Beneficial effect of shallot (Allium ascalonicum L.) extract on cyclosporine nephrotoxicity in rats.

The clinical use of an immunosuppressive cyclosporine A (CsA) is limited by its serious nephrotoxic effect. Evidences have suggested the role of oxidative stress in its pathogenesis. Shallot (Allium ascalonicum L.) has recently been shown to possess antioxidative and free radical scavenging abilities. The present study was undertaken to investigate the possible beneficial effect of shallot extract on renal injury caused by CsA. Male Wistar rats were treated orally with vehicle, CsA (25 mg/kg), shallot extract (1 g/kg), and CsA plus shallot extract for 21 days. Renal function, histopathology, tissue malondialdehyde (MDA) and glutathione (GSH) levels were evaluated 24 h after the last treatment. CsA-induced nephrotoxicity was evidenced by increased blood urea nitrogen and serum creatinine, but decreased urea and creatinine clearance. The kidney of CsA treated rats exhibited severe vacuolations and tubular necrosis. CsA also induced oxidative stress, as indicated by increased renal MDA and reduced GSH concentrations. Administration of shallot extract along with CsA counteracted the deleterious effects of CsA on renal dysfunction, oxidative stress markers, and morphological changes. These data indicate the protective potential of shallot extract against CsA nephrotoxicity and suggest a significant contribution of its antioxidant property to this beneficial effect.

Use of PCR-PIRA for screening of a point mutation at codon 12 in K-ras oncogene obtained from paraffin embedded tissue sections.

Among various methods which have been developed for facilitating the screening of point mutations in human genomic DNA, PCR-Primer Introduced Restriction Analysis (PCR-PIRA) is of particular interest due to its practicality and short procedure allowing detection of point mutations by simple restriction enzyme digestion directly after PCR amplification. However, one limitation of PCR-PIRA method is the absence of restriction sites in the region of detection, thus creation of the recognition site in primers has been introduced. Detection of a point mutation at codon 12 in K-ras oncogene by BstNI requires one base change in the primer sequence so that only the normal but not mutant PCR product will be digested by the enzyme. However, false positive results generated from undigested normal DNA sequence are always obtained. This effect is compounded when it is used to analyse mixed cell populations in paraffin embedded section of cancer cells. Assay of a mutant band generated from normal DNA by densitometric quantitation enabled the determination of background values and thereby eliminated false positive results. Samples with higher ratios between mutant and normal bands than the background one after the first PCR-PIRA would be subjected to the second PCR-PIRA in order to confirm the results. Screening of such mutations in cervical carcinomas from paraffin embedded sections using the above criteria should reduce misinterpretation of PCR-PIRA results.

Salmonella aortitis at Ramathibodi Hospital, Bangkok, Thailand.

Two cases of Salmonella aortitis were diagnosed by positive culture from the wall of aortic aneurysm. Common features in both cases included fever, abdominal/back pain and pulsatile abdominal mass with underlying severe atherosclerosis. The fatal outcome of both cases despite surgical and medical treatment was discussed and guideline of management was proposed.

Histopathologic spectrum of AIDS-associated lesions in Maharaj Nakorn Chiang Mai Hospital.

The histopathological alterations in various organs and the presence of AIDS-associated lesions were studied in 86 biopsy and 29 necropsy specimens of AIDS patients. The most common cancer seen in this study were malignant lymphomas (4% of cases) with development of extensive extranodal lymphomatous involvement from the outset. Although a preponderance of high grade B-cell pathologic subtypes is found in AIDS-associated lymphoma, we also report the first case of T-lymphoblastic lymphoma with a picture of acute lymphoblastic leukemia (T-ALL). Tuberculosis (34% of cases) was the most common opportunistic infection presented in tissue sections, and the majority of tissue biopsies revealed poorly organized granulomas and extensive necrosis with numerous bacilli. Penicilliosis (20% of cases) appeared to be the most common cutaneous lesion with multiple organ involvement. The involved organs showed a partially anergic tissue reaction characterized by poorly formed granulomas with diffuse infiltrate of fungi-laden macrophages and lymphoid cell depletion. This organism has to be distinguished from Histoplasma capsulatum and other yeast-form fungi. Co-existing cytomegalovirus and P. carinii infections were the predominant findings in lung necropsy specimens from pediatric patients who died from AIDS. A major pathologic feature in this group was diffuse alveolar damage stage II to III with heavy loads of organism and extensive lymphoplasmacytic infiltration.

Detection of various types of human papillomaviruses in premalignant and malignant cervical lesions using DNA-DNA in situ hybridization.

