Dynamic cooling strategy based on individual animal response mitigated heat stress in dairy cows.

Technological progress enables individual cow's temperatures to be measured in real time, using a bolus sensor inserted into the rumen (reticulorumen). However, current cooling systems often work at a constant schedule based on the ambient temperature and not on monitoring the animal itself. This study hypothesized that tailoring the cooling management to the cow's thermal reaction can mitigate heat stress. We propose a dynamic cooling system based on in vivo temperature sensors (boluses). Thus, cooling can be activated as needed and is thus most efficacious. A total of 30 lactating cows were randomly assigned to one of two groups; the groups received two different evaporative cooling regimes. A control group received cooling sessions on a preset time-based schedule, the method commonly used in farms; and an experimental group, which received the sensor-based (SB) cooling regime. Sensor-based was changed weekly according to the cow's reaction, as reflected in the changes in body temperatures from the previous week, as measured by reticulorumen boluses. The two treatment groups of cows had similar milk yields (44.7 kg/d), but those in the experimental group had higher milk fat (3.65 vs 3.43%), higher milk protein (3.23 vs 3.13%), higher energy corrected milk (ECM, 42.84 vs 41.48 kg/d), higher fat corrected milk 4%; (42.76 vs 41.34 kg/d), and shorter heat stress duration (5.03 vs 9.46 h/day) comparing to the control. Dry matter intake was higher in the experimental group. Daily visits to the feed trough were less frequent, with each visit lasting longer. The sensor-based cooling regime may be an effective tool to detect and ease heat stress in high-producing dairy cows during transitional seasons when heat load can become severe in arid and semi-arid zones.

Effect of forage-to-concentrate ratio on production efficiency of low-efficient high-yielding lactating cows.

Feed is usually the costliest input in lactating cow's farms. Therefore, the developing of methods for a better adjustment of feed intake to cow's energetic needs in order to improve efficiency is desired. The aim of this study was to improve feed efficiency of low-efficient (LE) cows through a moderate increase in diet forage-to-concentrate ratio. We studied the effects of replacing 8.2% corn grains in a control low-fiber (LF) diet that contained 17.5% forage neutral detergent fiber (NDF) with 7.5% wheat straw +0.7% soybean meal for a high-fiber (HF) diet that contained 23.4% forage NDF. Based on efficiency data of individual cows from the Agricultural Research Organization's herd measured in our previous study, 15 pairs of pre-classified LE multiparous mid-lactating Israeli Holstein dairy cows were selected, each pair with similar performance, intake, and efficiency data; each member of a pair was then adapted for 2 weeks to one or the other dietary treatment. Traits examined during the 5 weeks of the experiment were DM intake (DMI), eating behavior, milk production, in vivo digestibility, and estimation of feed efficiency [energy-corrected milk (ECM)/DMI and energy balance]. Cows fed the HF diet showed slower eating rate, smaller visit and meal sizes, longer daily eating time, higher visit frequency, and longer meal duration, compared to those fed the LF diet. The DMI of cows fed the HF diet was 9.1% lower, their DM digestibility decreased from 65.7 to 62.2%, and their ECM yield was 7.0% lower than in cows fed the LF diet. Feed efficiency, measured as net energy captured/digestible energy intake, improved in the cows fed the HF vs. LF diet while feed efficiency measured as ECM/DMI remained similar. Our results thus show the potential of improving feed efficiency for milk production in LE cows by increasing the forage-to-concentrate ratio.

Seasonal heat load is more potent than the degree of body weight loss in dysregulating immune function by reducing white blood cell populations and increasing inflammation in Holstein dairy cows.

