Analysis of canine myeloid-derived suppressor cells (MDSCs) utilizing fluorescence-activated cell sorting, RNA protection mediums to yield quality RNA for single-cell RNA sequencing.

Fluorescence-activated cell sorting (FACS) is a branch of flow cytometry that allows for the isolation of specific cell populations that can then be further analyzed by single-cell RNA sequencing (scRNA-seq). When utilizing FACS for population isolation prior to sequencing, it is essential to consider the protection of RNA from RNase activity, environmental conditions, and the sorting efficiency to ensure optimum sample quality. This study aimed to optimize a previously published MDSC flow cytometry strategy to FACS sort canine Myeloid-Derived Suppressor Cells (MDSC) with various permutations of RNAlater ™ and RiboLock™ before and after FACS sorting. Concentrations of RNAlater™ greater than 2 % applied before flow analysis affected cell survival and fluorescence, whereas concentrations ≤ 2 % and time ≤ 4 h had little to no effect on cells. To shorten the procedural time and to enhance the sorting of rare populations, we used a primary PE-conjugated CD11b antibody and magnetic column. The combination of RiboLock™ pre- and post-sorting for FACS provided the best quality RNA as determined by the RNA integrity number (RIN ≥ 7) for scRNA-seq in a normal and dog and a dog with untreated oral melanoma dog. As proof of principle, we sequenced two samples, one from a normal dog another from a dog with untreated oral melanoma. Applying scRNA-Seq analysis using the 10X Genomic platform, we identified 6 clusters in the Seurat paired analysis of MDSC sorted samples. Two clusters, with the majority of the cells coming from the melanoma sample, had genes that were upregulated (> log2); these included MMP9, MMP1, HPGD, CPA3, and GATA3 and CYBB, CSTB, COX2, ATP6, and COX 17 for cluster 5 and 6 respectively. All genes have known associations with MDSCs. Further characterization using pathway analysis tools was not attempted due to the lower number of cells sequenced in the normal sample. The benefit deriving from the results of the study helped to gain data consistency when working with cells prone to RNase activity, and the scRNA-seq provided data showing transcriptional heterogeneity in MDSC populations and potentially identifying previously unreported or rare cell populations.

Exercise and insulin resistance in PCOS: muscle insulin signalling and fibrosis.

OBJECTIVE: Mechanisms of insulin resistance in polycystic ovary syndrome (PCOS) remain ill defined, contributing to sub-optimal therapies. Recognising skeletal muscle plays a key role in glucose homeostasis we investigated early insulin signalling, its association with aberrant transforming growth factor ß (TGFß)-regulated tissue fibrosis. We also explored the impact of aerobic exercise on these molecular pathways. METHODS: A secondary analysis from a cross-sectional study was undertaken in women with (n = 30) or without (n = 29) PCOS across lean and overweight BMIs. A subset of participants with (n = 8) or without (n = 8) PCOS who were overweight completed 12 weeks of aerobic exercise training. Muscle was sampled before and 30 min into a euglycaemic-hyperinsulinaemic clamp pre and post training. RESULTS: We found reduced signalling in PCOS of mechanistic target of rapamycin (mTOR). Exercise training augmented but did not completely rescue this signalling defect in women with PCOS. Genes in the TGFß signalling network were upregulated in skeletal muscle in the overweight women with PCOS but were unresponsive to exercise training except for genes encoding LOX, collagen 1 and 3. CONCLUSIONS: We provide new insights into defects in early insulin signalling, tissue fibrosis, and hyperandrogenism in PCOS-specific insulin resistance in lean and overweight women. PCOS-specific insulin signalling defects were isolated to mTOR, while gene expression implicated TGFß ligand regulating a fibrosis in the PCOS-obesity synergy in insulin resistance and altered responses to exercise. Interestingly, there was little evidence for hyperandrogenism as a mechanism for insulin resistance.

Characterization of myeloid-derived suppressor cells and cytokines GM-CSF, IL-10 and MCP-1 in dogs with malignant melanoma receiving a GD3-based immunotherapy.

