Equine Preparations Research Articles

Human versus equine intramuscular antitoxin, with or without human intrathecal antitoxin, for the treatment of adults with tetanus: a 2 × 2 factorial randomised controlled trial

SummaryBackgroundIntramuscular antitoxin is recommended in tetanus treatment, but there are few data comparing human and equine preparations. Tetanus toxin acts within the CNS, where there is limited penetration of peripherally administered antitoxin; thus, intrathecal antitoxin administration might improve clinical outcomes compared with intramuscular injection.MethodsIn a 2 × 2 factorial trial, all patients aged 16 years or older with a clinical diagnosis of generalised tetanus admitted to the intensive care unit of the Hospital for Tropical Diseases, Ho Chi Minh City, Vietnam, were eligible for study entry. Participants were randomly assigned first to 3000 IU human or 21 000 U equine intramuscular antitoxin, then to either 500 IU intrathecal human antitoxin or sham procedure. Interventions were delivered by independent clinicians, with attending clinicians and study staff masked to treatment allocations. The primary outcome was requirement for mechanical ventilation. The analysis was done in the intention-to-treat population. The study is registered at ClinicalTrials.gov, NCT02999815; recruitment is completed.Findings272 adults were randomly assigned to interventions between Jan 8, 2017, and Sept 29, 2019, and followed up until May, 2020. In the intrathecal allocation, 136 individuals were randomly assigned to sham procedure and 136 to antitoxin; in the intramuscular allocation, 109 individuals were randomly assigned to equine antitoxin and 109 to human antitoxin. 54 patients received antitoxin at a previous hospital, excluding them from the intramuscular antitoxin groups. Mechanical ventilation was given to 56 (43%) of 130 patients allocated to intrathecal antitoxin and 65 (50%) of 131 allocated to sham procedure (relative risk [RR] 0·87, 95% CI 0·66–1·13; p=0·29). For the intramuscular allocation, 48 (45%) of 107 patients allocated to human antitoxin received mechanical ventilation compared with 48 (44%) of 108 patients allocated to equine antitoxin (RR 1·01, 95% CI 0·75–1·36, p=0·95). No clinically relevant difference in adverse events was reported. 22 (16%) of 136 individuals allocated to the intrathecal group and 22 (11%) of 136 allocated to the sham procedure experienced adverse events related or possibly related to the intervention. 16 (15%) of 108 individuals allocated to equine intramuscular antitoxin and 17 (16%) of 109 allocated to human antitoxin experienced adverse events related or possibly related to the intervention. There were no intervention-related deaths.InterpretationWe found no advantage of intramuscular human antitoxin over intramuscular equine antitoxin in tetanus treatment. Intrathecal antitoxin administration was safe, but did not provide overall benefit in addition to intramuscular antitoxin administration.FundingThe Wellcome Trust.

All things being equal: spatiotemporal differences between Open and Women’s 16-goal Polo

ABSTRACTPolo is an equestrian team sport, consisting of Open and Women’s only handicapping systems. Equine activities may differ in distribution and their affect upon match outcome in Women’s Polo compared to Open Polo, potentially impacting equine preparation and management. We aimed to quantify spatiotemporal differences between Open and Women’s Polo when matched for handicap and assess their interaction with chukka and match outcomes. Distance, speed and high-intensity activity data were collected via player-worn global positioning system (GPS) units during 16-goal Open and Women’s Polo tournaments. Notational analysis quantified chukka duration and chukka and game outcomes. Spatiotemporal metric differences between Open and Women’s Polo were small to large (ES: 0.54–1.81). In Open Polo, players covered moderately more distance in games won (mean: 429.0 m; 95% CI: 238.9 m to 619.0 m), with small to large increases in high-intensity activities also performed. Whereas in Women’s Polo, moderately higher maximum speeds were attained in games won (17.13 km.h−1; 11.86 km.h−1 to 22.40 km.h−1) and a small increase in accelerations performed (5.1; 0.2 to 10.0). Open and Women’s Polo, when matched for handicap, present with small to large spatiotemporal differences that are likely of practical significance, and may influence game outcome differently between codes.

