Literature Highlights.

Literature Highlights is a digest of notable papers recently published in the leading respiratory journals. Coverage includes shorter regimen for TB treatment; mapping the geographical evolution of TB incidence; diagnostic packages for active case finding for TB; TB burden estimation in settings with high levels of HIV; and digital approaches for TB treatment adherence.

Antifungal effect of (R) and (S)-citronellal enantiomers and their predictive mechanism of action on Candida albicans from voriconazole-resistant onychomycoses.

Onychomycosis is the most common disease affecting the nail unit and accounts for at least 50% of all nail diseases. In addition, Candida albicans is responsible for approximately 70% of onychomycoses caused by yeasts. This study investigated the antifungal effect of (R) and (S)-citronellal enantiomers, as well as its predictive mechanism of action on C. albicans from voriconazole-resistant onychomycoses. For this purpose, in vitro broth microdilution and molecular docking techniques were applied in a predictive and complementary manner to the mechanisms of action. The main results of this study indicate that C. albicans was resistant to voriconazole and sensitive to the enantiomers (R) and (S)-citronellal at a dose of 256 and 32 µg/mL respectively. In addition, there was an increase in the minimum inhibitory concentration (MIC) of the enantiomers in the presence of sorbitol and ergosterol, indicating that these molecules possibly affect the integrity of the cell wall and cell membrane of C. albicans. Molecular docking with key biosynthesis proteins and maintenance of the fungal cell wall and plasma membrane demonstrated the possibility of (R) and (S)-citronellal interacting with two important enzymes: 1,3-ß-glucan synthase and lanosterol 14α-demethylase. Therefore, the findings of this study indicate that the (R) and (S)-citronellal enantiomers are fungicidal on C. albicans from onychomycoses and probably these substances cause damage to the cell wall and cell membrane of these micro-organisms possibly by interacting with enzymes in the biosynthesis of these fungal structures.

Literature Highlights.

Literature Highlights is a digest of notable papers recently published in the leading respiratory journals. Coverage includes clinical trials to investigate the diagnostic and clinical effect of trial of antibiotics on TB; a Phase 3 trial to assess if glucocorticoids decrease mortality among patients with pneumonia; a Phase 2 trial on pretomanid use for treating drug-susceptible TB; contact investigation for TB in China; and post-TB sequelae after TB treatment in children.

Prevalence and clinical characteristics of pulmonary TB among pregnant and post-partum women.

SETTING: Antenatal care (ANC) and postpartum care (PPC) clinic in Manhiça District, Mozambique.OBJECTIVE: To estimate the prevalence of TB among pregnant and post-partum women and describe the clinical characteristics of the disease in a rural area of Southern Mozambique.METHODS: We conducted a cross-sectional TB prevalence study among pregnant and post-partum women recruited from September 2016 to March 2018 at the Manhiça Health Care Center (MHC). We recruited two independent cohorts of women consecutively presenting for routine pregnancy or post-partum follow-up visits.RESULTS: A total of 1,980 women from the ANC clinic and 1,010 from the PPC clinic were enrolled. We found a TB prevalence of 505/100,000 (95% CI: 242-926) among pregnant women and 297/100,000 (95% CI: 61-865) among post-partum women. Among HIV-positive pregnant women, TB prevalence was 1,626/100,000 (95% CI: 782-2,970) and among postpartum HIV-positive women, TB prevalence was 984/100,000 (95% CI: 203-2,848).CONCLUSIONS: The burden of TB was not higher in postpartum women than in pregnant women. Most TB cases were detected in HIV-positive women. TB screening and diagnostic testing among pregnant and postpartum women attending ANC and PPC clinics in Manhiça District is acceptable and feasible.

Breast and ovarian cancer predisposition due to de novo BRCA1 and BRCA2 mutations.

BRCA1 and BRCA2 are the two major genes predisposing to breast and ovarian cancer. Whereas high de novo mutation rates have been demonstrated for several genes, only 11 cases of de novo BRCA1/2 mutations have been reported to date and the BRCA1/2 de novo mutation rate remains unknown. The present study was designed to fill this gap based on a series of 12 805 consecutive unrelated patients diagnosed with breast and/or ovarian cancer who met the inclusion criteria for BRCA1/2 gene analysis according to French guidelines. BRCA1/2 mutations were detected in 1527 (12%) patients, and three BRCA1 mutations and one BRCA2 mutation were de novo. The BRCA1/2 de novo mutation rate was estimated to be 0.3% (0.1%; 0.7%). Although rare, it may be useful to take the possibility of de novo BRCA1/2 mutation into account in genetic counseling of relatives and to improve the understanding of complex family histories of breast and ovarian cancers.