Melanoma in humans and canines is an aggressive and highly metastatic cancer. The mucosal forms in both species share genetic and histopathologic features, making dogs a valuable spontaneous disease animal model. Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of cells of myeloid origin with immunosuppressive capabilities, which are increased in many human cancers and contribute to tumor immune evasion. They are a possible target to improve immunotherapy outcomes. Current information regarding MDSCs in canines is minimal, limiting their use as translational model for the study of MDSCs. The objective of this study was to characterize major MDSCs subsets (monocytic and polymorphonuclear) and the cytokines granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin 10 (IL-10) and monocyte chemoattractant protein-1 (MCP-1) in canines with malignant melanoma and to evaluate changes in MDSCs and the cytokines over time in response to a GD3-based active immunotherapy. Whole blood and serum collected from 30 healthy controls and 33 patients enrolled in the University of Florida melanoma vaccine trial were analyzed by flow cytometry with canine specific CD11b, MHCII and anti-human CD14 antibodies to assess ostensibly polymorphonuclear-MDSC (CD11b+ MHCII- CD14-) and monocytic-MDSC (CD11b+ MHCII- CD14+) subsets. IL-10, MCP-1 and both MDSCs subsets were significantly elevated in melanoma dogs versus controls. Both MDSCs subsets decreased significantly following GD3-based immunotherapy administration but no significant changes in cytokines were seen over time. To our knowledge, this is the first report documenting increased monocytic-MDSCs in canine melanoma. This is consistent with human malignant melanoma data, supporting dogs as a valuable model for therapeutic intervention studies.

An evaluation of rabies vaccination rates among canines and felines involved in biting incidents within the Wellington-Dufferin-Guelph Public Health Department.

The objectives of this study were to determine the rate of animal bite incidents occurring in the human population of a local health department, and to determine the proportion of these canines and felines that were not up to date on their rabies vaccination at the time the incident occurred. Data were obtained from animal bite incidents reported to Wellington-Dufferin-Guelph Public Health during 2010 and 2011. Descriptive statistics of 718 eligible reports revealed the average rate of animal biting was 1.55 bites per 1000 residents per year. Approximately 54% of these animals were vaccinated against rabies, 32% were not up to date with their rabies vaccination, and the remaining 14.5% were of unknown status. The unit of analysis was the municipality, and the four outcomes of interest were: (i) number of animal bite incidents per 1000 residents, (ii) number of dog bite incidents per 1000 residents, (iii) proportion of animals involved in bite incidents that were not up to date with their rabies vaccination, and (iv) proportion of dogs that were not up to date. Associations between the outcomes and selected demographic variables were investigated using regression analysis. The number of veterinary clinics per 10,000 residents, and whether the municipality was urban or rural were identified as significant predictors for the number of animal bites per 1000 residents, and the number of dog bites. There were no significant predictors for the proportion of unvaccinated animals or dogs. Spatial clustering and the location of spatial clusters were assessed using the empirical Bayes index and spatial scan test. This analysis identified five municipalities within the health department that have a high rate of biting incidents and a high proportion of animals that were not up to date on their rabies vaccination. Such municipalities are ideal for targeted educational campaigns regarding the importance of vaccination in pets.

Effect of exercise training on insulin sensitivity, mitochondria and computed tomography muscle attenuation in overweight women with and without polycystic ovary syndrome.

