A candidate gene approach identifies variants in SLC45A2 that explain dilute phenotypes, pearl and sunshine, in compound heterozygote horses.

Variations in the SLC45A2 gene are responsible for the dilution phenotypes cream and pearl in domestic horses. Cream dilution is inherited in an incomplete dominant manner, diluting only red in the heterozygous state but both red and black pigments when two alleles are present. The pearl dilution is recessive and dilutes only the red and black pigment in the homozygous state or when paired with a cream allele. Horses that inherit one copy of pearl (Cprl ) and one copy of the dominant cream allele (CC r ) display a dilution phenotype similar to that of homozygous cream, suggesting that pearl is the result of a different variation in the same gene responsible for cream. We sequenced SLC45A2 in two 'false double dilute' horses that appeared phenotypically homozygous cream but tested as possessing only a single CC r allele. We also sequenced one known pearl carrier to screen for putative causal variants. The missense variant ECA21:SLC45A2:c.985G>A; p.Ala329Thr (Cprl ) was present in one false double dilute and the pearl carrier and was also genotyped in an additional 126 horses for statistical evaluation. The genotype matched the expected phenotype in all horses (P-value = 6.5 × 10-41 ) and is identical to a pearl variant found previously. The second false double dilute horse and one non-dilute offspring genotyped as heterozygous for a novel missense variant ECA21:SLC45A2:c.568G>A (p.Gly190Arg), the proposed Csun variant (for the name of the horse). This variant produces a recessive dilution similar to pearl and indicates that multiple alleles of SLC45A2 result in dilution phenotypes in the domestic horse.

Complementary therapy use amongst Emergency Medicine patients.

OBJECTIVES: To determine the percentage of Emergency Medicine patients using Complementary Therapies (CTs), to investigate what types of CTs are used most commonly, and to gauge the acceptability of studies of CT effectiveness in a hospital setting. METHOD: Prospective cross sectional survey using a convenience sample of adult patients, presenting to the Emergency Care Centre at North Shore Hospital in Auckland, New Zealand between December 2004 and March 2006. The survey questionnaire collected demographic data, information on CT usage and attitudes to studies of CTs. Descriptive statistics with 95% confidence intervals are reported. Comparisons of proportions were made using Chi-Square or Fisher exact. RESULTS: 56.1% of participants had used a CT. Vitamins and minerals (21.2%), massage (13.4%), acupuncture (10.6%) were the most commonly used CTs. 53.3% of those using herbs or complementary medicines had not told their doctor. When compared to those who had not used CTs, respondents who had previously used CTs were both more likely to follow their doctor's advice to use a CT (71.7% vs. 55.3%, p=0.0035), and to participate in a study of CT effectiveness (65.0% vs. 45.4%, p=0.0007). CONCLUSIONS: More than half the patients surveyed had used CT in the past and more than half had not told their doctor. The majority of Emergency Medicine patients would follow the advice of their doctor if a CT was advised, and would agree to participate in a study of CT effectiveness, suggesting that such studies are practical in a hospital setting.

Chronic myeloid leukemia-associated membranoproliferative glomerulonephritis that responded to imatinib mesylate therapy.

There is no known clinical association between chronic myelogenous leukemia (CML) and membranoproliferative glomerulonephritis (MPGN). We present a patient who was followed in the renal clinic for proteinuria of unknown etiology (3.2 g/24 h) and normal renal function who was diagnosed with CML as well as MPGN and acute renal failure at the same time. The patient's renal function and proteinuria improved when his CML was treated with imatinib mesylate, suggesting that CML either caused or exacerbated existing MGPN. To the best of our knowledge, this is the first reported case of MPGN associated with CML that improved with imatinib mesylate therapy.

Isolation and characterization of structural components of Aloe vera L. leaf pulp.

