The effects of differing nutritional levels and body condition score on scrotal circumference, motility, and morphology of bovine sperm.

Bulls often experience various levels of nutrient availability throughout the year. Nutritional management is a critical factor on overall ejaculate composition and the ability to get females pregnant. We hypothesized that differing nutritional levels and body condition score (BCS) affect reproductive fertility parameters in bulls. Mature Angus bulls (n = 11) were individually housed and randomly assigned to one of two dietary regimens: 1) over-fed (n = 5) or 2) restricted (n = 6). Bulls were fed the same ration at different volumes to achieve desired effects resulting in eight individual treatments: gain to an over-fed body condition score ([BCS]; GO), gain after nutrient restriction (GR), loss after an over-fed BCS (LO), loss from nutrient restriction (LR), maintenance at ideal adiposity (BCS = 6) after overfeeding (IMO), maintenance at ideal adiposity after nutrient restriction (IMR), maintenance at an over-fed BCS (BCS = 8; MO), and maintenance at a restricted BCS (BCS = 4; MR). Body weight (BW) and BCS were recorded every 2 wk to monitor bull weight and BCS changes. Scrotal circumference was measured every 28 d. Body fat and sperm motility and morphology were evaluated every 84 d. Scrotal circumference, motility, and morphology were normalized to the initial value of each bull. Thus, allowing the individual bull to serve as a control. Statistical analyses were conducted with PROC GLIMMIX of SAS as a complete randomized design to determine if treatment influenced BW, BCS, scrotal circumference, motility, morphology, and adipose thickness. Scrotal circumference (P < 0.001) had the least amount of deviation from initial during the LR (0.29 ±â€…0.44) treatment and the greatest during the MO (3.06 ±â€…0.44), LO (2.28 ±â€…0.44), MR (2.43 ±â€…0.44), GR (3.03 ±â€…0.44), and IMR (2.91 ±â€…0.44) treatments. Sperm motility was not affected by nutritional treatments (P = 0.55). Both head and total defects of sperm differed (P = 0.02) due to nutritional treatments. Increased head abnormalities occurred during the LO (37.60 ±â€…8.61) treatment, with no differences between the other treatments. Total defects increased during the LO (43.80 ±â€…9.55) treatment with similar increases in bulls during the GR (29.40 ±â€…9.55) and IMR (35.60 ±â€…9.55) treatments. In conclusion, male fertility was impacted when a deviation from a BCS of 6 occurred which could be detrimental to reproductive and beef production efficiency.

Intravenous infusion of kisspeptin increased serum luteinizing hormone acutely and decreased serum follicle stimulating hormone chronically in prepubertal bull calves.

Kisspeptin (KP) is a hypothalamic neuropeptide that stimulates the secretion of gonadotropin releasing hormone. To determine the acute and chronic effects of KP on serum concentrations of luteinizing hormone (LH) and follicle stimulating hormone (FSH), prepubertal bull calves [12 ± 1 (SD) weeks of age; 96.5 ± 14.5 kg BW] were administered one of four treatments [0.0 (control; CON), 0.125 (L-KP), 0.25 (M-KP), or 0.5 (H-KP) µg of KP/kg BW/hour] by intravenous infusion for 76 h. Blood samples were collected every 15 min for the first (acute; 1-6 h; Day 1) and last (chronic; 71-76 h; Day 4) 6 h of the intravenous infusions. Serum concentrations of LH and FSH were determined by radioimmunoassay. For each day, effects of treatment, time, and interactions on LH and FSH concentrations and pulse parameters were analyzed using procedures for repeated measures with JMP Software (SAS Inst. Inc., Cary, NC). There was a treatment effect (P = 0.002) and a treatment × time interaction during Day 1 (P = 0.02) such that LH concentrations were greatest following administration of all doses of KP when compared to CON. However, there was no treatment effect (P = 0.57) or a treatment × time interaction during Day 4 (P = 0.20) on serum LH concentrations. There was a treatment by day interaction (P = 0.02) on mean serum FSH concentrations. Most notably, on Day 4 mean serum FSH concentrations during intravenous infusion of M-KP and H-KP doses were less than that of CON. There was a treatment by day interaction (P = 0.0054) on FSH pulse amplitude concentrations, such that intravenous infusion of all doses of KP on Day 4 decreased FSH pulse amplitudes. In conclusion, acute infusion of KP increased LH concentrations and chronic infusion of KP decreased FSH concentrations. Despite the potential suppression of the hypothalamic-pituitary-gonadal axis with chronic infusion of KP, there are likely applications of KP, KP analogs, or KP receptor agonists to hasten the onset of puberty in livestock.

