Non-Invasive Immunofluorescence Assessment of Lycopene Supplementation Status in Skin Smears.

Circulating lycopene level is negatively associated with the prevalence of cardiovascular disease, cancers (prostate and breast), type 2 diabetes mellitus, and aging. Traditionally, lycopene is measured in biological specimens by a combination of high-performance liquid chromatography (HPLC) and mass spectrometry methods. Moreover, as we recently reported, tissue/cell lycopene depositions can be observed by the immunohistochemistry method with a newly developed monoclonal antibody (mAb) against lycopene. A main objective of this study is to evaluate the performance of a new noninvasive immunofluorescence (IF) lycopene quantification skin test with mAbs against lycopene versus HPLC lycopene assay of serum lycopene in volunteers subjected to lycopene supplementation which represents a novel approach to lycopene measurement methodology. For this purpose, 32 healthy volunteers, 30-40 years old, were supplemented with lycopene (n = 15) or placebo (n = 17) for a period of 4 weeks. It was found that lycopene supplementation leads to a significant increase in serum lycopene concentration after 2 and 4 weeks by 2.6- and 3.4-fold over control, respectively. This was accompanied by a concordant step-wise rise in IF staining of skin corneocytes and sebum, quantifiable by arbitrary IF scores. Placebo supplementation did not affect serum lycopene values or intensity of IF staining of the skin samples. There was 86.6% agreement in paired HPLC/IF variants for the intermediate time point and 80.0% agreement at the end of the study in the lycopene group. Intraclass correlation between paired values in this group was +0.49 for the 2-week time point and +0.63 for the end point. These results indicate that the new antibody-based skin assay can be used for rapid detection of lycopene deficiencies. Moreover, the noninvasive nature of the skin swab test would allow using it to monitor, optimize, and personalize lycopene supplementation protocol of risk groups in the general population.

Polymerase Chain Reaction (PCR) as a Potential Point of Care Laboratory Test for Leprosy Diagnosis-A Systematic Review.

Leprosy is an infectious disease caused by Mycobacterium leprae and mainly affects skin, peripheral nerves, and eyes. Suitable tools for providing bacteriological evidence of leprosy are needed for early case detection and appropriate therapeutic management. Ideally these tools are applicable at all health care levels for the effective control of leprosy. This paper presents a systematic review analysis in order to investigate the performance of polymerase chain reaction (PCR) vis-à-vis slit skin smears (SSS) in various clinical settings and its potential usefulness as a routine lab test for leprosy diagnosis. Records of published journal articles were identified through PubMed database search. Twenty-seven articles were included for the analysis. The evidence from this review analysis suggests that PCR on skin biopsy is the ideal diagnostic test. Nevertheless, PCR on SSS samples also seems to be useful with its practical value for application, even at primary care levels. The review findings also indicated the necessity for improving the sensitivity of PCR and further research on specificity in ruling out other clinical conditions that may mimic leprosy. The M. leprae-specific repetitive element (RLEP) was the most frequently-used marker although its variable performance across the clinical sites and samples are a matter of concern. Undertaking further research studies with large sample numbers and uniform protocols studied simultaneously across multiple clinical sites is recommended to address these issues.

Utility of Bacillary Index in Slit Skin Smears in Correlation with Clinical and Histopathological Alterations in Hansen's Disease: An Attempt to Revive a Simple Useful Procedure.

Leprosy, a relatively common chronic contagious disease having diverse modes of clinical presentation, can mimic a variety of unrelated diseases. For proper and adequate treatment, the diagnosis must be made accurately with subtyping which should be done with the help of bacillary index, histopathological features, and clinical correlation. This is extremely important in patient care as paucibacillary and multibacillary types have different modes of treatment. Our aim is to categorize leprosy into various types based on bacillary index, morphological findings both in slit skin smears, and biopsy along with clinical correlation.

Comparison of bacillary index on slit skin smear with bacillary index of granuloma in leprosy and its relevance to present therapeutic regimens.

