Shigeo Nonaka

Detection of new endemic areas of cutaneous leishmaniasis in Pakistan: a 6‐year study

Background Cutaneous leishmaniasis (CL) is endemic in Pakistan and is widely spreading. Recently, an outbreak of the disease was observed in the region. We report some new endemic areas of CL in the country.

Usefulness of sampling with cotton swab for PCR-diagnosis of cutaneous leishmaniasis in the New World

In this study, we tested the polymerase chain reaction (PCR)-method to diagnose cutaneous leishmaniasis (CL) by taking exudate materials from lesions with cotton swabs, using our previously tested (PCR) panel comprised of Leishmania (Viannia) panamensis, L. (V.) braziliensis, L. (V.) guyanensis, L. (Leishmania) mexicana and L. (L.) amazonensis. The objectives of the present study were to improve the sampling method convenient for the patients and to test the usefulness of samples taken with cotton swabs. Sixteen patients were clinically diagnosed to have CL including one case of diffuse cutaneous leishmaniasis (DCL) in Ecuador and the causative Leishmania parasites were identified by PCR. All the 12 samples from CL patients of La Mana, positive for Leishmania DNA, were identified as L. (V.) panamensis, while two from CL of Huigra and one from DCL of San Ignacio were L. (L.) mexicana. In the field condition, taking biopsy material is not only painful but sometimes causes iatrogenic bacterial infections. Considering the sensitivity of the test, and convenient sampling procedure, it may be suggested that collection of exudates using cotton swabs may be a better alternative to biopsy sample for PCR-diagnosis of CL.

Rapid identification of Leishmania species from formalin-fixed biopsy samples by polymorphism-specific polymerase chain reaction

The precise identification and classification of Leishmania species is important for public health surveillance since different species cause different clinical features of the disease. A highly specific polymerase chain reaction (PCR) panel was developed to enable the identification of the five major Leishmania species that cause New World cutaneous leishmaniases. The primers used for this panel were designed to distinguish the polymorphism in sequences of commonly amplified DNA bands of the parasites produced by arbitrarily primed PCR. These polymorphism-specific PCR diagnoses were performed with formalin-fixed biopsy specimens of the leishmanial lesions from four patients in Ecuador and one hamster skin lesion, and these lesions were determined to be caused by Leishmania (Viannia) panamensis, L. (Leishmania) mexicana, and L. (L.) amazonensis. The PCR panel may offer an important and practical approach to the standardized identification of Leishmania species in field examinations.

Clinical and epidemiological observations of cutaneous tuberculosis in Larkana, Pakistan.

Background Cutaneous tuberculosis is widespread in Pakistan but has not been fully documented. This study was conducted to determine the clinical pattern, nature and existence of the disease in Larkana, Sindh province, Pakistan.

Association of human papillomavirus type 6 with a verruciform xanthoma.

We report a case of verruciform xanthoma (VX) associated with human papillomavirus (HPV) in a 67-year-old male. The patient had a pale-reddish, granular and verrucous tumor on the right side of his scrotum for four years. Histopathologic examination showed typical features of VX. HPV was detected by immunohistochemistry, electron microscopy, and PCR examinations. Ultrastructural examination revealed virus-like particles of 40–50 nm in the nucleus of the upper epidermal keratinocytes. HPV type 6a DNA was detected in lesional tissue by polymerase chain reaction and sequence analysis. To the best of our knowledge, this is the first case report of VX associated with HPV.

Mast Cell “Densities” in Vascular Proliferations: A Preliminary Study of Pyogenic Granuloma, Portwine Stain, Cavernous Hemangioma, Cherry Angioma, Kaposi's Sarcoma, and Malignant Hemangioendothelioma

