An association between the R990G polymorphism of the CaSR gene, coding for calcium-sensing receptor, and primary hypercalciuria was found in kidney stone formers. To confirm this relationship, we investigated hypercalciuric women without stones and studied the effect of CaSR gene in human embryonic kidney cells (HEK-293). We genotyped for CaSR A986S, R990G, and Q1011E polymorphisms, 119 normocalciuric and 124 hypercalciuric women with negative history of kidney stones. Homozygous (n=2) or heterozygous (n=21) women for the 990G allele considered as one group had an increased risk to be hypercalciuric (odds ratio=5.2; P=0.001) and higher calcium excretion (P=0.005) in comparison with homozygous women for the 990R allele (n=220). HEK-293 cells were transfected with the variant allele at the three CaSR gene polymorphisms and with the most common allele with no variants. The transient increment of intracellular calcium caused by the stepwise increase of extracellular calcium was evaluated in stable transfected cells loaded with fura-2 AM. The extracellular calcium concentration producing the half-maximal intracellular calcium response was lower in HEK-293 cells transfected with the 990G allele than in those transfected with the wild-type allele (P=0.0001). Our findings indicate that R990G polymorphism results in a gain-of-function of the calcium-sensing receptor and increased susceptibility to primary hypercalciuria.

Abstract. Calcium-sensing receptor (CaSR) is a plasma membrane protein that regulates tubular reabsorption of Ca. To establish its role in idiopathic hypercalciuria, the association of urinary Ca excretion with the polymorphisms of CASR gene has been studied in healthy subjects and in hypercalciuric and normocalciuric Ca stone formers. CASR exon 7 single nucleotide polymorphisms (SNP), G/T at codon 986, G/A at codon 990, and C/G at codon 1011, were evaluated by PCR amplification and direct sequencing in 97 normocalciuric stone formers, 134 hypercalciuric stone formers, and 101 normocalciuric healthy controls. Four haplotypes were defined on the basis of CASR gene SNP: haplotype 1 was characterized by the most frequent sequence; haplotypes 2, 3, or 4 by the presence of a single polymorphic variant at codon 986, 990, or 1011, respectively. The relative risk of hypercalciuria was calculated with multinomial logistic regression and was significantly increased only in individuals carrying haplotype 3 (Odds ratio, 13.0 [95% confidence interval,

A low-salt diet can reduce calcium excretion in hypercalciuric stone formers. This trial was registered at clinicaltrials.gov as NCT01005082.

Context: Primary hyperparathyroidism (PHPT) shows a great variability in clinical course and severity. Data concerning the association between polymorphic variants of the gene encoding the calciumsensing receptor (CaSR) and clinical characteristics of PHPT are not conclusive. Objective: To evaluate the frequency of three polymorphisms; A986S, R990G, and Q1011E of CaSR in patients with PHPT and to correlate the genotypes with clinical and biochemical parameters. Patients and methods: The study included 94 consecutive unrelated patients referred to our Departments for PHPT diagnosis and management between 2000 and 2005 and 137 age and sexmatched healthy subjects. Patients and controls were genotyped according to standard procedures. Due to the rarity of 990G allele, homozygous and heterozygous subjects were grouped in R/GCG/G set. All PHPT patients were studied for calcium metabolism parameters and renal and bone complications. Results: The proportion of CaSRvariants was similar in PHPT patients and controls. In PHPT patients, only R990G polymorphism was associated with disease parameters; in comparison with R/R, R/GCG/G patients showed lower mean serum parathyroid hormone (PTH) and phosphate levels (139.9G62.2 vs 199.9G136.3 pg/ml, P!0.05 and 0.69G0.12 vs 0.81G0.18 mmol/l, PZ0.031 respectively), higher mean 24-h urine calcium concentration and calcium excretion (9.05G2.05 vs 6.77G4.31 mmol/24 h, PZ0.012 and 67G20 vs 51G26 mmol/l GF, PZ0.039), and increased prevalence of nephrolithiasis (90.0 vs 44.2%, PZ0.007). Conclusions: The study showed that patients with PHPT, bearing the 990G allele, had lower serum PTH levels and higher urinary calcium excretion in comparison with the other genotype, suggesting an increased sensitivity of the variant receptor to extracellular calcium. Since this variant was associated with increased occurrence of nephrolithiasis, analysis of this polymorphism might help to predict renal complication of the disease.European Journal of Endocrinology 155 687-692

Currently, no animal-based protocol for on-farm welfare assessment of commercial turkeys is available. The birds’ size and flighty nature make obtaining a representative sample using traditional methods difficult. The transect walks (TW) approach provides a potential alternative for on-farm assessments of turkey welfare. We compared the TW approach with a traditional method, and data collected as the birds were moved out of the house during the load out process (L). Ten commercial 19- to 20-week-old Hybrid turkey flocks were evaluated (1 flock/house/farm). Half of the flocks were housed on farms deemed as “faring well” by the company, the other half were on “suboptimal” farms. Each house was subdivided longitudinally into 4 transects. Two observers walked the transects in random order, recording the total number of birds per transect that were immobile; lame; aggressive towards a mate; interacting with humans; with visible head, vent, or back wounds; engaging in mounting behaviors; small; featherless; dirty; sick; terminal; or dead. Flocks were re-evaluated on the same day using the individual sampling method (S), where randomly selected birds were scored as they took 10 steps. Flocks were re-assessed within 48 h of the transect evaluation, as birds were funneled out of the house during load out. Using ANOVAs we determined the effects of observers, method, management, and their interactions on proportions of turkeys per house within each category. Outcome parameters were not affected by management (P > 0.05 for all) or observer (P > 0.05 for most), but an assessment method effect was detected (P < 0.05). S differed from the 2 other methods (P < 0.05) for most parameters except aggression towards a mate, back wounds, dirty, sick, and vent wounds. Differences were not detected between data collected using TW and during L, except for dead (P = 0.0007) and immobile (P = 0.007). Results suggest that the TW method is a promising tool for on-farm turkey welfare assessment as it produced results similar to those obtained at L when all birds could be scored individually.