Comparison of plant- and egg yolk-based semen diluents on in vitro sperm kinematics and in vivo fertility of frozen-thawed bull semen.

Diluents using components of plant origin have been developed as an alternative to animal based extenders for the dilution of bull semen, however, it is unclear if use of these diluents results in in vivo fertility rates similar to those that occur with use of traditional egg yolk-based diluents. The aim of this study was to assess the effect of semen diluent on 60-day non-return rate (NRR) following artificial insemination (AI) with frozen-thawed bull semen. The effect of semen dilution in one of three different commercial diluents (BullXcell - egg yolk-based, OptiXcell - plant-based or AndroMed - plant-based) on post-thaw total and progressive motility as well as kinematic parameters (Experiment 1) and field fertility (Experiment 2, n = 1,480 inseminations) was assessed. Semen stored in OptiXcell had greater post-thaw total and progressive motility than AndroMed (P < 0.05) but did not differ from BullXcell. Semen stored in BullXcell had a greater beat cross frequency and straight line velocity compared to semen stored in AndroMed (P < 0.05) but did not differ when compared with use of OptiXcell; while values for these variables when using OptiXcell and AndroMed did not differ from each other (P > 0.05). There was no difference in any other sperm kinematic parameters (P > 0.05). There was no effect of diluent on 60-day NRR (71.5%, 67.8% and 70.6% for BullXcell, OptiXcell and AndroMed, respectively). In conclusion, while diluent significantly affected post-thaw sperm motility and kinematics, no effect on 60-day NRR was observed. Given that OptiXcell and AndroMed are animal protein-free media these diluents may be a suitable alternative to BullXcell for the storage of frozen-thawed bull semen.

Effect of storage temperature, nitrogen gassing and sperm concentration on the in vitro semen quality and in vivo fertility of liquid bull semen stored in INRA96.

The aim of this study was to assess the effect of storage temperature, nitrogen (N2) gassing and sperm concentration on in vitro characteristics and calving rate (CR) following artificial insemination (AI) of liquid bull semen stored in INRA96. In Experiment 1 the effect of liquid bull semen diluted in either N2 bubbled or non-bubbled INRA96 at a concentration of 5 × 106 sperm per 0.25 mL insemination dose and stored at 5 or 15 °C was assessed subjectively for total and progressive motility on Days 0, 1, 2, 3 and 4 post collection. In Experiment 2a, the effect of stored liquid semen at three sperm concentrations (3, 4 or 5 × 106 sperm per 0.25 mL insemination dose) on total and progressive motility was assessed subjectively on Days 0, 1 and 2 post collection. In Experiment 2b, the field fertility of liquid semen stored at ambient temperature at a concentration of 3, 4 or 5 × 106 sperm per 0.25 mL dose and inseminated on Days 1 or 2 post collection was assessed in comparison to frozen-thawed semen (total of n = 5742). In Experiment 1, total and progressive motility decreased with increased duration of storage (P < 0.01); however, there was no effect of N2 bubbling on motility on Days 0, 1, 2, 3 and 4 of storage. There was an effect of temperature on total and progressive motility, regardless of treatment, as semen stored at 15°C recorded higher motility values than semen stored at 5°C (P < 0.01). In Experiment 2a, there was no effect of sperm concentration on total or progressive motility on Days 0, 1 or 2 of storage. There was a linear decrease in motility with increased duration of storage (P < 0.01); however, there was no sperm concentration by day interaction. In Experiment 2b, there was an effect of sperm concentration on CR (P < 0.01); semen diluted to 3 and 4 × 106 sperm per dose resulted in a lower CR after 2 days of storage (41.1 and 44.7%, respectively) in comparison to frozen-thawed semen (55.2%) but did not differ to CR of semen diluted to 5 × 106 sperm per dose on Day 2 of storage. There was an effect of parity, fertility sub-index and days in milk (DIM) at AI on CR (P < 0.01). In conclusion, N2 bubbling and sperm concentration had no effect on in vitro sperm motility of liquid semen, but this study demonstrated a reduction in CR on Day 2 of storage at lower sperm concentrations in comparison to frozen-thawed semen.

