The Spectrum of Genetic Defects in Congenital Adrenal Hyperplasia in the Population of Cyprus: A Retrospective Analysis.

Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency (21-OHD) is caused by mutations in the CYP21A2 gene. The study refers to CAH patients of Greek-Cypriot ancestry between years 2007 and 2018. One hundred and twenty patients with various degrees of CAH were categorized and genotyped. The patients were categorized in 4 mutation groups based on their clinical and biochemical findings. The majority of patients (85.0%) belonged to the non-classic (NC)-CAH form and the disorder was more often diagnosed in females (71.7%). The most severe classic salt-wasting (SW) form was identified in 11 neonates (9.2%). Seven (5.8%) children were also identified with the simple virilizing (SV) form and a median presentation age of 5 years [interquartile range (IQR) 3.2-6.5]. In the 240 nonrelated alleles, the most frequent mutation was p.Val281Leu (60.0%) followed by c.655 A/C>G (IVS2-13A/C>G) (8.8%), p.Pro453Ser (5.8%), DelEx1-3 (4.6%), p.Val304Met (4.6%), and p.Gln318stop (4.2%). Other less frequent mutations including rare deletions were also identified. Following our recent report that the true carrier frequency of CYP21A2 in Greek-Cypriots is 1:10, this study reports that the CAH prevalence is predicted around 1.7 cases per 10 000 people. Therefore, the up-to-date 120 CAH patients identified by our group make only the 6.9% of the ones estimated (approximately 1750) to exist in the Greek Cypriot population. The compiled data from a coherent population such as the Greek-Cypriot could be valuable for the antenatal diagnosis, management and genetic counselling of the existing and prospect families with CAH.

Insulin resistance is associated with at least threefold increased risk for prothrombotic state in severely obese youngsters.

UNLABELLED: Obesity in childhood increases the risk for early adult cardiovascular disease. However, the underlying mechanism is not fully known. The aims of this study were to measure levels of prothrombotic factors and examine their possible association with obesity and insulin resistance in obese children and adolescents. A total of 313 obese children and adolescents were recruited. In a cross-sectional design, we measured anthropometric parameters, plasminogen activator inhibitor-1-antigen (PAI-1-Ag), von Willebrand factor-antigen (vWF-Ag), fibrinogen (FB), lipids, fasting glucose, and insulin (FI) levels. Insulin resistance was estimated using the homeostasis model assessment for insulin resistance (HOMA-IR) index. Boys presented significantly higher PAI-1-Ag levels than girls (82.6 vs. 71.3 ng/ml, p = 0.01). Higher levels of PAI-1-Ag (96.8 vs. 69 ng/ml, p < 0.001), vWF-Ag (123.5 vs. 107.6%, p = 0.004) but not FB (353.1 vs. 337.6 mg/dl, p = 0.137) were found in insulin-resistant (IR) participants after adjusted for age, gender, and pubertal stage. IR patients were at 2.98 (CI: 1.084-8.193) and 4.86 (CI: 1.119-15.606) times greater risk for high PAI-1-Ag and vWF-Ag levels, respectively. All three prothrombotic factors were positively correlated with body mass index (BMI) and FI levels (p < 0.05), but only PAI-1-Ag and vWF-Ag were significantly correlated with HOMA-IR index (p ≤ 0.001). After adjustment for confounding factors, both BMI and HOMA-IR indices remained significantly associated with PAI-1-Ag (r2 = 0.225, p < 0.001) and vWF-Ag levels (r2 =0.077, p = 0.003). CONCLUSION: This study shows that obesity in youngsters, when accompanied with insulin resistance, is associated with at least threefold increased risk for elevated levels of prothrombotic factors, contributing to the early development of atherothrombosis. This impaired prothrombotic state may partially explain the increased risk for developing cardiovascular disease later in adulthood.

Nutrition transition and health status of Cretan women: evidence from two generations.

