Disability Competent Care in Pediatric Milestones.

This cross-sectional study examines how disability competencies are addressed in the Accreditation Council for Graduate Medical Education's milestones for pediatric specialties.

Biosimilars in psoriasis: Clinical practice and regulatory perspectives in Latin America.

Latin American countries view biosimilar agents as an effective approach to curtail health-care expenditures while maintaining the safety and efficacy profile of their branded innovator comparators. To understand the complexities of the regulatory landscape and key therapeutic issues for use of biosimilars to treat moderate to severe psoriasis in Latin America, the International Psoriasis Council convened dermatology experts from Argentina, Brazil, Chile, Colombia and Mexico in October 2015 to review the definition, approval, marketing and future of biosimilars in each country and develop a consensus statement. The regulatory framework for marketing approval of biosimilars in Latin America is currently a mosaic of disparate, country-specific, regulatory review processes, rules and standards, with considerable heterogeneity in clarity and specificity. Regulations in Argentina, Brazil, Chile and Mexico have undergone multiple refinements whereas Colombia is finalizing draft guidelines. Verification of the similarity in quality, safety and efficacy of biosimilars to the innovator biologic remains a key challenge for policy makers and regulatory authorities. Other key regulatory challenges include: naming of agents and traceability, pharmacovigilance, extrapolation of indications, and interchangeability and substitution. An urgent need exists for more Latin American countries to establish national psoriasis registries and to integrate their common components into a multinational psoriasis network, thereby enhancing their interpretative power and impact. A Latin American psoriasis network similar to PSONET in Europe would assist health-care providers, pharmaceutical companies, regulators and patients to fully comprehend specific products being prescribed and dispensed and to identify potential regional trends or differences in safety or outcomes.

Differences in the glucuronidation of resveratrol and pterostilbene: altered enzyme specificity and potential gender differences.

Resveratrol, a natural polyphenol found in grapes, berries and other plants, has been proposed as an ideal chemopreventative agent due to its plethora of health promoting activities. However, despite its lofty promise as a cancer prevention agent its success in human clinical trials has been limited due to its poor bioavailability. Thus, interest in other natural polyphenols is intensifying including the naturally occurring dimethylated analog of resveratrol, pterostilbene. The UDP-glucuronosyltransferase (UGT) family of enzymes plays a vital role in the metabolism of both resveratrol and pterostilbene. The current study sought to elucidate the UGT family members responsible for the metabolism of pterostilbene and to examine gender differences in the glucuronidation of resveratrol and pterostilbene. We demonstrate that UGT1A1 and UGT1A3 are mainly responsible for pterostilbene glucuronidation although UGT1A8, UGT1A9 and UGT1A10 also had detectable activity. Intriguingly, UGT1A1 exhibits the highest activity against both resveratrol and pterostilbene despite altered hydroxyl group specificity. Using pooled human liver microsomes, enzyme kinetics were determined for pterostilbene and resveratrol glucuronides. In all cases females were more efficient than males, indicating potential gender differences in stilbene metabolism. Importantly, the glucuronidation of pterostilbene is much less efficient than that of resveratrol, indicating that pterostilbene will have dramatically decreased metabolism in humans.

Pathogenicity, Virulence, and Vegetative Compatibility Grouping of Verticillium Isolates from Spinach Seed.