We investigated the effects of seasonal heat stress and the degree of body weight (BW) loss postpartum (PP) on immune and inflammatory markers in transition dairy cows. Blood sampled twice weekly during the transition period (3 wk prepartum to 3 wk PP) from 12 peripartum Holstein cows in summer and 12 in winter was analyzed for plasma nonesterified fatty acids, the oxidative stress marker malondialdehyde (MDA), haptoglobin, and tumor necrosis factor-α (TNF-α). Complete blood counts and white blood cell (WBC) subpopulations (CD4, CD8, CD25, WC1, and CD335) were examined weekly during the transition period. Adipose tissue biopsies were taken at 7 d PP from summer-calving cows. In cows calving during seasonal heat stress, plasma TNF-α concentrations were 4.9-fold higher prepartum and 5.1-fold higher PP, and MDA concentrations were higher prepartum and PP than in winter. Hematocrit was lower prepartum and tended to be lower PP in summer compared with winter, and after adjusting to hematocrit values fewer WBC prepartum (-17%) and PP (-22%), lower monocytes prepartum (-37%) and PP (-49%), and fewer neutrophils (-25%) PP were found in summer compared with winter-calving cows. The percentages of cytotoxic T cells (CD8) and natural killer cells (CD335) were lower, as well as prepartum gamma delta-T cells (WC1) in summer compared with winter-calving cows. Moreover, lower red blood cell counts prepartum and mean platelet volume and red blood cell distribution width prepartum and PP, as well as higher hemoglobin prepartum and PP, were found in summer than winter-calving cows. In summer, cows that lost more BW PP had higher plasma MDA and TNF-α concentrations, fewer basophil numbers prepartum, a decrease in the percentage of cells expressing the IL-2 receptor (CD25), increased ratio of T helper cells (CD4):CD8, and a tendency for increased WC1 percentages in blood prepartum, as well as a higher percentage of WC1 PP and higher abundance of TNF-α and lower lipopolysaccharide-binding protein in adipose tissue compared with cows that lost less BW PP. In conclusion, seasonal heat stress during the peripartum period was associated with lower WBC counts coupled with increased levels of subacute inflammation, indicating dysregulation of immune function, whereas the degree of BW loss had minor effects. This suggests that heat load is a more potent stressor associated with immune function than the variation in the degree of BW loss PP in high-yielding peripartum dairy cows.

Saturated fat supplemented in the form of triglycerides decreased digestibility and reduced performance of dairy cows as compared to calcium salt of fatty acids.

The two most popular rumen-protected fatty acid supplements in dairy cow rations are calcium salts of palm oil fatty acid calcium salts of palm oil fatty acid (CSFA) and prilled saturated fatty acids (SFAs). The objectives of this study were to determine the effects of supplementing SFA in the form of triglycerides (TSFA), as compared to CSFA, on yields, efficiency and diet digestibility in high-yielding dairy cows. Twenty-eight (14 cows in each group) multiparous cows were fed a basal diet supplemented (on DM basis) with either 12 g/kg TSFA (~350 g/cow per day - contained 980 g/kg fat; 882.3 g/kg SFAs) or 14 g/kg CSFA (~440 g/cow per day - contained 800 g/kg fat; 566.4 g/kg SFAs). The supplement amounts in the diet were balanced according to fat content. Rumen samples were taken for measurements of ammonia and volatile fatty acids concentrations, and fecal samples were taken for digestibility measurements. The CSFA cows produced 3% higher milk yields (47.6 v. 46.2 kg/day; P < 0.0001) and 4.7% higher 4% fat-corrected milk (FCM; 44.7 v. 42.7 kg/day; P = 0.02) than the TSFA cows. No difference in milk-fat content was observed, but milk-protein content was higher in the TSFA than CSFA cows. Yields of fat and protein were similar, but lactose yields were higher in TSFA cows. There were no differences in dry matter intake or efficiency calculations between groups. The ruminal ammonia concentrations were similar between groups, whereas acetate concentrations and acetate : propionate ratio were greater for CSFA than TSFA cows. The apparent total-tract digestibility of dry (P < 0.0007) and organic matters (P < 0.0003), fat (P < 0.0001), NDF and ADF (P = 0.02) were lower in the TSFA v. CSFA cows. In conclusion, the CSFA-supplemented cows produced 3% higher milk and 4.7% higher 4% FCM than the TSFA cows. However, TSFA supplementation did not depress milk-protein content. The apparent total-tract digestibility was lower for all dietary components in the TSFA cows, which was probably due to the effects of both degree of saturation and triglyceride form of the TSFA supplement. Considering that diets were balanced according to the fat content of the supplements, the lower yields of milk and FCM observed in the TSFA than CSFA cows were likely due to the lower digestibility of the fat and other nutrients in the TSFA cows, which might have negatively influenced the dietary energy content.

Reducing milking frequency from thrice to twice daily in early lactation improves the metabolic status of high-yielding dairy cows with only minor effects on yields.

