CC chemokine receptor 5 Δ32 polymorphism: association analysis and allele distribution among cutaneous leishmaniasis patients from Pakistan.

BACKGROUND: Human immunodeficiency virus (HIV)/leishmaniasis coinfection is a matter of deep concern worldwide. CC chemokine receptor 5 (CCR5) functions as a co-receptor for HIV entry into host immune cells with an elevated expression observed during leishmaniasis, promoting parasite persistence. A 32 bp deletion (Δ32) in the CCR5 gene provides protection against HIV infection and increased resistance to Leishmania infection. METHODS: In this study, CCR5-Δ32 distribution within Pakistani population with cutaneous leishmaniasis was investigated to evaluate genetic susceptibility to HIV infection. CCR5-Δ32 polymorphism was analyzed in 276 leishmaniasis patients and 119 uninfected healthy controls. Genotypic and allelic frequencies were evaluated and tested for Hardy-Weinberg equilibrium (HWE). RESULTS: The overall Δ32 allele frequency was 6.58% of the population (n = 395). There was a significant difference (p < 0.05) in the geographical distribution of Δ32 allele which was higher in the northern region of the country when compared with the south. Five individuals were identified to be homozygous for the Δ32 allele which has not been reported before from Pakistan. However, no significant association was observed between CCR5-Δ32 and cutaneous leishmaniasis. CONCLUSION: The higher frequency of CCR5 wild-type allele among leishmaniasis patients may suggest an increased risk of HIV infection and also support its facilitative role in Leishmania infection.

Mutations in PGRN gene associated with the risk of psoriasis in Pakistan: a case control study.

BACKGROUND: Psoriasis is a chronic, autoimmune, papulosquamous skin disorder, characterized by the formation of drop-like papules and silvery-white plaques surrounded by reddened or inflamed skin, existing predominantly on the scalp, knees and elbows. The characteristic inflammation and hyperproliferation of keratinocytes in psoriasis is regulated by progranulin (PGRN), which suppresses the expression and release of inflammatory cytokines, such as TNF-α. METHODOLOGY: In this study mutation analysis of the PGRN gene was performed by extracting the genomic DNA from blood samples of 171 diagnosed psoriasis patients and controls through standard salting-out method, followed by amplification and sequencing of the targeted region of exon 5-7 of PGRN gene. RESULTS: Three single nucleotide polymorphisms, rs25646, rs850713 and a novel point mutation 805A/G were identified in the PGRN gene with significant association with the disease. The variant alleles of the polymorphisms were significantly distributed among cases and controls, and statistical analysis suggested that the mutant genotypes conferred a higher risk of psoriasis development and progression. Multi-SNP haplotype analysis indicated that the CAA (OR = 8.085, 95% CI = 5.16-12.66) and the CAG (OR = 3.204, 95% CI = 1.97-5.21) haplotypes were significantly associated with psoriasis pathogenesis. CONCLUSIONS: These findings demonstrate that polymorphisms in PGRN might act as potential molecular targets for early diagnosis of psoriasis in susceptible individuals.

Molecular studies of ichthyosis vulgaris in Pakistani families.

OBJECTIVE: To target and amplify a 1.5 kb FLG gene fragment known to carry R501X mutation responsible for causing ichthyosis vulgaris. STUDY DESIGN: A case series. PLACE AND DURATION OF STUDY: Centre for Molecular Genetics, University of Karachi and Dermatology Department, Jinnah Postgraduate Medical Centre (JPMC), Karachi, from October 2007 to December 2008. METHODOLOGY: Clinically examined seven ichthyosis vulgaris families were included in this study. The 1.5 kb FLG gene fragment was located in the genomic DNA of both the affected (patients) and unaffected (normal, controls) members of the families by PCR amplification using known primers FilF3 and RPTIP6. RESULTS: Amplification of 1.5 kb FLG gene fragment was successful in four families while one family showed amplification of the gene fragment in 3 members (one affected and two normal). Two families showed no amplification. CONCLUSION: The results obtained during this study suggested the possibility of the R501X mutation as being one of the major causes of ichthyosis vulgaris in Pakistan. In addition, the study also revealed the possibility of the presence of novel FLG gene mutations in our population.

Factors associated with onychomycosis in nail psoriasis: a multicenter study in Pakistan.