In 2005, Verticillium dahliae was first reported to be pathogenic to spinach seed crops in the Pacific Northwest, with symptoms only developing after initiation of the reproductive stage of plant growth, and to be prevalent on commercial spinach seed lots produced in Denmark, The Netherlands, and the United States. In this study, the genetic diversity, pathogenicity, and virulence were examined for a collection of isolates of Verticillium spp. from spinach as well as other hosts (alfalfa, cotton, lettuce, mint, peppermint, potato, radish, and tomato) from various countries and from different vegetative compatibility groups (VCGs). Of a total of 210 isolates of V. dahliae obtained from spinach seed produced in Denmark, the Netherlands, New Zealand, or the United States, 128 were assigned to VCG 4B (89% of 91 U.S. isolates, 86% of 42 isolates from the Netherlands, 19% of 43 Denmark isolates, and 8% of 13 New Zealand isolates), 65 to VCG 2B (92% of the New Zealand isolates, 79% of the Denmark isolates, 14% of the Netherlands isolates, and 9% of the U.S. isolates), and 3 to VCG 2A (2% of each of the Denmark and U.S. isolates, and 0% of the Netherlands and New Zealand isolates); 14 isolates could not be assigned to a VCG. Although little variation in the sequence of the internal transcribed spacer (ITS) region of ribosomal DNA was observed among isolates within each Verticillium sp., the ITS region readily differentiated isolates of the species V. dahliae, V. tricorpus, and Gibellulopsis nigrescens (formerly V. nigrescens) obtained from spinach seed. Greenhouse pathogenicity assays on spinach, cotton, lettuce, and tomato plants using isolates of V. dahliae (n = 29 to 34 isolates), V. tricorpus (n = 3), G. nigrescens (n = 2), and V. albo-atrum (n = 1) originally obtained from these hosts as well as from alfalfa, mint, peppermint, potato, and radish, revealed a wide range in virulence among the isolates. Isolates of V. tricorpus and G. nigrescens recovered from spinach seed and an isolate of V. albo-atrum from alfalfa were not pathogenic on spinach. In addition, isolates of V. dahliae from mint and peppermint were not pathogenic or only weakly virulent on the hosts evaluated. Although there was a wide range in virulence among the isolates of V. dahliae tested, all of the V. dahliae isolates caused Verticillium wilt symptoms on spinach, lettuce, tomato, and cotton. None of the isolates of V. dahliae showed host specificity. These results indicate that Verticillium and related species associated with spinach seed display substantial variability in virulence and pathogenicity to spinach and other plants but the V. dahliae isolates were restricted to three VCGs.

NADPH oxidase 1 overexpression enhances invasion via matrix metalloproteinase-2 and epithelial-mesenchymal transition in melanoma cells.

NADPH oxidase 1 (Nox1) is a member of the NADPH oxidase family that has not been well characterized in the melanocytic cell lineage. Here we demonstrated that Nox1 and Nox4 were detected in melanocytic lineage, with only Nox1 detected in normal human melanocytes and Nox4 in a subset of metastatic melanoma cell lines. The protein level and enzymatic activity of Nox1 was elevated in all melanoma cells as compared with normal melanocytes. Overexpression of GFP-Nox1 protein in Wm3211 primary melanoma cells increased invasion rate by 4- to 6-fold as measured by Matrigel invasion assay, whereas knocking down or inhibiting Nox1 decreased invasion by approximately 40-60% in Wm3211 and SK-Mel-28 cells. Matrix metalloproteinase-2 (MMP-2) was increased by Nox1 overexpression at the mRNA, protein, and activity levels, and decreased by Nox1 knockdown. MMP-2 promoter activity was also regulated by Nox1 knockdown. In addition, stable clones overexpressing Nox1 exhibited an epithelial-mesenchymal transition (EMT) as examined by cell morphology and EMT markers; knockdown or inhibiting Nox1 led to a reversal of EMT. Supplementing MMP-2 to culture media did not induce EMT, suggesting that EMT induction by Nox1 was not through MMP-2 upregulation. In summary, Nox1 was overexpressed in all melanoma cell lines examined, and enhanced cell invasion by MMP-2 upregulation and EMT induction.

Inpatient rehabilitation challenges in a quadrimembral amputee after bilateral hand transplantation.

Bilateral forearm and hand transplantation poses unique challenges especially in the setting of bilateral lower limb amputations. A 57-yr-old man with bilateral transradial amputations and bilateral transtibial amputations after remote streptococcal sepsis was admitted for inpatient rehabilitation because of severe debilitation after forearm/hand transplantations. He required 6 wks of bed rest to allow the healing of the allografts but developed profound deconditioning. Because of weight-bearing precautions and other complications such as femoral neurapraxia, he required the use of body weight-support apparatus to ambulate with lower limb prostheses, keeping weight off the allografts. He progressed to walking 600 ft using a platform-wheeled walker at a modified independent level, to climbing four stairs with minimal assistance, and to being able to toss a small football using his right hand, indicating improved flexor function in this hand. Tacrolimus levels were maintained without clinical evidence of acute rejection. Through an individualized therapy regimen, careful monitoring of the allografts and dedicated support staff, rehabilitation training of a previous quadrimembral amputee after bilateral hand transplantations can be successful.

The frequency of lymphedema in an adult spina bifida population.