Reducing milk production during early lactation might be of interest to improve the energy balance (EB) of high-yielding dairy cows. Therefore, the objective of this study was to determine how reducing the milking frequency (MF) of high-yielding dairy cows from thrice to twice a day during the first 30 d in milk (DIM) affects yields, intake, efficiency, metabolic status, and carryover effects. To this end, 42 multiparous cows were divided into 2 groups according to their previous lactation performance, parity, and body weight. The control cows were milked 3 times a day (3ML) and the treated cows were milked twice a day (2ML) until 30 DIM and then both groups were milked 3 times a day. Milk samples were taken twice a week from 2 or 3 consecutive milkings until 45 DIM for analysis of milk solids, and both groups were followed until 100 DIM to determine the carryover effects of MF until 30 DIM. Individual dry matter intake (DMI), milk yield, and body weight were recorded daily. Blood samples were taken 3 times weekly from 14 d prepartum until 45 DIM. Milk yield during the first 30 DIM was 8.6% higher (49.3 and 45.4 kg/d, respectively), milk fat percentage was lower (3.96 and 4.27%, respectively), and the yields of all milk solids were higher in the 3ML cows than in the 2ML cows. Dry matter intake and 4% fat-corrected milk were similar between groups. The EB during the first 30 DIM was lower in the 3ML cows than in the 2ML cows, and milk yield, but not 4% fat-corrected milk yield, per unit of DMI was higher in the 3ML cows. No differences were observed between groups from 31 to 100 DIM in milk yield (∼56.3 kg/d for both groups), milk solids yield, DMI, or milk/DMI; however, fat percentage was lower and EB was higher in the 3ML cows. Blood glucose concentrations between 0 and 30 DIM were lower and ß-hydroxybutyrate concentrations were higher in the 3ML cows than in the 2ML cows, but nonesterified fatty acids concentrations were lower, which may be attributed to the lower clearance frequency of nonesterified fatty acids from the blood stream in the 2ML cows. A lower proportion of the 3ML cows (10%) ovulated ≤15 DIM compared with the 2ML cows (40%), with no beneficial effects on preovulatory follicle characteristics. Reducing the MF from thrice to twice a day during the first 30 DIM improved EB and metabolic status, with only minor effects on production.

Dietary restriction improved feed efficiency of inefficient lactating cows.

The aim of this study was to reduce voluntary dry matter intake (DMI) to increase feeding efficiency of preclassified inefficient (INE) dairy cows through restricted feeding. We studied the effects of dietary restriction on eating behavior, milk and energy-corrected milk (ECM) production, in vivo digestibility, energy balance, and measures of feed efficiency [residual feed intake (RFI) and ECM/DMI]. Before the experiment, 12 pairs of cows were classified as INE. The 2 dietary treatments consisted of ad libitum feeding versus restricted feeding of the same total mixed ration containing 36.5% roughage. Inefficient cows fed the restricted total mixed ration had a shorter eating time and lower meal and visit frequency, but a similar rate of eating, meal size, and meal duration compared with INE cows fed ad libitum. Compared with the INE cows fed ad libitum, restricted INE cows had 12.8% lower intake, their dry matter and neutral detergent fiber digestibility remained similar, and their ECM yield was 5.3% lower. Feed efficiency, measured as RFI, ECM/DMI, and net energy retained divided by digestible energy intake, was improved in the restricted INE cows as compared with the ad libitum cows. Our results show that moderate DMI restriction has the potential to improve feed efficiency of preclassified INE cows.

Effect of lactation trimester and parity on eating behavior, milk production and efficiency traits of dairy cows.

There is absence knowledge about the effects of lactation trimester and parity on eating behavior, production and efficiency of dairy cows. Objective of this study was to identify and characterize in 340 dairy cows, the 20% high efficient (HE), 20% low efficient (LE) and 60% mid efficient (ME) cows according to their individual residual feed intake (RFI) values, within and between lactation trimesters and between 1st and 2nd parities. Efficiency effect within each lactation trimester, was exhibited in daily dry matter intake (DMI), eating rate and meal size, that were the highest in LE cows, moderate in the ME cows and lowest in the HE group. Daily eating time, meal frequency, yields of milk and energy-corrected milk (ECM) and BW were similar in the three efficiency groups within each trimester. The lower efficiency of the LE cows in each trimester attributes to their larger metabolic energy intake, heat production and energy losses. In subgroup of 52 multiparous cows examined along their 1st and 2nd trimesters, milk and ECM production, DMI, eating behavior and efficiency traits were similar with high Pearson's correlation (r=0.78 to 0.89) between trimesters. In another subgroup of 42 multiparous cows measured at their 2nd and 3rd trimesters, milk and ECM yield, DMI and eating time were reduced (P<0.01) at the 3rd trimester, but eating rate, meal frequency and meal size remained similar with high Pearson's correlation (r=0.74 to 0.88) between trimesters. In subgroup of 26 cows measured in 1st and 2nd parities, DMI, BW, milk and ECM yield, and ECM/DMI increased in the 2nd lactation, but eating behavior and RFI traits were similar in both parities. These findings encourage accurate prediction of DMI based on a model that includes eating behavior parameters, together with individual measurement of ECM production. This can be further used to identify HE cows in commercial herd, a step necessary for potential genetic selection program aimed to improve herd efficiency.

