Plasmapheresis in life-threatening verapamil intoxication.

Verapamil intoxications are life-threatening conditions with a far too often fatal outcome. In 2 patients, severe suicidal intoxication by 2.4 g and 9.6 g of verapamil orally resulted in life threatening hypotension and bradycardia with the need of heart-pacing and resuscitation. Plasmapheresis was started within less then 4 hours after intoxication and seemed to reduce the verapamil plasma concentration to less then 40%. A dramatic improvement of cardiovascular stability was already observed during plasmapheresis. In-vitro plasmapheresis was performed to verify the effectiveness of the extracorporeal detoxification. Verapamil was removed out of the blood by a clearance of 29.2 ml/min at blood flow of 200 ml/min. In conclusion, severe verapamil poisoning should be treated by early aggressive gut decontamination and an appropriate management of the haemodynamic complications. In case of lack of effectiveness for stabilisation, plasmapheresis can reduce verapamil related life threatening symptoms and bridge the time for the hepatic detoxification.

Severe coagulopathy after a bite of a green bush viper (Atheris squamiger): case report and biochemical analysis of the venom.

A 34 year old male bitten by an adult Atheris squamiger snake developed symptoms of nausea, vomiting, diarrhea which were followed by drowsiness and impaired breathing. Local hemorrhage, edema and pain at the bite-site occurred, but no systemic bleeding or hemorrhagic diathesis developed. All clinical and laboratory parameters were in the normal range except for afibrinogenemia, thrombocytopenia and slight proteinuria. Replacement therapy (fibrinogen and platelet concentrates) and treatment of shock stabilized the patient within 2d and coagulation returned to normal. Atheris squamiger venom was subjected to biochemical and biological analysis. The LD50 of the venom was 5 mg/kg (mice, s.c.). It produced local hemorrhage corresponding to about 25% of the activity of puff adder venom (Bitis arietans). In vitro the venom had a fibrinogen-converting activity, it did not activate purified prothrombin but very likely contained a F V and Ca2+-dependent prothrombin activator. The venom exhibited strong platelet-aggregating activity, which was not inhibited by protease inhibitors and by EDTA or EGTA. The venom also aggregated acetylsalicylic acid treated platelets indicating, that the arachidonic acid pathway was not essential for activation. Rat serum rapidly inhibited the platelet-aggregating activity of the venom; human serum, however, had only a partial inhibitory effect. Preliminary experiments showed that platelet-aggregating activity may be separated from fibrinogen-converting activity by anion-exchange chromatography.

The responsibilities of investigators, monitors, and sponsors of clinical studies supporting new animal drug applications. Center for Veterinary Medicine, FDA.

Clinical studies evaluating the effectiveness of investigational new animal drugs are conducted to support approval of a new animal drug application. These studies must be conducted in a manner that ensures the integrity and validity of the collected data. On November 2, 1992, the Center for Veterinary Medicine announced the availability of a guideline entitled "Conduct of Clinical Investigations: Responsibilities of Clinical Investigators and Monitors for Investigational New Animal Drug Studies." This guideline, which provides recommendations on the conduct of clinical investigations and the responsibilities of monitors and investigators, is presented in this manuscript. Additionally, the responsibilities of sponsors as delineated by the Food and Drug Administration's laws and regulations are also summarized.

Follicular fluid administration delayed, but did not prevent, the hemicastration-induced increase in follicle-stimulating hormone secretion and compensatory testicular hypertrophy in ram lambs.