The “densities” of mast cells (MCs) in six kinds of vascular proliferation, pyogenic granuloma, portwine stain, cavernous hemangioma, cherry angioma, Kaposis sarcoma, and malignant hemangioendothelioma (MHE), measured per mm2 were studied using respective specimens prepared with tryptase stain and a personal computer. The average densities of MCs in pyogenic granuloma and MHE were 103.5 ± 25.2/mm2 (n=10) and 106.3 ± 40.2/mm2 (n=10) [mean ± standard deviation (SD)]; that in normal skin was 6.85 ± 4.9/mm2 (n=20) (mean ± SD). is a significant difference [t‐test (p<0.0001) and Wilcoxon‐test (p<0.01)]. The results in portwine stain (n=4), cavernous hemangioma (n=9), cherry angioma (n=4), and Kaposis sarcoma (n=4) were 68.6 ± 28.9/mm2, 105.7 ± 56.9/mm2, 85.3 ± 45.6/mm2, 82.2 ± 28.4/mm2 (mean ± SD), respectively, all of which were greater than that in normal skin by a simple comparison. The results of immunofluorescence microscopy were positive with basic fibroblast growth factor staining in the tissues of pyogenic granuloma, Kaposis sarcoma and MHE. These facts may morphologically indicate a role of MCs in the angiogenesis of these vascular tumors.

Leishmaniasis recidiva cutis due to Leishmania (Viannia) panamensis in subtropical Ecuador: isoenzymatic characterization

Background  Information regarding leishmaniasis recidiva cutis (LRC), a clinical variant of cutaneous leishmaniasis, in the New World is scarce. LRC is characterized by slowly progressing lesion(s) that appear after a variable period of time, from months to years, in or around the scar of an apparently clinically healed sore.

Phacomatosis Pigmentovascularis Type IIb Associated with Sturge‐Weber Syndrome and Pyogenic Granuloma

A case of phacomatosis pigmentovascularis (PPV) in a 6‐year‐old girl with Sturge‐Weber syndrome, pyogenic granuloma, and other complications is described. It is relatively rare that a complete form of Sturge‐Weber syndrome was associated with PPV. A review of the literature on PPV, focusing on total number of reported cases and etiological speculations, is presented. To our knowledge, a total of 118 cases of PPV, including the present one, have been reported to date. Regardless of many speculations, the true etiology remains unknown. The average “density” of mast cells (MCs) per mm2 appearing in the central region of the pyogenic granuloma was calculated to be 86.3/mm2 and that in the adjacent nevus flammeus was 37.9/mm2. The “density” of mast cells in pyogenic granuloma separately calculated from ten other cases was 105.5 ± 28.6/mm2 (mean ± SD), compared with that in normal skin, 6.85 ± 4.9/mm2 (n=20). There was a significant difference between the two, indicating that MCs are closely associated with angiogenesis in pyogenic granuloma.

The Effects of Sunscreens on UVB Erythema and Langerhans Cell Depression

Langerhans cells (LCs) are epidermal antigen‐presenting cells capable of initiating a specific T lymphocyte‐mediated immune response. It is a well known fact that ultraviolet light B (UVB) suppresses LC number and function.

Comparative Studies of the Detection Rates of Leishmania Parasites from Formalin, Ethanol‐fixed, Frozen Human Skin Specimens by Polymerase Chain Reaction and Southern Blotting

In this study, detection rates of Leishmania parasites from human skin were compared among three different types of specimens, formalin‐fixed, ethanol‐fixed, and frozen, by polymerase chain reaction (PCR) and Southern blotting. For this purpose, we used biopsy specimens collected from 19 leishmaniasis patients and performed PCR and Southern hybridization with the probe specific for Leishmania (Viannia) braziliensis complex. Among these 19, 16 specimens were from cutaneous leishmaniasis (CL), one, diffuse cutaneous leishmaniasis (DCL) and 2, mucocutaneous leishmaniasis (MCL) and were formalin‐fixed and paraffin‐embedded. The causative agents for one case of CL and one case of DCL were already identified as L. (Leishmania) complex. Six specimens of CL were preserved in 100% ethanol. Two specimens of MCL were frozen tissues. PCR using the formalin‐fixed and paraffin‐embedded specimens revealed positive bands at 70bp in 9 (47.4%) out of 19 specimens of CL, MCL and DCL. Southern blotting detected the signals in 12 (63.2%) out of the 19. PCR using the 100% ethanol‐fixed specimens revealed positive bands in 4 (66.7%) out of 6, and Southern blotting also detected the signals in 4 (66.7%) out of the 6. PCR and Southern blotting using 2 frozen specimens of MCL were always positive (100%). Although we failed to detect significant differences by Chi‐square test between the results from the formalin‐fixed, paraffin‐embedded specimens and those from 100% ethanol‐fixed ones, we concluded that ethanol‐fixed specimens, convenient for transportation and storage, would be more useful for diagnosis of leishmaniasis by PCR in a developing country.