AIMS/HYPOTHESIS: Polycystic ovary syndrome (PCOS) is an insulin resistant (IR) state. Increased skeletal muscle lipid content and impaired mitochondrial biogenesis have been implicated in the pathogenesis of IR. We investigated whether differences in these variables explain the IR of women affected by PCOS and whether improvements in IR with exercise are reflected by changes in these variables. METHODS: Sixteen PCOS and 13 non-PCOS overweight women were assessed, and eight PCOS and seven non-PCOS women were reassessed after 12 weeks of moderate and vigorous exercise training. Outcomes included insulin sensitivity (glucose infusion rate [GIR]), skeletal muscle gene expression and protein abundance, enzyme activity of selected mitochondrial components, and computed tomography (CT) attenuation-estimated muscle lipid. RESULTS: GIR was lower in women with PCOS versus those without (p = 0.01) and increased with exercise in both groups. Baseline CT muscle attenuation suggested a trend to less muscle lipid in PCOS, which increased with exercise training, with a difference in the change in muscle lipid (p = 0.01, age-corrected), compared with non-PCOS women. GIR correlated with PGC1A gene expression across the whole group; skeletal muscle expression of mitochondrial biogenesis markers was not different between groups at baseline, or after training. Neither lipid changes nor mitochondrial changes correlated with changes in GIR. CONCLUSIONS/INTERPRETATION: Differences in IR in women with and without PCOS were not explained by differences in skeletal muscle lipid or mitochondrial parameters. Improvements in IR with exercise were dissociated from mitochondrial parameters. CT muscle attenuation suggested a differential capacity of PCOS muscle to store lipid compared with non-PCOS. TRIAL REGISTRATION: Clinicaltrials.gov ISRCTN84763265. FUNDING: National Health & Medical Research Council (Grant number 606553), Monash University and The Jean Hailes Foundation.

Psychological distress and unmet supportive care needs in cancer patients and carers who contact cancer helplines.

Cancer information services are a highly accessible source of support for people affected by cancer. To date the nature and extent of distress experienced by such callers and their unmet support needs have not been well described. A cross-sectional survey of 354 cancer patients and 336 carers who reported elevated distress on contact with a cancer information service assessed socio-demographic variables; anxiety, depression and somatization; unmet supportive care needs; cancer-specific distress; presenting problems; post-traumatic growth. Adjustment to cancer was most commonly reported; followed by anxiety. In all, 53.4% of patients and 45.2% of carers reached caseness in anxiety, depression or somatization. Carers had higher distress ratings and intrusive thinking compared to patients; whereas patients had higher somatization. For patients, most unmet supportive care needs were psychological; for carers unmet needs were related to health care services and information related to the person diagnosed with cancer. Being single, unemployed, in treatment, having higher initial distress scores, higher intrusion and avoidance predicted poorer outcomes. Information service frameworks should include distress screening and clear triage and referral processes for psychological care.

Exercise decreases anti-müllerian hormone in anovulatory overweight women with polycystic ovary syndrome: a pilot study.

Polycystic ovary syndrome (PCOS) is a common condition in women associated with menstrual irregularity and anovulation. While obesity worsens and weight loss or exercise improves reproduction function in PCOS, the mechanism for this is unclear. The aim of this study was to examine the effect of exercise on ovarian hormones [anti-Müllerian hormone (AMH)] and menstrual and ovulatory function in women with and without PCOS. Overweight women with (n=7) and without (n=8) PCOS of comparable age, weight and BMI undertook a 12-week intensified endurance exercise training program (1 h 3 times/week) with no structured energy restriction. Primary outcomes were AMH, ovulation (weekly urinary pregnanediol) and menstrual regularity. Secondary outcomes were insulin resistance (euglycemic hyperinsulinemic clamp) and body composition (computed tomography and dual X-ray absorptiometry). Exercise decreased BMI, total and android fat mass and improved insulin sensitivity for all women. AMH was significantly higher in women with PCOS compared to controls before (p<0.001) and after exercise (p=0.001). There was a significant interaction between AMH changes with exercise and PCOS status (p=0.007) such that women without PCOS had no change in AMH (+1.4±5.2 pmol/l, p=0.48) while women with PCOS had a decrease in AMH (- 13.2±11.7 pmol/l, p=0.025). Exercise is associated with improvements in ovarian hormones in women with abnormal ovarian function. This suggests that mechanisms associated with ovarian dysfunction can be improved by exercise in PCOS.

Novel inflammatory markers in overweight women with and without polycystic ovary syndrome and following pharmacological intervention.

