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Biomedical subjects

S Hurwitz

Publications and source records attributed to S Hurwitz.

At least 55 records · Page 3Linked to original sources
PubMed1997

Cloning, expression, and tissue localization of the calcium-sensing receptor in chicken (Gallus domesticus).

In previous studies, we characterized an extracellular Ca2+ (Cao(2+))-sensing receptor (CaR) that plays a central role in regulating parathyroid hormone secretion in mammals by sensing Cao2+. In the present study, we have cloned and characterized the chicken (Gallus domesticus) homolog of the CaR. The chicken parathyroid CaR shares a high degree of homology (84% amino acid identity) with the human CaR and displays a similar topology. Moreover, amino acid residues where mutations cause disorders of Cao(2+)-sensing in the human CaR share the wild-type human sequence in the chicken CaR. However, a single region in the extracellular domain of the chicken CaR differs substantially from its mammalian homologs. Xenopus laevis oocytes injected with chicken CaR cRNA respond to elevated ambient levels of Cao2+, extracellular Mg2+, or extracellular Gd3+ with the characteristic activation of inositol trisphosphate-dependent, intracellular Ca(2+)-induced Cl- currents elicited by mammalian CaRs as well as by G protein-linked receptors coupled to activation of phospholipase C. By in situ hybridization, clusters of cells in chicken parathyroid glands were shown to express CaR messenger RNA. Northern analysis and immunohistochemistry demonstrated expression of receptor transcripts and/or protein in kidney tubules and intestine as well as in brain. The close conservation of the amino acid sequence of the chicken CaR with its mammalian homologs as well as its similar tissue distribution suggest that the receptor may also play an important role in avian calcium homeostasis.

Amino Acid Sequence↗
PubMed1997

Cloning and characterization of a calcium-sensing receptor from the hypercalcemic New Zealand white rabbit reveals unaltered responsiveness to extracellular calcium.

The extracellular Ca2+ (Ca(0)2+)-sensing receptor (CaR) recently cloned from mammalian parathyroid, kidney, brain, and thyroid plays a central role in maintaining near constancy of Ca(0)2+. We previously showed that the hypercalcemia normally present in New Zealand white rabbits is associated with an elevated set point for Ca(02+)-regulated PTH release (the level of Ca(0)2+ half-maximally inhibiting hormonal secretion). This observation suggested an alteration in the Ca(02+)-sensing mechanism in the rabbit parathyroid, a possibility we have now pursued by isolating and characterizing the rabbit homolog of the CaR. The cloned rabbit kidney CaR (RabCaR) shares a high degree of overall homology (> 90% amino acid identity) with the bovine, human, and rat CaRs, although it differs slightly in several regions of the extracellular domain potentially involved in binding ligands. By Northern analysis and/or immunohistochemistry, a similar or identical receptor is also expressed in parathyroid, thyroid C cells, small and large intestine, and in the thick ascending limb and collecting ducts of the kidney. When expressed transiently in HEK293 cells and assayed functionally through CaR agonist-evoked increases in Ca(i)2+, the rabbit CaR shows apparent affinities for Ca(0)2+, Mg(0)2+, and Gd(0)3+ that are indistinguishable from those observed in studies carried out concomitantly using the human CaR. Therefore, at least as assessed by its ability to increase Ca(i)2+ when expressed in HEK293 cells, the intrinsic functional properties of the rabbit CaR cannot explain the hypercalcemia observed in vivo in the New Zealand white rabbit.

Amino Acid Sequence↗
PubMed1996

Trials of 9-amino-20(S)-camptothecin in Boston.