Phase II Study of Thymoglobulin, Cyclosporine and G-CSF for Initial Treatment of Aplastic Anemia and Low Risk Myelodysplastic Syndrome

Aplastic anemia (AA) is treated with equine anti-thymocyte globulin (hATG) and cyclosporine with response rates up to 65%. Rabbit anti-thymocyte globulin (rATG, Thymoglobulin®) has been used successfully in relapsed and refractory patients with AA, reducing the risk associated with readministration of equine preparations. Immunosuppressive therapy is also effective in some subsets of patients with Myelodysplastic syndrome (MDS).To evaluate if the combination of rATG, cyclosporine and G-CSF is safe and effective as first-line treatment of AA and low-risk MDS.In this single-arm, phase II study, we investigated the efficacy (primary endpoint) and safety (secondary endpoint) of combined rATG, cyclosporine and G-CSF as first-line therapy for patients with AA and Hypoplastic MDS. After 8 patients were enrolled, the study was modified to also include patients with low/intermediate-1 per the International Prognostic Scoring System. Thymoglobulin 3.5 mg/kg (or 2.5 mg/kg/day for patients ≥55 years) was given for 4 days in the first 10 patients and for 5 days in the remaining patients. Methylprednisolone (1 mg/kg/day) was given for 5 days followed by a tapering dose of oral prednisone. G-CSF (5 μg/kg) was given subcutaneously daily for up to 3 months starting after thymoglobulin. Cyclosporine (5 mg/kg) was given daily and continued for 6 months or longer at the discretion of the treating physician. All patients received prophylactic broad-spectrum antimicrobials. Responses were assessed about 3 months after initiating therapy.Thirty-six patients have been enrolled on study with 3 patients choosing alternate therapies and 1 patient dying before initiation of therapy. Among the thirty-two patients treated, 4 have been on study for less than 2 months; therefore 14 patients with AA and 14 with MDS were evaluable for a response. The median age was 62 years (20–83) for patients with AA and 63 (42–80) for patients with MDS. Thirteen of the fourteen (93%) patients with AA responded (5 CR, 7 PR, 1 HI-N), while four of 14 (29%) patients with MDS responded (1 CR, 3 PR). For patients with AA, the median time to response (TTR) was 93 days (79–623). For patients with MDS, the median TTR was 111 days (77–139). The median response duration for both groups has not been reached. Side effects were listed for 30 of 36 patients on treatment. The main grade 3/4 toxicities included thrombocytopenia in 20 patients (67%), anemia in 8 (27%), leukopenia in 17 (57%), and neutropenic fever in 4 (13%). Other grade 3/4 toxicities included renal failure, hypertension, pneumonia, urinary tract infection, fever, leukocytosis, increased liver function tests, hyperglycemia, and syncope (in 1 patient each).The combination of rabbit thymoglobulin (rATG), cyclosporine and G-CSF is safe and effective in first-line treatment of AA and has significant activity in low-risk MDS.

Phase II study of thymoglobulin, cyclosporine and G-CSF for initial treatment of aplastic anemia and low risk myelodysplastic syndrome

7093 Background: Aplastic anemia (AA) is commonly treated with equine anti-thymocyte globulin (ATG) and cyclosporine with response rates up to 65%. Thymoglobulin has been used successfully in relapsed/refractory AA, reducing the risk associated with re- administration of equine preparations. Immunosuppressive therapy is also effective in some patients (pts) with myelodysplastic syndrome (MDS). Methods: In this single-arm, phase II study, we investigated efficacy (primary endpoint) and safety (secondary endpoint) of combined thymoglobulin, cyclosporine and G-CSF as a first-line therapy for pts with AA (goal: 30 pts) or low-risk MDS (goal: 30 pts, low or intermediate-1 International Prognostic Scoring System). Thymoglobulin 3.5 mg/kg (or 2.5mg/kg/d for pts ≥55 years) was given for 4 days in the first 10 pts and for 5 days in the remaining pts. Methylprednisone (1 mg/kg/d) was given for 5 days, followed by a tapering dose of oral prednisone. Cyclosporine (5 mg/kg), and G-CSF (5 μg/kg) were also given daily and continued for 6 months or longer. All pts received prophylactic broad-spectrum antibiotics. Responses were assessed approximately 1 to 3 months after therapy. Results: Thirty pts have been enrolled on study; 3 pts chose alternative therapies and 1 pt died before initiation of treatment. Thus 26 (12 with AA and 14 with MDS) have received treatment. Of these, 1 pt was lost to follow-up and 3 have been on study for ≤ 2 months; therefore 10 with AA and 12 with MDS were evaluable. Median age was 65 years (21 - 82) for AA pts and 63 years (47 - 78) for MDS pts. All 10 (100%) AA pts responded (4 CRs, 5 PRs, 1 HI-N). Three of 12 (21%) MDS pts responded (1 CR, 2 PRs). For AA pts median time to response (TTR) was 92 days (30–346) and median response duration was 282 days (14–827). For MDS pts median TTR was 122 days (77–139) and median response duration was 33 days (19–476). Grade 3/4 toxicities were mainly cytopenias and neutropenic fever, occurring in 19 pts (86%). Other grade 3 toxicities included pneumonia, hyperglycemia, elevated ALT and abdominal pain (in 1 patient each). Conclusions: The combination of thymoglobulin, cyclosporine and G-CSF is safe and effective in first-line treatment of AA and has significant activity in low-risk MDS. Author Disclosure Employment or Leadership Consultant or Advisory Role Stock Ownership Honoraria Research Expert Testimony Other Remuneration Genzyme Genzyme