BACKGROUND: Polycystic ovary syndrome (PCOS) is associated with reproductive and metabolic abnormalities. AIM: The aim of this study was to assess novel inflammatory markers [adipokines leptin, adiponectin, and leptinadiponectin ratio (L/A)] in overweight women with and without PCOS and to examine alterations in these markers [aldosterone, leptin, adiponectin, and L/A] with pharmacological interventions modulating insulin resistance (IR) in PCOS. MATERIALS/SUBJECTS AND METHODS: Overweight age, and body mass index (BMI)-matched women with (no.=80) or without PCOS (no.=27) were assessed cross-sectionally. Subjects with PCOS were then randomised to 6 months metformin (1 g b.d, no.=26) or oral contraceptive pill (OCP) (35 g ethinyl estradiol/2 mg cytoproterone acetate, no.=30). Outcome measures were leptin, adiponectin, L/A, aldosterone, highly sensitive C-reactive protein, lipid profile, IR, and androgen levels. RESULTS: Leptin levels were lower (156.4+/-85.9 vs 208.5+/-105.2 ng/ml, p=0.015) while adiponectin and L/A were not different between women with and without PCOS. Following intervention, IR increased for the OCP and decreased for metformin, however leptin and aldosterone decreased equivalently with the OCP and metformin with no difference between each treatment (p=0.583 and p=0.801, respectively). There was no change in adiponectin or L/A with the OCP or metformin. On multiple regression, the only baseline predictor of leptin was BMI (r(2)=0.485, p<0.001) and the strongest predictor of change in leptin was change in weight (r(2)=0.402, p<0.001). CONCLUSIONS: Alterations in leptin between women with and without PCOS and following pharmacological interventions are primarily related to adiposity and not IR. Aldosterone was reduced equivalently with metformin and the OCP despite differential effects on IR.

Recording of clinical information in a Scotland-wide drug deaths study.

The aim of this study was to analyse the nature and extent of data extracted from case files of deceased individuals in contact with services 6 months prior to drug deaths in Scotland during 2003. A cross-sectional descriptive analysis of 317 case notes of 237 individuals who had drug-related deaths was undertaken, using a data linkage process. All contacts made with services in the 6 months prior to death were identified. Information on clinical and social circumstances obtained from social care, specialist drug treatment, mental health, non-statutory services, the Scottish Prison Service and Criminal Records Office was collated. More than 70% (n = 237) were seen 6 months prior to their drug death. Sociodemographic details were reported much more frequently than medical problems, for example, ethnicity (49%), living accommodation (66%), education and income (52%) and dependent children (73%). Medical and psychiatric history was recorded in only 12%, blood-borne viral status in 17% and life events in 26%. This paucity of information was a feature of treatment plans and progress recorded. The 237 drug deaths were not a population unknown to services. Highly relevant data were missing. Improved training to promote in-depth recording and effective monitoring may result in better understanding and reduction of drug deaths.

Imaging T-cell movement in the brain during experimental cerebral malaria.

T-cells are known to play a role in the pathology associated with experimental cerebral malaria, although it has not previously been possible to examine their behaviour in brain. Using multiphoton laser scanning microscopy, we have examined the migration and movement of these cells in brain tissue. We believe that this approach will help define host-parasite interactions and examine how intervening in these relationships affects the development of cerebral pathology.

Assessment of iron status and the role for iron-replacement therapy in anaemic cancer patients under the care of a specialist palliative care unit.

Anaemia is common in advanced cancer, may develop for several reasons, and is not always symptomatic. Our observations of the seemingly indiscriminate prescription of iron-replacement therapy (IRT) for anaemic palliative care patients, and our practice of discontinuing IRT in patients with normal red-cell indices, prompted a study to determine (1) the prevalence of anaemia in our patients, (2) what proportion had iron deficiency, (3) the prevalence and benefits of IRT and (4) the prevalence of side effects attributable to IRT. The prevalence of anaemia was 65%. We found a 9% prevalence of iron deficiency, and suggestive but inconclusive evidence of iron deficiency in a further 41%, but only three (27%) of these patients had typical iron deficiency red-cell indices. Only two patients within the study population were taking IRT. Haemoglobin increased significantly in one, but fell in the other, and both experienced side effects attributable to iron. IRT should neither be indiscriminately prescribed nor withheld for anaemic palliative care patients, and the decision should not be based on red-cell indices alone. When symptomatic anaemia is found in patients whose general condition indicates that IRT would be acceptable, iron status should be fully assessed. A therapeutic trial of IRT may be justified where ferritin is elevated, but other parameters suggest iron deficiency.