OBJECTIVE: Research has suggested the abandonment of the traditional Mediterranean diet (MD) by youngsters for a more Westernized regime. The present study aimed (i) to delineate differences in the consumption of traditional Cretan dishes and key nutrients in the MD between two contiguous generations of women and (ii) define how these dietary changes contribute to the morbidity of the younger generation. DESIGN: Cross-sectional. SETTING: The island of Crete, Greece. SUBJECTS: Eighty pairs of mothers and daughters, all living in their own households, were recruited from Crete. Information regarding the consumption of traditional Cretan dishes and two previous-day recalls were collected. The health status of the participants was also recorded. Simple correspondence analysis (SCA) was used to assess associations between differences in the food intake of daughters compared with their mothers and the prevalence of disease. RESULTS: The younger women showed increased intakes of rusks and meat dishes and decreased consumption of green vegetables, pulses and wine compared with their mothers. When nutrients were accounted for, only sugar consumption of the younger women surpassed the intake reported by their mothers. SCA associated hypercholesterolaemia in the daughters with increased cheese and meat intakes; allergies and respiratory problems were associated with lower cheese, fish and oil intakes; being overweight was associated with higher baked goods, oils, desserts, fish and alcohol intakes; and Fe-deficiency anaemia was associated with lower consumption of green vegetables. CONCLUSIONS: Results showed that, although a trend towards a more Westernized diet was noted in the younger women, the differences between the two generations appear to be small. Therefore, in Crete, the MD is still the primary diet regime.

Rhabdomyolysis, renal failure, pericardial effusion, and acquired von Willebrand disease resulting from hypothyroidism in a 10-year-old girl.

A 10-year-old girl manifested unexplained muscle aches and high creatine phosphokinase (CPK) concentrations attributed to rhabdomyolysis in association with severe hypothyroidism due to autoimmune thyroiditis. The response to therapy strongly suggested that hypothyroidism was the cause of rhabdomyolysis. Hypothyroidism is a rare cause of rhabdomyolysis. It should always be considered in a patient with muscular symptoms and elevated CPK concentrations. In addition, the patient developed other uncommon manifestations of hypothyroidism such as pericardial effusion, acute renal failure, and acquired von Willebrand disease. After thyroxine replacement, the symptoms and abnormal findings disappeared. The patient was also diagnosed as having celiac disease, which is often associated with autoimmune thyroiditis. Conditions accompanying autoimmune thyroid disease may result from altered thyroid function and from the presence of other autoimmune diseases. The butterfly-shaped thyroid gland has a tremendous impact on metabolism, which may be compared to a phenomenon termed the "Butterfly Effect".

Growth, the Mediterranean diet and the buying power of adolescents in Greece.

Background The aim of the present cross-sectional study was to evaluate associations between pocket money, Mediterranean diet (MD) adherence and growth among Greek adolescents. Methods A total of 319 (157 boys and 162 girls) Greek adolescents, aged 10-18 years participated in the study. Pocket money was recorded, MD adherence was assessed with the KIDMED score and growth was evaluated using the World Health Organization (WHO) growth charts. Results Participants receiving pocket money exceeding 6.0€ daily demonstrated increased fast-food consumption and breakfast skipping. Overall, a negative relationship was revealed between pocket money and obesity. However, lower allowance receivers were less likely to be obese, consume fruit per day and more likely to consume breakfast and sweets, compared to average pocket money receivers. Increased MD adherence was associated with a lower risk of overweight and as expected, unhealthy eating habits were observed among obese adolescents. Conclusions Interrelationships tend to exist between MD adherence, pocket money and growth among adolescents.

Ghrelin and leptin levels in young adults with cystic fibrosis: relationship with body fat.

BACKGROUND: Ghrelin and leptin are hormones implicated in energy balance coordination and body weight regulation. There are conflicting data regarding the levels and role of leptin while ghrelin has not been studied in CF. The aim of this study was to investigate fasting serum ghrelin and leptin levels in CF adolescents as compared to healthy controls and analyze their association with body fat. METHODS: Fourteen CF adolescents having pancreatic insufficiency and twenty healthy adolescents were enrolled in the study. Diabetic patients were excluded. In all participants' height, weight, body mass index (BMI) and body fat % (BF %) were estimated. Ghrelin and leptin levels were determined after an overnight fast. RESULTS: Weight, BMI and BF% were significantly lower in CF adolescents than those of controls. Fasting leptin levels in CF were significantly higher in CF patients (p=0.030), compared to controls and significantly lower in CF males as compared to CF females (p=0.01). Fasting ghrelin levels were significantly lower in CF males as compared to male controls (p<0.001) and comparable in females. CONCLUSIONS: As the overall clinical outcome of CF patients is related to the nutritional status and body weight, the role of ghrelin and leptin in these patients needs to be elucidated.

Ghrelin serum levels during oral glucose tolerance test in prepubertal obese children with insulin resistance.