Effect of increasing equilibration time of diluted bull semen up to 72 h prior to freezing on sperm quality parameters and calving rate following artificial insemination.

An equilibration period of approximately 3-4 h prior to semen cryopreservation is standard practice for maintaining membrane integrity and motility of bull sperm. However, a number of studies indicate that an overnight equilibration period prior to freezing results in improved post-thaw semen quality thus optimising pregnancy rates. The aim of this study was to assess the effect of increasing the equilibration time of bull semen up to 72 h before freezing on sperm quality parameters and calving rate (CR) following artificial insemination (AI) with frozen-thawed semen. The effect of holding semen at 4 °C for 6, 24, 48 or 72 h post dilution before freezing on subsequent post-thaw total and progressive motility (Experiment 1) and field fertility (n = 1640 inseminations, Experiment 2) of frozen-thawed semen was assessed. Equilibration time did not affect post-thaw total and progressive motility (P > 0.05). In addition, there was no effect (P > 0.05) of equilibration time on field fertility with a CR of 53.3, 50.5, 51.3 and 47.3 for the 6, 24, 48 and 72 h treatments, respectively. In conclusion, increasing the equilibration time of diluted bull semen from 6 to 72 h had no significant effect on CR, within the expected range of fertility outcomes, thus providing semen processing centres with flexibility in the time which semen can be held prior to freezing.

Effect of seminal plasma from high- and low-fertility bulls on cauda epididymal sperm function.

The aim of this study was to characterise the effect of seminal plasma (SP) from bulls of high or low fertility on sperm function. First, the effect of SP on the motility of fresh cauda epididymal spermatozoa (CES) and frozen-thawed ejaculated spermatozoa was assessed (Experiment 1a). Seminal plasma was then collected from bulls of known high and low fertility. Pooled CES were incubated in the SP from each bull, diluted and assessed for motility and viability on Days 1, 2, 3 and 5 after packaging as fresh semen (Experiment 1b). Also assessed were motility, kinematics, viability and mitochondrial membrane potential after thawing (Experiment 1c) as well as hypotonic resistance (Experiment 2) and fertilisation potential using in vitro fertilisation (Experiment 3). Seminal plasma increased the motility of CES (P<0.05); however, there was no effect of SP on the motility and viability of fresh CES or on CES post-thaw motility, viability and mitochondrial membrane potential (P>0.05). The hypotonic resistance of CES was reduced by SP (P<0.05), irrespective of whether the SP was from high- or low-fertility bulls. Seminal plasma from high- or low-fertility bulls had no effect on cleavage or blastocyst rates (P>0.05). In conclusion, SP affects the physiological function of CES but there is no difference between SP from high- or low-fertility bulls.

The effect of dietary supplementation of algae rich in docosahexaenoic acid on boar fertility.

The objective of this study was to assess the effects of dietary supplementation of a commercial algal product rich in docosahexaenoic acid (DHA) on boar fertility as assessed in vitro and in vivo. Boars were fed one of three experimental diets for 19 weeks: (i) Control (Ctl) diet (n = 31), (ii) Ctl diet plus 75g All-G-Rich per day (n = 31) or (iii) Ctl diet plus 150g All-G-Rich per day (n = 30). Parameters assessed were (i) raw semen quality; volume, sperm concentration, total motility and morphology (ii) liquid semen quality; progressive motility, viability, hypotonic resistance and acrosomal integrity (iii) frozen-thawed semen quality; motility, thermal stress, viability, membrane fluidity and mitochondrial activity (iv) sperm and seminal plasma (SP) fatty acid composition (FAC) (v) total antioxidant capacity (TAC) of SP and (vi) farrowing rates and litter sizes of sows (n = 1158) inseminated with liquid semen. Boars consuming 75g All-G-Rich had a larger semen volume (P < 0.05) and a higher total sperm number (P < 0.01) than the Ctl treatment, however, there was no effect of treatment on any other semen quality parameter (P > 0.05). There was no effect of dietary treatment on the FAC and TAC of SP or on farrowing rate and litter size (P > 0.05). There was an effect of dietary treatment on the FAC of sperm, represented by an 1.72 and 1.60 fold increase in the DHA content for 75 and 150g treatments, respectively, compared to the Ctl treatment. In conclusion, a significant increase in semen volume and total sperm number in boars supplemented 75g All-G-Rich daily, resulted in an increase in production of 3 to 4 more doses per ejaculate, thus, indicating that the feeding regime described within this study has the potential for increasing the output of boar studs.