An investigation of the impact of the location and timing of antigen-specific T cell division on airways inflammation.

It is widely accepted that allergic asthma is orchestrated by T helper type 2 lymphocytes specific for inhaled allergen. However, it remains unclear where and when T cell activation and division occurs after allergen challenge, and whether these factors have a significant impact on airways inflammation. We therefore employed a CD4-T cell receptor transgenic adoptive transfer model in conjunction with laser scanning cytometry to characterize the location and timing of T cell division in asthma in vivo. Thus, for the first time we have directly assessed the division of antigen-specific T cells in situ. We found that accumulation of divided antigen-specific T cells in the lungs appeared to occur in two waves. The first very early wave was apparent before dividing T cells could be detected in the lymph node (LN) and coincided with neutrophil influx. The second wave of divided T cells accumulating in lung followed the appearance of these cells in LN and coincided with peak eosinophilia. Furthermore, accumulation of antigen-specific T cells in the draining LN and lung tissue, together with accompanying pathology, was reduced by intervention with the sphingosine 1-phosphate receptor agonist FTY720 2 days after challenge. These findings provide greater insight into the timing and location of antigen-specific T cell division in airways inflammation, indicate that distinct phases and locations of antigen presentation may be associated with different aspects of pathology and that therapeutics targeted against leukocyte migration may be useful in these conditions.

Interactive effects of dietary fat source and slaughter weight in growing-finishing swine: I. Growth performance and longissimus muscle fatty acid composition.

Crossbred pigs (n=288) were used to test the interactive effects of dietary fat source and slaughter weight on live performance, carcass traits, and fatty acid composition of the LM. Pigs were blocked by initial BW, and, within each of 9 blocks, pens (8 pigs/pen) were randomly assigned to either control corn-soybean meal grower and finisher diets devoid of added fat (Ctrl) or diets formulated with 5% beef tallow (BT), poultry fat (PF), or soybean oil (SBO). Immediately after treatment allotment, as well as at mean block BW of 45.5, 68.1, 90.9, and 113.6 kg, 1 pig was randomly selected from each pen, slaughtered, and allowed to chill for 48 h at 1 degrees C. Backfat was measured on the right sides, and a sample of the LM was removed for fatty acid composition analysis. Regardless of source, inclusion of fat in swine diets did not (P >or= 0.349) affect ADG, ADFI, or G:F. Furthermore, carcasses from pigs fed diets formulated with 5% fat had greater (P=0.013) average backfat depths than those from pigs fed the Ctrl diet. Body weight, carcass weight, and backfat depths increased (P<0.001) as slaughter weight increased from 28.1 to 113.6 kg. The proportion of SFA in the LM increased (P<0.001) with increasing slaughter weight from 28.1 to 68.1 kg, but SFA percentages were similar between 68.1 and 113.6 kg, and pigs fed the Ctrl diet had greater (P=0.032) proportions of SFA than pigs fed the SBO and PF diets. Moreover, the proportion of all MUFA increased (P<0.001) by 9.4 percentage units from 28.1 to 113.6 kg; however, only pigs fed the SBO diet had reduced (P=0.004) MUFA percentages than those fed the Ctrl, BT, and PF diets. Even though the proportion of PUFA in the LM decreased with increasing slaughter weight, pigs fed SBO had greater PUFA percentages, a greater PUFA-to-SFA ratio, and greater iodine values than pigs fed all other dietary treatments when slaughtered at BW of 45.5 kg or greater (fat source x slaughter weight, P < 0.001). Results of this study indicate that fat source had little to no impact on live pig performance, but feeding a polyunsaturated fat source altered the fatty acid profile of the LM within the first 17.4 kg of BW gain; more specifically, including 5% SBO in swine diets could lead to economical ramifications associated with soft pork or fat.

