Relationship between mean blood glucose and glycated haemoglobin in Type 2 diabetic patients.

AIMS: To correlate the values of MBG to HbA(1c) in Greek patients with Type 2 diabetes and/or metabolic syndrome. METHODS: We followed up 140 Greek adult patients: 92 patients with Type 2 diabetes treated with insulin or oral glucose-lowering medication, and 48 patients with newly diagnosed Type 2 diabetes or metabolic syndrome not receiving any treatment. MBG was calculated for each patient from self-measurements of blood glucose using a portable glucometer, made six times a day (before eating and 2 h after a meal), three times a week for 1 month. HbA(1c) was determined by HPLC at 0 and 12 weeks. RESULTS: HbA(1c) at 0 (x) and 12 weeks (y) correlated strongly (y = 0.790x + 1.115, r = 0.92), confirming that the patient's glycaemic status remained stable during the whole period of follow-up. Linear regression was performed on MBG values; HbA(1c) at 12 weeks, sex, age, body mass index (BMI) and patient status (Type 2 diabetes treated or not) were used as independent variables. None of the independent variables reached statistical significance in the model, with the exception of HbA(1c) at 12 weeks. The final model was: MBG (mg/dl) = (34.74 x HbA(1c)) - 79.21, r = 0.93; or MBG (mmol/l) = 1.91 x HbA(1c) - 4.36, r = 0.93. CONCLUSIONS: Our results establish for the first time a strong correlation between MBG and HbA(1c) in Type 2 diabetic patients and support the idea of expressing HbA(1c) results as MBG. This will help patients to gain a clearer interpretation of the result, with less confusion. This simplification will allow every person with diabetes using home glucose-monitoring to understand his or her own target level.

Pretreatment with aspirin and ticlopidine confers lower thrombogenic potential of radiofrequency catheter ablation.

In the present study we pretreated 31 patients undergoing radiofrequency catheter ablation (RFA) with combined aspirin and ticlopidine for 3 days before the procedure, whereas 37 patients did not receive pretreatment. D-dimer levels reflecting the thrombogenic potential of RFA were significantly lower in the pretreated group at each stage before, during, and after the procedure, whereas there were no significant differences between the 2 groups in the number of RFA lesions or duration of the procedure.

Detection of hepatitis-B virus-DNA at high-frequency in liver neoplasias using a PCR technique.

Hepatocellular carcinoma is the most frequent liver cancer and hepatitis B virus is included among the risk factors for the development of this type of neoplasia. Direct detection of this virus is difficult due to the lack of a simple tissue culture system for growing the virus. Amplification of HBV nucleic acid sequences with the Polymerase Chain Reaction technique leads to the direct detection of the virus, but involves several critical steps and it is prone to false positive results due to inter sample contaminations. We overcame these shortcomings by using a simple boiling method for extracting DNA from histological slides of tissues coupled with double ('nested') PCR amplification. In this study we evaluated the frequency of presence of HBV nucleic acid sequences in samples of neoplastic liver tissues from patients in Greece. We studied 20 DNA samples from hepatocellular carcinomas and we found 11 positive for HBV DNA and 8 DNA samples from hepatoblastomas and we found 3 positive for the viral DNA.

H-ras oncogene mutations in the urine of patients with bladder-tumors - description of a noninvasive method for the detection of neoplasia.

Bladder cancers are usually curable, by surgical or transurethral excision, if diagnosed at an early stage. Tumor derived mutations in oncogenes potentially provide specific markers for the detection of surgically resectable tumors. The detection of point mutations of H-ras oncogene correlated with this disease. DNA sequences produced by the Polymerase Chain Reaction (PCR) can be considered for this application, because theoretically bladder tumors should shed cells containing this mutation into the urine. We examined urine from 21 individuals with bladder cancer before any treatment, as well as tissue specimens from the excised tumor and we found 10 mutations of the H-ras gene at codon 12 in the urine (47.61%) and 14 mutations in the tumor specimens (66.66%). We were able to detect nearly 50% of the patients with bladder tumors using this method. We also studied two relapses; in one case (which presented the mutation in the original tumor and the urine) the relapse grade had progressed from II to III. In the other case the relapse grade stayed at III but it presented for the first time the studied mutation in the urine. These results provide the theoretical and technical basis for the detection of bladder tumors by a non-invasive method and possibly for the evaluation of the invasiveness of the disease.

K-ras oncogene mutations in neoplasias of kidney transplanted patients: preliminary results with a new technique.

K-ras oncogene activations by point mutations are frequent in many forms of human cancers but there is a special category of cancers occurring in immunosuppressed patients after kidney transplantation in which the frequency of K-ras oncogene activation has not been fully studied. We used a new sensitive and easy method for the detection of this mutation, and in 8 DNA samples studied from various neoplasias of 8 patients after kidney transplantation, we found 4 mutations. Our preliminary results indicate that the activation of K-ras oncogene at codon 12, is a common event among the kidney transplanted patients who present a neoplasia, even in the least aggressive forms of the disease, contrary to the sporadic cases.

Detection of the cytomegalovirus by the polymerase chain reaction DNA amplification in a kidney transplanted patient.