BACKGROUND: Treatment of psoriatic nail disease is challenging, and dystrophic psoriatic nails can get secondarily infected with fungi. METHODS: This 2-year, matched case-control study was conducted at three tertiary care centers of Karachi, Pakistan. Data were collected from patients with nail psoriasis as cases with age- and gender-matched controls. A detailed questionnaire was filled for all study participants. Nail Psoriasis Severity Index (NAPSI) scoring tool was used to assess dystrophy. Fungal infection was inferred by nail clippings for fungal hyphae and culture. RESULTS: Among 477 participants, 159 cases and 318 controls completed the study. Their mean age was 44 years, and one-third were female. Fungal culture positivity was statistically significant in cases as compared to the control group (P < 0.001). The most frequent species identified was Candida parapsilosis in both cases and controls. Body mass index, NAPSI scoring, socioeconomic status, elevated diastolic blood pressure, smoking status psoriasis among first-degree relatives, and longstanding disease of more than 10 years were significant factors in univariable analysis. Multivariable logistic regression identified independent factors like low to middle socioeconomic status, history of psoriasis in first-degree relative, current smoker, and obesity. CONCLUSION: We found nearly one-third of the psoriatic patients with nail involvement having concomitant fungal infection. We emphasize that nail clipping for fungal smear and culture should be advised to those patients with coexisting factors found significant in our study results. This opinion can be incorporated in psoriasis management guidelines for improving treatment of psoriatic nails.

SLC11A1 polymorphisms and host susceptibility to cutaneous leishmaniasis in Pakistan.

BACKGROUND: The vector-borne cutaneous leishmaniasis (CL) is endemic in several regions of Pakistan mainly affecting poor populations. Host genetic factors, particularly SLC11A1 (solute carrier transmembrane protein) within macrophages, play a crucial role in disease pathology and susceptibility. Association of SLC11A1 with cutaneous leishmaniasis, a neglected tropical disease, is not well established. Inconsistencies have been observed within different populations worldwide with respect to genetic susceptibility. This study was designed to investigate genetic variation(s) in SLC11A1 and to assess possible association with cutaneous leishmaniasis in Pakistan. RESULTS: Eight polymorphisms (rs2276631, rs3731864, rs2290708, rs2695342, rs201565523, rs17215556, rs17235409, rs17235416) were genotyped across SLC11A1 in 274 patients and 119 healthy controls. Six polymorphisms were studied by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing. Two single nucleotide polymorphisms were analyzed with newly designed semi-nested PCR assays. Case-control analysis showed no association between selected polymorphisms in SLC11A1 and cutaneous leishmaniasis. No significant difference was observed in the distribution of alleles between leishmaniasis patients and healthy individuals. Strong pairwise linkage disequilibrium was observed between rs2276631 and rs2290708 (r 2 = 64); and rs17235409 and rs17235416 (r 2 = 78). CONCLUSIONS: This study shows that genetic variations in the candidate gene SLC11A1 do not affect susceptibility to cutaneous leishmaniasis in the sample population from Pakistan.

Association of lichen planus with hepatitis C virus infection.

BACKGROUND: An association between HCV infection and lichen planus is uncertain because the prevalence of HCV infection in patients with lichen planus varies considerably from one geographic area to another. The purpose of this study was to determine the frequency of anti-HCV antibodies and its association with various clinical types of lichen planus in Mekkah, Saudi Arabia. METHODS: A total of 114 cases of lichen planus were selected for the study. These were divided into four categories, including patients with skin lesions, skin and oral lesions, and oral or genital lesions alone. The sera of these patients were tested for HCV antibodies by means of a third-generation ELISA and serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were simultaneously determined. A group of 65 volunteers served as a control group. RESULTS: Of the 114 patients with lichen planus, 30 had HCV antibodies (26.3%). In the 65 control group subjects anti-HCV antibodies were observed in 3 volunteers (4.6%). There was a significant difference between the two groups (P<0.0001). The ALT was raised in 22 patients and the AST level was elevated in 14 of the 114 cases of lichen planus. In the control group, the AST level was raised in 3 of the 65 controls while 2 had an elevated ALT level. The number of patients with an abnormal transaminase level also significantly differed in the two groups. CONCLUSION: A high prevalence of HCV infection was detected in patients with lichen planus. These results support a possible relationship between lichen planus and hepatitis C.

Dermatophyte growth and degradation of human stratum corneum in vitro (pathogenesis of dermatophytosis).

