[Non-enzymatic glycation of proteins: from diabetes to cancer].

Incubation of proteins with glucose leads to their non-enzymatic glycation and formation of Amadori products known as an early glycation product. Oxidative cleavage of Amadori products is considered as a major route to advanced glycation endproducts (AGEs) formation in vivo. Nonenzymatic glycation of proteins or Maillard reaction is increased in diabetes mellitus due to hyperglycemia and leads to several complications such as blindness, heart disease, nerve damage and kidney failure. Accumulation of the early and advanced glycation products in plasma and tissues of diabetic patients and causes production of autoantibodies against corresponding products. The advanced glycation products are also associated with other diseases like cancer. This review summarizes current knowledge of these stage specific glycated products as common and early diagnostic biomarkers for the associated diseases and the complications with the aim of a novel therapeutic target for the diseases.

Evidence for involvement of TNFR1 and TIMPs in pathogenesis of post-kala-azar dermal leishmaniasis.

Semi-quantitative RT-PCR was exploited to analyse the intralesional cytokine gene expression in 14 post-kala-azar dermal leishmaniasis (PKDL) and 10 kala-azar (KA) patients. The data provided evidence for both inflammatory and non-inflammatory responses, as reflected by elevated tumour necrosis factor (TNF)-alpha and interleukin (IL)-10 in PKDL lesions compared with normal skin tissue (n = 6). The ratio of TNF-alpha : IL-10 message was 2.66 in PKDL cases, substantially higher than in KA (1.18). Investigation of TNF-alpha receptors (TNFR1 and TNFR2) revealed a significant down-regulation of TNFR1 transcript in both PKDL and KA compared with control. In the presence of elevated levels of TNF-alpha transcript, interference with type 1 effector activity in PKDL may be due to minimal expression of the TNFR1 gene. Investigation of matrix metalloproteinases, known to be induced by TNF-alpha, and the tissue inhibitors of matrix metalloproteinases (TIMPs), provided evidence for the roles of TIMP-1 and TIMP-3 in the pathogenesis of PKDL.

Elevated levels of IgG3 and IgG4 subclass in paediatric cases of kala azar.

Visceral leishmaniasis (VL) or Kala azar (KA) is a systemic disease caused by the parasites of the Leishmania donovani complex. Control measures rely on treatment with antileishmanial agents, however, fraught with problems such as toxicity or drug resistance. The incidence rate is on the rise for children, for reasons yet undefined. Previously we have shown significantly elevated level of IL-10 in children compared to adult KA cases. Here, antileishmanial antibody and C-reactive protein (CRP) levels were investigated in paediatrics and adult patients of KA and post-KA dermal leishmaniasis (PKDL). Level of IgG4 was significantly elevated in PKDL compared to KA, although total IgG and IgG1 were significantly lower. The antileishmania antibody levels of subclass IgG3 and IgG4 were found significantly elevated in paediatrics, however, levels of IgG, IgG1, IgG2 and CRP were comparable in paediatric and adult KA cases. In case of PKDL, levels of IgG and it subclass were similar in the two groups. No significant difference in antileishmanial antibody level was noticed between macular or polymorphic cases of PKDL. The differential antibody intensity in paediatric cases, together with significant level of circulating IL-10, could be considered as a marker of differential disease susceptibility.

Nested PCR assay for detection of Leishmania donovani in slit aspirates from post-kala-azar dermal Leishmaniasis Lesions.

A nested PCR assay to detect parasite DNA in slit aspirates from skin lesions of patients with post-kala-azar dermal lesihmaniasis (PKDL) is described. PCR results were positive in 27 of 29 (93%) samples by nested PCR assay, while only 20 of 29 (69%) were positive in a primary PCR assay. The nested PCR assay allowed reliable diagnosis of PKDL in a noninvasive manner.

Antigenaemia and antibody response to Leishmania donovani stage-specific antigens and rk39 antigen in human immunodeficiency virus-infected patients.

In order to define the possible markers for the early diagnosis of asymptomatic visceral leishmaniasis in human immunodeficiency virus (HIV)-infected individuals, the antigenaemia and antibody response to stage-specific Leishmania donovani and rk39 antigens is assessed by enzyme-linked immunosorbent assay (ELISA) and immunoreactivity to stage-specific antigens analysed by Western blot. Serum samples from two out of 100 HIV-infected individuals were found positive for antigenaemia, antibody response to stage-specific L. donovani antigens and rk39 antigen, and one sample was also positive for antigenaemia and antibody response to L. donovani antigens, while antibody detection to rk39 antigen was not carried on this sample. Additionally, one sample was found positive for amastigote antigenaemia and antibody response to amastigote antigen, while in this patient promastigote antigenaemia and antibody response to promastigote L. donovani and rk39 antigen could not be detected. One sample was found positive for antigenaemia, antibody response to amastigote antigen and negative for antibody response to promastigote antigen, while in this patient response to rk39 antigen was borderline. Although antibody response to rk39 antigen could be detected in 9/88 (10%) HIV-infected individuals, in six of these nine patients neither antigenaemia nor antibody response to stage-specific L. donovani antigens could be detected. All 10 confirmed visceral leishmaniasis and HIV-negative control patients had positive antigenaemia and antibody response to L. donovani amastigote and promastigote antigens, while all the normal healthy individuals were negative. The study indicated that detection of antibody response to rk39 antigen, amastigote antigenaemia and antibody response to amastigote antigen may prove to be better markers than detection of promastigote antigenaemia, antibody response to promastigote antigen and immunoblot reactivity.