BACKGROUND: As the world moves toward elimination of leprosy, persistence of infective cases in endemic pockets remains a significant problem. The use of clinical criteria to decide the paucibacillary (PB) versus multibacillary (MB) regimens has greatly simplified therapy at the field setting. However, a small but significant risk of under-treatment of so-called "PB" cases which actually have significant bacillary load exists. This study was undertaken to assess this risk and compare two methods of assessment of bacillary load, namely bacillary index on slit skin smear (BIS) versus bacillary index of granuloma (BIG). AIMS: To compare BIS with BIG on skin biopsy in consecutive untreated cases of leprosy. MATERIALS AND METHODS: This prospective study was conducted over a period of 12 months, wherein new untreated patients with leprosy were consecutively recruited. After a thorough clinical examination, each patient underwent slit skin smear (SSS) where the BIS was calculated. The same patient also underwent a skin biopsy from a clinical lesion where, the BIG was calculated. SSS and skin biopsy for BIS and BIG respectively were repeated for all patients at the end of therapy for comparison. All patients received therapy according to World Health Organization-Multidrug Therapy Guidelines. RESULTS: The BIG was positive in all cases where the BIS was positive. Significantly, BIG was positive in three cases of borderline tuberculoid leprosy with <5 lesions who received PB regimen, whereas the BIS was negative in all three cases. CONCLUSION: This study suggests that BIG may be a better indicator of the true bacillary load in leprosy as compared to BIS. Its role in management is significant, at least in tertiary care centers to prevent "under-treatment" of so called PB cases, which may actually warrant MB regimens.

Mycobacterium leprae in the periodontium, saliva and skin smears of leprosy patients / Mycobacterium leprae no periodonto, saliva e raspados intradérmicos de sujeitos com hanseníase

Purpose: To verify the presence of M. leprae in the periodontium, saliva and skin slit smears of leprosy patients. To correlate bacteriological and molecular findings with clinical data and compare laboratory techniques. Methods: A cross-sectional study was designed to use bacteriological (baciloscopy) and molecular (PCR) parameters to detect M. leprae in exudates of the gingival sulcus/periodontium pocket, saliva and skin slit smears from multiple clinical forms of leprosy patients without previous treatment. Results: The study included 48 leprosy patients with 15 multibacillary (MB) cases and 33 paucibacillary (PB) cases. The diagnosis of MB was confirmed through bacteriological examination and PCR results from skin slit smears. A total of 16 (48.5%) PB patients were PCR positive only. Four PB patients with negative PCR skin smears were PCR positive for the periodontium and saliva, with 2 cases and 1 case, respectively. No periodontium or saliva samples had positive bacteriological results. Conclusion: There was no correlation between periodontal disease and the presence of M. leprae. Bacteriological examination did not prove to be an efficient technique for the analysis of saliva and periodontium samples. PCR analysis of skin smears was more efficient at diagnosing PB patients than bacteriological examination. PCR positive results for the detection of M. leprae in PB patients can be increased by collecting slit skin smears, periodontium and saliva samples.

Objetivo: verificar através da baciloscopia e da reação em cadeia da polimerase (PCR) a presença do M. leprae no periodonto, saliva e raspados intradérmicos de pacientes com hanseníase. Metodologia: Realizou-se um estudo transversal do tipo detecção de casos numa instituição referência de hanseníase no Amazonas. Resultados: Foram avaliados 48 pacientes, sendo 15 multibacilares (MB) e 33 paucibacilares (PB). Os pacientes MB tiveram o diagnóstico confirmado pela baciloscopia e PCR dos raspados intradérmicos, enquanto que 16 (48,5%) dos PB foram positivos apenas na PCR. Quatro pacientes PB negativos na PCR de raspados intradérmicos foram positivos no periodonto e na saliva, 1 positivo na saliva e 2 no periodonto. Nenhuma amostra do periodonto e da saliva foi positiva na baciloscopia. Conclusão: Não houve relação entre a doença periodontal e a presença do M. leprae; a baciloscopia não mostrou ser uma técnica eficiente para análise da saliva e periodonto; a técnica de PCR de raspado dérmico mostrou ser um método mais eficaz no diagnóstico dos PB do que a baciloscopia; a positividade da PCR para detecção do M. leprae nos PB pode ser aumentada coletando raspado intradérmico, periodonto e saliva.