BACKGROUND: An involvement of ghrelin in glucose metabolism has been suggested; nevertheless, the relationship between ghrelin and insulin resistance (IR) remains unclear. AIMS: 1. To investigate the effect of glucose loading on ghrelin in prepubertal obese children with IR. 2. To assess possible correlations between IR and changes in circulating ghrelin. PATIENTS AND METHODS: Twenty prepubertal obese, insulin-resistant and 18 age- and sex-matched lean children were studied. Fasting glucose, insulin and ghrelin levels were measured. In the obese group, measurements were repeated during an OGTT. RESULTS: Ghrelin levels were decreased at 60 min, but thereafter increased to baseline values. The fall in circulating ghrelin was negatively correlated with IR and the respective rise in insulin levels. CONCLUSIONS: In prepubertal, insulin-resistant obese children, ghrelin is significantly suppressed shortly after glucose intake. It is possible that the above effect is attenuated by IR and the resultant increase in insulin levels.

Ghrelin and leptin levels in obese adolescents. Relationship with body fat and insulin resistance.

OBJECTIVE: Ghrelin and leptin levels are influenced by body fat (BF%), pubertal stage and possibly insulin resistance (IR). The aim of our study was: 1) To compare fasting ghrelin and leptin levels between obese and non-obese, adolescents, 2) to investigate possible correlations of these hormones with BF %, as well as IR. DESIGN: Twenty obese insulin resistant (IR) adolescents, twenty obese non IR (NIR) and fifteen healthy non-obese, age-matched adolescents were studied. In all participants, height, weight, body mass index (BMI) and BF % were measured. Fasting glucose, insulin, ghrelin and leptin levels were determined. IR was assessed using HOMA-IR index. RESULTS: BMI, BF %, insulin and HOMA-IR values were positively correlated with leptin and negatively with ghrelin levels. A negative correlation between circulating leptin and ghrelin levels was found. A suggestive positive correlation between leptin levels and BF %, independent of BMI, was also observed (P=0.075). Ghrelin levels were significantly correlated with insulin levels and HOMA-IR, independent of BMI (P=0.077). CONCLUSIONS: Obesity and IR may play an important role in the release of ghrelin as well as in the negative correlation between ghrelin and leptin.

Genetic screening of non-classic CAH females with hyperandrogenemia identifies a novel CYP11B1 gene mutation.

OBJECTIVE: Congenital adrenal hyperplasia (CAH) is an endocrine autosomal recessive disorder with various symptoms of diverse severity. Mild hyperandrogenemia is the most commonclinical feature in non-classic CAH patients and 95% of the cases are identified by mutations in the CYP21A2 gene. In the present study, the second most common cause for non-classic CAH (NC-CAH), 11ß-hydroxylase deficiency due to mutations in the CYP11B1 gene, is investigated. DESIGN: Screening of the CYP21A2 and CYP11B1 genes by direct sequencing was carried out for the detection of possible genetic defects in patients with suspected CAH. RES ULTS: It wasobserved that CYP11B1 variants co-exist only in rare cases along with mutations in CYP21A2 in patients clinically diagnosed with CAH. A total of 23 NC-CAH female patients out of 75 were identified with only one mutation in the CYP21A2 gene. The novel CYP11B1 gene mutation, p.Val484Asp, was identified in a patient with CAH in the heterozygous state. The structural characterization of the novel p.Val484Asp was found to likely cause distortion of the surrounding beta sheet and indirect destabilization of the cavity that occurs on the opposite face of the structural elements, leading to partial impairment of the enzymatic activity. CONCLUSIONS: CYP21A2 gene mutations are the most frequent genetic defects in cases of NC-CAH even when these patients are in the heterozygous state. These mutations have a diverse phenotype giving rise to a variable extent of cortisol synthesis impairment; it is also clear that CYP11B1 mutants are a rare type of defects causing CAH.

Association between magnesium concentration and HbA1c in children and adolescents with type 1 diabetes mellitus.

BACKGROUND: Magnesium levels may be decreased in patients with type 1 diabetes mellitus (T1DM), influencing disease control. Relevant studies concern mainly adults and there are few data from the pediatric population. The aim of the present study was to evaluate magnesium levels and examine their possible association with glycemic control in youths with T1DM. METHODS: In all, 138 children and adolescents with T1DM aged between 1.9 and 20.3 years were recruited to the study. Using a cross-sectional design, we measured anthropometric parameters, HbA1c, serum magnesium, ionized and total calcium, phosphorus, potassium, sodium, and urinary albumin (UA). Estimated glomerular filtration rate (eGFR), based on serum creatinine concentrations, was also calculated. RESULTS: Lower levels of magnesium were found in subjects with poor versus good glycemic control (0.79 ± 0.09 vs 0.82 ± 0.09 mmol/L, respectively; P = 0.002). Serum magnesium levels were negatively correlated with HbA1c (P < 0.001) and positively correlated with UA, calcium, phosphorus, and potassium levels (P < 0.05). After adjustment for confounding factors, only magnesium levels remained significantly associated with HbA1c (adjusted r(2) = 0.172; P = 0.004). The odds ratio for poor glycemic control, indicated by HbA1c >7.5%, between the highest and lowest magnesium concentration quartiles was 0.190 and amounted to a decrease of 1.7% in the HbA1c level. CONCLUSIONS: The present study shows that low serum magnesium levels in children and adolescents with T1DM are associated with an increased risk of poor glycemic control, potentially contributing to the early development of cardiovascular complications.