Prevalence of Tritrichomonas foetus in tennessee bulls.

The prevalence of bovine trichomonosis (BT) in TN bulls was estimated through both active screening of bulls and review of previous laboratory records. During the active bull screening, preputial smegma specimens were collected from 458 TN beef bulls at 2 cattle slaughterhouses and 2 stockyards, which serve most beef bulls in TN, between March 2014 and June 2015. Each specimen was cultured for Tritrichomonas foetus (T. foetus) as well as evaluated microscopically every other day for seven days for any protozoa resembling T. foetus. An aliquot of the culture media from each specimen was used for DNA extraction and subsequent qPCR testing. Two specimens were considered suspect on microscopic evaluation, but all specimens were negative for T. foetus on qPCR. This suggests that the 2 specimens were most likely contaminated by fecal trichomonads. Retrospectively, 1979 T. foetus test records from 2 major TN diagnostic laboratories were reviewed between October 2013 and September 2016. True prevalence of BT in TN beef bulls was estimated at <0.01% from the laboratory records, although the county prevalence differed in 2 TN counties (Marshal: 0.09% and Bedford: 0.5%). Overall, the prevalence of BT in TN is low, and the current screening efforts to help control BT disease in TN are acceptable. Future efforts should focus on educating cattle stakeholders on the importance of optimal specimen collection and handling as well as routine testing for BT before cattle movement. In addition, cattle producers should be reminded of leading risk factors associated with BT in cattle.

Changes in microtubular tau protein after morphine in a cultured human neuroblastoma cell line.

1. Cultured human SH-SY5Y adrenergic neuroblastoma cells were used to examine the action of morphine sulfate on microtubular tau protein. 2. After 48 hr treatment morphine sulfate (200 microM) reduced tau protein in the cytoplasmic (supernatant) fraction of undifferentiated cells, and in the cytoplasmic as well as membrane (pellet) fractions of differentiated cells. 3. A 71% increase (P < 0.05) in total protein in the membrane (pellet) fraction of undifferentiated cells and a 188% increase (P < 0.01) in that of differentiated cells accompanied the decrease in tau protein. 4. A 51% reduction (P < 0.01) in the number of undifferentiated (but not differentiated) cells was seen after this drug (200 microM).

Diurnal variations in tear glycoproteins: evidence for an epithelial origin for the major non-reducible > or = 450 kDa sialoglycoprotein(s).