Cytomegalovirus injection is common in kidney transplanted patients. Viremia is the only marker of active CMV infection, but the use of cell culture for the direct detection of the virus is time-consuming and not very sensitive, while the detection of CMV by measuring the titre of antibodies is difficult due to the immunosuppression these patients undergo. Thus the ability to amplify CMV DNA by the polymerase chain reaction from blood or urine samples of the patients becomes a valuable diagnostic tool for the detection of CMV in the early stages of the infection. Using a set of primers specific for the amplification of a 435 bp region of the IE-1 gene, we detected CMV DNA in blood leucocytes of a kidney transplanted patient who received the transplant from a CMV-seropositive donor, 45 days after the operation, while the antibody titre showed no evidence of active CMV infection.

Detection of minority point mutations by modified PCR technique: a new approach for a sensitive diagnosis of tumor-progression markers.

The detection of point mutations correlated with diseases, in enzymatically amplified DNA sequences (Polymerase Chain Reaction), is currently performed by digestion of PCR products when an existing restriction site disappears at least in one allele of the amplified mutated sequence or by allele specific radiolabeled probes in all other cases. These methods are the most sensitive but they cannot detect a mutation if it is present in less than 5% of the studied cells. We describe here a method based on the introduction of an artificial restriction site, using a modified primer during the PCR, which creates a RFLP indicative of the studied mutation. This RFLP is detected by a radiolabeled oligonucleotide probe which is not related to the mutation. Our approach multiplies the sensitivity by a factor of 1000 and it is practical for use in screening purposes and the detection, after treatment, of the residual disease in human malignancies. Using this method we detected 20% more mutations at codon 12 in the Ki ras oncogene in DNA from colorectal cancers that were undetectable with all the previous methods.

Frequency of the major CF mutation in French CF patients.

We have studied 124 patients of French origin, whose CF status had already been clearly established. These children belong to families previously tested with restriction fragment length polymorphism (RFLP) markers in our laboratory for genetic counselling. The most common mutation (delta F508) accounts for 67% in this population sample.

Usefulness of linkage disequilibrium of KM-19 and XV-2c DNA probes for genetic counselling in a high-risk CF family.

A French couple with an individual risk of carrying the cystic fibrosis (CF) mutation of 1/2 sought genetic counselling. From the DNA haplotypes generated by XV-2c and KM-19 RFLPs, it could be deduced that only one subject was a carrier, lowering the risk of having a CF baby from 1/16 to 1/200. The strong linkage disequilibrium between these RFLPs and the CF allele observed in France reduced the risk to 1/1600.

C-Reactive protein concentrations and angiographic characteristics of coronary lesions.

BACKGROUND: C-Reactive protein (CRP) is a strong predictor of clinical outcome in coronary artery disease (CAD), and inflammation has been implicated in the process. We aimed to evaluate whether CRP concentrations measured with a new, automated particle-enhanced immunoturbidimetric method for high-sensitivity CRP may be related to specific high-risk angiographic features of coronary lesions. METHODS: In a cross-sectional study, we examined 103 consecutive patients undergoing cardiac catheterization for suspected CAD. We assessed the association of preprocedural CRP concentrations with clinical presentation (unstable angina) and angiographic features of coronary lesions. RESULTS: Twenty patients had unstable angina. Independent predictors of unstable angina included increased CRP [odds ratio (OR), 2.93 per 10-fold increase in CRP; 95% confidence interval (CI), 1.28-6.69; P = 0.01] and the presence of macroscopic thrombus (OR, 7.08; 95% CI, 1.33-37.8; P = 0.02). Thirty-two culprit lesions had macroscopic thrombus or eccentric/irregular discrete morphology without total occlusion. Increased CRP was the strongest predictor of such features (OR, 2.04 per 10-fold increase in CRP; 95% CI, 1.03-4.04; P = 0.04), and the effect was independent of the presence of unstable angina. CONCLUSIONS: Among patients with suspected CAD undergoing coronary angiography, increased CRP is strongly associated with unstable angina and with specific high-risk features of the culprit coronary lesions.

Ras activation in myelodysplastic syndromes: clinical and molecular study of the chronic phase of the disease.

We studied N-ras and Ki-ras point mutations respectively at codons 12-13 and 12 in 15 patients with myelodysplastic syndromes (MDS) using the polymerase chain reaction (PCR) method for DNA amplification, and slot blot hybridization to allele specific oligonucleotide (ASO) probes. We analysed peripheral blood and bone marrow samples collected at diagnosis and repeatedly during the chronic phase of the disease to define when the activation occurred and in which haemopoietic cell populations, in order to establish possible relationships between clinical and molecular features. In three cases the N-ras oncogene was mutated at codon 12 in every cell population, both at diagnosis and throughout the chronic phase. Point mutations were not seen at the 12 codon of the Ki-ras oncogene. In patients lacking activated ras oncogene at diagnosis, mutations were not discovered during the entire period of observation. Therefore in our cases disease progression and leukaemic transformation did not correlate with the presence of the activated N-ras. Our data suggest that ras activation occurs early in the pathogenesis of MDS and involves a haemopoietic progenitor with multiple differentiative capacity, without however conferring an apparent proliferative advantage on its progeny.