A systematic review and meta-analysis of weight status among adolescents in Cyprus: scrutinizing the data for the years 2000-2010.

OBJECTIVE: The aim of the present study was to evaluate by pooled and sensitivity analyses all available data on adolescent overweight/obesity in Cyprus. DESIGN: A thorough literature search determined the studies to be examined using Cypriot adolescent samples aged 10-18 years old, with weight status in each sex classified according to the IOTF criteria, published between the years 2001-2011. Eight studies were retrieved, but three fulfilled the criteria for the sensitivity analyses. RESULTS: The pooled prevalence of obesity was 9.8% in boys (n = 6081). The pooled analysis classified 6.1% (n = 3886) of girls as obese, whereas a higher prevalence was observed by the sensitivity analysis 6.4% (n = 1956, p ≤ 0.001). The boys' prevalence of overweight was 19.3% and the girls' 17.1%. Between sexes, boys demonstrated a higher prevalence of obesity and overweight (p ≤ 0.001 for both). The cumulative analyses demonstrated an increase in the prevalence of overweight/obesity until the year 2005 and thereafter a plateauing in boys and a slight decrease in girls in a non-linear manner. CONCLUSION: Approximately 1/3 of adolescent boys and 1/4 of adolescent girls in Cyprus were overweight/obese during the previous decade.

Ghrelin-leptin network influences serum chitinase 3-like protein 1 (YKL-40) levels in obese prepubertal children.

OBJECTIVE: This study aimed to investigate any possible interactions between hormonal regulators of weight gain and markers of subclinical inflammation in childhood obesity. Forty-one obese prepubertal children and 41 age- and gender-matched lean controls were included. Children were classified as obese or non-obese according to international age- and gender-specific body mass index (BMI) cutoff points defined by the International Obesity Task Force to define childhood obesity. Anthropometric measurements, serum insulin, chitinase 3-like protein (YKL-40), ghrelin and leptin levels as well as plasma glucose in the fasting state were determined. RESULTS: Obese children as compared with controls had higher YKL-40 (50.7±15.2 vs 41.0±10.5 ng/ml, p=0.003), higher leptin (33.8±16.0 vs 9.7±7.5 ng/ml, p<0.001) and lower ghrelin serum levels (871.4±368.0 vs 1417.6±387.3 pg/ml, p<0.001). The obese children with ghrelin levels above median (43.8±10.2 ng/ml) as compared to those with ghrelin below median (57.2±16.6 ng/ml) presented lower serum YKL-40 levels (p=0.009), indicating more severe inflammation with lower levels of ghrelin. By contrast, although the obese children with leptin levels above median (49.7±16.3 ng/ml) presented lower serum YKL-40 levels as compared to those with leptin levels below median (51.6±14.6 ng/ml), this difference did not reach the level of statistical significance (p=0.726). Moreover, serum YKL-40 levels were significantly correlated with ghrelin (r=-0.359, p=0.014) but not with leptin levels (r=0.169, p=0.261). A significant negative correlation between ghrelin and leptin levels was also found (r=-0.276, p=0.041). These findings remained unchanged for obese, when analyses were done separately, whereas the significance of correlations was lost for non-obese subjects. CONCLUSIONS: Ghrelin-leptin network had an impact on serum YKL-40 levels in obese prepubertal children; upregulation of YKL-40 secretion seems to be a consequence of reduced ghrelin rather than elevated leptin concentrations.

Elevated circulating levels of the serum acute-phase protein YKL-40 (chitinase 3-like protein 1) are a marker of obesity and insulin resistance in prepubertal children.

