EVALUATION OF THE ASSOCIATION OF PLASMA PENTRAXIN-3 LEVELS WITH CAROTID INTIMA-MEDIA THICKNESS AND HIGH-SENSITIVE CRP IN PATIENTS WITH SUBCLINICAL HYPOTHYROIDISM.

Context: Inflammation-related markers may predict cardiovascular diseases. Objective: In this study, it was aimed to assess pentraxin-3 (PTX-3) levels and its relationship with carotid intima-media thickness (CIMT) and high-sensitive C-reactive protein (hsCRP) in patients with subclinical hypothyroidism. Design: Prospective cross-sectional study. Methods: This study included 60 patients (aged 30-60 years) with subclinical hypothyroidism and 30 healthy volunteers as controls. The demographic characteristics and anthropometric measurements were performed in all patients and controls. In addition, sonographic carotid artery examination, thyroid functional tests, lipid profile, hsCRP, and PTX-3 levels of the participants were investigated. Results: The PTX-3, hsCRP levels and CIMT were higher in patients with subclinical hypothyroidism when compared to controls (p=0.008, p=0.001, p<0.001, respectively). The PTX-3 level was strongly correlated with hsCRP (r=0.865; p<0.001), but no such correlation was detected with CIMT (r=-0.255; p=0.50). In binominal logistic regression analysis, it was found that CIMT and serum uric acid levels were independent parameters associated with subclinical hypothyroidism. In ROC analysis, a cut-off value of >3.75 ng/mL for serum PTX-3 level predicted subclinical hypothyroidism with a sensitivity of 60% and specificity of 60.7% (AUC: 0.672, p=0.004). Conclusion: Showing inflammation and endothelial dysfunction, the PTX-3 may be a helpful marker in patients with subclinical hypothyroidism associated with increased risk for cardiovascular disease.

CUSHING'S SYNDROME DUE TO PRIMARY BILATERAL MACRONODULAR ADRENAL HYPERPLASIA AND METACHRONOUS PHEOCHROMOCYTOMA.

Introduction: Primary bilateral macronodular adrenal hyperplasia (PBMAH) is one of the uncommon causes of endogenous Cushing's syndrome (CS).Pheochromocytoma (PCC) is another adrenal tumor which is derived from neural crest arising in the adrenal medulla. Here we are reporting a case with recurrent overt CS due to PBMAH, 2 years after unilateral adrenalectomy, concomitant with recently developed PCC. Case Presentation: A 43-year-old woman was admitted to our clinic with a 30 kg weight gain, proximal muscle weakness, menstrual irregularity, easy bruising and excessive hair growth on face and body.The lab results were compatible with a diagnosis of solely ACTH-independent CS. Screening showed bilateral macronodular lesions and she underwent right adrenalectomy. Postoperatively, she had lost weight and her well-being had improved; 2 years later, she developed CS and paroxysmal hypertension. The left adrenal gland was laparoscopically removed. Histopatologically, the lesion was reported as a typical PCC and macronodular-micronodular hyperplasia of the adrenal tissue surrounding that lesion. Conclusions: Pheochromocytoma with synchronous ACTH-independent CS originating from the same adrenal gland is very rare. To the best of our knowledge,our case is the first one describing the coexistence of overt ACTH-independent CS due to PBMAH and metachronous PCC.The importance of detailed re-evaluation of patients with recurrent ACTH-independent CS is highlighted here.

Effects of Nigella sativa oil on ovarian volume, oxidant systems, XIAP and NF-kB expression in an experimental model of diabetes.

