Development of a fast LC-MS/MS protocol for combined measurement of six LSDs on dried blood spot in a newborn screening program.

New treatment options and improved strategies for Lysosomal Storage Disorders (LSDs) diagnosis on dried blood spot (DBS) have led to the development of several pilot newborn screening programs. Building on a previously published protocol, we devised a new 6-plex assay based on a single DBS punch incubated into a buffer containing a combination of substrates (GAA, GLA, ASM, GALC, ABG and IDUA). This new protocol incorporates a new trapping and clean-up procedure using perfusion chromatography connected on-line with an analytical column for analyte separation, after enzymatic reaction. Results are available after 4.5 min. Several incubation times were tested in order to reduce sample preparation times and to improve accuracy and reproducibility, also regarding the quenching of the reaction within the time window of linear product accumulation. The collected data demonstrate that an incubation time of 4 h is enough to achieve good reaction efficiency without any impact on sensitivity. The method proved versatile and robust for various instrument configurations. The fast sample preparation and running times allow a high sample throughput; an advantage in newborn screening procedures. This method can also be used for diagnostic purposes, allowing a rapid diagnosis in a few hours.

Reducing the False-Positive Rate for Isovalerylcarnitine in Expanded Newborn Screening: The Application of a Second-Tier Test

Abstract The isodecyl neopentanoate is an ingredient used in the cosmetic industry to prepare a nipple fissure balm. We report on 12 newborns that showed elevated C5-acylcarnitine levels upon newborn screening following treatment with balm. The first 3 neonates were immediately recalled for confirmatory tests and resulted negative for both isovaleric acidemia and short/branched chain acyl-CoA dehydrogenase deficiency. In the other 9 cases, the immediate recall was avoided by applying a new second-tier test able to confirm the presence of pivaloylcarnitine. The concentration of C5-acylcarnitine was measured in the days following the suspension of balm application. Abnormal concentrations of C5-acylcarnitine did not seem to be associated with free carnitine deficiency, probably due to the short time of exposure. A direct correlation between balm ingestion and the elevation in pivaloylcarnitine has been demonstrated in 10 adult volunteers. The commercial balm containing a pivalic acid derivative is causal of false-positive results during newborn screening, and it could have the potential to cause secondary carnitine deficiency when used chronically.

[Solar ultraviolet radiation risk in outdoor workers: a specific project of Tuscany Region (Italy)]. / Rischio da radiazione solare ultravioletta nei lavoratori outdoor: piano mirato della Regione Toscana.

OBJECTIVE: The aims of Tuscany Regional project were: to study the sun protection attitude of outdoor workers; to measure solar ultraviolet (UV) exposure in work environment; to describe the frequency of photoaging, precancerous lesions, and skin cancers in outdoor workers; to collect information on solar ultraviolet radiation exposure from incident cases of Non-Melanoma Skin Cancer (NMSC) recruited from Tuscany Cancer Registry. DESIGN: Outdoor workers completed a questionnaire devoted to collect information on sun protection attitudes during a typical summer working week. Environmental and personal measurements were carried out. Expert dermatologists examined outdoor workers to assess the frequency of photoaging, precancerous lesions, and skin cancer. A structured questionnaire was mailed to incident cases of NMSC. Information were collected on personal habits and working history, focusing on solar ultraviolet radiation exposure. SETTING AND PARTICIPANTS: Agriculture, construction, quarrying and fishing activities were considered: 292 employees responded to questions about the type of clothing used in the morning and in the afternoon,while working outdoors; 637 outdoor workers underwent skin examination. We contacted 743 cases of NMSC occurred in 2004; 498 subjects accepted to participate in this study. RESULTS: The clothing worn by surveyed subjects was often inadequate compared to the high level of exposure to UV. The skin examination of 637 outdoor workers highlighted 2 melanomas, 7 epitheliomas and 35 actinic keratoses. Among the 498 cases of NMSC, 135 (27%) were diagnosed in outdoor workers. Most represented economic activity sectors were: agriculture, construction, transport, sports. CONCLUSION: The characterization of outside workers revealed unsatisfactory sun protection behaviours. Moreover, previously undetected skin cancers were diagnosed. The study on MNSC confirms the complexity of studying the exposure to UV radiation. The Tuscany Regional project provided useful information on the risk of solar ultraviolet radiation in outdoor workers. Prevention programs are needed.