To determine whether the post-hemicastration rise in serum FSH is related to compensatory testicular hypertrophy, 4-mo-old ram lambs were hemicastrated and injected s.c. with 5 ml of saline (HC; n = 5) or steroid-free follicular fluid (HC + FF; n = 5) twice daily for 28 days. Sham-operated intact lambs, injected with saline, served as controls (INT; n = 5). Blood samples were collected frequently during the first 48 h, daily for the remainder of the first week, and then three times/wk for the next 11 wk. Serum FSH concentrations in HC lambs increased above presurgery levels by 9 h, peaked at 13 days, and returned to baseline as compared to INT controls by 5 wk. Follicular fluid administration delayed (p < 0.05) the hemicastration-induced rise in FSH, but only for 14 of the 28 days of treatment; after this time, concentrations increased to the levels observed in HC rams. Within 2 days after the last follicular fluid injection (Day 30), FSH concentrations increased again to levels greater (p < 0.05) than those observed in HC animals. FSH secretion in HC + FF rams then declined to basal values by 7 wk, 2 wk later than in HC rams. Although the time course differed, the duration of elevated FSH did not differ between HC and HC + FF rams (36.4 +/- 2.0 vs. 37.8 +/- 4.7 days, respectively; p > 0.05). Testicular volume, determined on the last day of treatment (Day 28), was 70% greater (p < 0.05) in HC than in INT or HC + FF lambs; volume did not differ (p > 0.05) between INT and HC + FF rams.(ABSTRACT TRUNCATED AT 250 WORDS)

Increased bioactivity of serum follicle-stimulating hormone, but not luteinizing hormone, following hemicastration in ram lambs.

This study evaluated the short-term effects of hemicastration on levels of bioactive (bio-) and immunoactive (immuno-) FSH and LH in 3.5-mo-old ram lambs. Biological activities of FSH and LH were determined using in vitro rat granulosa cell aromatase and mouse Leydig cell testosterone assays, respectively. Four rams each were either hemicastrated (HC) or left intact (INT) and blood samples were collected over a 2-wk period. In HC rams, serum bio- and immuno-FSH concentrations increased several fold within 5-8 h after surgery and remained elevated over INT controls for 7 and 14 days, respectively. The increase in bio-FSH was greater than that for immuno-FSH, resulting in an increase in the ratio of biological to immunological activity (B/I ratio) through Day 7. Neither immuno-FSH, bio-FSH, nor the B/I ratios varied throughout the 2-wk study in INT rams. In contrast to FSH, serum concentrations of immuno- and bio-LH and the B/I ratios were similar between HC and INT rams. For both gonadotropins, biological activity was positively correlated to immunological activity. These results demonstrate that (1) immuno- and bio-FSH concentrations are elevated after hemicastration; (2) the FSH biopotency (B/I ratio) also is increased, at least temporarily; and (3), despite these changes in FSH production, neither immuno- nor bio-LH secretion is altered by hemicastration.

Consequences of severe copper deficiency are independent of dietary carbohydrate in young pigs.

The ability of carbohydrates (CHO), such as fructose and sucrose, to aggravate copper deficiency in rats and the recent dietary trends of Western human populations led to the suggestion that the Cu X CHO interaction may be pertinent to public health. This hypothesis was tested with pigs because their cardiovascular and gastrointestinal systems closely resemble those of humans. Weanling pigs were fed a diet containing either 59% sucrose or cornstarch with either deficient (0.8 mg/kg diet) or adequate (6.4 mg/kg) copper for 10 wk. Plasma and tissue copper, the activities of plasma ceruloplasmin ferroxidase and erythrocyte Cu,Zn-superoxide dismutase, hematocrits, and serum cholesterol and triglyceride were all decreased (p less than 0.05) and relative cardiac mass was increased (p less than 0.05) by severe dietary copper deficiency. The type of dietary CHO did not differentially influence the values of these variables. Thus, these data fail to support the hypothesis that the Cu X CHO interaction observed in rats represents a health risk for humans.

Consequences of copper deficiency are not differentially influenced by carbohydrate source in young pigs fed a dried skim milk-based diet.