The clear pulp, also known as inner gel, of Aloe vera L. leaf is widely used in various medical, cosmetic and nutraceutical applications. Many beneficial effects of this plant have been attributed to the polysaccharides present in the pulp. However, discrepancies exist regarding the composition of pulp polysaccharide species and an understanding of pulp structure in relation to its chemical composition has been lacking. Thus, we examined pulp structure, isolated structural components and determined their carbohydrate compositions along with analyzing a partially purified pulp-based product (Acemannan hydrogel) used to make Carrisyn hydrogel wound dressing. Light and electron microscopy showed that the pulp consisted of large clear mesophyll cells with a diameter as large as 1000 microm. These cells were composed of cell walls and cell membranes along with a very limited number of degenerated cellular organelles. No intact cellular organelles were found in mesophyll cells. Following disruption of pulp by homogenization, three components were isolated by sequential centrifugation. They were thin clear sheets, microparticles and a viscous liquid gel, which corresponded to cell wall, degenerated cellular organelles and liquid content of mesophyll cells based on morphological and chemical analysis. These three components accounted for 16.2% (+/-3.8), 0.70% (+/-0) and 83.1% of the pulp on a dry weight basis. The carbohydrate composition of each component was distinct; liquid gel contained mannan, microparticles contained galactose-rich polysaccharide(s) and cell walls contained an unusually high level of galacturonic acid (34%, w/w; Gal A). The same three components were also found in Acemannan Hydrogel with mannan as the predominant component. Thus, different pulp structural components are associated with different polysaccharides and thus may potentially be different functionally. These findings may help lay a basis for further studies and development of better controlled processing methods and applications for this well-accepted medicinal plant.

Medical care at the Sweetwaters Music Festival.

AIMS: To describe medical cover and medical presentations at the four-day 1999 Sweetwaters Music Festival, and make comparisons with other festivals. METHODS: All medical contacts were counted, and patients presenting to the medical tent were included in the study. Case records were studied to determine demographic data, nature of complaint, treatment and disposition. A Medline literature search was performed to obtain information on other festivals. RESULTS: There were 2,231 medical contacts overall (8.9% of estimated attendees) and 217 presentations to the medical tent (0.9% of estimated attendees). 53% of patients presenting to the medical tent were men and the mean patient age was 25 years. Lacerations (16%), intoxication (13%), local infections (12%) and soft tissue injuries (9%) were the most common problems. There were no deaths or cardiac arrests. CONCLUSION: Problems encountered were similar to other music festivals, with minor injuries predominant.

"Herbal Ecstasy": a case series of adverse reactions.

AIMS: To report five cases of adverse effects associated with the ingestion of "Herbal Ecstasy" tablets and discuss possible mechanisms of toxicity. METHOD: Composition of the "Herbal Ecstasy" tablets was determined by discussions with distributors and with MEDSAFE: Reference to relevant texts and an internet and Medline literature search was used to identify articles of interest RESULTS: Three patients complained of minor symptoms such as perceptual disturbances, anorexia, inability to sleep, dizziness, palpitations and paresthesia. A fourth patient presented with palpitations and ventricular bigeminy, and a fifth patient presented with headache, vomiting, and a hypertensive crisis. Different brands of "Herbal Ecstasy" have different compositions, some containing caffeine and kava, and one contained ephedrine. CONCLUSION: These five patients presented with adverse reactions to "Herbal Ecstasy" ranging from minor to major. Knowledge of the possible mechanisms of toxicity will help plan therapy in cases of major toxicity.

Mechanisms of Ca2+ antagonism in imipramine-induced toxicity of isolated adult rat cardiomyocytes.

Classic explanations of cyclic antidepressant toxicity often focus on Na+ channel blockade; however, cyclic antidepressant toxicity often causes decreased myocardial contractile function. The present experiments first examine inhibition of cytosolic Ca2+ signals by imipramine. Second, the experiments test if alkalinization prevents the inhibition of Ca2+ signals. Cardiomyocytes from adult rat hearts were loaded with fura-2 dye, and intracellular calcium, [Ca2+]i, was quantified using ratio fluorescence techniques. Changes in [Ca2+]I were induced by electrical pacing, depolarization with KCl (84 mM), or treatment with caffeine (10 mM). Imipramine (10-30 microM) inhibited [Ca2+]i transients in electrically paced cardiomyocytes. Imipramine (7.5-30 microM) also inhibited Ca2+ signals in KCl depolarized cells. These inhibitory effects were similar to those observed with nisoldipine (100-2000 nM), a selective L-channel blocker. The rise in [Ca2+]i that was triggered with caffeine (10 mM) was not significantly changed by imipramine (30 microM). Inhibition of KCl-induced Ca2+ signals by imipramine was prevented by alkalinization of the medium (tris(hydroxymethyl)aminomethane, pH 7.6), but not by elevation of extracellular sodium to 170 mM. Alkalinization was effective in the presence of HOE642, a selective Na+/H+ (NHE) subtype 1 inhibitor. These data show that imipramine causes Ca2+ antagonism in heart cells which is independent of sarcoplasmic reticulum Ca2+, and that alkaline treatment prevents this Ca2+ antagonism rather than stimulating an alternate source of Ca2+ via Na+/H+ and subsequent Na+/Ca2+ exchange.