OBJECTIVE: In the United States, there are more than 100,000 people with spina bifida. There have been very few studies to date documenting the occurrence of lymphedema in the spina bifida population, despite a case series in 2001 that suggested that the occurrence may be higher than in the general population. Currently, approximately 1 million people have lymphedema in the United States. The purpose of this study was to document the occurrence of lymphedema and associated medical factors in a regional adult spina bifida population. DESIGN: A total of 240 electronic medical records from the Adult Spina Bifida Clinic from January 2005 to August 2008 were retrospectively reviewed. Subjects were divided into two groups based on the presence or absence of lymphedema. χ² analyses were used to compare lymphedema groups with respect to history of medical comorbidities and ethnicity. Fisher exact tests were used to compare groups with respect to mobility status and the presence of power wheelchair seat functions. Mann-Whitney U tests were used to compare groups with respect to age, anatomic lesion level, employment level, and income. RESULTS: Twenty-two (9.2%) patients had lymphedema. Mean ± SD population age was 35.1 ± 11.1 yrs. Lymphedema was associated with a history of trauma (P = 0.044), cellulitis (P < 0.001), cancer (P = 0.038), obesity (P < 0.001), wounds (P < 0.001), hypertension (P = 0.036), higher lesion level spina bifida (P = 0.049), and mobility status (P = 0.007). Hypertension and obesity were present in 38.3% and 37.5% of the total study population, respectively. CONCLUSIONS: This is the first study to document the occurrence of lymphedema in a spina bifida patient population, which was almost 100 times higher than that in the general patient population. We also documented a high occurrence of hypertension and obesity in the total study population. These findings may help guide further prospective studies to more clearly delineate the risk factors for the development of lymphedema and to determine the appropriate therapies. Better screening, prevention and treatment algorithms are needed for hypertension and obesity in the spina bifida population.

Large-scale RT-PCR recovery of full-length cDNA clones.

Pseudogenes, alternative transcripts, noncoding RNA, and polymorphisms each add extensive complexity to the mammalian transcriptome and confound estimation of the total number of genes. Despite advanced algorithms for gene prediction and several large-scale efforts to obtain cDNA clones for all human open reading frames (ORFs), no single collection is complete. To enhance this effort, we have developed a high-throughput pipeline for reverse transcription PCR (RT-PCR) gene recovery. Most importantly, novel molecular strategies for improving RT-PCR yield of transcripts that have been difficult to isolate by other means and computational strategies for clone sequence validation have been developed and optimized. This systematic gene recovery pipeline allows both rescue of predicted human and rat genes and provides insight into the complexity of the transcriptome through comparisons with existing data sets.

Enfermedad de Behçet / Behçet´s disease

La enfermedad de Behçet (EB) es un desorden complejo, caracterizado por aftas orales, erupción en piel, artritis, manifestaciones oculares como uveitis, vasculitis y en algunos casos afección de sistema nervioso y grandes vasos. Son múltiples las teorías sobre su etiopatogenia, entre las que se resaltan la presencia del HLA-B51, agentes infecciosos, disregulación inmune, daño endotelial, entre otros. Se piensa que algunos agentes infecciosos pueden disparar una respuesta por parte de las células mononucleraes y endoteliales, que en un individuo susceptible por la presencia del HLA-B51, llevaría como paso final a la generación de una respuesta inflamatoria y finalmente la vasculitis. Los tratamientos son tan variados, como las manifestaciones clínicas, incluyen colchicina, talidomida, esteroides y agentes inmunosupresores, según la severidad del compromiso, y se requiere de un grupo interdisciplinario para llegar a un adecuado control de la enfermedad (AU)

Concatenation cDNA sequencing for transcriptome analysis.

We describe a high-throughput cDNA sequencing pipeline (http://www.hgsc.bcm.tmc.edu/projects/cdna) built in response to the emerging need for rapid sequencing of large cDNA collections. Using this strategy cDNA inserts are purified and joined through concatenation into large molecules. These 'pseudo-BACs' are subjected to random shotgun sequencing whereby the majority of cDNA inserts in the pool are sequenced. Using this concatenation cDNA sequencing platform, we have contributed more than 13000 full-length cDNA sequences from human and mouse to the Mammalian Gene Collection (MGC).

Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences.

The National Institutes of Health Mammalian Gene Collection (MGC) Program is a multiinstitutional effort to identify and sequence a cDNA clone containing a complete ORF for each human and mouse gene. ESTs were generated from libraries enriched for full-length cDNAs and analyzed to identify candidate full-ORF clones, which then were sequenced to high accuracy. The MGC has currently sequenced and verified the full ORF for a nonredundant set of >9,000 human and >6,000 mouse genes. Candidate full-ORF clones for an additional 7,800 human and 3,500 mouse genes also have been identified. All MGC sequences and clones are available without restriction through public databases and clone distribution networks (see http:mgc.nci.nih.gov).