Carbohydrates (CHO) such as fructose (FR) or sucrose, but not starch (ST), aggravate the consequences of dietary copper (Cu) deficiency in rats. To evaluate whether this Cu X CHO interaction is pertinent to human health, the pig was used as an animal model. In two studies, 66 weanling pigs were fed dried skim milk (DSM)-based diets for 10 wk with 20% of the total calories provided as either FR, glucose, or ST and containing either deficient (1.0-1.3 micrograms/g diet) or adequate (7.1 micrograms/g) levels of Cu. Plasma and tissue levels of Cu, the activities of plasma ceruloplasmin ferroxidase and erythrocyte Cu, Zn-superoxide dismutase, and hematocrits were lower (p less than 0.05) in animals fed Cu-deficient diets. The relative cardiac mass of all Cu-deficient groups was greater (p less than 0.05) than that of animals fed Cu-adequate diets. These effects were in general unaffected by type of CHO. For comparison, weaned male rats were also fed DSM-based containing diets ST or FR with adequate or deficient Cu for as long as 10 wk. Rats consuming the Cu-deficient diets were characterized by significantly lower hematocrits, decreased tissue Cu levels, and enlarged hearts, regardless of the CHO source. Together, these data demonstrate that DSM-based diets are not suitable for delineation of potential Cu X CHO interactions, and one or more components of DSM may exacerbate the consequences of dietary Cu deficiency.

Influence of naloxone and yohimbine administration on pulsatile LH secretion in luteal phase beef cows.

This study examined the effects of two specific neurotransmitter receptor antagonists, naloxone (NAL; mu-opioid) and yohimbine (YOH; alpha(2)-adrenergic), on pulsatile luteinizing hormone (LH) release during the luteal phase (Day 10; Day 0 = estrus) of beef cows. Treatments were saline i.m. (C; n = 4); 1mg/kg NAL i.m. followed 3 h later by two 0.5 mg/kg injections spaced 2.5 h apart (N; n = 4); 0.2 mg/kg YOH i.v. (Y; n = 3); or combined N and Y regimens, with Y preceding N by 30 min (NY; n = 4). Blood samples were collected for 8 h before (Period I) and after (Period II) initiation of treatment. Respiration rates of Y cows were similar to C cows during Period II. However, respiration rates of N and NY animals increased 70% within 30 min of the first NAL injection. Acute LH release was not observed in response to either NAL or YOH. Pulsatile LH secretion was unchanged in N, Y and NY cows during Period II when compared with Period I. In contrast, basal and pulsatile LH secretion was inhibited in C cows during Period II. The inhibition of LH secretion in C animals following NAL indicate that the cows were under stress during Period II. Thus, these data suggest that the inhibition of LH release in stressed animals can be overcome by pharmacologic attenuation of inhibitory (N) or accentuation of stimulatory (Y) signals to LHRH-containing neurons.

Cardiac and splenic levels of norepinephrine and dopamine in copper deficient pigs and rats.

1. Copper deficiency decreased the concentration and content of norepinephrine in the hearts of pigs and rats. 2. Concentration, but not content, of norepinephrine was decreased in spleen of copper-deficient pigs, while splenic norepinephrine levels in rats were not altered by copper deficiency. 3. Cardiac and splenic concentrations and contents of dopamine were elevated in copper-deficient pigs and rats. 4. Tissue concentrations of catecholamines and the magnitude of change due to copper deficiency were greater in pigs than rats.

Nutritional anestrus in beef cows: concentrations of glucose and nonesterified fatty acids in plasma and insulin in serum.

Twenty-two nonlactating Hereford cows exhibiting normal estrous cycles were fed either maintenance (M) or restricted (R) diets until most of the R cows became anestrus; R cows then were fed 160% of the M diet until estrous cycles resumed. Concentrations of progesterone, glucose, insulin and nonesterified fatty acids (NEFA) were determined in weekly blood samples. Blood also was collected frequently, before and after i.v. infusion of 300 ml of a 40% glucose solution, to evaluate responses in blood concentrations of glucose and insulin when cows were exhibiting normal estrous cycles, when R cows were initiating anestrus, during anestrus, and at the reinitiation of estrous cycles. Losses in BW and body condition score in R cows were associated with reduced (P less than .01) concentrations of glucose and insulin and greater (P less than .01) concentrations of NEFA in blood plasma compared with those of M cows. During normal estrous cycles, disappearance of infused glucose from plasma and concentrations of insulin in serum were similar for R and M cows. Glucose disappearance from plasma was retarded and serum concentrations of insulin remained increased for a longer time after glucose infusion in R at the start of anestrus compared with M cows (P less than .01). Similarly, during anestrus, the rate of glucose disappearance was slower for R cows (P less than .01). During refeeding of R cows, disappearance of infused glucose was similar for R and M cows. In conclusion, reduced concentrations of glucose and insulin and increased concentrations of NEFA in blood were associated with nutritional anestrus and the glucoregulatory effects of insulin were compromised during nutritional anestrus.