We investigated the effects of Nigella sativa oil on ovary volume, nuclear factor-kappaB (NF-κB), X-linked inhibitor of apoptosis protein (XIAP) expression, and serum malondialdehyde (MDA), superoxide dismutase (SOD), total antioxidant status (TAS) and total oxidant status (TOS) levels in diabetic rats. We divided 21 adult female rats into three groups: controls, diabetics and diabetics + N. sativa oil. The diabetics + N. sativa oil group was given 0.2 mg/kg/day N. sativa oil 6 days/week for 4 weeks. NF-κB and XIAP expression was assessed in ovarian sections using immunohistochemistry. The right and left ovary volumes were calculated using stereology. We also measured serum MDA, SOD, TAS and TOS levels. We found that N. sativa oil reduced hyperglycemia, but not to control levels. N. sativa oil also exhibited antioxidant properties as demonstrated by reduced serum TOS and MDA levels, and increased SOD and TAS levels compared to controls. We found no significant difference in total ovarian volume, XIAP or NF-κB expression among the groups, which may be due to the short study period. Our findings suggest that N. sativa oil may be useful for reducing blood glucose levels and elevated oxidant activity in diabetic patients.

ASSOCIATION OF THE HUMAN PPARγ2 PRO12ALA POLYMORPHISM WITH OBESITY IN A POPULATION FROM TURKEY.

BACKGROUND: There have been a number of reports on the relationship between the PPARγ2 Pro12Ala genotype and the development of obesity. OBJECTIVE: A case-control survey was designed to investigate the potential association between a Pro12Ala polymorphism in the PPARγ2 gene and obesity and/or obesity-related phenotypes in a population from Turkey. MATERIALS AND METHODS: The polymerase chain reaction and restriction enzyme digestion were used to genotype the Pro12Ala polymorphism of the PPARγ2 gene in 149 unrelated obese and 105 non-obese control subjects from Turkey. The data were analyzed statistically. RESULTS: We found that the overall minor allele frequency was 0.12 in cases and 0.095 in controls. In terms of genotype distribution and allele frequencies among the cases versus controls in the population studied, only the gender-stratified analysis revealed a significantly higher frequency of Pro/Ala genotype within males. The polymorphism was associated with significantly higher weight, height, waist circumference, central adiposity (waist-to-hip ratio, WHR), lean body weight as well as dry body weight, but not overall adiposity (total body fat percentage, TBF) in cases carrying Ala allele (Pro/Ala or Ala/Ala). However, in the subjects carrying Ala allele of the control group, WHR values were found significantly lower. CONCLUSION: Our results showed that the Pro12Ala polymorphism in the PPARγ2 gene is associated with obesity in the studied adult population from Turkey. These data suggest that the Pro12Ala polymorphism in PPARγ2 may be a potential genetic risk factor for central obesity.

Vitamin E and selenium treatment of monocrotaline induced hepatotoxicity in rats.

Monocrotaline (MCT) is a hepatotoxic pyrrolizidine alkaloid that is derived from plants; exposure may occur by consumption of contaminated grains, herbal teas and medicines. MCT can cause liver damage. We investigated the antioxidant effects of selenium (Se) and vitamin E against the toxic effects of MCT. Female Wistar albino rats were divided into four groups: a control group, an MCT group, an MCT + Se group, and an MCT + vitamin E group. Liver tissues were harvested, fixed, processed to paraffin and sections were cut. Anti-von Willebrand factor (vWF) immunohistochemistry, terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL), and hematoxylin and eosin staining were performed. Serum and liver tissue glutathione (GSH), catalase (CAT), and glutathione peroxidase (GPx) levels were measured. Histopathological and TUNEL data showed significantly increased liver damage in the MCT group compared to controls. Histopathological and TUNEL staining indicated significant improvements in the MCT + vitamin E and MCT + Se groups compared to the MCT group. MCT significantly reduced the serum GSH level and GPx activity, and liver GPx activity. Biochemical data indicated a significant improvement in serum GSH level in the MCT + vitamin E group compared to the MCT group. We suggest that vitamin E and Se afford limited protection against MCT hepatotoxicity.

Effects of curcumin on methyl methanesulfonate damage to mouse kidney.