Dissecting the contribution of innate and antigen-specific pathways to the breach of self-tolerance observed in a murine model of arthritis.

BACKGROUND: The relative roles of innate immunity and antigen-specific T cells in rheumatoid arthritis remain controversial. Previous studies demonstrated that T-helper type 1 cells of irrelevant antigen specificity (ovalbumin) induced a transient arthritis in BALB/c mice, which recapitulates many of the pre-articular and articular features of human disease and is associated with the emergence of autoreactive T and B-cell responses to joint-specific antigens. However, the mechanisms underlying this phenomenon were unclear. OBJECTIVES: The aim of this study was to dissect the relative contribution of innate and heterologous antigen-specific pathways to the breach of self-tolerance and pathology observed in this model and how this may result from modified T and B-cell interactions. METHODS: To address this issue, experimental arthritis was elicited either by a non-specific inflammatory stimulus alone, by activation of T cells of an irrelevant specificity or a combination of both. RESULTS: The non-specific inflammatory response generated by lipopolysaccharide led to articular inflammation and cartilage erosion, but did not break tolerance to joint-specific antigens. In contrast, local activation of T cells of an irrelevant specificity produced a similar pathological picture but, in addition, induced T-cell responses to unrelated joint-specific antigens with associated activation of autoreactive B cells. These effects could be further potentiated by the addition of lipopolysaccharide. CONCLUSION: These data demonstrate that non-specific inflammation alone is insufficient to breach self-tolerance. In contrast, T cells of an irrelevant specificity, when triggered locally in an antigen-specific manner, can breach self-tolerance leading to arthritis and autoantibody production, which can then be amplified in a non-specific manner.

Tumour necrosis factor-alpha blockade suppresses murine allergic airways inflammation.

Asthma is a heterogeneous disease that has been increasing in incidence throughout western societies and cytokines, including proinflammatory tumour necrosis factor alpha (TNF-alpha), have been implicated in the pathogenesis of asthma. Anti-TNF-alpha therapies have been established successfully in the clinic for diseases such as rheumatoid arthritis and Crohn's disease. TNF-alpha-blocking strategies are now being trialled in asthma; however, their mode of action is poorly understood. Based on the observation that TNF-alpha induces lymph node hypertrophy we have attempted to investigate this as a mechanism of action of TNF-alpha in airway inflammation by employing two models of murine airway inflammation, that we have termed short and long models, representing severe and mild/moderate asthma, respectively. The models differ by their immunization schedules. In the short model, characterized by eosinophilic and neutrophilic airway inflammation the effect of TNF-alpha blockade was a reduction in draining lymph node (DLN) hypertrophy, eosinophilia, interleukin (IL)-5 production and immunoglobulin E (IgE) production. In the long model, characterized by eosinophilic inflammation, TNF-alpha blockade produced a reduction in DLN hypertrophy and IL-5 production but had limited effects on eosinophilia and IgE production. These results indicate that anti-TNF-alpha can suppress DLN hypertrophy and decrease airway inflammation. Further investigations showed that anti-TNF-alpha-induced inhibition of DLN hypertrophy cannot be explained by preventing l-selectin-dependent capture of lymphocytes into the DLN. Given that overall TNF blockade was able to suppress the short model (severe) more effectively than the long model (mild/moderate), the results suggest that TNF-alpha blocking therapies may be more effective in the treatment of severe asthma.

Chemical composition and in situ dry matter and fiber disappearance of sorghum x Sudangrass hybrids.