Cana-de-açúcar em substituição ao feno de capim-tifton 85 em rações para cabras Saanen / Sugar cane replacing grass hay tifton 85 in rations for Saanen goats

Avaliou-se o consumo e digestibilidade dos nutrientes em dietas para cabras da raça Saanen arranjadas em um quadrado latino 5x5, alimentadas com cana-de-açúcar em substituição (0, 25, 50, 75 e 100%) ao feno de capim-tifton 85. O experimento foi constituído de cinco períodos de 15 dias (10 dias para adaptação dos animais às dietas experimentais e 5 dias para coleta de dados e amostras). O consumo voluntário foi calculado pela diferença entre o ofertado e as sobras. Utilizou-se o óxido crômico para estimativa da produção de matéria seca fecal. A digestibilidade aparente da matéria seca, matéria orgânica, extrato etéreo e fibra em detergente neutro foi influenciada negativamente com a substituição, enquanto a digestibilidade da proteína e dos carboidratos não fibrosos não sofreu influência. O baixo consumo de matéria seca de rações com cana-de-açúcar torna-se um fator limitante para cabras de média a alta produção de leite.

The intake and digestibility of nutrients in the diets of Saanen goats arranged in a 5x5 Latin square design, fed with sugar cane replacement (0, 25, 50, 75 and 100%) hay Tifton 85 were evaluated. The experiment consisted of 5 periods of 15 days (10 days for animal adaptation to experimental diets and 5 days for data collection). The voluntary intake was calculated as the difference between the offered and leftovers. Digestibility chromic oxide was used to estimate the fecal dry matter production. The digestibility of dry matter, organic matter, ether extract and neutral detergent fiber was negatively influenced, whereas the digestibility of protein and non-fiber carbohydrates were not affected. The low dry matter intake of diets with sugar cane becomes a limiting factor for goats medium for high milk production.

Effects of Bermudagrass hay and soybean hulls inclusion on performance of sheep fed cactus-based diets.

The objective of this experiment was to determine the effects of replacing corn with soybean hulls (SH) or Bermudagrass tifton hay (TH) on performance of sheep fed cactus-based diets. Three ruminally fistulated sheep were used in a 3 x 3 Latin square experiment with 21-day periods. All diets contained 75% spineless cactus (dry matter basis, DM) and formulated to be isonitrogenous. Fiber source had no influence on nutrient intakes except for the intake of neutral detergent fiber (NDF) which was lower (p<0.05) for animals fed corn relative to those fed SH or TH. Time expended in rumination and total chewing time were higher (p<0.05) for animals fed TH than those fed SH or corn. In vivo nutrient digestibilities were similar for all dietary treatments and averaged 69.6%, 74.8%, 69.9%, and 61.8% for DM, organic matter, crude protein, and NDF, respectively. Feeding SH relative to TH and corn decreased ruminal pH (p<0.05) and increased concentration of total volatile fatty acids (p<0.05). However, ruminal NH3-N concentration was higher (p<0.05) for animal fed TH than for those fed SH or corn. Abdominal distension and ruminal biofilm production were greater (p<0.05) in animals fed corn or SH than in those fed TH. It was concluded that replacing corn with SH or TH up to 15% of the diet DM in a cactus-based diet had no effect on nutrient intakes or total tract nutrient utilization. Changes in ruminal fermentation parameters reflected differences in ruminal degradability between the two fiber sources. Bermudagrass tifton hay was more effective than SH in reducing the risk of bloat associated with feeding high levels of spineless cactus to ruminants.

Syndromes of disordered chromatin remodeling.

Syndromes of disordered 'chromatin remodeling' are unique in medicine because they arise from a general deregulation of DNA transcription caused by mutations in genes encoding enzymes which mediate changes in chromatin structure. Chromatin is the packaged form of DNA in the eukaryotic cell. It consists almost entirely of repeating units, called nucleosomes, in which short segments of DNA are wrapped tightly around a disk-like structure comprising two subunits of each of the histone proteins H2A, H2B, H3 and H4. Histone proteins are covalently modified by a number of different adducts (i.e. acetylation and phosphorylation) that regulate the tightness of the DNA-histone interactions. Mutations in genes encoding enzymes that mediate chromatin structure can result in a loss of proper regulation of chromatin structure, which in turn can result in deregulation of gene transcription and inappropriate protein expression. In this review we present examples of representative genetic diseases that arise as a consequence of disordered chromatin remodeling. These include: alpha-thalassemia/mental retardation syndrome, X-linked (ATR-X); Rett syndrome (RS); immunodeficiency-centromeric instability-facial anomalies syndrome (ICF); Rubinstein-Taybi syndrome (RSTS); and Coffin-Lowry syndrome (CLS).