Methylmethane sulfonate (MMS) is an alkylating agent that may react with DNA and damage it. We investigated histological changes and apoptosis caused by MMS and the effects of curcumin on MMS treated mouse kidneys. Twenty-four mice were divided into four equal groups: controls injected with saline, a group injected with 40 mg/kg MMS, a group injected with 40 mg/kg MMS and given 100 mg/kg curcumin by gavage, and a group given 100 mg/kg curcumin by gavage. MMS caused congestion and vacuole formation, and elevated the apoptotic index significantly, but had no other effect on kidney tissue. Curcumin improved the congestion and vacuole formation caused by MMS and decreased the apoptotic index. Curcumin administered with MMS appears to decrease the deleterious effects of MMS on the kidney.

Low vitamin D levels are associated with increased risk for fatty liver disease among non-obese adults.

Recent data have revealed an inverse relationship between insulin resistance, which is associated with fatty liver disease, and blood 25-hydroxy-vitamin D (25(OH)D) levels. The aim of the present study was to determine the association of vitamin D levels with the presence and stage of fatty liver disease among non-obese subjects and to determine the effect of vitamin D status on fatty liver disease development. A total of 613 non-obese (body mass index <30 kg/m(2)) gastroenterology and internal medicine outpatients (472 women and 141 men) were enrolled in the study. The patients' laboratory values, including liver function tests, lipid profiles, C-reactive protein, fasting blood glucose, insulin, calcium and 25(OH)D levels were studied. Low vitamin D levels, higher triglyceride levels and higher alanine aminotransferase levels were found to be the significant determinants for non-alcoholic fatty liver disease. When the patients were evaluated as low or normal vitamin D groups, low vitamin D levels was determined to be a risk factor for fatty liver disease, with an odds ratio of -1.59 (confidence interval -1.22 to -1.97). The increased risk for fatty liver disease among patients with low vitamin D status may be suggestive of mechanisms promoting fat flow and accumulation in the liver. Molecular studies are warranted to elucidate the action of vitamin D on the liver with respect to fat metabolism.

Genetic and molecular characterization of uveal melanoma cell lines.

The recent identification of frequent activating mutations in GNAQ or GNA11 in uveal melanoma provides an opportunity to better understand the pathogenesis of this melanoma subtype and to develop rational therapeutics to target the cellular effects mediated by these mutations. Cell lines from uveal melanoma tumors are an essential tool for these types of analyses. We report the mutation status of relevant melanoma genes, expression levels of proteins of interest, and DNA fingerprinting of a panel of uveal melanoma cell lines used in the research community.

The involvement of distinct neural systems in patients with obsessive-compulsive disorder with autogenous and reactive obsessions.

OBJECTIVE: To investigate the regional metabolite abnormalities and changes after treatment in patients with OCD with autogenous and reactive obsessions. METHOD: We assessed right anterior cingulate cortex (ACC) and amygdala-hippocampal region (Am + Hpp) N-acetyl-aspartate (NAA), choline (Cho) and creatine (Cr) concentrations and NAA/Cr and Cho/Cr ratios using single-voxel proton magnetic resonance spectroscopy in 15 patients with autogenous obsessions (OCD-A), 15 patients with reactive obsessions (OCD-R) and 15 healthy controls (HC). Measurements were repeated after 16 weeks of fluoxetine treatment. RESULTS: Baseline ACC NAA/Cr ratios of both OCD groups were significantly lower than HC. OCD-A group had significantly lower baseline NAA/Cr ratios in the Am + Hpp than other groups. These differences were more likely to be explained by higher Cr levels in ACC. We found no significant differences and changes for Cho levels and Cho/Cr ratios between groups and within groups. Significant increase in NAA/Cr ratios of OCD-A group found in the Am + Hpp was more likely to be explained by increased NAA levels. No significant changes were found in ACC NAA/Cr ratios. CONCLUSION: While disturbed energy metabolism in ACC might reflect a common pathology in patients with OCD regardless of symptom dimension, alterations in mesiotemporal lobe are more likely for autogenous obsessions.