Detection of human papillomavirus (HPV) in formalin-fixed paraffin embedded tissue sections by in situ hybridization technique employing biotinylated (HPV) DNA probe 6/11, 16/18 and 31/33/35 was done retrospectively in 25 cases of cervical dysplasia, 32 cases of cervical squamous cell carcinoma in situ, 52 cases of invasive cervical squamous cell carcinoma and 7 cases of adenocarcinoma. HPV could be demonstrated in 10 cases (40.00%) of cervical dysplasia, 8 cases (25.00%) of cervical squamous cell carcinoma in situ and 31 cases (59.61%) of invasive cervical squamous cell carcinoma and 4 cases (57.14%) of adenocarcinoma. Among the dysplastic cases, 4 cases showed HPV 31/33/35, 4 cases showed HPV 16/18 together with HPV 31/33/35, 1 case showed mixed typing of HPV 6/11 and 31/33/35, and 1 case showed mixed typing of HPV 6/11, 16/18 and 31/33/35. In 32 cases of squamous cell carcinoma in situ, 1 case of HPV 6/11; 3 cases of HPV 16/18, 3 cases of HPV 31/33/35 and 1 case of mixed typing of HPV 16/18 and 31/33/35 were present. A case of HPV 6/11, 10 cases of HPV 16/18 and 9 cases of HPV 31/33/35 could be detected among the cases of invasive squamous cell carcinoma. Mixed typing of HPV 6/11 and 16/18; HPV 6/11 and 31/33/35; HPV 16/18 and 31/33/35; HPV 6/11, 16/18 and 31/33/35 were revealed in 2, 1, 3 and 5 cases of invasive squamous cell carcinoma, respectively. In 7 cases of adenocarcinoma, 1, 2 and 1 cases exhibited positivity for HPV 16/18; HPV 6/11 and HPV 16/18; and HPV 16/18 and HPV 31/33/35.(ABSTRACT TRUNCATED AT 250 WORDS)

Disseminated cutaneous cytomegalic inclusion disease resembling prurigo nodularis in a HIV-infected patient: a case report and literature review.

Cytomegalovirus infection is common disease but most of the healthy patients are asymptomatic. Cytomegalovirus infection is a serious problem in immunocompromised patients particularly in acquired immunodeficiency syndrome. About 90 per cent of patients with AIDS develop active cytomegalovirus infection and up to 25 per cent may experience life threatening infection due to the virus. However, cytomegalovirus cutaneous lesion have rarely been reported. A 40-year-old Thai man presented with multiple prurigo nodularis-like lesions on both legs and feet. The investigation revealed consistent with acquired immunodeficiency syndrome associated with cytomegalovirus cutaneous infection.

Immunohistochemical analysis of non-Hodgkin's lymphomas with the monoclonal antibody to follicular center lymphocytes reactive in paraffin sections.

The monoclonal antibody designated LN-1 was used in an attempt to identify the antigen in follicular center cell lymphomas using tissue sections fixed in formalin. The LN-1 antibody has been shown in previous studies to identify follicular center cells and give reproducible results in tissue fixed in B5. We used the ABC peroxidase technique to examined formalin-fixed, paraffin embedded sections representing 52 cases of various histologic subgroups of non-Hodgkin's lymphomas based upon the Lukes-Collins classification. Following immunostaining with LN-1 using overnight incubation of the antibody and papain treated sections, 37 cases, and all of the 38 cases previously diagnosed as follicular center cell lymphomas, gave a positive reaction to the LN-1 monoclonal antibody.

Fine needle aspiration of the thyroid gland.

Fine needle aspiration (FNA) has become a major diagnostic triage for clinical management of patients with a thyroid mass. Diagnostic accuracy of this procedure can be evaluated by cyto-histologic correlation. Out of 1,761 fine needle aspirates of thyroid glands performed from January 1996 to December 1999, 230 satisfactory specimens contained respective histologic correlates. Using histologic diagnoses as the gold standard, the overall accuracy, sensitivity and specificity for cytologic diagnoses of thyroid neoplasm were 90.4 per cent, 85.7 per cent, and 92.5 per cent respectively. The positive predictive value for diagnosing thyroid neoplasm was 83.3 per cent while the negative predictive value was 93.7 per cent. The common pairs of cyto-histological diagnostic discrepancy included adenomatous goiter vs follicular neoplasm, adenomatous goiter vs papillary carcinoma, and thyroiditis vs follicular neoplasm. In conclusion, FNA is a rapid, reliable and safe diagnostic tool to distinguish non-neoplastic from neoplastic lesions in patients with thyroid masses.

Ubiquinol supplementation protects against renal ischemia and reperfusion injury in rats.