Eating behavior, milk production, rumination, and digestibility characteristics of high- and low-efficiency lactating cows fed a low-roughage diet.

This study aimed to identify individual characteristics differing among high-efficiency (HEf; upper 20%, n = 31), low-efficiency (LEf; lower 20%, n = 31), and mid-efficiency (MEf, 60% n = 93) lactating cows. Primiparous (37) and multiparous (118) high-producing milking cows at 30 to 180 d in milk were fed individually a low-roughage diet [31.6% of dry matter (DM)] for 4 wk. Daily average DM intake, rate of eating, visit duration, meal size, and daily rumination time were higher in LEf compared with HEf cows. On the other hand, HEf cows exhibited higher digestibility of DM, crude protein, and neutral detergent fiber than the LEf cows. Daily eating time was similar in the HEf and LEf groups and higher than that of the MEf cows. Visit and meal frequency, average visit and meal duration, daily lying time, and pedometer activity were similar in the HEf, LEf, and MEf groups. The HEf cows produced 1.75% more milk, but similar energy-corrected milk compared with the LEf cows. Milk fat and protein content were lower by 1.8 and 3.8%, respectively, in the HEf cows than in the LEf group. Body weight (BW) and BW gain were similar in the 3 efficiency groups. Diurnal distribution of DM intake showed 6 distinct major meals, each composed of 1.1 to 1.6 visits. Higher intake peaks (greater meal size) were found in the LEf cows compared with the HEf group. Daily DM intake was highly correlated (affected) with energy-corrected milk production (r = 0.61), BW (r = 0.4), eating rate (r = 0.57), and visit size (r = 0.54). Energy balance showed that the lower efficiency of the LEf cows was attributed to their excess heat production and energy loss.

Mid-term outcomes of routine proximal row carpectomy compared with proximal row carpectomy with dorsal capsular interposition arthroplasty for the treatment of late-stage arthropathy of the wrist.

AIMS: The aims of this study were to compare the mid-term outcomes of patients with late-stage arthritis of the wrist treated with proximal row carpectomy (PRC) and dorsal capsular interposition (DCI) arthroplasty with a matched cohort treated with routine PRC alone. PATIENTS AND METHODS: A total of 25 arthritic wrists (24 patients) with pre-existing degenerative changes of the proximal capitate and/or the lunate fossa of the radius were treated with PRC + DCI over a ten-year period. This group of patients were matched 1:2 with a group of 50 wrists (48 patients) without degenerative changes in the capitate or lunate fossa that were treated with a routine PRC alone during the same period. The mean age of the patients at the time of surgery was 56.8 years (25 to 81), and the demographics and baseline range of movement of the wrist, grip strength, Quick Disabilities of the Arm, Shoulder, and Hand (QuickDASH) score, and Patient-Rated Wrist Evaluation (PRWE) score were similar in both groups. RESULTS: At a mean follow-up of 5.9 years (1.8 to 11.8), significant improvements in mean grip strength, the flexion-extension arc of movement of the wrist, QuickDASH, and PRWE scores were seen in both groups. There was no diifference between the groups for any of the outcomes. One patient in the PRC + DCI group required additional surgery for a deep infection, while two in the PRC group had complications (one wound dehiscence requiring revision closure, one transient radial sensory neuritis). One patient in each group required total arthrodesis of the wrist for progressive degenerative radiocarpal changes. A total of 70 patients (93%) were satisfied with the outcomes. CONCLUSION: PRC with DCI is an effective form of treatment for late-stage arthritis of the wrist involving the capitolunate joint, with mid-term outcomes that are similar to those in patients without degenerative changes affecting the capitate or lunate fossa who are treated with a routine PRC alone. Cite this article: Bone Joint J 2018;100-B:197-204.