Mutations in GNA11 in uveal melanoma.

BACKGROUND: Uveal melanoma is the most common intraocular cancer. There are no effective therapies for metastatic disease. Mutations in GNAQ, the gene encoding an alpha subunit of heterotrimeric G proteins, are found in 40% of uveal melanomas. METHODS: We sequenced exon 5 of GNAQ and GNA11, a paralogue of GNAQ, in 713 melanocytic neoplasms of different types (186 uveal melanomas, 139 blue nevi, 106 other nevi, and 282 other melanomas). We sequenced exon 4 of GNAQ and GNA11 in 453 of these samples and in all coding exons of GNAQ and GNA11 in 97 uveal melanomas and 45 blue nevi. RESULTS: We found somatic mutations in exon 5 (affecting Q209) and in exon 4 (affecting R183) in both GNA11 and GNAQ, in a mutually exclusive pattern. Mutations affecting Q209 in GNA11 were present in 7% of blue nevi, 32% of primary uveal melanomas, and 57% of uveal melanoma metastases. In contrast, we observed Q209 mutations in GNAQ in 55% of blue nevi, 45% of uveal melanomas, and 22% of uveal melanoma metastases. Mutations affecting R183 in either GNAQ or GNA11 were less prevalent (2% of blue nevi and 6% of uveal melanomas) than the Q209 mutations. Mutations in GNA11 induced spontaneously metastasizing tumors in a mouse model and activated the mitogen-activated protein kinase pathway. CONCLUSIONS: Of the uveal melanomas we analyzed, 83% had somatic mutations in GNAQ or GNA11. Constitutive activation of the pathway involving these two genes appears to be a major contributor to the development of uveal melanoma. (Funded by the National Institutes of Health and others.).

Discrimination of 2n = 60 Spalax leucodon cytotypes (Spalacidae, Rodentia) in Turkey by means of classical and molecular cytogenetic techniques.

Comparative studies among seven populations of 2n = 60 S. leucodon employing classic cytogenetics (G- bands, C-bands, AgNOR-staining), fluorochrome staining, and fluorescence in situ hybridization of telomeric and rDNA probes are reported here for the first time. The studied specimens were assigned to two cytotypes: 2n = 60W and 2n = 60R. The basic karyotype of both cytotypes consisted of eight pairs of subtelocentric and 21 pairs of acrocentric autosomes, subtelocentric X and acrocentric Y chromosomes. Both cytotypes had variable numbers of B-chromosomes (1-3) and variable numbers of autosomal arms (NFa = 74-76) caused by amplification (deletion) of heterochromatin short arms in the second pair. The short arms of subtelocentric chromosomes were comprised of heterochromatin in both cytotypes. Nucleolar organizer regions (NORs) and rDNA clusters were detected at telomeric sites of the short arms in pairs Nos. 3, 5, 6, 9, and 13 in cytotype W, and in the short arms of pair No. 6, 8, 12, 13, and 16 in cytotype R. Different locations of rDNA clusters allowed unambiguous discrimination between two S. leucodon cytotypes possessing the same 2n = 60 and similar NFa (74-76) variability. Our findings suggest a high level of chromosomal divergence, which means that it is possible to consider these cytotypes as a well-differentiated, chromosomal lineage within the leucodon group.

Multiplex ARMS analysis to detect 13 common mutations in familial hypercholesterolaemia.