9-Amino-20(S)-camptothecin (9-AC) is an analog of camptothecin with limited water solubility which has shown significant preclinical activity in a variety of human solid tumor xenografts. A Phase I trial using a soluble formulation of 9-AC, given as a 72-hour continuous infusion, has been completed. Thirty-one patients with resistant cancers received 5-60 micrograms/M2/h at three week intervals. The Maximum Tolerated Dose (MTD) was 45 micrograms/M2/hour. Neutropenia was the dose limiting toxicity, with few significant non-myelosuppressive toxicities. Minor responses were seen in 3/31 patients. Pharmacokinetic studies of 9-AC lactone (closed ring) showed substantial interpatient variability with a predicted half-life of 36 hours. A phase I/II trial of the same formulation of 9-AC is ongoing in refractory leukemia. Stomatitis and diarrhea are the non-myelosuppressive dose limiting toxicities. Evidence of antineoplastic activity has been seen in 3/15 patients. A Phase II trial in previously untreated metastatic breast cancer is also underway. A Phase I trial of a colloidal dispersion formulation, not yet completed, is better tolerated with a MTD > 45 micrograms/M2/h as a 72-hour continuous infusion. Evidence of antineoplastic activity has also been demonstrated.

Adult↗
PubMed1996

Pilot study of topical trifluridine for the treatment of acyclovir-resistant mucocutaneous herpes simplex disease in patients with AIDS (ACTG 172). AIDS Clinical Trials Group.

SUMMARY: Twenty-six AIDS patients were enrolled in an open label pilot study to evaluate the efficacy and toxicity of topical 1 percent ophthalmic trifluridine solution for the treatment of chronic mucocutaneous herpes simplex virus disease unresponsive to at least 10 days of acyclovir therapy. Susceptibility testing to acyclovir, trifluridine, and foscarnet was determined by plaque reduction assay. Twenty-four patients were evaluable for efficacy and 25 for toxicity analyses. Seven patients (29 percent) had complete healing of lesions. The overall estimated median time to complete healing was 7.1 weeks. An additional seven patients had > or = 50 percent reduction in lesion area. The overall estimated median time to 50 percent healing was 2.4 weeks. Ten (42 percent) patients discontinued treatment for reasons other than primary treatment failure and seven (29 percent) for failure to respond to therapy. Baseline patient characteristics associated with greater reduction in lesion area included higher Karnofsky score (p = 0.05), fewer lesions (p = 0.07), smaller lesion area (p = 0.11), and trifluridine susceptibility (p = 0.07). Eight (33 percent) patients developed new lesions outside of the treatment area while on study, reflecting the local nature of this therapy. No dose-limiting toxicity attributable to trifluridine was reported. Given the limited options for the treatment of acyclovir-resistant herpes simplex disease, topical trifluridine may be a useful alternative in selected patients.

AIDS-Related Opportunistic Infections↗
PubMed1996

Immunolocalization of osteonectin in avian tibial dyschondroplastic cartilage.

Osteonectin is an acidic calcium-binding protein found in cartilage, bone matrix, vascular endothelium, and areas of tissue repair. Using immunocytochemistry, osteonectin has been localized in all zones of the normal avian epiphyseal growth plate with notably high amounts in the hypertrophic zone. In the proximal portion of this zone the staining was intracellular, while in the distal calcifying portion of the hypertrophic zone staining was both intracellular and extracellular. Osteonectin was also detected in the growth plate associated with lesions of chickens with tibial dyschondroplasia (TD). Intense intracellular staining was observed in hypertrophic chondrocytes proximal to the lesion; staining was markedly diminished in the TD lesion; extracellular matrix was devoid of staining. Staining intensity was high along the peripheral edges of the lesion that were undergoing vascularization and resorption. This was the only area in the dysplastic cartilage where staining was observed in the extracellular matrix as well as intracellularly. Similar patterns were viewed in all TD lesions examined, whether they were spontaneous or induced by dietary treatments or genetic selection.

Animals↗
PubMed1996

Effects of diurnally cycling versus constant temperatures on chicken growth and food intake.

1. Male broiler chickens were exposed in separate experiments to: (a) constant ambient temperatures (Ta) ranging between 10 and 35 degrees C; (b) 12 h:l2 h diurnal high-low temperature of 10:30 degrees C, 15:35 degrees C and 15:30 degrees C in different trials; and (c) 12 h of 30 or 35 degrees C and 12 h of variable lower temperature. 2. A progressive decline in weight gain and food intake was obtained as Ta increased from 18 to 35 degrees C. Under diurnally cycling temperature, weight gain and food intake were lower than in the average corresponding constant temperature, with the exception of chickens exposed to 15 : 30 degrees C, where weight gain and food intake were not significantly different from those of the constant average temperature. 3. As the diurnal cold period was made colder, chickens exposed during half of the diurnal cycle to high temperature (30, 35 degrees C) demonstrated a significant (P <= 0.05) increase in weight gain and food intake. 4. There was a good correlation between plasma triiodothyronine (T3) and Ta. The correlation between T3 and food intake was highly significant, but that between T3 and weight gain was poor. 5. The results indicate the difficulties in predicting the performance of broilers exposed to diurnally cycling temperatures from knowledge of the average temperature.