Pathology and causes of death in a group of 128 predominantly HIV-positive patients in Botswana, 1997-1998.

BACKGROUND: Little is known about causes of death in countries of southern Africa seriously affected by the HIV/AIDS epidemic. METHODS: After obtaining informed consent, autopsies were performed on 128 mainly hospitalised adults in Francistown, Botswana, between July 1997 and June 1998. Criteria for case selection included those who died before a diagnosis could be established, those whose condition deteriorated unexpectedly during hospitalization, and those who had respiratory disease. This represented 14% of adult medical patients who died in hospital during the study period. RESULTS: Of the 128 patients, 104 (81%) were HIV-positive. Among HIV-positive patients, the most common pathologic findings were tuberculosis (TB) (40%), bacterial pneumonia (23%), Pneumocystis carinii pneumonia (11%), and Kaposi's sarcoma (11%); these conditions were the cause of death in 38%, 14%, 11%, and 6%, respectively. Of the 40 pulmonary TB cases, 90% also had disseminated extra-pulmonary TB. Chest radiology could not reliably distinguish the pathologies pre-mortem. CONCLUSIONS: TB was the leading cause of death in our series of HIV-positive adults in Botswana, selected towards those with chest disease; in most, it was widely disseminated. Bacterial pneumonia also played an important role in mortality. Pneumocystis carinii pneumonia was present, but relatively uncommon.

Diagnosis of visceral leishmaniasis: comparative potential of amastigote antigen, recombinant antigen and PCR.

Development of simple, economical and non-invasive tests for the early diagnosis of visceral leishmaniasis (VL) or kala-azar (KA) remains a challenge, and serological studies based on antigen prepared from the amastigote stage of Leishmania donovani, the stage that causes infection, are lacking. In the present study, circulating antibodies to total antigen isolated from the promastigote and amastigote stages of the parasite, as well as to recombinant K39 (rK39) antigen, are measured by enzyme-linked immunosorbent assay (ELISA) and the results compared with a polymerase chain reaction (PCR) test for KA diagnosis. In 116 samples of KA examined, the amastigote antigen gave significantly higher mean absorbance values in ELISA than did the promastigote antigen. The sensitivity for KA detection was significantly higher using the amastigote antigen (94%) than the promastigote antigen (90.5%). Analysis in 91 controls showed that specificity was higher with amastigote antigen (92.3%) than with promastigote antigen (86.8-89.0%). Reliability of ELISA diagnosis with amastigote antigen was only marginally lower than that with rK39 ELISA or with the PCR test. Easy availability and low cost of indigenous amastigote antigen, together with the simplicity of ELISA compared with PCR, make ELISA based on amastigote antigen a promising choice for the diagnosis of KA.

Diagnosis of visceral leishmaniasis: comparative potential of amastigote antigen, recombinant antigen and PCR.

Development of simple, economical and non-invasive tests for the early diagnosis of visceral leishmaniasis (VL) or kala-azar (KA) remains a challenge, and serological studies based on antigen prepared from the amastigote stage of Leishmania donovani, the stage that causes infection, are lacking. In the present study, circulating antibodies to total antigen isolated from the promastigote and amastigote stages of the parasite, as well as to recombinant K39 (rK39) antigen, are measured by enzyme-linked immunosorbent assay (ELISA) and the results compared with a polymerase chain reaction (PCR) test for KA diagnosis. In 116 samples of KA examined, the amastigote antigen gave significantly higher mean absorbance values in ELISA than did the promastigote antigen. The sensitivity for KA detection was significantly higher using the amastigote antigen (94%) than the promastigote antigen (90.5%). Analysis in 91 controls showed that specificity was higher with amastigote antigen (92.3%) than with promastigote antigen (86.8-89.0%). Reliability of ELISA diagnosis with amastigote antigen was only marginally lower than that with rK39 ELISA or with the PCR test. Easy availability and low cost of indigenous amastigote antigen, together with the simplicity of ELISA compared with PCR, make ELISA based on amastigote antigen a promising choice for the diagnosis of KA.

Effect of anxiolytics on blood sugar level in rabbits.

Anxiety disorders are more prevalent not only in normal individuals but also in diabetes mellitus. Diazepam, a benzodiazepine, and buspirone, an azaspirodecanedione, are the most often prescribed anxiolytics. Present study was aimed to investigate the effect of diazepam and buspirone on the blood sugar levels in rabbits. Buspirone (0.5 mg/kg/day p.o.) and diazepam (0.6 mg/kg/day p.o.) did not affect the glucose levels in rabbits even after one month of treatment. Present findings suggest that these two anxiolytics have minimal effect on blood sugar control.