DNA analysis and mutation identification is useful for the diagnosis of familial hypercholesterolaemia (FH), particularly in the young and in other situations where clinical diagnosis may be difficult, and enables unambiguous identification of at-risk relatives. Mutation screening of the whole of the three FH-causing genes is costly and time consuming. We have tested the specificity and sensitivity of a recently developed multiplex amplification refractory mutation system assay of 11 low-density lipoprotein receptor gene (LDLR) mutations, one APOB (p.R3527Q) and one PCSK9 (p.D374Y) mutation in 400 patients attending 10 UK lipid clinics. The kit detected a mutation in 54 (14%) patients, and a complete screen of the LDLR gene using single-stranded conformation polymorphism/denaturing high performance liquid chromatography identified 59 different mutations (11 novel) in an additional 87 patients, for an overall detection rate of 35%. The kit correctly identified 38% of all detected mutations by the full screen, with no false-positive or false-negative results. In the patients with a clinical diagnosis of definite FH, the overall detection rate was higher (54/110 = 49%), with the kit detecting 52% of the full-screen mutations. Results can be obtained within a week of sample receipt, and the high detection rate and good specificity make this a useful initial DNA diagnostic test for UK patients.

A rare mutation in AgRP, +79G>A, affects promoter activity.

The agouti-related protein is a powerful orexigenic peptide. A rare mutation, +79G>A, was identified in its minimal promoter in two white carriers. Comparison of the 45-year-old male proband, who was also a carrier of the common Ala67Thr polymorphism, with an age- and weight-matching wild-type population showed marginal differences for resting metabolic rate (RMR) and body mass index. The second carrier however was an obese 57-year-old female with reduced RMR. Functional analysis in hypothalamus- and periphery-derived cell lines showed reduced promoter activity for the +79A allele in the adrenocortical cells only, suggesting that it could affect the peripheral expression levels of AgRP. The +79G>A mutation could predispose to body weight gain (as suggested by the phenotype of the second carrier), but it could only affect the proband at an older age as he may be protected by the Ala67Thr polymorphism that is associated with resistance to late-onset fatness.

Long-term results of pediatric kidney transplantation at one center in Turkey.

Kidney transplantation is considered the treatment of choice for children with end-stage renal disease (ESRD). From November 1975 to June 2004, 80 of a total of 1477 kidney transplantations were performed in 78 pediatric patients. We retrospectively reviewed the records of 42 boys and 36 girls. Patient age ranged from 8 to 16 years (mean, 14.9 +/- 2.2). Sixty-three (78.7) grafts were from living donors and 17 (21.3%) from deceased donors. Two patients (2.6%) underwent preemptive transplantation, while 76 had preoperative renal replacement therapy with either hemodialysis in 62 (79.5%) or peritoneal dialysis in 14 (17.9%). Although the cause of ESRD could not be established in 30 cases, the other 48 showed the most common etiologies to be reflux nephropathy and glomerulonephritis. In conclusion, despite relatively poor socioeconomic conditions and health care problems in our country, the overall outcomes for pediatric patients at our transplantation center are good. We seek to perform more preemptive kidney transplantations in children with ESRD, and to increase our efforts to educate the Turkish public about organ transplantation and donation.

The molecular basis of familial hypercholesterolaemia in Turkish patients.

Familial hypercholesterolaemia (FH) is an autosomal dominant disorder of lipoprotein metabolism. In the majority of patients FH is caused by mutations in the gene for the low-density lipoprotein receptor (LDLR), and to date more than 700 mutations have been reported worldwide. In this study, 36 paediatric patients with a clinical diagnosis of FH (20 homozygous and 16 heterozygotes) were screened for mutations in the LDLR gene. Each exon, with intron-exon junctions, was screened by capillary fluorescent SSCP (F-SSCP) and heteroduplex analysis. Samples showing different band patterns were sequenced. Ten novel (including three frame shift small deletions or insertions) and seven known mutations were detected. A total of 37 out of the predicted 56 FH-causing alleles were identified (66.1%). No patients with the R3500Q mutation in the APOB gene were found. W556R was the most common mutation, explaining 21.4% of the predicted defective LDLR alleles. The novel sequence changes were deemed to be pathogenic if they altered a conserved amino acid (L143P, D147E, Q233H-C234G, C347G) or occurred in or close to a splice site (IVS 16+5) and were absent in DNA from 50 healthy Turkish subjects. These data confirm the genetic heterogeneity of FH in Turkey, and demonstrate the usefulness of F-SSCP for mutation detection.