Adipose Tissue↗
PubMed1996

Homeostatic control of plasma calcium concentration.

Due to the importance of Ca2+ in the regulation of vital cellular and tissue functions, the concentration of Ca2+ in body fluids is closely guarded by an efficient feedback control system. This system includes Ca(2+)-transporting subsystems (bone, and kidney), Ca2+ sensing, possibly by a calcium-sensing receptor, and calcium-regulating hormones (parathyroid hormone [PTH], calcitonin [CT], and 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]). In humans and birds, acute Ca2+ perturbations are handled mainly by modulation of kidney Ca2+ reabsorption and by bone Ca2+ flow under PTH and possibly CT regulation, respectively. Chronic perturbations are also handled by the more sluggish but economic regulatory action of 1,25(OH2)D3 on intestinal calcium absorption. Peptide hormone secretion is modulated by Ca2+ and several secretagogues. The hormones' signal is produced by interaction with their respective receptors, which evokes the cAMP and phospholipase C-IP3-Ca2+ signal transduction pathways. 1,25 (OH)2D3 operates through a cytoplasmic receptor in controlling transcription and through a membrane receptor that activates the Ca2+ and phospholipase C messenger system. The calciotropic hormones also influence processes not directly associated with Ca2+ regulation, such as cell differentiation, and may thus affect the calcium-regulating subsystems also indirectly.

Algorithms↗
PubMed1996

Induction of thermotolerance in male broiler chickens by temperature conditioning at an early age.

Chicks were conditioned by exposure to heat stress (36 +/- 1 C; to 80% RH) for 24 h at the age of 5, or 5 and 7 d. During conditioning, weight gain was depressed. Due to accelerated growth during the postconditioning period, a complete compensation for lost weight gain was achieved by the group conditioned at 5 d, but only a partial compensation was obtained in the group conditioned at both 5 and 7 d. At the age of 42 d, challenge with acute heat stress (35 +/- 1 C; 20 to 30% RH) resulted in a large increase in cloacal temperature of the controls and a more moderate increase in the conditioned chickens. Mortality during the thermal challenge was significantly higher in the control than that of the previously exposed (conditioned) groups. Conditioning at an early age resulted in hemodynamic changes (significant decrease in heart weight and hematocrit) and reduced plasma triiodothyronine (T3) concentration. The results suggest that reduced T3 and hemodynamic changes may be part of the mechanism associated with improved thermotolerance by early age temperature conditioning.

Adaptation, Physiological↗
PubMed1996

Developmental changes in skin collagen biosynthesis pathway in posthatch male and female chickens.

The developmental changes in skin collagen biosynthesis pathway in male and female chickens were evaluated. Concentration of collagen, levels of mRNA for collagen type I subunits and for lysyl hydroxylase, and the level of three lysyl oxidase-derived cross-links: dehydro-dihydroxylysinonorleucine (DHLNL), dehydro-hydroxylysinonorleucine (HLNL), and dehydro-histidinohydroxymerodesmosine (HHMD) were determined during 4 wk posthatching. Skin collagen content increased with age and was higher in males than in females. In both sexes, the expression of the genes coding for alpha 1 and alpha 2 of collagen type I decreased with age: alpha 1(I) gene expression decreased from Day 3 onwards, whereas the reduction in alpha 2(I) gene expression started 1 wk later. At all ages examined, the expression of both genes was higher in male than in female skin. Males and females lysyl hydroxylase gene expression remained low until Day 16, after which an increase in the enzyme gene expression was observed. An increase in skin HLNL content was observed from Day 3 in both sexes reaching a peak in males at Day 9 and in females 1 wk later. The DHLNL content, which was higher in males than in females at all ages tested, dramatically decreased in both male and female skin from 3 d of age, reaching its lowest level at Day 16, and remained at that low level thereafter. The skin content of HHMD in males and females followed an oscillatory behavior with higher peaks in the male skin. The results suggest that the higher tensile strength of male skin than female skin may be due to the elevated skin collagen content that resulted from increased expression in collagen type I genes on the one hand, and from the higher amounts of various collagen cross-links on the other.

