). A daily supply of 0.9 g nitrogen/kg of a well balanced amino acid solution was administered to all rats. Growth, nitrogen balance, net nitrogen utilization, and blood status were evaluated. Carcass, liver, and muscle composition were investigated with respect to protein, fat, and water content. Liver and body composition was similar between groups at the same energy level. At the higher energy level the fat deposition was increased. At the low energy level, rats from no fat and low fat groups lost weight during the first 3 days of the experimental period. The medium fat groups showed the highest weight gain at each energy level. This suggests the importance of the presence of fat in total parenteral nutrition programs. During the last days of the experimental period, all rats gained weight, and showed a positive nitrogen balance. All groups given 350 kcal/kg/day gained weight more rapidly and showed a better nitrogen balance and utilization than the corresponding group given 270 kcal/kg/day. This study emphasizes that a well-balanced proportion of energy and substrates is important.