Cytoreductive surgery and intraperitoneal chemotherapy for peritoneal mesothelioma.

AIMS: Peritoneal mesothelioma is a rare disease and traditionally has been associated with a gloomy prognosis. The present study aimed to report the outcomes following surgery and intraperitoneal chemotherapy in selected patients with peritoneal mesothelioma. METHODS: Clinicopathological features, operative procedures, early outcomes and survival were analysed for 17 consecutive patients who underwent surgery for peritoneal mesothelioma between 1998 and 2007. Seventeen consecutive patients who underwent surgery for peritoneal mesothelioma between 1998 and 2007 were analysed for clinicopathological features, operative procedures, early outcomes and survival. RESULTS: Seventeen patients underwent 18 laparotomies. Most presented with abdominal distension (71%) and abdominal pain or discomfort (53%). Complete cytoreduction was achieved in 8 patients, major debulking in 8, and 1 patient had an exploratory laparotomy only due to extensive disease. One patient died on day 30 postoperatively due to a chest infection and pulmonary embolism. The median survival for 8 patients who underwent complete cytoreduction was 3.7 years (range, 0.7-6.9), whereas that for 8 patients with palliative debulking was 1.0 years year (range, 0.3-5.7). Among the 12 patients who had significant ascites as a presenting symptom, 10 reported good palliation of ascites. CONCLUSIONS: Cytoreductive surgery combined with intraperitoneal chemotherapy appears to be the optimal treatment for selected patients with peritoneal mesothelioma. Increased familiarity with this condition's presentation and natural history, and knowledge of available treatment options, will hopefully facilitate treatment of these patients and expedite speedy referral to appropriate treatment centres.

Quantification of subclasses of human colonic myenteric neurons by immunoreactivity to Hu, choline acetyltransferase and nitric oxide synthase.

An accurate method to count human enteric neurons is essential to develop a comprehensive account of the classes of nerve cells responsible for gut function and dysfunction. The majority of cells in the enteric nervous system utilize acetyl choline, or nitric oxide, or a combination of these, as neurotransmitters. Antisera raised against the RNA-binding protein Hu, were used to identify nerve cell bodies in whole mounts of the myenteric plexus of human colon, and then were utilized to analyse cells immunoreactive for combinations of choline acetyltransferase and nitric oxide synthase. Antisera to Hu provided a reliable means to count apparently all enteric nerve cell bodies, revealing 10% more cell bodies than labelling with neuron specific enolase, and no labelling of glial cells as revealed by S100. ChAT+/NOS- neurons accounted for 48% (+/-3%) of myenteric neurons and ChAT-/NOS+ neurons accounted for 43% (+/-2.5%). ChAT+/NOS+ neurons comprised 4% (+/-0.5) of the total number of neurons, and a novel class of small ChAT-/NOS- neurons, making up 5% (+/-0.9%) of all cells, was described for the first time.

Management of an unexpected appendiceal neoplasm.

BACKGROUND: Appendiceal neoplasms are rare and most present unexpectedly as acute appendicitis. The classification and management are confusing, and there are few substantial reports in the literature. METHODS: A systematic literature review was performed to access relevant publications on the presentation, pathology and management of appendiceal tumours. RESULTS: Appendiceal tumours account for 0.4 to 1 per cent of all gastrointestinal tract malignancies and are found in 0.7 to 1.7 per cent of appendicectomy specimens. Carcinoid tumours are most common. Most are cured by simple appendicectomy if the tumour is less than 2 cm in size and does not involve the resection margin or mesoappendix. Epithelial tumours may present with, or in time develop, pseudomyxoma peritonei, the optimal management of which involves complete tumour resection and intraperitoneal chemotherapy, usually available only in specialized centres. CONCLUSION: Suggested algorithms for the management of unexpected appendiceal tumours are provided. Recommendations are made for follow-up of patients with a perforated appendiceal epithelial tumour.