Long-term results of pediatric renal transplantation at one center in Turkey.

Kidney transplantation is more frequently indicated in children than in adults because growth retardation is an additional problem associated with chronic kidney disease in the pediatric age group. This study retrospectively analyzed the data from 75 kidney transplantations performed on 73 children (38 male and 35 female) at a center in Turkey from late 1975 through 2003. The aim of the study was to investigate the case characteristics and the long-term outcomes in this patient group. Patient ages ranged from 8 to 16 years (mean, 14.9 +/- 2.2 years). Sixty (82.1%) children were on hemodialysis, and 12 (16.4%) on peritoneal dialysis prior to transplantation. Pre-emptive transplantation was performed for one (1.4%) patient. Fifty-nine transplantations used organs from live donors (78.7%) and 16 cadaver transplants (21.3%). The mean cold ischemia time for the cadaveric transplantations was 38.6 hours (range, 23-56 hours). All recipients were placed on a low-dose immunosuppressive regimen. One graft was lost due to hyperacute rejection. Twenty-one patients (28.8%) experienced a total of 24 acute rejection episodes. The follow-up period ranged from 1 to 190 months (mean, 44.1 +/- 31.8 months). Concerning postoperative complications, four patients (5.5%) developed a lymphocele; there were two cases each (2.7% each) of distal ureteral stricture, perirenal hematoma, or renovascular stenosis; and one patient (1.4%) developed a urine leakage. Two patients (2.7%) developed Kaposi's sarcoma at 17 and 3 months after transplantation. Six recipients died (mortality 8%), four of whom had a functioning graft. Two patients (2.7%) underwent retransplantation at 4 and 2 years after the initial operation. The 1-, 3-, and 5-year graft survival rates for living-related transplantations were 92%, 81%, and 70%, and the corresponding patient survival rates were 98%, 93%, and 92%, respectively. The 1-, 3-, and 5-year graft survival rates for cadaveric transplantations were 90%, 78%, and 68%, and patient survival rates 98%, 91%, and 90%, respectively. These results indicate that kidney transplantation is successful in pediatric end-stage renal disease patients particularly from living-related donors.

Three novel mutations in the apolipoprotein E gene in a sample of individuals with type 2 diabetes mellitus.

BACKGROUND: Apolipoprotein E (apoE) is found in association with triglyceride-rich lipoproteins and is the ligand for the removal of these particles from the plasma. Genetic variations in exon 4 lead to three common gene variants: E2, E3, and E4. METHODS: We performed apoE genotyping in 765 individuals with type 2 diabetes. RESULTS: We identified three new variant heteroduplex patterns. Sequencing of these variants revealed three novel mutations that were related to biochemical and clinical characteristics. One mutation produced a frameshift at amino acid position 166, which predicted termination of protein synthesis. This individual had a heteroduplex pattern and sequence of E3E3, which was associated with a change in the plasma isoelectric focusing pattern and a 70% lower plasma concentration of apoE compared with healthy individuals. The other mutations were both single base changes. A CGC>CAC change at amino acid position 150 predicted a substitution of Arg>His. This individual had a heteroduplex pattern and sequence of E2E2, which was not associated with major changes in plasma lipids or apoE concentration. The third individual had a CGC>CCC base change at amino acid position 114, which predicted an Arg>Pro change. This person had a heteroduplex pattern and sequence of E3E3, higher plasma total cholesterol, and moderately decreased plasma apoE. CONCLUSIONS: The frequency of new mutations in this sample (1 in 255) is higher than that of a healthy population (1 in 7900). Further screening for common apoE gene variants in individuals at risk for dyslipidemia may reveal abnormal heteroduplex patterns and uncover further mutations in this important lipid-regulating gene.