BACKGROUND: This study was carried out to determine growth of dermatophytes using human stratum corneum in vitro and the degrading effect of Keratinases (Proteinases) on stratum corneum for a complete understanding of the host parasite relationship. METHOD: Trichophyton rubrum isolates derived from patients with tinea cruris infections were obtained from the Department of Medical Microbiology, University Hospital of Wales, U.K. Human stratum corneum sterilized with ethylene oxide was used as a nitrogen source in agar culture medium plates. RESULT: Fungal growth took place in plates which contained human stratum corneum particles while there was no growth in the plates without stratum corneum at three weeks after initiation. There was a gradual disappearance of the particles of stratum corneum from the plates at the end of the third week CONCLUSION: The growth of organisms in plates with human stratum corneum and their disappearance at third week suggested that stratum corneum was not only source of nutrition for the dermatophytes, but also the growing fungal mycelia and the proteinases induced by them were playing a part in the digestion of granules and thus may have an important role in the pathogenesis of dermatophyte infections.

Ectodermal dysplasia with acanthosis nigricans (Lelis' syndrome).

Autosomal recessive ectodermal dysplasia with acanthosis nigricans is termed Lelis' syndrome. It is a rare condition and one case is described with overall clinical, dermatological and dental findings.

Spectrum of skin disorders presenting to King Abdul Aziz Hospital during Hajj season-2000.

BACKGROUND: The pilgrimage (Hajj) to the holy mosque in the city of Makkah takes place once every year and during this huge gathering skin diseases are quite common due to hot weather and over crowding. The aim of this study was to collect and report data regarding different dermatological problems occurring during the holy month of Zil-hajj. METHODS: Data regarding skin diseases was collected from pilgrims which were examined and diagnosed clinically at the Department of Dermatology King Abdul Aziz Hospital, during the month of Zil-Hajja of the year 2000. These patients were referred from various primary health centers, medical hajj missions of various countries and Children and Maternity hospital in Makkah. RESULTS: During the month of Zil-hajja of the year 2000, 1510 cases were seen, of these 1143 were males and 367 were females. The criteria for diagnosis for most of the cases were mainly clinical. The highest number of patients was in the age group of 20-50 years. Eczemas of different types were the most common skin disease observed in these pilgrims, intertrigo was the next most common presenting condition this was followed by fungal and bacterial infections. CONCLUSION: A high frequency of skin diseases such as eczemas, intertrigo, pyoderma and fungal infections was found among the pilgrims. More detailed studies regarding skin conditions during this season would enable us to have better understanding of skin problems there management and prevention in full.

Detection of filaggrin gene mutation (2282del4) in Pakistani Ichthyosis vulgaris families.

The aim of this study was to detect an 811 bp filaggrin (FLG) gene fragment known to carry a mutation 2282del4 which causes ichthyosis vulgaris. Seven clinically examined ichthyosis vulgaris families were included in this study. An 811 bp FLG gene fragment was targeted in the genomic DNA of all the members of the seven families by PCR amplification using known primers RPT1P7 and RPT2P1. Successful amplification of an 811 bp FLG gene fragment in all the families suggested the possible role of the 2282del4 mutation in causing ichthyosis vulgaris in Pakistani population.

Homozygous frame shift mutation in ECM1 gene in two siblings with lipoid proteinosis.

BACKGROUND: The extracellular matrix protein 1 (ECM1) is a glycoprotein, expressed in skin and other tissues. Loss-of-function mutation in ECM1 causes a rare autosomal recessive disorder called lipoid proteinosis. Lipoid proteinosis is presented by varying degrees of skin scars, beaded papules along the eyelid margins, variable signs of hoarseness of voice and respiratory disorders. More than 250 cases of this disorder have been described in the literature, but occurrence of lipoid proteinosis in siblings is very rare. This study was designed to investigate the possible mutation causing lipoid proteinosis in a Pakistani family and to elaborate the scope of possible genetic changes, causing the genodermatosis in Pakistan. MAIN OBSERVATIONS: In this study, two siblings (12 and 9-years sisters) were presented with scaly itchy lesions on whole body, hoarse voice and macroglossia. Their deceased father had similar clinical manifestations but mother and younger brother were unaffected. Blood samples from clinically affected and unaffected family members were collected with informed consent. The coding region of ECM1 gene containing 10 exons were amplified and sequenced. Both the affected siblings were shown to have homozygous frame shift mutation by deletion of the nucleotide T at 507, codon 169, exon 6. This resulted in a frame shift from codon 169 and appearance of a premature stop codon at 177, causing formation of a mutated protein (176 amino acids) instead of normal ECM1 protein (540 amino acids). CONCLUSION: A case of homozygous 62-bp insertion in ECM1 gene causing lipoid proteinosis has been reported in another Pakistani family. The current study presents a homozygous frame shift mutation supporting an unusual function of ECM1 protein and broadens the spectrum of disease-linked mutations in this rare case of genodermatosis in this region.