Hematopoietic augmentation by a beta-(1,4)-linked mannan.

CARN 750 (injectable acemannan) is a polydispersed beta-(1,4)-linked acetylated mannan isolated from the Aloe barbadensis plant. It has multiple therapeutic properties including activity in wound repair and as a biological agent for the treatment of neoplasia in animals as well as the ability to activate macrophages. We report herein that CARN 750 directly or indirectly has significant hematoaugmenting properties. We observed that the subcutaneous administration of CARN 750 significantly increases splenic and peripheral blood cellularity, as well as hematopoietic progenitors in the spleen and bone marrow as determined by the interleukin-3-responsive colony-forming unit culture assay and the high-proliferative-potential colony-forming-cell (HPP-CFC) assay (a measure of primitive hematopoietic precursors) in myelosuppressed (7 Gy) C57BL/6 mice. The greatest hematopoietic effect was observed following sublethal irradiation in mice receiving 1 mg CARN 750/ animal, with less activity observed at higher or lower doses. Further, CARN 750, following daily injection, has activity equal to or greater than the injection of an optimal dose of granulocyte-colony-stimulating factor (G-CSF) in myelosuppressed mice. In this comparison, significantly greater activity was observed in the splenic and peripheral blood cellularity, and in the frequency and absolute number of splenic HPP-CFC as compared to the mice receiving G-CSF at 3 micrograms/animal. CARN 750, when administered to myelosuppressed animals, decreased the frequency of lymphocytes with a concomitant significant increase in the frequency of polymorphonuclear leukocytes (PMN). However, owing to the increased cellularity, a significant increase in the absolute number of PMN, lymphocytes, monocytes and platelets was observed, suggesting activity on multiple cell lineages. The latter is the primary difference in activity as compared to G-CSF which has activity predominantly on PMN.

Studies on optimal dose and administration schedule of a hematopoietic stimulatory beta-(1,4)-linked mannan.

Several complex carbohydrates have been found to significantly stimulate hematopoiesis. CARN 750, a polydispersed beta-(1,4)-linked acetylated mannan isolated from the Aloe vera plant, has been shown to have activity in wound repair, to function as a antineoplastic, and to activate macrophages. We report, herein, the hematoaugmenting properties of CARN 750 and its optimal dose and timing of administration in an animal model of irradiation-induced myelosuppression. We observed that subcutaneous injections of 1 mg/animal of CARN 750 had equal or greater stimulatory activity for white blood cell (WBC) counts and spleen cellularity as well as on the absolute numbers of neutrophils, lymphocytes, monocytes and platelets than did higher or lower doses of CARN 750 or an optimal dose of granulocyte-colony stimulating factor (G-CSF). Hematopoietic progenitors, measured as interleukin-3-supported colony forming units-culture (CFU-C) and high proliferative potential colony-forming cells (HPP-CFC) assays, were similarly increased by CARN 750 in the spleen but not in the bone marrow. The frequency of splenic HPP-CFCs and absolute number of splenic HPP-CFCs and CFU-Cs were optimally increased by 1 mg/animal of CARN 750. In contrast, bone marrow cellularity, frequency and absolute number of HPP-CFCs and CFU-Cs had as a dosage optimum 2 mg/animal of CARN 750. These parameters were similarly increased by G-CSF. In studies to determine the optimal protocol for the administration of CARN 750 we found that the hematopoietic activity of CARN 750 increased with the frequency of administration. The greatest activity in myelosuppressed mice was observed for all hematopoietic parameters except the platelet number in mice receiving daily administration of 1 mg/animal of CARN 750 with activity equal to or greater than G-CSF.