Nutritional anestrus in beef cows: body weight change, body condition, luteinizing hormone in serum and ovarian activity.

Multiparous Hereford cows (n = 22) with moderate to good body condition scores (BCS) were randomly allotted to maintenance (M) or restricted (R) diets under drylot conditions. Cows on M diets received adequate feed to maintain initial BW, whereas R cows were fed to lose 1% of their initial BW weekly until luteal activity ceased. When most of the R cows became anestrous, their diet was increased to allow BW gain and resumption of ovarian cyclicity. Body weights and BCS were recorded weekly and luteal activity was assessed by weekly determination of progesterone in plasma. Concentrations of LH in serum were quantified in weekly samples and in samples obtained frequently at four selected times. Restricted cows had reduced BW (P less than .01) by 5 wk and reduced BCS (P less than .01) by 15 wk compared with M cows. Luteal activity ceased after 26 +/- 1 wk of reduced nutrient intake in 91% of the R cows; R cows had lost 24.0 +/- .9% of their initial BW and had a BCS of 3.5 +/- .3. Lack of luteal activity was associated with absence of behavioral estrus. Estrous cycles resumed 9 +/- 2 wk after the diet of R cows was increased, at which time R cows weighed 12 +/- 3% less than at the start of the experiment and had a BCS of 4.6 +/- .2. Concentrations of LH in serum samples obtained weekly were reduced (P less than .01) in R cows compared with M cows. In addition, LH pulse frequency was reduced (P less than .05) when R cows were initiating anestrus.(ABSTRACT TRUNCATED AT 250 WORDS)

Short-term pituitary-testicular responses of prepubertal ram lambs to hemicastration.

To examine the short-term effects of hemicastration on pituitary-gonadal responses, 12 ram lambs were anesthetized and hemicastrated at 4 mo of age and killed (n = 4) at 2 (HC2), 7 (HC7), or 14 (HC14) days following surgery. Four intact (INT) rams were killed 14 days following anesthesia. Testis and pituitary weights were similar between HC and INT rams. Serum follicle-stimulating hormone (FSH) in HC rams increased within 6 h, peaked at 12 h, and remained elevated above INT levels throughout the study. Overall mean serum testosterone levels in HC rams were lower than in INT rams for the first 48 h, but were similar by 3 days post-surgery. Pulsatile luteinizing hormone (LH) and testosterone secretion was suppressed for the first 9.5 h following anesthesia and/or surgery in both HC and INT animals. A single LH pulse and succeeding testosterone pulse occurred in 10/12 HC and 4/4 INT rams between 10 and 14 h post-surgery, both of which were lower in amplitude in HC than INT animals. However, on Day 7, pulsatile secretory patterns of LH and testosterone were similar, suggesting compensatory androgen secretion had occurred in HC rams. Pituitary LH content was unaffected by hemicastration. In contrast, pituitary FSH content was greater in HC7 and HC14 compared to HC2 and INT animals. Pituitary gonadotropin hormone-releasing hormone (GnRH) receptor concentrations were similar in INT, HC7, and HC14 rams, but were slightly reduced in HC2 rams. Neither testicular LH nor FSH receptor concentrations were altered by hemicastration at any time.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of FSH treatment on LH and FSH receptors in chronic cystic-ovarian-diseased dairy cows.