Inositol phosphate stimulation by LH requires the entire α Asn 56 oligosaccharide

Lentil lectin-bound, fucose-enriched hTSH was reported to stimulate both cAMP and inositol phosphate (IP) intracellular signalling pathways, whereas fucose-depleted hTSH stimulated only the cAMP pathway. Gonadotropins activate the cAMP pathway and in several studies higher concentrations activate the IP pathway. Since only the 10% of α subunit Asn 56 oligosaccharides (Asn 52 in humans) are fucosylated, the higher glycoprotein hormone concentrations required for IP pathway activation might be related to the abundance of competent hormone isoforms. Lentil lectin-fractionated equine (e)LHα and eFSHα preparations were combined with a truncated, des(121–149)eLHβ preparation. All four hybrid hormone preparations induced IP accumulation in porcine theca cells, suggesting that activation of the IP pathway was not dependent on fucosylation at α subunit Asn 56. However, the presence of Asn 56 carbohydrate was necessary for increased IP accumulation. Intact, rather than Asn 56-deglycosylated eLH preparations provoked a biphasic steroidogenic response by rat testis Leydig cells, suggesting that Gα i stimulation was also sensitive to loss of Asn 56 carbohydrate. While rat granulosa cells responded to human FSH preparations in a biphasic manner, a classical sigmoidal response was obtained to eFSH and Asn 56-deglycosylated eFSH, suggesting that the equine preparations did not activate Gα i. Purified oLHα Asn 56 oligosaccharides inhibited FSH-stimulated steroidogenesis in rat granulosa cell cultures indicating a direct role for carbohydrate in FSH action. The same carbohydrate preparation inhibited hCG-stimulated fluorescence energy transfer suggesting oligosaccharide involvement in activated LH receptor self-association.

Comparative Expression of Liver Cytochrome P450-dependent Monooxygenases in the Horse and in other Agricultural and Laboratory Species

The apoprotein expression and the catalytic activities of cytochrome P450s involved in the biotransformation of xenobiotics were investigated in horse liver microsomes and compared with those of food producing (cattle, pigs, broiler chicks, and rabbits) and laboratory species (rats). Western blot analysis revealed the presence of proteins immunorelated to rat CYP 1A, CYP 2B, CYP 2E, and CYP 3A subfamilies in hepatic microsomes from horses and from any other examined species. With the exception of the N-demethylation of N-nitrosodimethylamine in broiler chicks, all the recorded interspecies differences were quantitative in nature. Equine preparations proved the most active in the biotransformation of the CYP 1A substrates ethoxy- and methoxyresorufin and the least active in the metabolism of aminopyrine and ethoxycoumarin. On a comparative basis, large differences were observed in the rate of the in vitro metabolism of model substrates between “minor” (rabbits, horses) and “major” food producing species. Taken in due consideration the limitations of the in vitro approach, results from this study reinforce the conclusion that studies on drug efficacy and residue depletion should be performed in each target species.

Vasomotor effects of histamine on bovine and equine basilar arteries in vitro.