PURPOSE: To characterize the nature and origin of changes in tear glycoproteins accompanying eye closure. METHODS: Reflex (R) and overnight closed (C) eye tears collected by capillary tubes were centrifuged with the resulting R pellets (primarily desquamated epithelial cells) and C pellets (primarily PMN and some epithelial cells) extracted in acidic PBS. Extracts and supernatants were separated by size-exclusion HPLC and/or SDS-PAGE. Gels were stained or blotted and immune- or lectin-probed. An HPLC glycoprotein fraction of > or = 450 kDa isolated from all four sources was characterized before and after partial deglycosylation, using antibodies specific to known mucin and carbohydrate epitopes. Immunofluorescence microscopy was carried out on human conjunctiva, using as probe a MAb to salivary mucin specific for a sialyl Lea epitope, which was found to cross-react specifically with the major non-reducible high molecular weight sialoglycoproteins (SGs) in tears. These SGs were immunoprecipitated and blot-probed along with tissue extracts. RESULTS: R fluid contained minor amounts of numerous glycoproteins, including probably several of inducible lacrimal secretory origin. Results confirmed sIgA as the principal source of the intense reducible glycoprotein bands common to C fluid. Smaller amounts of free secretory component and serum glycoproteins were also visualized. The HPLC fraction (> or = 450 kDa) consisted of four major non-reducible glycoproteins. In R fluid, this fraction (< 1% total protein) consisted primarily of two entities: a 450-500 kDa SG and a larger asialoglycoprotein. The SG accounts for as much as 85% of the total protein in the R pellet extract. C fluid was associated with a selective increase in SGs and a shift in distribution to two SGs > 500 kDa. All SGs exhibited a common antigenicity reacting specifically with the MAb for the sialyl Lea epitope. SGs indistinguishable in size and antigenicity were recovered in epithelial extracts. Immunofluorescence microscopy revealed that reactivity was localized to the epithelial plasma membrane, increasing in intensity from basal to apical cells. Although these SGs exhibited some properties in common with MUC1, immunological and other data suggest a unique SG. CONCLUSIONS: Tear glycoproteins are derived from four principal sources. In R fluid, an inducible lacrimal secretion predominates. In C fluid, a constitutive sIgA secretion predominates, augmented by a serum exudate and SGs derived at least in part from the epithelium. In R fluid and pellet extracts, the SGs consist primarily of a 450-500 kDa species that is most probably derived from the plasma membrane. Larger antigenically related SGs are prevalent in C fluid.

Tau protein after delta-9-tetrahydrocannabinol in a human neuroblastoma cell line.

1. A human neuroblastoma cell line, SH-SY5Y, was used to determine the effects of delta-9-tetrahydrocannabinol (THC) on microtubule-associated tau protein. 2. After 48-hr treatment, THC (10(-9) M) decreased 50 kD tau protein in the cytoplasmic (supernatant) fraction, and in the membrane (pellet) fraction the drug (10(-7) M) also decreased 50 kD tau protein. 3. This reduction in tau protein was accompanied by a 27% reduction (P < 0.05) in the membrane (pellet) total protein after (10(-7) M) THC and a 28% increase (P < 0.02) in cytoplasmic (supernatant) total protein after 10(-9) M THC.

D-lysergic acid reduces microtubule-associated tau protein in SH-SY5Y human neuroblastoma cells.

1. SH-SY5Y, an adrenergic human neuroblastoma cell line, was used to examine the hypothesis that D-lysergic acid (LSD) affects the metabolism of microtubule-associated tau protein, thus affecting microtubule assembly and the transport of neurotransmitters. 2. After 48 hr treatment LSD (10(-5) and 10(-7) M) decreased 50 kDa tau protein in the membrane (pellet) fraction. The drug (10(-5) M) also decreased in the cytoplasmic (supernatant) fraction. 3. This reduction in tau protein was accompanied by a 65% increase (P < 0.05) in total protein after LSD (10(-7) M) in the cytoplasmic fraction.

Helicobacter pylori infection does not reduce the viscosity of human gastric mucus gel.

The mechanism by which Helicobacter pylori undermines host defence mechanisms is unclear. Several in vitro studies using soluble mucins have suggested that H pylori may compromise mucus function. Gastric mucus gel was obtained from 13 H pylori infected patients; six untreated subjects and seven after eradication of the infection. Gastric mucus is a non-Newtonian substance in that its viscosity changes with changing rates of shear, requiring mucus viscosity to be measured in a rotational cone-plate microviscometer. Viscosity was measured at shear rates varying from 1.15 s-1 to 46 s-1. The gastric mucus viscosity was significantly higher in patients infected with H pylori compared with mucus gel obtained after eradication of the infection. The results of our study suggest that the previous studies using in vitro methods involving soluble mucins or its components may have lead to erroneous conclusions about the in vivo interactions of H pylori and gastric mucus gel. The present findings argue against the hypothesis that degradation of gastric mucus by H pylori is important in the pathogenesis of peptic ulcer.