This experiment was conducted to 1) determine whether chronic cystic-ovarian-diseased (CCOD) cows fail to respond to luteinizing hormone (LH) treatment because of a lack of adequate ovarian LH receptors and 2) determine the effect of follicle stimulating hormone (FSH) treatment on ovarian LH and FSH receptors in ovaries of CCOD cows. The CCOD cows were those that did not resume cyclic ovarian activity after repeated treatment with human chorionic gonadotropin (hCG) and(or) LH-releasing hormone (LHRH) and were considered chronic by veterinarians. Thirteen CCOD cows were purchased from producers; six of them were injected with 5 mg FSH twice daily for 3 or 5 d (TCCOD) and the remaining seven remained untreated. Seven control (noncystic) cows in the luteal phase of the estrous cycle were injected with Lutalyse approximately 48 to 50 h before slaughter so they would be in the follicular phase (FP) of the cycle at the time of slaughter. Analysis of serum and pituitaries showed no differences (P greater than .05) in mean concentrations of serum or pituitary LH and FSH or pituitary LHRH receptor concentration and affinity among FP, CCOD and TCCOD cows. Ovarian follicle wall concentrations of receptors for LH (3.2 +/- .6; 13.0 +/- 2.5; 22.4 +/- 5.1 fmol/mg protein) and FSH (10 +/- 2.6; 43 +/- 7.2; 29 +/- 6.7 fmol/mg protein) were lower (P less than .05) in CCOD cows compared with FP and TCCOD cows, respectively. The same pattern was observed for concentrations of granulosa cell LH and FSH receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Elevation of serum testosterone during chronic LHRH agonist treatment in the bull.

The objective of this study was to try to depress serum testosterone (T) in bulls by prolonged treatment with a potent luteinizing hormone-releasing hormone (LHRH) agonist. Eight sexually mature bulls (325 to 475 kg) were assigned to treatment or control groups. Treatment consisted of 150 micrograms nafarelin acetate 6-D-2-naphthyl-alanine-LHRH (LHRH-A) injected im every 6 h for 15 d. Bovine serum albumin (BSA, .01%) in a carrier solution was injected at the same times in control bulls. Serial 15-min blood samples were collected via jugular cannula during the initial 36 h of treatment and during 6-h windows on d 4, 8 and 14. Bulls were slaughtered and pituitaries and testes collected on d 15. Serum luteinizing hormone (LH), follicle stimulating hormone (FSH) and T were elevated after initial injection of LHRH-A, but returned to basal concentrations by 12, 5 and 17 h, respectively. Prolonged LHRH-A treatment prevented pulsatile LH and T secretion compared with control bulls. Mean serum LH did not differ from that of controls on d 4, 8 and 14 of LHRH-A treatment, while serum T was elevated (P less than .01) during the same time periods. Oscillating patterns and mean concentrations of serum FSH were not different between control and LHRH-A-treated bulls. Fifteen days of LHRH-A treatment depressed pituitary LHRH receptor numbers (P less than .05) and pituitary LH (P less than .01) and FSH (P less than .05) concentrations. Testicular LH receptor numbers were elevated (P less than .01), but testicular FSH receptor numbers were not altered.(ABSTRACT TRUNCATED AT 250 WORDS)

Pituitary luteinizing hormone-releasing hormone receptors in ovariectomized cows after challenge with ovarian steroids.

Acute changes of bovine pituitary luteinizing hormone-releasing hormone (LHRH) receptors in response to steroid challenges have not been documented. To investigate these changes 96 ovariectomized (OVX) cows were randomly allotted to one of the following treatments: 1) 1 mg estriol (E3); 2) 1 mg 17 beta-estradiol (E2); or 3) 25 mg progesterone (P) twice daily for 7 days before 1 mg E2 and continuing to the end of the experiment. Serum was collected at hourly intervals from 4 animals in each group for 28 h following estrogen treatment. Four animals from each treatment were killed at 4-h intervals from 0 to 28 h after estrogen injection to recover pituitaries and hypothalami. Treatment with E3 or E2 decreased serum luteinizing hormone (LH) within 3 h and was followed by surges of LH that were temporally and quantitatively similar (P greater than 0.05). Progesterone did not block the decline in serum LH, but did prevent (P less than 0.05) the E2-induced surge of LH. Serum follicle-stimulating hormone (FSH) was unaffected (P less than 0.05) by treatment. Pituitary concentrations of LH and FSH were maximal (P less than 0.001) at 16 h for E3 and 20 h for E2, whereas P prevented (P greater than 0.05) the pituitary gonadotropin increase. Concentrations of LHRH in the hypothalamus were similar (P greater than 0.05) among treatments. Pituitary concentrations of receptors for LHRH were maximal (P less than 0.005) 12 h after estrogen injection (approximately 8 h before the LH surge), even in the presence of P. This study demonstrated that in the OVX cow: 1) E2 and E3 increased the concentration of receptors for LHRH and this increase occurred before the surge of LH; and 2) P did not block the E2-induced increase in pituitary receptors for LHRH but did prevent the surge of LH.