Severe hemolytic anemia associated with Hb Volga [beta27(B9)Ala-->Asp]: GCC-->GAC at codon 27 in a Turkish family.

A boy presented at age 4 years with severe congenital hemolytic anemia characterized by highly elevated reticulocyte count (30-50%) and prominent basophilic stippling. Hb had been 4 g/dL at age 7 months. The patient was on a monthly transfusion regimen up to the age of 7 years, when he underwent splenectomy. After removal of the spleen, his Hb stabilized at 11 g/dL. No abnormal pattern was detected in hemoglobin electrophoresis at pH 9 and 6. In-vitro globin synthesis revealed the presence of an abnormal beta-chain in front of the gamma-chain. The beta(A)/beta(X) ratio was 0.77 at 30 min and 0.74 at 2 hr of incubation. Molecular analysis revealed that the patient had GCC-->GAC alteration at codon 27 (beta27(B9)Ala-->Asp) causing the abnormal hemoglobin Volga. The beta-cDNA derived from the beta-Hb Volga allele could be differentiated from HbA beta-cDNA on silver-stained gel. No imbalance in the mRNA of beta(A)/beta(Hb Volga) ratio was observed.

The TRC8 hereditary kidney cancer gene suppresses growth and functions with VHL in a common pathway.

VHL is part of an SCF related E3-ubiquitin ligase complex with 'gatekeeper' function in renal carcinoma. However, no mutations have been identified in VHL interacting proteins in wild type VHL tumors. We previously reported that the TRC8 gene was interrupted by a t(3;8) translocation in a family with hereditary renal and non-medullary thyroid cancer. TRC8 encodes a multi-membrane spanning protein containing a RING-H2 finger with in vitro ubiquitin ligase activity. We isolated the Drosophila homologue, DTrc8, and studied its function by genetic manipulations and a yeast 2-hybrid screen. Human and Drosophila TRC8 proteins localize to the endoplasmic reticulum. Loss of either DTrc8 or DVhl resulted in an identical ventral midline defect. Direct interaction between DTrc8 and DVhl was confirmed by GST-pulldown and co-immunoprecipitation experiments. CSN-5/JAB1 is a component of the COP9 signalosome, recently shown to regulate SCF function. We found that DTrc8 physically interacts with CSN-5 and that human JAB1 localization is dependent on VHL mutant status. Lastly, overexpression of DTrc8 inhibited growth consistent with its presumed role as a tumor suppressor gene. Thus, VHL, TRC8, and JAB1 appear to be linked both physically and functionally and all three may participate in the development of kidney cancer.

beta-Thalassaemia intermedia in a Turkish girl: homozygosity for G-->A substitution at +22 relative to the beta-globin cap site.

We provide the first description of a homozygote patient for the G-->A substitution in the 5' UTR of the beta-globin gene. The proband was a 17-year-old girl with beta-thalassaemia intermedia who had never received a blood transfusion. The physical examination revealed a well-developed women with no facial or bony abnormalities. There was mild paleness and mild splenomegaly which was 2 cm below the costal margin. The haemoglobin (Hb) was 7.6 g/dl, Hb A(2) 5.4% and Hb F 14.6% of the total Hb. The Hb A(2) of both parents was 3.5%. The Hb F level in the mother and father were 0.9, 1.2% and the mean cell volume (MCV) value was 70 and 72 fl respectively. DNA analysis of the beta-gene region of the propositus revealed homozygosity for a G-->A substitution at nucleotide +22 relative to the beta-gene cap site, within a functional downstream region that was referred to as the DCE (downstream core element). In addition to the data obtained previously from in vitro transcription assays, clinical findings and in vivo expression studies gave some valuable clues about the effect of +22 G-->A mutation on the expression of beta-gene. Phenotypic expression of this homozygous patient is highly suggestive that G-->A substitution at nt +22 confers a relatively mild (silent) beta(+)-thalassaemia phenotype.