Preovulatory follicle characteristics and oocyte competence in repeat breeder dairy cows.

The varied and elusive etiology of repeat breeding (RB) in dairy cows necessitates evaluation of oocytes and follicles, which have not previously been assessed together. Accordingly, we evaluated characteristics of preovulatory follicles and the competence of oocytes in control (CTL) and RB cows. The estrous cycles of 35 cows (18 CTL and 17 RB) were synchronized using PGF2α and estrus detection. Cows with a corpus luteum were treated with PGF2α and, 14 to 15 d after a visible behavioral estrus, they were administered a second PGF2α, followed 48 h later by follicular fluid (FF) aspiration of the preovulatory follicles. Estradiol (E2)-active preovulatory follicles did not differ in diameter between the 2 groups of cows. However, FF of RB cows had higher E2 concentrations than that of CTL cows: 1,854.9 and 1,073.6 ng/mL, respectively, but similar androstenedione and progesterone concentrations. In the second part of the study, 14 consecutive ovum pick-ups (OPU) were performed in 5 CTL and 5 RB cows, at 3- to 4-d intervals. The RB and CTL cows did not differ in average numbers of follicles available per cow per session (7.1 and 7.3, respectively), oocyte recovery rates (42.2 and 44.1%, respectively), or cleavage rates (57.6 and 63.4%, respectively), but blastocyst production was markedly less in RB than in CTL cows (12.5 and 29.2%, respectively). We conclude that part of the RB cows' etiology occurs at an earlier phase of folliculogenesis, thereby impairing oocyte competence, and subsequently reducing the probability of normal fertilization, which diminish embryo vitality and development.

Global HIV-1 transmitted drug resistance in the INSIGHT Strategic Timing of AntiRetroviral Treatment (START) trial.

OBJECTIVES: HIV-1 transmitted drug resistance (TDR) in treatment-naïve individuals is a well-described phenomenon. Baseline genotypic resistance testing is considered standard of care in most developed areas of the world. The aim of this analysis was to characterize HIV-1 TDR and the use of resistance testing in START trial participants. METHODS: In the Strategic Timing of AntiRetroviral Treatment (START) trial, baseline genotypic resistance testing results were collected at study entry and analysed centrally to determine the prevalence of TDR in the study population. Resistance was based on a modified 2009 World Health Organization definition to reflect newer resistance mutations. RESULTS: Baseline resistance testing was available in 1946 study participants. Higher rates of testing occurred in Europe (86.7%), the USA (81.3%) and Australia (89.9%) as compared with Asia (22.2%), South America (1.8%) and Africa (0.1%). The overall prevalence of TDR was 10.1%, more commonly to nonnucleoside reverse transcriptase inhibitors (4.5%) and nucleoside reverse transcriptase inhibitors (4%) compared with protease inhibitors (2.8%). The most frequent TDR mutations observed were M41L, D67N/G/E, T215F/Y/I/S/C/D/E/V/N, 219Q/E/N/R, K103N/S, and G190A/S/E in reverse transcriptase, and M46I/L and L90M in protease. By country, the prevalence of TDR was highest in Australia (17.5%), France (16.7%), the USA (12.6%) and Spain (12.6%). No participant characteristics were identified as predictors of the presence of TDR. CONCLUSIONS: START participants enrolled in resource-rich areas of the world were more likely to have baseline resistance testing. In Europe, the USA and Australia, TDR prevalence rates varied by country.

Immediate and carryover effects of Gram-negative and Gram-positive toxin-induced mastitis on follicular function in dairy cows.

