Small marker chromosome identification in metaphase and interphase using centromeric multiplex fish (CM-FISH)

Multicolor karyotyping procedures, such as multiplex fluorescence in situ hybridization (M-FISH), spectral karyotyping, or color-changing karyotyping, can be used to detect chromosomal rearrangements and marker chromosomes in prenatal diagnosis, peripheral blood cultures, leukemia, and solid tumors, especially in cases where G-banding is not sufficient. A regular M-FISH analysis requires relatively large amounts of labeled DNA (microgram quantities), is not informative in interphase nuclei, hybridization can take up to 2 to 3 days, and unlabeled human chromosome-painting probes are not available commercially. Unique probes (plasmids, PAC), specific for centromeric or subtelomeric chromosomal regions, can replace the painting probes in M-FISH to address specific issues, such as the identification of marker chromosomes and aneuploidies. A set of plasmid probes carrying repetitive sequences specific for the alpha-satellite region of all human chromosomes were combined in a metaphase assay and an interphase assay, allowing identification of aneuploidies in one hybridization step, on a single cytogenetic slide. The fluorophore-dUTP and the labeled antibodies required to label and detect the DNA probes can be prepared in any laboratory. All DNA probes can be easily isolated and labeled using common molecular cytogenetic procedures. Because of the repetitive nature of the probes, hybridization time is short, usually less than 1 hour, and the analysis can be performed with nonspecialized image-processing software.     

Conventional risk factors and acute coronary syndrome during a period of socioeconomic transition: population-based case-control study in Tirana, Albania

Aim

To assess the association between conventional risk factors and acute coronary syndrome in Albania, a transitional country in Southeast Europe.

Methods

A population-based case-control study was conducted in Tirana in 2003-2006. A total of 467 consecutive patients with nonfatal acute coronary syndrome were recruited. There were 370 men with mean ± standard deviation (SD) age of 59.1 ± 8.7 years and 97 women with mean±SD age of 63.3 ± 7.1 years. The control group comprised a population-representative sample of Tirana residents. In the control group, there were 469 men with mean±SD age of 53.1 ± 10.4 years and 268 women aged 54.0 ± 10.9 years. A structured questionnaire on demographic, socioeconomic, psychosocial factors, and health behaviors was administered. Physical measurements included anthropometrics and blood pressure. Venous blood and adipose tissue aspirations from the gluteal region were frozen-stored for future analysis. Multivariable-adjusted logistic regression was used to assess the independent associations of conventional risk factors with acute coronary syndrome.

Results

Upon adjustment for covariates, family history of coronary heart disease was found to be a strong predictor of acute coronary syndrome in both men (odds ratio [OR], 3.70; 95% confidence interval [CI], 2.58-5.30) and women (OR, 4.53; 2.40-8.57). Waist-to-hip ratio in men (OR, 4.03; 2.83-5.73) and obesity in women (OR, 3.31; 1.54-7.14) were strongly associated with acute coronary syndrome. In men, but not in women, there was a significant association with hypertension and current smoking (P = 0.011 and P<0.001, respectively). Diabetes was not significantly independently associated in either sex.