Identification of Bacillus thuringiensis subsp. kurstaki strain HD1-Like bacteria from environmental and human samples after aerial spraying of Victoria, British Columbia, Canada, with Foray 48B.

Aerial applications of Foray 48B, which contains Bacillus thuringiensis strain HD1, were carried out on 9 to 10 May, 19 to 21 May, and 8 to 9 June 1999 to control European gypsy moth (Lymantria dispar) populations in Victoria, British Columbia, Canada. A major assessment of the health impact of B. thuringiensis subsp. kurstaki was conducted by the Office of the Medical Health Officer of the Capital Health Region during this period. Environmental (air and water) and human (nasal swab) samples, collected before and after aerial applications of Foray 48B, both in the spray zone and outside of the spray zone, were analyzed for the presence of strain HD1-like bacteria. Random amplified polymorphic DNA analysis, cry gene-specific PCR, and dot blot DNA hybridization techniques were used to screen over 11,000 isolates of bacteria. We identified bacteria with genetic patterns consistent with those of B. thuringiensis subsp. kurstaki HD1 in 9,102 of 10,659 (85.4%) isolates obtained from the air samples, 13 of 440 (2.9%) isolates obtained from the water samples, and 131 of 171 (76.6%) isolates from the nasal swab samples. These analyses suggest that B. thuringiensis subsp. kurstaki HD1-like bacteria were present both in the environment and in the human population of Victoria prior to aerial applications of Foray 48B. The presence of B. thuringiensis subsp. kurstaki HD1-like bacteria in human nasal passages increased significantly after the application of Foray 48B, both inside and outside the spray zone.

Selectively labeling the heterologous protein in Escherichia coli for NMR studies: a strategy to speed up NMR spectroscopy.

Nuclear magnetic resonance is an important tool for high-resolution structural studies of proteins. It demands high protein concentration and high purity; however, the expression of proteins at high levels often leads to protein aggregation and the protein purification step can correspond to a high percentage of the overall time in the structural determination process. In the present article we show that the step of sample optimization can be simplified by selective labeling the heterologous protein expressed in Escherichia coli by the use of rifampicin. Yeast thioredoxin and a coix transcription factor Opaque 2 leucine zipper (LZ) were used to show the effectiveness of the protocol. The (1)H/(15)N heteronuclear correlation two-dimensional NMR spectrum (HMQC) of the selective (15)N-labeled thioredoxin without any purification is remarkably similar to the spectrum of the purified protein. The method has high yields and a good (1)H/(15)N HMQC spectrum can be obtained with 50 ml of M9 growth medium. Opaque 2 LZ, a difficult protein due to the lower expression level and high hydrophobicity, was also probed. The (15)N-edited spectrum of Opaque 2 LZ showed only the resonances of the protein of heterologous expression (Opaque 2 LZ) while the (1)H spectrum shows several other resonances from other proteins of the cell lysate. The demand for a fast methodology for structural determination is increasing with the advent of genome/proteome projects. Selective labeling the heterologous protein can speed up NMR structural studies as well as NMR-based drug screening. This methodology is especially effective for difficult proteins such as hydrophobic transcription factors, membrane proteins, and others.

Detection of p53 mutations in benign and dysplastic nevi.

We have examined melanocytic cells derived directly from fresh biopsy tissue for the presence of p53 mutations. Using selective media that permits growth of melanocytes and inhibits growth of fibroblasts and keratinocytes, we established short-term, primary cultures of melanocytes from skin biopsies of common acquired nevi, dysplastic nevi, and from metastatic melanoma. Using PCR-single-stranded conformational polymorphism analysis, we have detected p53 mutations in 2 of 11 benign compound nevi and 2 of 5 dysplastic nevi. All nevi positive for p53 mutations were derived from patients who previously had cutaneous moles and three of the four had a family and/or personal history of melanoma.