Endocinch therapy for gastro-oesophageal reflux disease: a one year prospective follow up.

BACKGROUND: Gastro-oesophageal reflux disease (GORD) is, in the main, treated with either proton pump inhibitor (PPI) drugs or a Nissen fundoplication operation. Recently, BARD developed Endocinch, a device used to place sutures just below the oesophagogastric junction (OGJ) to treat GORD. AIM: To evaluate the long term benefit of the Endocinch technique in patients seen up to 12 months post procedure. PATIENTS: Twenty six patients with symptoms of GORD were recruited and had the procedure performed. Four patients were lost to follow up. METHODS: Twenty two patients completed their one year follow up. Pre procedure and post procedure (up to 12 months) assessments included symptom scoring (DeMeester), upper intestinal endoscopy, oesophageal manometry and 24 hour oesophageal pH, and completion of quality of life (QOL) questionnaires. RESULTS: Mean age was 39 years (range 22-62). Heartburn symptom score was reduced from a mean value of 19.22 at baseline to 7.5 at 12 months (n=22) (p<0.0001). Regurgitation score reduced from a mean of 2.27 at baseline to 0.86 at 12 months (n=22) (p<0.001). Mean (SEM) pH DeMeester acid score was reduced from 44.1 (4.3) to 33.32 (4.73) (p=0.028) at three month post procedure. Percentage upright acid exposure and number of reflux episodes were also reduced significantly. Use of PPIs was reduced by 64% at 12 months post procedure. All QOL assessments showed significant improvement (p=0.01). All transient post procedure complaints resolved within 72 hours. CONCLUSION: The Endocinch procedure is an effective and safe outpatient procedure that offers GORD patients significant improvement in symptomatology, QOL, and reduced requirements for PPIs over at least a one year period.

Characterization and origin of dissolved organic carbon in Yegua ground water in Brazos County, Texas.

Dissolved organic carbon (DOC) concentrations in five shallow (< 20 m) and three deeper wells (27 to 30 m) in the Eocene Yegua Formation (Brazos County in east-central Texas) ranged from 92 to 500 microns. Characterization of high, intermediate, and low molecular weight DOC fractions (HMW > 3000 amu, IMW 1000 to 3000 amu, and LMW 500 to 1000 amu) and combined neutral sugar analyses provide information on organic matter sources in the Yegua aquifers. Combined neutral sugars ranged in concentration from 0.6 to 2.7 mumol/L and comprised 0.8% to 6.7% of DOC in ground water. Glucose was the most abundant neutral sugar, followed by xylose and galactose, arabinose, mannose, rhamnose, and fucose. These combined neutral sugars were more diagenetically altered in shallow, oxic ground water as indicated by high mole % fucose + rhamnose and low neutral sugar yield. The precursors for neutral sugars are most probably angiosperm leaves, which show a similar distribution pattern of neutral sugars. Ground water DOC was depleted in 13C relative to soil-zone organic matter (OM) (-16@1000 to -19@1000). The delta 13C values of bulk DOC and HMW DOC ranged from -24@1000 to -32@1000, whereas LMW and IMW DOC ranged from -32@1000 to -34@1000 and -16@1000 to -28@1000, respectively. This variability in delta 13C values is probably related to microbial processes and selective preservation of OM. Carbon isotope analyses in bulk and different molecular weight DOC fractions imply a predominantly C3 OM source and a low contribution of soil-zone OM to DOC.

Stochastic-convective transport with nonlinear reaction and mixing: application to intermediate-scale experiments in aerobic biodegradation in saturated porous media.

Aerobic biodegradation of benzoate by Pseudomonas cepacia sp. in a saturated heterogeneous porous medium was simulated using the stochastic-convective reaction (SCR) approach. A laboratory flow cell was randomly packed with low permeability silt-size inclusions in a high permeability sand matrix. In the SCR upscaling approach, the characteristics of the flow field are determined by the breakthrough of a conservative tracer. Spatial information on the actual location of the heterogeneities is not used. The mass balance equations governing the nonlinear and multicomponent reactive transport are recast in terms of reactive transports in each of a finite number of discrete streamtubes. The streamtube ensemble members represent transport via a steady constant average velocity per streamtube and a conventional Fickian dispersion term, and their contributions to the observed breakthroughs are determined by flux-averaging the streamtube solute concentrations. The resulting simulations were compared to those from a high-resolution deterministic simulation of the reactive transport, and to alternative ensemble representations involving (i) effective Fickian travel time distribution function, (ii) purely convective streamtube transport, and (iii) streamtube ensemble subset simulations. The results of the SCR simulation compare favorably to that of a sophisticated high-resolution deterministic approach.