Aging↗
PubMed1996

Intestinal uptake and metabolism of fatty acids in the chick.

Uptake and metabolism of fatty acids in the duodenal mucosa of the chick was studied. In vitro incubations of a mucosal homogenate with 14C-oleic acid indicated that together with glycerides, water-soluble 14C compounds and 14CO2 were formed; the latter comprised approximately one quarter of metabolites formed. Studies in chickens introducing 14C-oleic acid into the duodenum and determining appearance of label in the cannulated pancreaticoduodenal vein and brachial artery indicated that similar processes occur in vivo, with both labeled glycerides and water-soluble compounds appearing in the blood. Quantitatively, metabolism of 14C-oleic acid to nonlipid compounds amounted to some 15% of the dose after 60 min. These findings suggest that a considerable portion of ingested fatty acids are metabolized before transport to the circulation.

Animals↗
PubMed1996

A novel 29-kDa chicken heat shock protein.

The family of small heat shock proteins is the more variable among the highly conserved superfamily of heat shock proteins (HSP). Using a metabolic labeling procedure with tissue explants, we have detected in chickens a new member of the small HSP family with an apparent molecular weight of 29-kDa. This protein was induced in broiler chickens' heart muscle and lungs following an in vivo heat stress. The 29-kDa band appears after 3 h of heat stress, much later than the induction of HSP 90, HSP 70, and HSP 27. The late onset of induction suggests that HSP 29 plays a more specific role of a "second stage defense protein".

Animals↗
PubMed1995

Growth hormone inhibits differentiation of avian epiphyseal growth-plate chondrocytes.

The effect of chicken growth hormone (cGH) on the proliferation and differentiation of avian growth-plate chondrocyte was evaluated in culture. In culture, addition of ascorbic acid to the culture media caused cell differentiation. Treatment of proliferating chondrocytes with cGH caused a time-dependent increase in collagen type II gene expression together with a decrease in the appearance of osteopontin (OPN) in the medium. In addition, the ascorbic acid-dependent increase in alkaline phosphatase (AP) activity was inhibited by cGH. IGF-I, on the other hand, caused an increase in AP activity in the ascorbic acid-treated chondrocytes. In the presence of ascorbic acid, cGH did not affect collagen type II gene expression or the appearance of OPN in the medium. Proliferation of avian growth-plate chondrocytes, in contrast to mammalian chondrocytes, was not stimulated by GH alone, although the presence of cGH was essential for chondrocyte survival in long-term culture. cGH in combination with epidermal growth factor (EGF) stimulated cell proliferation. These results suggest that GH inhibits differentiation in avian growth-plate chondrocytes, thereby sustaining their proliferative state and maintaining their sensitivity to growth factors such as EGF.

Alkaline Phosphatase↗
PubMed1995

Synthesis and phosphorylation of osteopontin by avian epiphyseal growth-plate chondrocytes as affected by differentiation.