Keratotic horn on left fifth fingertip: congenital ectopic nail misdiagnosed as a common wart.

The term "ectopic nail" refers to nail tissue found in a location other than in the normal nail bed. Here we report a 9-year-old girl with an asymptomatic keratotic "horn" on the tip of her left fifth finger. Present since the age of 1 month, it was misdiagnosed as a common wart and treated using liquid nitrogen cryotherapy without benefit.

Digital image management project for dermatological health care environments: a new dedicated software and review of the literature.

BACKGROUND/PURPOSE: Because the skin is the only organ completely accessible to visual examination, digital technology has therefore attracted the attention of dermatologists for documenting, monitoring, measuring and classifying morphological manifestations. To describe a digital image management system dedicated to dermatological health care environments and to compare it with other existing softwares for digital image storage. METHODS: We designed a reliable hardware structure that could ensure future scaling, because storage needs tend to grow exponentially. For the software, we chose a client-web server application based on a relational database and with a 'minimalist' user interface. RESULTS: We developed a software with a ready-made, adaptable index of skin pathologies. It facilitates classification by pathology, patient and visit, with an advanced search option allowing access to all images according to personalized criteria. The software also offers the possibility of comparing two or more digital images (follow-up). The fact that the archives of years of digital photos acquired and saved on PCs can easily be entered in the program distinguishes it from the others in the market. This option is fundamental for accessing all the photos taken in years of practice in the program without entering them one by one. The program is available to any user connected to the local Intranet and the system may directly be available in the future from the Internet. CONCLUSIONS: All clinics and surgeries, especially those that rely on digital images, are obliged to keep up with technological advances. It is therefore hoped that our project will become a model for medical structures intending to rationalise digital and other data according to statutory requirements.

Venous ulcers of the lower limbs due to congenital thalidomide-related valve defect.

A 44-year-old woman with fetal thalidomide syndrome and congenital pseudoainhum of the left big toe had a 5-year history of painful nonhealing ulcers in the left malleolar region. Venous Doppler ultrasonography showed hypoagenesis of the valve flaps of the deep and superficial venous circuit. To our knowledge, this is the first description of congenital pseudoainhum associated with fetal thalidomide syndrome. The coexistence of cutaneous ulcers in our case might be coincidental but may be related to a congenital valve defect (hypoagenesis) caused by thalidomide.

Cutaneous metastases from adenocarcinoma of the rete testis.

Adenocarcinoma of the rete testis (ACRT) is extremely rare and has only been the subject of sporadic case reports, in most of which the neoplasm manifested as a scrotal mass with diffuse enlargement of the testis. Only a few cases of scrotal infiltration by a contiguous ACRT have been described. To our knowledge, none have reported distant skin metastases. We report a case of ACRT presenting with suprapubic skin metastases. The diagnosis was based on clinical and histopathological findings and supported by the results of immunohistochemical and ultrastructural studies. We discuss the differential diagnosis to this rare entity, which include metastatic adenocarcinoma, serous tumor of the testis, and mesothelioma of the tunica vaginalis.

Acral malignant melanoma and striated palmoplantar keratoderma (Brunauer-Fohs-Siemens syndrome): a fortuitous association?

BACKGROUND: Striated palmoplantar keratoderma or Brunauer-Fohs-Siemens syndrome is a very rare, focal, nonepidermolytic palmoplantar keratoderma with autosomal inheritance. Unlike other palmoplantar keratodermas, no association with visceral or skin cancer has ever been reported. OBJECTIVE: We report a case of malignant melanoma arising in the hyperkeratotic lesions on the right heel of a patient with striated palmoplantar keratoderma. The lesion was completely excised; our patient also underwent sentinel lymph node biopsy and then was treated with high-dose interferon adjuvant therapy. METHODS: Sentinel lymph node biopsy incision was made in elliptical fashion, long enough to harvest a full-thickness skin graft to cover the wide local excision defect. The skin graft was defatted by sharp dissection. Several perforations were made in graft and it was secured in place with sutures and bolster dressing. RESULTS: At follow-up, the grafted skin showed hyperkeratotic changes but no local or systemic signs of the disease was observed. CONCLUSION: The association between striated palmoplantar keratoderma and acral melanoma is discussed.