The influence of restricted calorie intake on peritoneal macrophage function.

Malnutrition leads to immune dysfunction with greatly increased morbidity. However, restrictive dietary regimens are also known to preserve immune function in autoimmune-susceptible mice. The macrophage (Mø) is central to both immune effector and autoregulatory functions and is critical to host-defense mechanisms. The aim of this study was to investigate the effect of calorie restriction on Mø functions in mice. Female, 6- to 8-wk-old, Swiss Webster mice were randomized to ad libitum feeding for 7 or 21 d (n = 10 mice/group), restricted feeding (13.5 to 14.0 g/cage/d; n = 10) for 7 d, or restricted feeding (16.5 to 17.0 g/cage/d; n = 10) for 21 d. These restrictions were equivalent to a decrease in calorie intake of 21.9% and 5.1%, respectively, over 7 and 21 d. All mice were allowed free access to water. On days 8 and 22, respectively, the mice were killed, and peritoneal Møs were isolated by lavage and adhered to 96-well polystyrene tissue-culture-treated plates. After stimulation with lipopolysaccharide, supernatant prostaglandin E2 and interleukin-6 levels were measured by enzyme-linked immunosorbent assay. Supernatant NO2- in response to stimulation with lipopolysaccharide and interferon-gamma was determined by the Greiss reaction. Prostaglandin E2 production was significantly elevated in peritoneal Møs from the calorie-restricted mice compared with the ad-libitum-fed mice after 7 d. After 21 d, production of both prostaglandin E2 and nitric oxide was significantly increased (P < 0.05) in peritoneal Møs from the restricted mice compared with the ad-libitum-fed mice. These results indicate that calorie restriction influences immune function by altering prostaglandin E2 and nitric oxide generation by Møs.

Prostaglandins regulate melanoma-induced cytokine production in macrophages.

Tumor-secreted products can affect macrophage cytokine expression and in that way alter the immune response. Prostaglandins (PGs) are found in the tumor microenvironment and have been associated with local and regional immunosuppression. We investigated whether tumor-secreted factors could induce PG synthesis in macrophages and whether these PGs could alter macrophage production of immunoregulatory cytokines. In both murine and human models, melanoma conditioned medium (MCM) induced macrophage production of PGE(2), IL-6, and TNF-alpha. PGE(2) production increased over 24 h and was accompanied by an increase in cyclooxygenase-2 (COX-2) expression, while COX-1 expression remained unchanged. In the presence of 10 microM NS398, a selective COX-2 inhibitor, MCM-stimulated PGE(2) synthesis was almost completely suppressed, while production of IL-6 and TNF-alpha proteins and mRNA also was partially abrogated. In the murine model, 200 microM NS398 resulted in more significant inhibition of cytokine protein and mRNA production. Although MCM induced NFkappaB and NF-IL-6 activation, neither dose of NS398 altered this effect. We conclude that melanoma-secreted products stimulate COX-2 expression and PGE(2) synthesis in macrophages and that inhibition of COX-2-derived PG synthesis results in partial abrogation of macrophage cytokine production.

Rib lesions and tuberculosis: the palaeopathological evidence.

SETTING: Tuberculosis diagnosis in past populations relies on lesions in the spine and major weight bearing joints of the body. Bone formation on visceral surfaces of ribs has also been suggested to be the result of chronic pulmonary disease. OBJECTIVE: To test whether these lesions are the result of pulmonary infection (most likely tuberculosis), by reviewing past work, and to discuss whether these lesions could be considered another diagnostic criterion for pulmonary tuberculosis. DESIGN: A review of the literature on new bone formation on ribs, and consideration of further evidence from archaeological skeletal material from the UK. RESULTS: Results from modern studies suggest that bone formation on ribs is often the result of pulmonary tuberculosis, that lesions are relatively common in archaeological skeletal material, and that some skeletons have rib lesions plus pathognomonic changes of tuberculosis. CONCLUSION: Evidence suggests that new bone formation on visceral surfaces of ribs should be considered a possible indicator of tuberculosis. If accepted, historical evidence, when correlated with rib data, produces closer approximations to the frequency of the disease in the past. This study indicates the importance of palaeopathology in identifying sometimes subtle lesions that may not be noted by clinicians because of their non-visibility on radiographs.