This study compared immediate and carryover effects of mastitis induced by Gram-negative endotoxin (E. coli LPS) and Gram-positive exosecretions (Staph. aureus ex.) on preovulatory follicle function. Synchronized, uninfected cyclic lactating Holstein cows were treated with PGF(2α) on day 6 of the cycle and 36 h later, a dose of either E. coli LPS (n = 8), S. aureus ex. (n = 10), or saline (n = 9) was administered into the mammary gland. Follicular fluids and granulosa cells were aspirated 6 h later from the preovulatory follicles and cows were treated with GnRH. This (cycle 1; immediate effect) was repeated three times (excluding the mammary injections) to induce three 7 d cycles (cycles 2, 3, and 4; carryover effect). E. coli LPS increased body temperature, plasma cortisol concentration, and somatic cell count (SCC), whereas S. aureus ex. induced a minor, subclinical elevation of SCC and slight rise (NS) in body temperature and cortisol concentration. Follicular estradiol, androstenedione, and progesterone concentrations in the E. coli LPS group decreased (P < 0.05) in cycle 1 to about 40%, 13%, and 35%, respectively, of control levels, whereas in the S. aureus ex. group, only estradiol decreased (P < 0.05), to 56% of control concentrations. In cycles 3 and 4, follicular steroids in the E. coli LPS group returned to control concentrations, whereas in the S. aureus ex. group, follicular concentrations of estradiol and androstenedione were lower (P < 0.10) than in controls. In the control group, the concentrations of all follicular and circulating steroids remained stable (P > 0.05) throughout the study. Follicle size was similar in all groups, but the S. aureus ex. treatment caused a decrease (P < 0.02) in the number of follicles developed in cycles 3 and 4. The mRNA expression of steroidogenic genes and LHCGR in the granulosa cells was not affected (P > 0.05) by either treatment during the study, except for a tendency toward lower (P < 0.1) expression in cycle 1 and lower (P < 0.05) expression in cycle 4 of the latter in the S. aureus ex. group. Strain levels, such as SCC and body temperature, following toxin injection correlated well with the magnitude of the immediate decline in follicular steroids. As is typical for Gram-negative clinical events, E. coli LPS-induced acute mastitis caused immediate, short-term, but not long-term impairment of follicular responses, whereas the Gram-positive S. aureus ex.-induced subclinical mastitis exhibited both immediate and carryover disruptive effects on preovulatory follicle function.

Predictors of HIV disclosure to secondary partners and sexual risk behavior among a high-risk sample of HIV-positive MSM: results from six epicenters in the US.

This study sought to identify predictors of HIV disclosure and serodiscordant unprotected anal intercourse (SDUAI) among HIV-positive men who have sex with men (MSM). Between January 2005 and April 2006, 675 HIV-positive MSM were recruited into the Positive Connections intervention trial held in six US cities with intentional over-sampling of HIV-positive MSM of Color (74%) and men engaging in unprotected anal intercourse (UAI) in the previous year. Baseline survey data showed 30 and 31%, respectively, of participants disclosed to none or some of their secondary sex partners in the last 90 days. Greater disclosure to secondary partners was associated with having fewer sexual partners, being extremely out as MSM, longer HIV diagnosis, knowledge of CD4 count, detectable viral load and being white. Disclosure to all secondary partners was associated with lower SDUAI. Recommendations for prevention for HIV-positive MSM include the promotion of serodisclosure to all secondary partners and increasing comfort with, and outness about, one's sexuality.

Concentrations of ganglioside type M1 and immunoglobulin G in colostrum are inversely related to bacterial infection at early lactation in cows.

The levels of IgG and ganglioside type M1 in the colostrum of cows and heifers were analyzed to examine their utility in predicting acquisition of intramammary infection (IMI) during the first weeks postpartum. In general, high levels of IgG and ganglioside type M1 in cows were associated with lower new incidence of IMI, and linear discriminate analysis based on these 2 variables yielded 69.4% successful classification into cows that did or did not acquire new IMI. This analysis was less successful in heifers because a high proportion of them joined the herd when already infected with bacteria in their udders. It is suggested that application of a wider range of measures that reflect the immune status would enable the identification of most cows prone to new IMI.

Exposure to endotoxin during estrus alters the timing of ovulation and hormonal concentrations in cows.

The effect of intramammary (IMM) or intravenous (IV) administration of E. coli endotoxin (LPS), at the onset of estrus, at the time of ovulation was examined. Steroid and gonadotropin concentrations around ovulation were also determined. Lactating Holstein cows (n=33) were assigned to saline-controls (n=12) and treated with LPS-IV (0.5 microg/kg; n=13) or LPS-IMM (10 microg; n=8). Synchronized cows were observed continuously for estrus. LPS (or saline) was injected within 30 min from the onset of standing estrus, at peak estradiol concentrations. The typical rise of body temperature, somatic cell count, cortisol, and NAGase activity was noted. One-third of both LPS-IV- and LPS-IMM-treated cows were manifested by an extended estrus to ovulation (E-O) interval of around 75 h or did not ovulate, compared with about 30 h in the other 2/3 of LPS cows and all controls. Estradiol concentrations 24 h before and after LPS did not differ between groups. However, LPS-IV cows with extended intervals exhibited another estrus and an additional rise of estradiol followed by delayed ovulation. LPS-treated cows with a delayed E-O interval had low or delayed LH surge; two LPS-treated cows did not exhibit LH surge and did not ovulate. All control cows exhibited normal hormone levels. Delayed ovulation was associated with a delayed rise of luteal progesterone. The results indicated that exposing cows to endotoxin during estrus induced a decreased and delayed LH surge in one-third of the cows. This was associated with delayed ovulation, which reduces the chances of successful fertilization.