Clarithromycin, tetracycline, and bismuth: a new non-metronidazole therapy for Helicobacter pylori infection.

OBJECTIVE: Metronidazole resistance has become an increasing problem that has limited the usefulness of the original triple therapy. Our objective was to evaluate clarithromycin, a new macrolide compound active against Helicobacter pylori. METHODS: We evaluated a new clarithromycin triple therapy for H. pylori infection consisting of the combination of clarithromycin (500 mg t.i.d.), tetracycline (500 mg q.i.d.), and bismuth subsalicylate tablets (2 q.i.d.) for 14 days. Patients with ulcer also received concomitant ranitidine, 300 mg after the evening meal, for 6 wk. RESULTS: Thirty men with documented H. pylori infection were studied; 29 had peptic ulcer disease. Seven had previously failed antimicrobial therapy, including three with metronidazole-based triple therapy. H. pylori status was determined by histology. H. pylori status and ulcer status were evaluated 4 wk after the end of antimicrobial therapy. The ulcer was healed in 90%. The H. pylori infection was cured in 93%, including all three patients who previously failed metronidazole-based triple therapy. CONCLUSION: We conclude that the combination of clarithromycin, tetracycline, and bismuth is an effective new therapy for treatment of H. pylori infection.

Changes in microtubular tau protein after estrogen in a cultured human neuroblastoma cell line.

1. Cultured human SH-SY5Y neuroblastoma cells were used to determine whether 17-beta-estradiol affects the metabolism of microtubular tau protein. 2. After 24-hr treatment 17-beta-estradiol (10(-7) M) increased 50 kDa tau protein in the cytoplasmic (supernatant) fraction and decreased it in the membrane (pellet) fraction. 3. The increase in cytoplasmic tau was accompanied by increases in total protein in both cytoplasmic and membrane fractions, 50 and 70%, respectively. 4. The estrogen (10(-7) M) also caused a 31% reduction in the total number of cells.

Treatment of Helicobacter pylori reduces the rate of rebleeding in peptic ulcer disease.

BACKGROUND: We evaluated whether therapy designed to eradicate Helicobacter pylori infection resulted in a reduction in rebleeding in patients with peptic ulcer disease. Patients presenting because of major upper gastrointestinal hemorrhage from peptic ulcer and whose ulcers healed in a study in which they were randomized to receive ranitidine alone or triple therapy plus ranitidine were followed up regularly with endoscopy. No maintenance anti-ulcer therapy was given after ulcer healing. METHODS: Patients received ranitidine, 300 mg, or ranitidine plus triple therapy. Triple therapy consisted of tetracycline, 2 g; metronidazole, 750 mg; and bismuth subsalicylate, 5 or 8 tablets (151 mg bismuth per tablet), and was administered for the first 2 weeks of treatment; ranitidine therapy was continued until the ulcer had healed or 16 weeks had elapsed. After ulcer healing, no maintenance antiulcer therapy was given. Development of ulcer recurrence with or without recurrent upper gastrointestinal bleeding was evaluated. RESULTS: Thirty-one patients with major upper gastrointestinal bleeding from peptic ulcer were studied; 17 received triple therapy and 14 ranitidine alone. Major rebleeding occurred significantly (p = 0.031) more often in those in the ranitidine group (28.6%), compared with none (0%) in the triple therapy group. CONCLUSION: Eradication of H. pylori infection reduces the rate of ulcer recurrence and rebleeding in complicated ulcer disease.

Light therapy of late luteal phase dysphoric disorder: an extended study.

Nineteen patients with late luteal phase dysphoric disorder (LLPDD) and 11 healthy comparison subjects underwent a 3-month crossover trial of bright (more than 2500 lux) white morning, bright white evening, and placebo dim (less than 10 lux) red evening light, administered daily for 1 week during the premenstrual phase of the menstrual cycle. All light treatments significantly reduced depressive ratings from baseline levels.

Gastric mucosal hydrophobicity and Helicobacter pylori: response to antimicrobial therapy.