The vasomotor effects of histamine on isolated bovine and equine basilar arteries were examined. Histamine induced contractions in both these preparations. The maximal response to and pEC50 value for histamine of the equine artery were larger than those of bovine tissue. Similar results were obtained with endothelium-denuded basilar arteries. Diphenhydramine (H1-receptor antagonist) inhibited histamine-induced contractions of the basilar arteries from both species in a concentration-dependent manner and its pA2 values (with 95% confidence limits) were 7.61 (7.39-7.83) and 8.15 (8.01-8.29) for the bovine and equine preparations, respectively. Cimetidine (H2-receptor antagonist) slightly potentiated histamine-induced contractions of bovine, but not equine, basilar arteries. 2-Thiazolylethylamine (H1-receptor agonist) induced contractions in both preparations, whereas impromidine (H2-receptor agonist) induced weak relaxation of the bovine, but not the equine, tissue. These findings indicate that bovine basilar arterial smooth muscle cells possess H1- and H2-receptors. Stimulation of the former results in contraction, whereas stimulation of the latter results in weak relaxation. Equine basilar arterial smooth muscle cells possess H1-receptors, stimulation of which results in contraction.

Immunological studies of the luteinizing hormone: II. Comparative study of antigenic compounds

Comparison of the antigenic compounds in luteinizing hormone preparations of ovine, bovine, equine, and human origin was made using a hyperimmune rabbit antiserum to ovine luteinizing hormone. Methods involving agar double diffusion and immunoelectrophoresis revealed more precipitin lines in the ovine and bovine preparations than in those of either human or equine origin. The lines of the ovine and bovine hormones were fairly well balanced in electrophoretic mobility and similar to each other in appearance, whereas the lines of the equine and human hormones were predominantly anodic and differed in appearance from those of ovine and bovine preparations. The hemagglutination inhibition method showed that larger quantities of human and equine hormones than of ovine and bovine hormones were needed to inhibit the interaction between the hyperimmune antiserum and its homologous hormone. There was no correlation between the results of these tests and biological potency. Contamination with follicle-stimulating hormone was demonstrated by immunoprecipitation tests in all five hormone preparations, suggesting a lack of species specificity for this hormone and pointing to the clear antigenic dissimilarity between luteinizing and follicle stimulating hormones. Contamination with thyroid-stimulating hormone was demonstrated serologically only in the extracts of ovine and bovine glands. The ovine, bovine, and equine preparations of luteinizing hormone contained antigenic components which overlapped 3 3 An overlapping component is defined as a component certain antigenic determinants of which are shared by another hormone. with pregnant mare's serum gonadotropin, and the existence of a component in ovine, bovine, and human extracts which overlapped with human chorionic gonadotropin was also established.

THE EFFECTS OF SOME URINARY EXTRACTS ON GASTRIC SECRETION IN THE SHAY OPERATED RAT

Following the general procedure of Risley, Raymond, and Barnes the effects of urinary extracts on gastric secretion have been studied in the Shay operated young, male, albino rat. The procedure consisted of ligating the pylorus following a 24 hr. fast. Immediately postoperatively the test material was administered. After a five hour period the stomach was removed, opened, and rinsed into 10 ml. of distilled water. An aliquot of this was titrated for its free and total acid content. Human chorionic gonadotrophin and preparations obtained by the benzoic acid absorption method from human and equine pregnancy urine were studied for their effects on gastric section using the above technique after subcutaneous, intraduodenal, and oral administration. It was found that chorionic gonadotrophin caused gastric secretory depression at a dose level as low as 10 mgm. per rat subcutaneously. Both the human and equine preparations were observed to reduce the gastric secretion proportionately to the dose when administered subcutaneously. When administered by intraduodenal injection at the time of operation, the equine preparation was active but required several times the subcutaneous dose to produce comparable results. Oral administration of both equine and human pregnancy urine extracts showed antisecretory activity only in the former at the dose levels employed. In addition, the effects of possible complicating factors such as estrogens, pyrogens, and nonspecific damage on gastric secretion have been investigated and found to play no appreciable part in the results produced by the urine fractions.

The use of bovine antitoxin for the prophylaxis of tetanus

1. 1. Bovine tetanus antitoxin has been used with safety in selected persons known to give severe anaphylactic reactions to horse serum. 2. 2. The only indication for the use of equine tetanus antitoxin occurs in those cases where the person is known to be, or can be demonstrated to be, sensitive to the bovine but not to the equine preparation. Thus far no such cases have been reported. 3. 3. Persons undergoing an allergic study should be tested routinely with equine and bovine sera. This should be done by the scratch method only to avoid as far as possible the sensitizing of the patient to the serum. If the patient is sensitive to both sera, he should be immunized by means of tetanus toxoid, the precautions suggested by Cooke and his associates 5 being carefully observed.