Conclusion

Classical risk factors predicted coronary heart disease in Albania, similarly as in the rest of the world, although associations with family history and anthropometric indices were stronger. These findings are resulting largely from the heterogeneous adoption of lifestyles conducive to increased coronary risk in transitional countries, and they point to the urgent need for targeted public health interventions.The relationship between coronary heart disease and risk factors such as smoking, high blood pressure, diabetes, obesity, and distribution of body fat is well established in developed countries (1-4). However, data on conventional risk factors, their patterns, determinants, and associations with coronary heart disease are scarce for the transitional countries of South East Europe. Unique among former communist countries in Southeast Europe, Albania was under Stalinist communist regime, and following the breakdown of this rigid government in 1990, it experienced a major social, cultural, and economic upheaval (5). Over the past 15 years changes in life-style (diet, tobacco, alcohol consumption, and physical exercise) have taken place, particularly in urban settings (6-8). The available evidence suggests an increase in alcohol intake, particularly among men (6). Smoking appears to have increased too, although smuggling makes it very difficult to validly estimate cigarette consumption in Albania (7). In 2001, the lifetime prevalence of smoking in Tirana was 61% in men and 24% in women (8). Furthermore, in 2001, 49% of men and 58% of women in Tirana reported largely sedentary leisure time activities such as reading and watching television (9). Although there is little information about cardiovascular disease occurrence in Albania, available data suggest that cardiovascular disease mortality may have increased over the past few years (10,11), and that cardiovascular disease morbidity (based on hospital admission counts) in both Tirana and the whole country has been increasing continually in the last decade (10).In this context, we undertook a population-based case-control study of coronary heart disease in Tirana between June 2003 and January 2006. Here, we report on the association of selected conventional risk factors such as smoking, anthropometric indices, hypertension, diabetes, and family history of coronary heart disease with acute coronary syndrome.     

The effects of one- and two-stage orchiopexy on postoperative serum testosterone levels and testicular volume in adult patients with bilateral nonpalpable testes

In this study, we investigated the effects of one- and two-staged orchiopexy on testicular volume and serum testosterone levels in cases with high, abdominally localized, bilateral nonpalpable testes. Between March 1996 and April 2001, orchiopexy was performed on 46 testes in 23 patients with bilateral nonpalpable testes. In 15 of the 23 patients, a two-stage Fowler-Stephens orchiopexy was performed, and in 8 of the 23 patients, a one-stage laparoscopic orchiopexy was performed. For one patient who lacked both testes, bilateral prosthetic testes were inserted. The patients' ages were between 20 and 23 years (average, 21 years). In the first stage of the two-stage orchiopexy, the spermatic artery was ligated laparoscopically. Six months later, open surgery orchiopexy was performed. Patients were followed for 2 to 16 months (average, 10 months) and were evaluated for testicular atrophy, serum testosterone levels, and complications. In all the patients, the preoperative secondary sexual characteristics and serum testosterone levels were normal. Among the postoperative controls, evaluated at months 3 and 6, one patient's testes were found at the pubic area. Testicular atrophy was not observed in any of the patients, and the serum testosterone levels were in the normal range. In the two-stage orchiopexy group, 5 (33%) of the 15 patients had an average 30% volume decrease, and in the one-stage group, 2 (25%) of the 8 patients had an average 40% volume decrease. In this study, it was shown that either two-stage Fowler-Stephens orchiopexy or laparoscopic orchiopexy can be successfully performed in patients with bilateral abdominal testes, and that the testes can be fixed to their palpable original positions without interfering with the secretion of testosterone. Although the results are similar, our experience suggests that single-stage laparoscopic orchiopexy is the preferable method.     

Lamivudine monotherapy in children with immune-tolerant chronic hepatitis B virus

We investigated the efficacy of lamivudine treatment in inducing a primary and sustained response in naive pediatric patients with immune-tolerant chronic hepatitis B viral (HBV) infection. Lamivudine was used (maximum 100 mg/day) for an average of 35 months (16-60 months) for 17 patients, while 6 patients were untreated. All the treated patients became HBV-DNA negative in the twelfth month of therapy with an average of 7 months, however none of them had a sustained virological response. They all became HBV-DNA positive by the twentieth month on average. One patient had significant improvement in liver histology. No side effects were observed. Spontaneous HBV-DNA clearance or HBe seroconversion was obtained in none of the untreated cases. This study suggests that lamivudine cannot induce complete viral control in HBV pediatric immune-tolerant patients.     

Abdominal tuberculosis in adolescents. Difficulties in diagnosis

Since the nature of abdominal tuberculosis is mimicking a number of diseases, this may cause delayed diagnosis resulting in evident increased morbidity and mortality. Most of the time, serologic and bacteriologic tools are not enough. We report 3 adolescents with distinct presentations, one mimicking Crohn's disease, one with hepatitis, and the last one with ascites. Terminal ileitis and mesenteric lymphadenitis were found in laparotomy of the first case mimicking Crohn disease. Granulomatous hepatitis was found in the liver biopsy of the second patient, and peritonitis was found by laparoscopy of the third patient. Tuberculosis could be diagnosed merely by histopathologic investigation. All were treated successfully without complication.     