3,3'-diindolylmethane and paclitaxel act synergistically to promote apoptosis in HER2/Neu human breast cancer cells.

BACKGROUND: HER2/neu positive breast tumors are difficult to treat. About 25 to 30% of invasive breast tumors overexpress the HER2/neu oncogene. These tumors are aggressive and become resistant to chemotherapeutic drugs. 3'3'-diindolylmethane (DIM), the active metabolite of indole-3-carbinol, a naturally occurring compound found in cruciferous vegetables, has been found to have anti-cancer properties in both humans and animals. DIM has been shown to induce cell cycle arrest and apoptosis in animal breast cancer models. Because HER2/neu overexpression confers resistance to paclitaxel, and DIM has anti-tumor effects, we hypothesized that DIM will enhance the cytotoxic effects of paclitaxel, a common taxane drug, on human Her2/neu breast cancer cells by potentiating its effect on cell cycle and stimulating apoptosis. METHODS: The MDA-MB-435eB1 human Her2/neu breast cancer cells were treated with varying concentrations of DIM and paclitaxel. The cells were analyzed at different time points (24, 48, and 72 h). Proliferation was measured by a commercial cell proliferation assay (Promega Procheck Assay). Cell-cycle analysis and apoptosis were determined by flow cytometry. Western blot analysis was performed on to determine the effect of DIM and/or paclitaxel on the proteins involved in apoptosis, and epidermal growth factor-induced activation of HER2/neu and ERK1/2 signaling proteins. RESULTS: Both DIM and paclitaxel exhibited time and concentration dependent inhibition of cell proliferation. TUNEL assay indicated that the combination also increased the number of apoptotic cells more than either agent alone. The presence of cleaved poly (ADP-Ribose) polymerase (PARP) significantly increased in the combination treatment, whereas Bcl-2 is decreased. DIM alone decreased the activation of the Her2/neu receptor; the combination decreased the activation of ERK1/ERK2. CONCLUSIONS: DIM in combination with paclitaxel synergistically inhibits growth of Her2/neu human breast cancer cells through G2M phase cell-cycle arrest and induction of apoptosis/necrosis. The Her2/neu receptor and its downstream signaling protein ERK1/2 appear to be involved in DIM's affect on cell growth and differentiation, whereas apoptosis appears to be mediated through the mitochondrial pathway (Bcl-2/PARP). It appears DIM, a naturally occurring, nontoxic compound, may be a beneficial addition to a traditional (taxane-based) chemotherapy regimen.

Nitric oxide is required for the induction and heterosynaptic spread of long-term potentiation in rat cerebellar slices.

1. In the cerebellar cortex, brief, 8 Hz activation of parallel fibres (PFs) induces a cyclic adenosine 3'5'-monophosphate (cAMP) and protein kinase A (PKA)-dependent form of long-term potentiation between PFs and Purkinje cells. 2. With 10 mM BAPTA in the recording pipette, potentiation evoked by raised frequency stimulation (RFS) to one of two, synaptically independent PF inputs to the same Purkinje cell did not remain input specific but consistently spread to synapses that did not receive RFS, up to the maximum distance tested of 168 microm. 3. LTP at activated and non-activated sites was accompanied by a decrease in paired pulse facilitation (PPF). The PKA inhibitor H-89 blocked both of these effects. Inhibition of nitric oxide synthase (NOS), either by 7-nitro-indazole (7-NI) or N (G)-nitro-L-arginine methyl ester (L-NAME), completely prevented heterosynaptic potentiation and associated reduction in PPF. LTP at distant synapses was selectively prevented by the nitric oxide scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO). Inhibition of soluble guanylate cyclase or protein kinase G had no effect on either pathway. 4. Synaptic potentiation at PF-PC synapses, induced by the adenylate cyclase activator forskolin, was also prevented by inhibition of NOS. Forskolin-induced increases in mEPSC frequency were similarly prevented by NOS inhibition and mimicked by the NO donor spermine NONOate. 5. These results are consistent with the notion that heterosynaptic potentiation is of pre-synaptic origin and dependent upon activation of cAMP/PKA and NO. Moreover, they suggest that cAMP/PKA activation stimulates NO production and this diffusible messenger facilitates pre-synaptic transmitter release at synapses within a radius of upwards of 150 microm, through a mechanism that does not involve cGMP.