The hydrophobic properties of the gastric mucosa are reduced by NSAIDs and by Helicobacter pylori infection. Our investigation was to determine whether this abnormality was due to the bacteria or to the inflammatory response. Contact angle measurements were made on gastric antral and corpus biopsies taken from 10 H. pylori-infected volunteers before eradication therapy, after 2 and 14 days of therapy, and 4 weeks after therapy. The contact angle improved steadily and statistically throughout the 2 weeks of therapy (for day 0, 3, 14, respectively) antral mucosa 54.2 +/- 2, 59.3 +/- 2, and 63.2 +/- 2; corpus mucosa 55 +/- 1, 57.8 +/- 3, and 66.6 +/- 1. After 2 days of therapy, H. pylori bacteria were no longer evident, and yet the contact angle continued to improve, suggesting that bacteria and bacterial products (e.g., lipases) may not be critical factors. H. pylori was eradicated in five and failed in five. One month after ending therapy, the contact angles of those with recrudescence of infection and those with eradication were similar and higher (p < 0.05) than before therapy (antrum: 69.8 +/- 1 vs. 71.1 +/- 2, corpus: 66.4 +/- 4 vs. 70.8 +/- 2) (p > 0.25 for both). We conclude that gastric surface hydrophobicity abnormalities do not appear to be directly related to the presence of H. pylori organisms or the histologic features of acute inflammation, but are responsive to antimicrobial therapy.

Helicobacter pylori infection and exaggerated gastrin release. Effects of inflammation and progastrin processing.

Helicobacter pylori infection is associated with exaggerated gastrin release. We investigated whether this abnormality was due to the bacteria or the immune response. Fasting and meal-stimulated 'total' and amidated gastrin were measured in 10 H. pylori-infected volunteers before eradication therapy, after 2 and 14 days of therapy, and 4 weeks after completion of therapy. The exaggerated meal-stimulated gastrin concentration remained unchanged after 2 days of therapy, although the polymorphonuclear cell infiltrate and H. pylori bacteria were no longer evident. The expected fall in gastrin concentration after 14 days of therapy was associated with a reduction in the density of mucosal mononuclear cells, suggesting exaggerated gastrin release was related to chronic inflammation or to H. pylori or its products. The effect of H. pylori on normal progastrin processing was also assessed; 2 control groups were included: 10 H. pylori-uninfected volunteers and 13 patients with H. pylori peptic ulcers. There was a significant difference in the proportion of circulating gastrins that were biologically active amidated gastrins between ulcer patients and uninfected controls (56.7 +/- 4% versus 33.8 +/- 4%, p < 0.001). The proportion of amidated to total gastrins did not increase after successful eradication.

Antral G-cell and D-cell numbers in Helicobacter pylori infection: effect of H. pylori eradication.

BACKGROUND: It has recently been recognized that Helicobacter pylori infection is associated with abnormalities in the regulation of gastrin secretion. We investigated whether there was a relationship between H. pylori infection and G-cell and D-cell numbers. METHODS: The numbers of antral G cells and D cells were compared between 20 patients with duodenal ulcer and 24 volunteers, 12 with and 12 without H. pylori infection. The effect of eradication of H. pylori infection on G-cell number was also evaluated. Antral mucosal biopsy specimens were examined using immunohistochemical techniques specific for the presence of gastrin and somatostatin. RESULTS: The number of G cells was significantly (P < 0.02) less in patients with duodenal ulcer than in either infected or uninfected controls (3.7 +/- 0.3 vs. 6.2 +/- 0.6 and 5.3 +/- 0.5 G cells per gland for infected and uninfected controls, respectively). The ratio of G-cells to D-cells was similar in duodenal ulcer patients (2.2) and uninfected controls (2.0). It was found that, although eradication of the H. pylori infection results in a dramatic reduction in stimulated gastrin secretion, it is not associated with a change in the numbers of antral G cells or D cells in patients with duodenal ulcer. CONCLUSIONS: It is concluded that H. pylori infection-associated increase in gastrin secretion appear to be related to local factors regulating G-cell function.