Three sorghum x Sudangrass hybrids were planted in twelve 0.2-ha plots to test the effect of date of harvest and hybrid on plant maturity, DM yield, chemical composition, and in situ DM and fiber disappearance. Sweet Sunny Sue (a non-brown midrib (BMR) hybrid; nonBMR), NutriPlus BMR (a BMR hybrid; NP-BMR), and Dry Stalk BMR (a BMR hybrid; DS-BMR) were planted on 26 June 2003 at 22.4 kg of seed/ha. Beginning 34 d after planting, plant height and phenological growth stage were assessed weekly in 10 random, 0.5-m(2) quadrats per plot. Plants were clipped to 2.5 cm in height and analyzed for CP, NDF, and ADF using near-infrared spectroscopy. Composite samples harvested from each plot on d 34, 48, and 63 were incubated in the rumen of 3 steers to determine the in situ disappearance of DM and NDF in a 3 x 3 Latin square. Forage yield was greater (P < or =0.02) for nonBMR than NP-BMR on d 41 and 55 and tended (P = 0.08) to be greater on d 48. The DS-BMR hybrid produced more (P = 0.04) forage DM than the NP-BMR on d 48. When DM yield was regressed on growth stage at harvest, BMR hybrids were predicted to produce 265 kg/ha more DM (P < 0.01) than nonBMR, at the late-boot stage. At all harvest dates, NDF concentrations were less (P < or =0.02) for BMR than nonBMR. The DS-BMR had greater (P < or =0.02) NDF concentrations than NP-BMR on d 41, 48, 55, and 63. Detergent fiber concentrations were predicted to be greater (P < 0.01) in nonBMR than BMR when regressed on growth stage at harvest, but the magnitude of the differences in fiber concentration diminished with growth stage. The A fractions of DM and NDF were greater (P < 0.01) and the C fraction was less (P < 0.01) for BMR hybrids than nonBMR. The B fraction of DM was not affected (P = 0.15) by hybrid type. The B fraction of NDF was not different (P = 0.28) on d 34 but was greater (P < 0.01) on d 48 and 63 for BMR than nonBMR. Effective degradability of NDF and DM was greater (P < 0.02) for BMR than nonBMR on all harvest dates. The A fraction of DM was less for DS-BMR than NP-BMR (P < 0.01), but the B and C fractions of NDF and DM did not differ (P > 0.13) between BMR hybrids. This research indicates that forage chemical composition and ruminal in situ disappearance are improved in the BMR sorghum x Sudangrass hybrids tested compared with the nonBMR. Yield reductions are commonly reported for BMR hybrids, but predicted DM yields in the current study were not reduced if harvested at a similar phenological growth stage.

Effect of crabgrass (Digitaria ciliaris) hay harvest interval on forage quality and performance of growing calves fed mixed diets.

Twelve 0.81-ha crabgrass (Digitaria ciliaris [Retz.] Koel.) hay fields were harvested at 21, 35, and 49 d of regrowth (average phonological growth stage of 30, 51, and 56, respectively). Increased harvest interval exhibited a linear decrease (P < 0.01) in CP (14.1, 13.7, and 10.6% of DM, respectively) and increase (P < 0.01) in NDF (65.3, 70.6, and 70.2% of DM, respectively) and ADF (35.7, 38.9, and 42.7% of DM, respectively). Hays were incorporated into 3 diets that contained 20% (DM basis) crabgrass hay, ground corn (33%), and soybean hulls (32%). Diets contained 14.4, 14.4, and 13.6% CP; 1.83, 1.72, and 1.81 Mcal of NE(m)/kg; and 1.21, 1.10, and 1.17 Mcal of NE(g)/kg; respectively. Diets were fed to beef calves in 12 pens at a rate of 2.3% (DM basis) of BW in 1 experiment (n = 120, initial BW 210 +/- 4.4 kg) and ad libitum in another experiment (n = 60, initial BW 207 +/- 4.4 kg). To measure passage rate of the hay and concentrate portions of the diets, 12 heifer calves (BW = 145 +/- 4.5 kg) were individually fed at 2.3% of BW for 14 d and dosed with Dy-labeled soybean hulls and Yb-labeled hay. In situ DM digestibility of the hays and diets were determined using 3 ruminally cannulated steers (BW = 584 +/- 10.4 kg). Harvest interval did not affect (P > or = 0.11) ADG of limit-fed calves during the diet acclimation or growing phases (average 0.32 and 0.80 kg, respectively) or ADG of calves fed ad libitum (average 1.21 kg). Dry matter intake of calves fed ad libitum averaged 7.9 kg/d (3.28% of BW) and was not affected (P > or = 0.22) by harvest interval. Gain:feed was not affected (P > or = 0.20) by harvest interval (0.13 and 0.15 for limit-fed and ad libitum-fed calves, respectively). Increased harvest interval linearly increased (P < 0.01) ruminal retention time of the hay and tended (P = 0.06) to linearly increase ruminal retention time of the concentrate portions of the diet. Harvest interval linearly decreased (P < or = 0.05) the extent of degradability and effective degradability of DM and NDF of hays, but DM disappearance of the total diet did not differ (P > or = 0.35). In the conditions of this study, increasing harvest interval of crabgrass hay from 21 to 49 d had no deleterious impact on animal performance or efficiency of gain when fed to growing calves in a high-concentrate mixture.