Molecular mapping of SSRs for Pgm1 and C8b in the vicinity of the rat fatty locus.

Recessive mutations at the rat fatty locus (fa, facp), which produce obesity, insulin resistance, and diabetes, provide useful experimental models for similar phenotypes in humans. The molecular pathogenesis of the metabolic phenotype in animals segregating for fa is unknown and difficult to study once the confounding metabolic effects of obesity are present. Although various experimental methods distinguish preobese from lean rats (phenotypic markers and molecular markers genetically linked to fatty), technical difficulties limit their utility. We report the identification of two (GT)n simple sequence repeats (SSRs) near the rat phosphoglucomutase gene (Pgm1) gene and two SSRs, (GA)n and (GT)n, near the rat complement component 8 beta gene (C8b). These SSRs map to an approximately 4-cM interval flanking the fatty locus on rat chromosome 5. Use of these molecular markers in combination offers an improved method for early assessment of gene dosage for fa and hence for studying the fundamental molecular physiology underlying the derangements of metabolism and behavior resulting from mutations in this gene.

Comparison between geochemical and biological estimates of subsurface microbial activities.

Geochemical and biological estimates of in situ microbial activities were compared from the aerobic and microaerophilic sediments of the Atlantic Coastal Plain. Radioisotope time-course experiments suggested oxidation rates greater than millimolar quantities per year for acetate and glucose. Geochemical analyses assessing oxygen consumption, soluble organic carbon utilization, sulfate reduction, and carbon dioxide production suggested organic oxidation rates of nano- to micromolar quantities per year. Radiotracer timecourse experiments appeared to overestimate rates of organic carbon oxidation, sulfate reduction, and biomass production by a factor of 10(3)-10(6) greater than estimates calculated from groundwater analyses. Based on the geochemical evidence, in situ microbial metabolism was estimated to be in the nano- to micromolar range per year, and the average doubling time for the microbial community was estimated to be centuries.

Geographic distribution of HTLV-I and identification of a new high-risk population.

Epidemiologic studies indicate that human T-cell lymphotropic virus type I (HTLV-I), the causative agent of most cases of adult T-cell leukemia/lymphoma (ATLL) in Southeast Japan and the Caribbean islands and the probable cause of a progressive neurological disorder often referred to as tropical spastic paraparesis, occurs with unusual geographic clustering. The current large-scale serosurvey was undertaken to improve our understanding of HTLV-I prevalence in different parts of the world. We analyzed 43,445 serum samples collected from various geographic locales worldwide; 76% of these sera came from clinically healthy donors. Samples were initially screened by an enzyme-linked immunosorbent assay (ELISA) and 4,353 were further evaluated by means of competition assays. In this study, which did not include sera from endemic areas of Japan, a high prevalence of infection was observed in several countries in the Caribbean basin. A significant age-sex difference was observed between populations in the Caribbean and non-endemic regions of Japan. The reason for the male excess in non-endemic areas of Japan will require further study, while the female excess in the Caribbean basin is compatible with the previously described pattern for other HTLV-I-endemic areas. A newly recognized area of possible endemicity was southern Florida, where evidence of infection with HTLV-I or a related virus was found in a group of native Americans whose sera were collected in 1968. In certain parts of the world, particularly sub-Saharan Africa, important problems in determining specificity of reactivity occurred, probably because of cross-reacting antibodies. No pattern was detected that could explain the cross-reactivity solely on the basis of geographic areas, specific patterns of non-viral parasitic infection, or methods of handling the specimens. It is possible that these cross-reactivities are antibodies to proteins from HTLV-I-related retroviruses yet to be discovered.