Changes in the gastric mucosa following eradication of Helicobacter pylori.

Although most studies reporting on the examination of Helicobacter pylori infection have focused on the clearance of the bacteria and the rapid disappearance of the neutrophil infiltrates, the evolution of inflammatory and architectural changes in the antral and corporal mucosa following the eradication of H. pylori has not been addressed systematically. This study examines in detail the histopathologic appearance of the antral and corporal mucosa in a group of patients infected with H. pylori and follows the spectrum of morphologic changes in each of them after the eradication of the infection. At least 11 biopsies ("gastric mapping") were obtained from the antrum and body of each of 15 patients with H. pylori. Complete mapping was then repeated 1, 4, and 10 to 12 mo after the eradication of H. pylori by a course of "triple therapy." Each biopsy was assessed in a semi-quantitative fashion for presence of H. pylori, neutrophils, eosinophils, lymphocytes, lymphoid follicles, and intestinal metaplasia. Other features (integrity of surface epithelium, architecture, fibrosis) were evaluated descriptively. Results were compared with those obtained from a control group of 16 uninfected, healthy adult volunteers. H. pylori infection was eradicated in 11 subjects. The disappearance of neutrophils and the normalization of the surface epithelium closely paralleled that of H. pylori. Persistence of even small numbers of neutrophils was a predictor of relapse. Eosinophils and lymphocytes decreased slowly and did not return to normal levels within 1 yr. Lymphoid follicles decreased very slowly in all patients but were still present in all gastric locations at one year after treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Short report: a non-metronidazole triple therapy for eradication of Helicobacter pylori infection--tetracycline, amoxicillin, bismuth.

Triple therapies using bismuth, metronidazole and tetracycline or amoxicillin were the first truly successful anti-H. pylori therapies. Metronidazole resistance has become an increasing problem that has severely limited the usefulness of the original triple therapy. Resistance to tetracycline or amoxicillin has not been reported and both are effective against H. pylori. We therefore tested a new triple therapy consisting of 500 mg tetracycline, 500 mg amoxicillin, and 2 tablets of bismuth subsalicylate each administered four times daily (with meals and at bedtime) for 14 days during treatment with ranitidine 300 mg daily. H. pylori eradication was defined as no evidence of H. pylori one or more months after stopping therapy. H. pylori status was evaluated by a combination of urea breath test and histology. Sixteen patients with H. pylori infection and active peptic ulcers were enrolled. The new triple therapy was successful in only 7 individuals (43%). Metronidazole appears to be critical for the effectiveness of the original triple therapy. An alternative to metronidazole will be required for a new successful triple therapy.

P300, habituation, and response mode.

The P3(00) event-related brain potential (ERP) was elicited with an auditory discrimination paradigm under two response task conditions in which the probability of the target tone was always 0.20. Subjects either counted the occurrence of each target tone or pressed a button to the target in different conditions. No response was made to the standard tone. Consecutive blocks of trials were obtained to assess habituation of P3 amplitude. The count task demonstrated less habituation than the button-press task, with a strong interaction obtained between response mode and trial block for all electrode sites. The findings suggest that habituation of P3 amplitude is sensitive to the amount of attentional resources allocated to the processing of a target stimulus, with more resources required for a count task compared to fewer needed for a button-press response.

Microtubular tau protein after cocaine in cultured SH-SY5Y human neuroblastoma.

1. SH-SY5Y, a human neuroblastoma cell line, was used as a tissue culture model to examine the hypothesis that cocaine may affect the metabolism of tau protein which stabilizes microtubules and promotes microtubule assembly. 2. Cocaine hydrochloride (10(-9)-10(-3) M) caused dose-dependent reductions in cell number, with 10(-3) M causing 28% reduction after 48 hr. 3. This drug also decreased tau protein (50 Kd) in the cytoplasmic (supernatant) as well as the membrane (pellet) fraction after 48-hr treatment.