The regulation of synthesis and phosphorylation of osteopontin in relation to avian epiphyseal growth-plate chondrocyte differentiation was studied in situ and in culture. Osteopontin gene expression was evaluated in the tibia growth-plate of 3-week-old chickens by in situ hybridization. The gene was expressed mainly at the lower hypertrophic zone where cartilage matrix is calcified and endochondral bone formation is initiated. Within the hypertrophic region, a poorly labeled area separated the layer of osteopontin-positive hypertrophic chondrocytes from those associated with endochondral bone formation. In culture, proliferative chondrocytes show no alkaline phosphatase activity in contrast to ascorbic acid-treated chondrocytes which display the enzyme activity. Chondrocytes not treated with ascorbic acid, exhibited lower levels of osteopontin mRNA than the treated cells. The phorbol ester TPA--an activator of protein kinase C--and to a lesser extent FGF but not EGF, stimulated osteopontin gene expression. Chondrocytes secreted low levels of phosphorylated osteopontin to the medium. EGF treatment resulted in the appearance of phosphorylated osteopontin in the medium, without affecting the synthesis of other proteins. FGF and TGF beta, but not IGF-I or IGF-II, also caused phosphorylation of osteopontin. Ascorbic acid-treated chondrocytes secreted higher levels of phosphorylated osteopontin than the non-treated cells, but addition of FGF or TPA did not stimulate osteopontin phosphorylation any further. Parathyroid hormone caused a dose-dependent attenuation of osteopontin phosphorylation and inhibited the EGF-dependent osteopontin phosphorylation. The results suggest that osteopontin gene expression and phosphorylation in chondrocytes are regulated by separate mechanisms. The response to the various controlling agents varies with the state of differentiation. Both processes--the synthesis and phosphorylation of osteopontin--are under the control of local growth factors which are involved in bone growth and calcification.

Alkaline Phosphatase↗
PubMed1995

Osteopontin gene expression and alkaline phosphatase activity in avian tibial dyschondroplasia.

Osteopontin (OPN) gene expression and alkaline phosphatase activity were evaluated in the epiphyseal growth plates of normal chickens and in diet-induced tibial dyschdroplasia (TD)-afflicted chickens. In the normal growth plate, OPN gene was expressed by a) cells of the subperichondrial zone surrounding the articular cartilage, b) a narrow layer of hypertrophic chondrocytes at the hypertrophic zone, and c) lower hypertrophic chondrocytes at the zone of matrix calcification and endochondral bone formation. The latter two layers were separated by OPN-negative chondrocytes. Osteopontin gene was not expressed throughout the zone of articular cartilage in the nonhypertrophic or upper hypertrophic portions of the growth plate cartilage. Only at sites of calcification of the lower hypertrophic zone was the expression of the OPN gene associated with alkaline phosphatase activity. In all TD lesions, regardless of the induction procedure, the layer of chondrocytes of the lower hypertrophic zone expressing the OPN gene and the layer of OPN-negative cells separating the two areas of OPN-expressing cells were grossly enlarged. This resulted in a wide discontinuity between the chondrocytes of the lower hypertrophic zone expressing the OPN gene and the cells expressing the OPN gene that are associated with mineralization. In TD, no alkaline phosphatase activity was detected within the growth plate cartilage, but normal OPN gene expression was observed at the subperichondrium zone and at the zone of endochondral bone formation. The results of this study suggest that in the epiphyseal growth plate, OPN expression is not restricted to sites of bone calcification.

Alkaline Phosphatase↗
PubMed1995

Calcium metabolism and requirements of chickens are affected by growth.

The interaction between growth and calcium homeostasis was studied by comparing the responses of (a) fast-growing broiler chickens (Cobb) and slow-growing Leghorns, and (b) fast-growing chickens (Cobb) fed either high energy (12.13 kJ/g) or low energy (9.2 kJ/g) diets, to dietary calcium concentration ranging between 4 and 20 g/kg). Plasma calcium increased as dietary calcium increased, reaching an apparent plateau between 0.8 and 1.5% dietary calcium, regardless of basal growth rate. Dietary calcium levels of 1.5% and higher induced hypercalcemia and hypophosphatemia in fast- but not in slow-growing chickens. Weight gain was unaffected by dietary calcium in the slow-growing Leghorns, but followed a bell-shaped response pattern in the fast-growing Cobb chickens. Growth inhibition by feeding of low energy diets changed the response pattern from a quadratic form to that of an increase towards a plateau. The response of bone ash to dietary calcium was characterized as quadratic in fast-growing chicks, changing to a pattern of increase towards a plateau in slow-growing chicks. Intestinal calbindin was suppressed by dietary calcium and was higher in the fast-growing than in the slow-growing chicks. An increase in dietary phosphorus resulted in a shift in the response curves of weight gain and bone ash and an increase in the calcium requirements. The results indicate that the response of chicks to dietary calcium and calcium requirements is markedly modified by growth rate.

Analysis of Variance↗