Effect of supplemental iron on finishing swine performance, carcass characteristics, and pork quality during retail display.

Crossbred pigs (n = 185) were used to test the effects of dietary Fe supplementation on performance and carcass characteristics of growing-finishing swine. Pigs were blocked by BW, allotted to pens (5 to 6 pigs/pen), and pens (5 pens/block) were allotted randomly to either negative control (NC) corn-soybean meal grower and finisher diets devoid of Fe in the mineral premix, positive control (PC) corn-soybean meal grower and finisher diets with Fe included in the mineral premix, or the PC diets supplemented with 50, 100, or 150 ppm Fe from Availa-Fe (an Fe-AA complex). When the lightest block averaged 118.2 kg, the pigs were slaughtered, and bone-in pork loins were collected during fabrication for pork quality data. During the grower-I phase, there was a tendency for supplemental Fe to reduce ADG linearly (P = 0.10), whereas in the grower-II phase, supplemental Fe tended to increase ADG linearly (P = 0.10). Even though pigs fed NC had greater G:F during the finisher-I phase (P < 0.05) and across the entire trial (P = 0.07), live performance did not (P > or = 0.13) differ among dietary treatments. There were linear increases in 10th-rib fat depth (P = 0.08) and calculated fat-free lean yield (P = 0.06); otherwise, dietary Fe did not (P > 0.19) affect pork carcass muscling or fatness. Moreover, LM concentrations of total, heme, and nonheme Fe were similar (P > 0.23) among treatments. A randomly selected subset of loins from each treatment was further fabricated into 2.5-cm-thick LM chops, placed on styrofoam trays, overwrapped with polyvinyl chloride film, and placed in coffin-chest display cases (2.6 degrees C) under continuous fluorescent lighting (1,600 lx) for 7 d. During display, chops from NC-fed pigs and pigs fed the diets supplemented with 100 ppm Fe tended to have a more vivid (higher chroma value; P = 0.07), redder (higher a* value; P = 0.09) color than LM chops of pigs fed 50 ppm of supplemental Fe. Moreover, greater (P < 0.01) redness:yellowness ratios in chops from pigs supplemented with 100 ppm Fe indicated a more red color than chops from PC-fed pigs or pigs fed diets supplemented with 50 ppm Fe. In conclusion, however, increasing dietary Fe had no appreciable effects on performance, carcass, or LM characteristics, suggesting that current dietary Fe recommendations are sufficient for optimal growth performance, pork carcass composition, and pork quality.

Insulin resistance, the metabolic syndrome, diabetes, and cardiovascular disease risk in women with PCOS.

Polycystic ovary syndrome is the most common endocrinopathy of reproductive aged women affecting 6-10% of the population. Traditionally considered a reproductive disorder manifesting as chronic anovulation, infertility, and hyperandrogenism, management has primarily focused on short-term reproductive outcomes. Recently, however, significant metabolic aspects in conjunction with longer-term health sequealae of PCOS have been recognized. The metabolic features are primarily related to underlying insulin resistance (IR), which is now understood to play an important role in both the pathogenesis and long-term sequelae of PCOS.