Parenteral supplementation with a fish-oil emulsion prolongs survival and improves rat lymphocyte function during sepsis.

Nutritional intervention with omega-3 fatty acids during trauma and infection has been shown to improve the clinical outcome of patients and the survival rate in laboratory animals. This study evaluated the effects of parenteral administration of lipid emulsions containing fish oil (FO) or soybean oil (SBO) on survival and T-lymphocyte response during sepsis. Male Sprague-Dawley rats (250-275 g) were prepared for parenteral feeding 4 d before inducing sepsis by cecal ligation and puncture (CLP). Standard resuscitation was provided with normal saline. Thirty minutes after completing CLP, sham control or CLP rats were infused continuously with saline or a parenteral diet containing SBO or a 1:1 FO:SBO emulsion. The survival rate was significantly improved in rats receiving the FO-supplemented diet, with 50% alive by 120 h in comparison with the saline-infused, chow-fed rats (0% alive by 120 h) or the SBO-fed rats (12% alive at 120 h). The T-lymphocyte response was evaluated at 24 h after CLP. Sepsis led to a decline in lymphocyte proliferation in rats infused with saline or the SBO emulsion, which was associated with a greater release of splenocyte interleukin-10, transforming growth factor-beta and prostaglandin E2. Administering the 1:1 FO:SBO parenteral diet during sepsis improved the survival rate and prevented the sepsis-induced suppression of lymphocyte proliferation and interleukin-2 release. The FO effect on lymphocyte function was associated with decreased splenocyte release of transforming growth factor-beta and prostaglandin E2.

Intercellular action of nitric oxide increases cGMP in cerebellar Purkinje cells.

cGMP is thought to play a role in cerebellar signalling yet its production within Purkinje cells has never been detected. In the present study, the hydrolysis of a fluorescent substrate analogue, 2'-O-anthranyloyl cyclic GMP, by type 5 phosphodiesterase was monitored within Purkinje cells in slices and in culture. Nitric oxide, either endogenously released from adjacent neurons or pharmacologically applied, accelerated the rate of hydrolysis in a manner that was dependent on soluble guanylyl cyclase, demonstrating that nitric oxide triggers cyclic GMP production in Purkinje cells, which in turn activates type 5 phosphodiesterase. We conclude that NO acts as an intercellular messenger in the cerebellar cortex and that parallel fibre terminals are a probable source of nitric oxide.

Escherichia coli sepsis increases hepatic apolipoprotein B secretion by inhibiting degradation.

Sepsis leads to hypertriglyceridemia in both humans and animals. Previously, we reported that plasma very low density lipoprotein apolipoprotein (apo) B and hepatic production of apoB increased during Escherichia coli sepsis. The present experiments were undertaken to determine whether the altered hepatic secretion of apoB was associated with an increase in synthesis or a decrease in degradation rate. Sepsis was induced in male, Lewis rats (225-275 g) by intravenous injection of 3.8 x 10(8) live E. coli colonies/100 g body. Twenty-four hours later rats were sacrificed, and primary hepatocytes were prepared and incubated overnight with 35S-methionine. Hepatocytes from E. coli-treated rats secreted twice as much apoB-48 and total apoB than the hepatocytes from control rats. Escherichia coil sepsis increased cellular triglyceride mass by 86%, which was due to a stimulation in triglyceride synthesis from newly synthesized fatty acids, measured by 3H2O incorporation into triglycerides. The apoB synthesis rate, apoB mRNA levels, and apoB mRNA editing were not altered during E. coil sepsis. The pulse-chase experiments showed that the rate of apoB degradation decreased in E. coli-treated rats. These findings demonstrate that the secretion of apoB is regulated posttranslationally during E. coli sepsis by decreasing the degradation of newly synthesized apoB, which contributes to the development of hypertriglyceridemia.