LHRH receptor, LH and FSH concentrations in anterior pituitaries of cycling, noncycling and early pregnant heifers.

In domestic animals limited data are available concerning levels of pituitary luteinizing hormone-releasing hormone (LHRH) receptors during various physiological states. The objectives of this study were to quantify anterior pituitary gonadotropin and LHRH receptor concentrations in cycling, noncycling and early pregnant beef heifers. To accomplish these objectives, five heifers each were slaughtered, after synchronization with prostaglandin F2 alpha (PGF2 alpha), on d 0 (estrus), 7 and 14 of the estrous cycle and d 40 of pregnancy. Four heifers determined to be noncycling were also slaughtered. Pituitaries were collected and analyzed for LHRH receptor and gonadotropin concentrations. Pituitary luteinizing hormone (LH) concentrations were low on d 0 (1.4 +/- .2 micrograms/mg pituitary, mean +/- SE) and remained low on d 7 (1.4 +/- .1 micrograms/mg pituitary) before increasing (P less than .01) on d 14 (2.6 +/- .5 micrograms/mg pituitary). Luteinizing hormone concentrations, compared with d 0, were also elevated (P less than .01) in noncycling (NC; 2.6 +/- .2 micrograms/mg pituitary) animals and in 40-d pregnant (PG; 2.5 +/- .2 micrograms/mg pituitary) heifers. Pituitary follicle stimulating hormone (FSH) concentrations, though similar (P greater than .05) for all groups, paralleled changes in LH concentration. Pituitary LHRH receptor binding affinity did not differ (P greater than .05) among groups, with an overall Kd = .64 +/- .02 X 10(-9) M. Luteinizing hormone-releasing hormone receptor concentrations were highest on d 0 (1.09 +/- .12 fmol/mg pituitary) and fell (P less than .01) to low levels on d 7 (.75 +/- .11 fmol/mg pituitary).(ABSTRACT TRUNCATED AT 250 WORDS)

Nursing enhances the negative effect of estrogen on LH release in the cow.

Twenty-three crossbred beef cows between 4 and 5 yr of age were assigned at random to one of six treatments: (1) ovariectomized 4 d postpartum (OVX) with early weaning of calves 21 d postpartum (OVX-EW; n = 4), (2) OVX-EW and 17 beta-estradiol implants (OVX-E2-EW; n = 4), (3) OVX and normal nursing by calves throughout the experiment (OVX-NN; n = 3), (4) OVX-NN and 17 beta-estradiol implants (OVX-E2-NN; n = 4), (5) intact cows and early weaning of calves 21 d postpartum (EW), (6) intact cows and normal nursed (NN). Blood was collected at 15-min intervals over a 4-h period once weekly during the 12-wk postpartum period in the OVX cows. Early weaned intact cows exhibited estrus 23 d sooner (P less than .05) than normally nursed cows. A hormone level for each cow at each week was determined from the mean of the 17 samples collected over the 4 h period each week. There were no significant changes due to E2 treatment, for concentrations of LH, FSH or number of pulses during wk 1 through 3. However, during wk 4 through 12 the linear and quadratic contrasts of wk X estrogen X nursing were significant for serum LH, indicating there was no difference between the treatments for EW and NN without E2 but there was a large difference in the presence of E2. During nursing E2 suppressed serum LH below that of nonestrogen-treated cows while after weaning E2 stimulated LH release above that of nonestrogen-treated cows.(ABSTRACT TRUNCATED AT 250 WORDS)