Generation of toxic oxygen metabolites followed by oxidant- and inflammatory-mediated tissue injury plays a crucial role in the pathogenesis of ischemia and reperfusion (IR). Ubiquinol, the reduced form of coenzyme Q10, is recognized for its potent antioxidant and anti-inflammatory properties in biological membranes. The present study was established to examine the possible protective effect of ubiquinol against renal IR injury. Groups of male Wistar rats were assigned into sham, ubiquinol, IR (45-min bilateral renal ischemia followed by 24-h reperfusion), and ubiquinol+ IR (ubiquinol 300 mg/kg given orally for 7 consecutive days before IR induction). Renal morphology, function, oxidative stress, and inflammatory markers were evaluated at the end of reperfusion. IR caused renal dysfunction as shown by significant increases in blood urea nitrogen, plasma creatinine, and a decrease in creatinine clearance. Light and electron microscopic examinations exhibited severe tubular damages and abnormal mitochondrial structure. IR-induced renal injuries were associated with significant increases in malondialdehyde, nitric oxide, tumor necrosis factor-α, but decreases in antioxidant thiols and superoxide dismutase. Pretreatment with ubiquinol obviously attenuated all the changes caused by IR, whereas it had no considerable effect in the sham-operated rats. These findings indicate that supplementation of ubiquinol prior to IR incidence confers functional and morphological protection to the ischemic kidney by maintaining the redox balance and regulating the generation of inflammatory mediator. The outcomes suggest that ubiquinol may be a potential candidate to counteract organ dysfunction in conditions involving IR injury.

Osteoarticular penicilliosis marneffei. A report of eight cases and review of the literature.

Penicillium marneffei, a dimorphic fungus that is endemic in Southeast Asia and the southern part of China, can cause infections in both healthy and immunocompromised hosts. Bone and joint infections caused by this fungus have rarely been described. The clinical features of eight patients with bone and joint infections caused by this fungus are reported. The English literature on P. marneffei osteoarticular infection is reviewed.

Store-and-forward diagnostic telepathology of small biopsies by e-mail attachment: a feasibility pilot study with a view for future application in Thailand diagnostic pathology services.

Diagnostic telepathology by electronic mail (e-mail) attachment is relatively simple and incurs minimal cost. We assessed its accuracy and practical aspects in routine diagnostic pathology. Using 100 small biopsy specimens, a total of 1,488 images were digitized by one pathologist and sent as e-mail attachments from Nara Medical University, Japan, to a pathologist at Rajavithi Hospital, Thailand. His diagnoses were compared with his conventional light microscopy interpretation at a later date. The average total turnaround time spent on each case was 215 minutes, far less than the several days required by conventional post. There were two clinically significant errors. One was a diagnostically difficult case of colonic dysplasia, which was called carcinoma with telepathology. The other was a signet ring cell carcinoma of the stomach which was undetected with telepathology. Microscopy objective magnification and digital image quality may have played a role in impairing interpretation in both cases. Store-and-forward telepathology provides acceptable efficacy, a comparatively faster turnaround time than post and could be applied in routine work within Thai pathology services.

Studies of human leprosy lesions in situ using suction-induced blisters. 1. Cellular components of new, uncomplicated lesions.

The cellular contents of blisters induced by suction over new, uncomplicated leprosy lesions, and in the skin of cured, control patients, have been examined with enzyme- and immuno-histochemical staining over a period of 4 days. The total cellularity of the blisters varied over a wide range, not correlated with the type of leprosy. Mononuclear cells predominated at all times studied, with nearly equal percentages of monocytes and T lymphocytes. The T-helper: suppressor ratio was significantly greater in BT than in BL and LL lesions at 48 hr. Suction blisters offer a painless, quantitative, reproducible, multiple-sampling method for obtaining cells from the cutaneous infiltrates of leprosy for phenotyping or functional analysis.

Studies of human leprosy lesions in situ using suction-induced blisters. 2. Cell changes and soluble interleukin 2 receptor (Tac peptide) in reversal reactions.

To examine the pathogenesis of type 1 (reversal) reactions in leprosy, we studied cellular and soluble immunologic components of skin lesions in 10 patients with reactions, 24 active patients without reactions, and 33 control patients whose leprosy had been treated and cured. Cells and Tac-peptide levels were obtained from fluid aspirated from blisters induced by suction directly over representative skin lesions. During reversal reactions: a) the lesions contained an increased number and percentage of CD4+ (T-helper) cells; b) Tac-peptide levels were elevated in half of the lesions; c) the increases in Tac peptide and CD4+ cells were directly correlated; and d) systemic administration of corticosteroids appeared to cause a reduction in the intralesional CD4+ cell population. These findings were localized to the skin, and do not represent simple filtration of these components from the peripheral blood. We conclude that spontaneous lymphocyte activation in situ, primarily of CD4+ cells, is an important feature of reversal reactions, and may be an intermittent or cyclic phenomenon during the reaction. Findings in active patients without reactions are consistent with the hypothesis that differing states of immunologic equilibrium have been established in different portions of the leprosy spectrum. In reversal reactions we may, therefore, be examining immunologic processes set in motion when a pre-existing equilibrium has been upset by spontaneous, natural events. The mechanism of such spontaneous changes in immunity in leprosy is of considerable interest, not only to understand the reaction, but also to examine the underlying determinants of delayed-type hypersensitivity and cell-mediated immunity in leprosy and the potential for artificially manipulating these responses, as proposed with vaccines or immunotherapy.