Diagnosis of malignant mesothelioma by fine needle aspiration of a cervical lymph node. A case report.

BACKGROUND: Clinically documented distant metastases are rare in mesothelioma and tend to occur late in the course of the disease, well after the diagnosis has been made. In this instance, diagnosis was not made until a metastatic deposit was identified microscopically in the enlarged lymph node. CASE: A 65-year-old male with no definite history of occupational asbestos exposure presented with chest pain, pleural effusion and supraclavicular lymphadenopathy. Cytologic examination of material obtained by fine needle aspiration from his cervical lymph node revealed malignant mesothelioma. This was confirmed on histology. CONCLUSION: This was a particularly rare presentation and, as far as we are aware, was the first case in which mesothelioma was diagnosed by fine needle aspiration of a cervical lymph node. It serves to remind the pathologist that when confronted with a lymph node involved by tumor, the possibility of mesothelioma should be included in the differential diagnosis. The case also demonstrates the usefulness of fine needle aspiration in the diagnosis of metastatic tumor.

Investigation of an outbreak of gastroenteritis caused by Norwalk-like virus, using solid phase immune electron microscopy.

In February 1993, 95 persons (47 patients and 48 staff members) were affected by an hospital outbreak of viral gastroenteritis. Using direct electron microscopy (EM) the causative agent was identified as a small round structured virus. This was confirmed as a Norwalk-like virus using solid phase immune electron microscopy (SPIEM). Of 94 stool samples examined, 12 (13%) samples containing small round structured viruses (SRSV) were SPIEM positive for Norwalk-like virus. A further 25 (27%) samples contained small round featureless virus (SRFV) identified by direct EM and were negative on SPIEM. The illness was characterized by preceding influenza-like symptoms in 76% of cases followed by vomiting (76%), diarrhoea (79%) and abdominal pain (79%). One fatality was recorded. The outbreak lasted for 15 days, with a peak incidence of new cases amongst patients and staff occurring on day 5. It was controlled through a combination of ward closures, patient cohorting, suspension of duties for affected staff and disinfection procedures. Difficulties were encountered in the education of staff and in the implementation of environmental control measures. Screening of hospital catering services and a case control study, carried out among affected staff members, failed to identify a foodborne source. Consumption of tap water in the hospital was commoner among affected staff members than among controls, but this did not reach significance (P = 0.1).

Metastatic transitional cell carcinoma presenting as diffuse liver disease.

Three cases of transitional cell carcinoma (TCC) metastatic to the liver are reported. Each case presented as diffuse liver disease. This is a rare presentation, most liver metastases occurring as an isolated mass or multiple discrete masses.

Primary Hodgkin's disease of the lung.

Three cases of Hodgkin's disease presenting primarily in the lung are described. All 3 patients presented with respiratory symptoms and chest X-rays revealed discrete masses within the lung parenchyma. CT examination of the mediastinum did not reveal regional or generalized lymphadenopathy at the time of diagnosis in 2 of the cases. A diagnosis of Hodgkin's disease was made on open lung biopsies and despite aggressive chemotherapy, 2 patients died within 2 yrs. The histological and immunocytochemical features of these cases were typical of that expected in Hodgkin's disease. However, the stated indolent course of primary pulmonary Hodgkin's disease is not invariable. Those patients with bilateral interstitial disease and systemic symptoms have a poor prognosis. Furthermore, it is difficult to exclude definitely lymph node involvement (although not enlarged) at the time of diagnosis. Since several cases described in the literature have concurrent and/or subsequent nodal involvement, the entity of primary pulmonary Hodgkin's disease without lymph node involvement is exceedingly rare.

Solid-phase extraction and GC/MS quantitation of cocaine, ecgonine methyl ester, benzoylecgonine, and cocaethylene from meconium, whole blood, and plasma.

A selective solid-phase extraction technique has been applied to the analysis of cocaine and selected cocaine metabolites in meconium, whole blood, and plasma. This technique uses a mixed-mode Bond Elut Certify column that utilizes the characteristics of hydrophobic and polar interactions and ion exchange chromatography. Following extraction, cocaine, ecgonine methyl ester, benzoylecgonine, and cocaethylene were identified and quantitated using GC/MS. Linear quantitative response curves have been generated for the metabolites over a concentration range of 0-1000 ng/g for meconium and 0-1000 ng/mL for whole blood and plasma. The overall extraction efficiencies, depending on the metabolite, were between 58.1 and 99.7% for meconium, 95.6 and 124.0% for blood, and 86.9 and 128.9% for plasma. Linear regression analyses of the standard curve for the four analytes exhibited correlation coefficients ranging from 0.850 to 0.946 for meconium, 0.939 to 0.993 for whole blood, and 0.981 to 0.996 for plasma. Because of its capability to detect cocaethylene in meconium, blood, and plasma, the procedure can be used to determine if drug exposure occurred during the latter stages of gestation and if it involved only cocaine or a combination of cocaine and ethanol.