The effect of Acemannan Immunostimulant in combination with surgery and radiation therapy on spontaneous canine and feline fibrosarcomas.

Eight dogs and five cats with histopathologically confirmed fibrosarcomas were treated with Acemannan Immunostimulanta in combination with surgery and radiation therapy. These animals had recurring disease that had failed previous treatment, a poor prognosis for survival, or both. Following four to seven weekly acemannan treatments, tumor shrinkage occurred in four (greater than 50%; n = 2) of 12 animals, with tumors accessible to measurement. A notable increase in necrosis and inflammation was observed. Complete surgical excision was performed on all animals between the fourth and seventh week following initiation of acemannan therapy. Radiation therapy was instituted immediately after surgery. Acemannan treatments were continued monthly for one year. Seven of the 13 animals remain alive and tumor-free (range, 440+ to 603+ days) with a median survival time of 372 days. The data suggests that Acemannan Immunostimulant may be an effective adjunct to surgery and radiation therapy in the treatment of canine and feline fibrosarcomas.

Pilot study of the effect of acemannan in cats infected with feline immunodeficiency virus.

Acemannan, a complex carbohydrate shown to stimulate interleukin-1, tumor necrosis factor alpha and prostaglandin E2 production by macrophages, has also demonstrated antiviral activity in vitro against human immunodeficiency virus, Newcastle disease virus and influenza virus. A pilot study was undertaken to determine acemannan's effect in 49 feline immunodeficiency virus (FIV) infected cats with clinical signs of disease (Stage 3, 4 or 5), 23 of which had severe lymphopenia. Cats received acemannan either by intravenous (Group 1) or subcutaneous (Group 2) injection once weekly for 12 weeks, or by daily oral (Group 3) administration for 12 weeks. Upon entry into the study, cats were randomly assigned to one of the three groups. Laboratory analyses were performed at the beginning of the study and at Weeks 6 and 12. Cats were allowed to continue with a predetermined maintenance regimen of acemannan after completing the 12-week study. Thirteen cats died during the course of treatment. Upon necropsy, the most frequent histopathologic findings were neoplastic, kidney and pancreatic disease. Friedman's two-way ANOVA test showed no significant differences in efficacy among groups administered acemannan by the different routes. Therefore, groups were combined and a signed-ranks test was used to determine changes over time. A significant increase was seen in lymphocyte counts (P < 0.001). Neutrophil counts decreased significantly (P = 0.007), as did incidence of sepsis (P = 0.008). When cats entering with lymphopenia were analyzed separately, a much greater increase in lymphocyte counts was noted (235%) compared with non-lymphopenic cats (42%). A survival rate of 75% was found for all three groups. Thirty-six of 49 animals are alive 5-19 months post-entry. These results suggest that acemannan therapy may be of significant benefit in FIV-infected cats exhibiting clinical signs of disease.

Efficacy of acemannan in treatment of canine and feline spontaneous neoplasms.

Forty-three dogs and cats with spontaneous tumors were treated with the immunostimulating polysaccharide acemannan by intraperitoneal and intralesional routes of administration. Tumors from 26 of these animals showed histopathological evidence of immunological attack as shown by marked necrosis or lymphocytic infiltration. Thirteen showed moderate to marked tumor necrosis or liquefaction. Twenty-one demonstrated lymphoid infiltration, and seven demonstrated encapsulation. Twelve animals showed obvious clinical improvement as assessed by tumor shrinkage, tumor necrosis, or prolonged survival; these included five of seven animals with fibrosarcomas. It is believed that acemannan exerts its antitumor activity through macrophage activation and the release of tumor necrosis factor, interleukin-1, and interferon.

Synthesis and anticancer activity of nitrosourea derivatives of phensuximide.

Nitrosourea derivatives 9--13 which utilize phensuximide (1) as the carrier were synthesized as potential central nervous system antitumor agents. The N-(2-chloroethyl)-N-nitrosourea 13 was active in the mouse ependymoblastoma brain-tumor system, as well as the intraperitoneal L1210 leukemia system.