YKL-40 (chitinase 3-like protein 1) is a newly recognized protein that is secreted by activated macrophages and neutrophils and expressed in a broad spectrum of inflammatory conditions and cancers. It has also been associated with endothelial dysfunction and diabetes in adults. Its role in childhood obesity has not been evaluated yet. Our aim was to evaluate the associations of serum YKL-40 levels with markers of obesity, inflammation, and insulin resistance in children. Forty-one obese prepubertal children and 41 age- and sex-matched lean controls were included, and serum YKL-40 levels were determined. Body mass index (BMI), blood pressure (BP), body fat percentage, fasting glucose, insulin, homeostasis model assessment for insulin resistance (HOMA-IR) index, whole-body insulin sensitivity index, lipids, white blood cell (WBC) count, C-reactive protein, and fibrinogen levels were also assessed. Obese children had higher YKL-40 levels compared with controls (P = .003). Insulin-resistant individuals showed higher YKL-40 compared with non-insulin-resistant individuals after adjusting for age and BMI (adjusted P = .039). Serum YKL-40 levels were positively correlated with age, BMI, body fat percentage, fasting glucose and insulin, HOMA-IR index, whole-body insulin sensitivity index, systolic BP, mean BP, and WBC count (P < .05). After adjustment for age, sex, BMI, WBC count, and systolic BP, HOMA-IR index remained significantly associated with YKL-40 levels (P < .001). The study suggests that YKL-40 levels are elevated in obese youth and represent a marker of insulin resistance even in childhood. Prospective studies are needed to determine whether children with elevated YKL-40 levels are at higher risk for future cardiovascular disease.

Ghrelin and growth hormone serum levels during the clonidine test in children with short stature and variable growth hormone status.

OBJECTIVE: The exact role of ghrelin in the control of growth hormone (GH) secretion has not been completely clarified as yet. The aim of the present study was 1) to investigate the effect of a substance promoting GH secretion (clonidine) on ghrelin levels in children with short stature with growth hormone deficiency (GHD) and normal growth hormone (NGH), and 2) to assess possible correlations between GH and ghrelin values during the clonidine test. DESIGN: Eighteen prepubertal children with short stature were included in the study. Using the results of two GH-provocative tests (glucagon and clonidine), the participants were divided into two groups: GHD and NGH. In both groups, ghrelin levels were determined during the clonidine stimulation test. RESULTS: Different responses regarding ghrelin levels during the clonidine stimulation test were observed in the two study groups (GHD and NGH). A decrease in ghrelin levels was observed in the NGH children accompanied by a rise in the circulating GH levels, whereas the GHD children demonstrated a rise in both ghrelin and GH levels. CONCLUSIONS: The data indicate an inverse relationship between circulating ghrelin and GH in NGH children, suggesting the presence of a negative feedback loop between ghrelin and GH. Analogous changes were not observed in GHD children.

Growth hormone deficiency: an unusual presentation of floating harbor syndrome.

Floating-Harbor Syndrome (FHS) is a very rare condition of unknown etiology characterized by short stature, delayed bone age, characteristic facial features, delayed language skills and usually normal motor development. This syndrome has only once been associated with growth hormone deficiency and precocious puberty in the same patient. We describe a 5 4/12 year-old girl with the typical features of FHS in whom growth hormone deficiency was diagnosed and two years later central precocious puberty was noted. The patient showed a good response to human recombinant growth hormone as well as gonadotropin releasing hormone analogue treatment.

A preliminary case-control study on nutritional status, body composition, and glycemic control of Greek children and adolescents with type 1 diabetes.

BACKGROUND: Because scientific data on the diet of diabetic Greek youngsters are scarce, diabetic experts use findings from international studies. However, because of diet variations between countries, this may result in problems in diabetes control. The aim of the present pilot study was to assess body composition, nutritional status, and diabetes control in Greek youngsters with type 1 diabetes mellitus (T1DM). METHODS: Twenty-four children and adolescents with diabetes, aged 4-16 years, and the same number of age- and sex-matched controls participated in the study. Anthropometry included stature, weight, and body fat determined by bioelectrical impedance analysis. Body mass index (BMI), fat mass index (FMI), fat-free mass index (FFMI), and z-scores were calculated. Diabetes control was evaluated through glycosylated hemoglobin (HbA1c) and dietary intake was recorded for 3 days. RESULTS: The FFMI, BMI z-score and weight-for-age z-score were lower in controls compared with diabetic youngsters (P ≤ 0.001, P ≤ 0.02, and P ≤ 0.01, respectively). Three diabetic participants were overweight (12.5%) and two controls were underweight (8.3%). The energy and nutrient intake was similar between the two groups, and all participants consumed a diet high in fats and proteins at the expense of carbohydrates. Dietary fat was highly correlated with BMI in both groups. The consumption of vitamin D was inadequate in the diabetic participants, but they had a higher intake of antioxidant vitamins, vitamin B(6) , and folate compared with the control group. CONCLUSIONS: In conclusion, youngsters with T1DM failed to adhere to the macronutrient recommendations for diabetes, but dietary patterns were similar in both the diabetic and control groups. The control of diabetes was not associated with any nutrient or anthropometric variable.