Simultaneous bilateral microvascular tissue transfer for soft-tissue defects of the lower extremities in pyoderma gangrenosum

Pyoderma gangrenosum is a necrotizing skin disease of unknown etiology. The lesions are painful and occur mostly on the lower extremities, either single or multiple, but can appear in any region of the body. The authors report a case of pyoderma gangrenosum involving bilateral skin lesions over the anterior surface of the legs in a 13-year-old male patient. The lesions presented with ulcerative colitis, which is one of the most commonly associated diseases of pyoderma gangrenosum. After examination, systemic immunosuppressive treatment was administered and the progression of the disease was controlled. The lesions were treated with the simultaneous application of two free anterolateral thigh fasciocutaneous flaps after radical debridement. The procedure was successfully performed and no recurrence was observed. Although microvascular free tissue transfer cannot be performed in a standard fashion in pyoderma gangrenosum cases because of the risk of pathergy response, it should be considered as a surgical option in selected cases, otherwise difficult to manage, with the simultaneous application of appropriate systemic medical treatment.     

The importance of genomic copy number changes in the prognosis of glioblastoma multiforme

Glial tumors are the most common tumors of the nervous system, affecting individuals at any age. Since understanding of the molecular pathologies underlying human gliomas is still very poor, the treatment and therefore prognosis of this malignancy could not yet be improved. In order to determine whether different glioblastoma-associated genomic aberrations may serve as prognostic markers in combination with histopathological findings, 20 primary glioblastoma multiforme tumors were screened by comparative genomic hybridization, and the results were compared with histopathological and clinical features. All tumors showed genomic copy aberrations detected by comparative genomic hybridization. Regional and numerical increases in chromosome 7 copy number were the most frequently seen abnormality (10/20 tumors), followed by loss of chromosome 10 (8/20). Both of these aberrations were associated with shorter surveillance time. Chromosome 12q amplification was detected in seven tumors. Loss of 17p, 1p, and 19q in combination was seen in three cases. One of them was a giant cell GBM, whereas the remaining two cases were still alive. Combination of chromosome 1p and 19q deletions was also seen in a case with long surveillance. According to the preliminary findings of this study, in addition to the EGFR gene, amplification of other genes on chromosome 7 and the deletion of PTEN gene and other cancer-related genes on chromosome 10 appeared important to the development of glioblastoma multiforme and were associated with poor prognosis, whereas the combination of chromosome 1p and 19q deletions seems to be an informative molecular marker for better prognosis. The clinical features and genetic alterations of primary and secondary glioblastoma multiforme should be compared in large series to clarify the effective prognostic markers; and further molecular analyses focused on chromosomes 7 and 10 will be very helpful for understanding the molecular mechanisms underlying the progression of glioblastoma.     

Late diagnosis of severe colchicine intoxication

A 4-year-old Turkish girl was referred to our hospital with the findings of encephalopathy and pancytopenia. She had a history of severe abdominal cramps and gastrointestinal bleeding. A confused state, muscle pain and weakness, erythema-bullous and erythema-nodosum-like skin lesions, and alopecia were observed at her hospitalization. All of these symptoms resolved on follow-up. On laboratory investigation severe thrombocytopenia and leukopenia, mild anemia, a moderate increase in aspartate aminotransferase and alanine aminotransferase levels were detected. After reevaluating her medical history, it was learned that she had accidentally taken 1.3 to 1.5 mg/kg of colchicine 3 to 4 days before her first hospitalization. The possibility of misdiagnosis of colchicine intoxication should be borne in mind, and pediatricians must be aware of its toxic effects, especially in areas where patients with familial Mediterranean fever are present.