Metabolic and chromosomal changes in a Bacillus subtilis whiA mutant.

IMPORTANCE: WhiA is a conserved DNA-binding protein that influences cell division in many Gram-positive bacteria and, in B. subtilis, also chromosome segregation. How WhiA works in Bacillus subtilis is unknown. Here, we tested three hypothetical mechanisms using metabolomics, fatty acid analysis, and chromosome confirmation capture experiments. This revealed that WhiA does not influence cell division and chromosome segregation by modulating either central carbon metabolism or fatty acid composition. However, the inactivation of WhiA reduces short-range chromosome interactions. These findings provide new avenues to study the molecular mechanism of WhiA in the future.

Leiomioma vascular que afecta el labio: reporte de un caso y un análisis actualizado de los 78 casos reportados en la literature / Vascular leiomyoma affecting the lip: report of a case and an updated analysis of the 78 cases reported in the literature / Leiomioma vascular que afeta o lábio: relato de um caso e análise atualizada dos 78 casos relatados na literatura

Resumen El leiomioma vascular (LV) es un tumor benigno de tejido blando que rara vez se observa en los tejidos orales. El objetivo de este trabajo es presentar un nuevo caso de leiomioma vascular del labio (VLL), describiendo sus características clínicas, microscópicas e inmunohistoquímicas, y revisar la literatura sobre el tumor. Una mujer de 27 años presentó una pequeña lesión dolorosa en la mucosa del labio inferior. La biopsia escisional reveló una masa encapsulada bien circunscrita formada por células pequeñas y fusiformes. El análisis de inmunohistoquímica reveló una expresión intensa y difusa de actina del músculo liso (AME) dentro de las células tumorales y la inmunorreactividad CD34 de las células endoteliales que recubren los espacios vasculares, lo que indica la presencia de vasos sanguíneos. Se estableció un diagnóstico final de VLL. No se observaron signos de recurrencia después de cinco años de seguimiento. Aunque VLL es una lesión rara, debe considerarse en el diagnóstico diferencial de las lesiones nodulares en los labios inferior y superior.

Abstract Vascular Leiomyoma (VL) is a benign soft tissue tumor rarely observed in oral tissues. The aim of this paper is to present a new case of vascular leiomyoma of the lip (VLL), describing its clinical, microscopical, and immunohistochemical features, and review the literature on the tumor. A 27-year-old woman presented with a small painful lesion on lower lip mucosa. Excisional biopsy revealed a well-circumscribed, encapsulated mass formed by small and fusiform cells. Immunohistochemistry analysis revealed intense and diffuse expression of smooth muscle actin (SMA) within the tumor cells and CD34 immunoreactivity of the endothelial cells lining the vascular spaces, indicating the presence of blood vessels. A final diagnosis of VLL was established. No signs of recurrence were observed after five years of follow-up. Although VLL is a rare lesion, it must be considered in the differential diagnosis of nodular lesions on lower and upper lips.

Resumo O Leiomioma Vascular é um tumor benigno de tecidos moles raramente observado nos tecidos da cavidade bucal. O objetivo desse artigo é apresentar um novo caso de um leiomioma vascular no lábio (LVL), descrevendo seus achados clínicos, microscópicos e imunoistoquímicos, e revisar a literatura sobre o tumor. Uma mulher de 27 anos de idade se apresentou com uma pequena lesão dolorosa na mucosa do lábio inferior. A biópsia excisional revelou um massa encapsulada e bem circunscrita, formada por células pequenas e fusiformes. A análise imunoistoquímica revelou expressão intensa e difusa de actina musculo liso (AML) nas células tumorais e imunorreatividade para CD34 em células endoteliais que revestem espaços vasculares, indicando a presença de vasos sanguíneos. O diagnóstico final de LVL foi estabelecido. Não foram observados sinais de recorrência após cinco anos de acompanhamento. Embora o LVL seja uma lesão rara, deve ser considerada no diagnóstico diferencial de lesões nodulares nos lábios inferior e superior.

The experience and outcomes of a specialised preterm birth clinic in New Zealand.

BACKGROUND: A greater understanding of the risk factors for spontaneous preterm birth and the importance of risk stratification to guide interventions has led to the introduction of preterm birth prevention clinics. AIM: To evaluate the experience and outcomes of the first specialised preterm birth clinic in New Zealand. MATERIALS AND METHODS: This observational study reviewed pregnancies cared for in a preterm birth clinic from 2013 to 2018. Cases were identified and data collected from a maternity database and electronic medical records. Analysis was by referral type. RESULTS: A total of 423 cases were included; 309 elective and 22 acute referrals in pregnancy, and 92 consultations outside pregnancy. For those referred electively in pregnancy, 138/309 (44.7%) fulfilled multiple referral criteria, and 57/309 (18.4%) had ≥2 previous spontaneous preterm births or second trimester losses. Excluding five pregnancies with first trimester miscarriage, 77/304 (25.3%) were managed with a history-indicated cerclage (11 placed pre-conception) and 217/304 (71.4%) had cervical surveillance as primary management, of which 133 (61.3%) did not require treatment. The remaining had treatment for a short cervix; 37 (17.0%) received an ultrasound-indicated cerclage only, 21 (9.7%) vaginal progesterone only and 26 (12.0%) both. Five women (1.6%) had a second trimester loss at 13+0 -19+6 and 58/297 (19.5%) had a spontaneous preterm birth at 20+0 -36+6  weeks. The 'take home baby' rate was 95.4%. CONCLUSIONS: Pregnancy outcomes were similar to those reported by other preterm birth prevention clinics. The majority of women who received cervical surveillance as primary management were able to avoid additional treatment.

Mitotic instability in triploid and tetraploid one-year-old eastern oyster, Crassostrea virginica, assessed by cytogenetic and flow cytometry techniques.

For commercial oyster aquaculture, triploidy has significant advantages. To produce triploids, the principal technology uses diploid × tetraploid crosses. The development of tetraploid brood stock for this purpose has been successful, but as more is understood about tetraploids, it seems clear that chromosome instability is a principal feature in oysters. This paper is a continuation of work to investigate chromosome instability in polyploid Crassostrea virginica. We established families between tetraploids-apparently stable (non-mosaic) and unstable (mosaic)-and normal reference diploids, creating triploid groups, as well as tetraploids between mosaic and non-mosaic tetraploids. Chromosome loss was about the same for triploid juveniles produced from either mosaic or non-mosaic tetraploids or from either male or female tetraploids. However, there was a statistically significant difference in chromosome loss in tetraploid juveniles produced from mosaic versus non-mosaic parents, with mosaics producing more unstable progeny. These results confirm that chromosome instability, as manifested in mosaic tetraploids, is of little concern for producing triploids, but it is clearly problematic for tetraploid breeding. Concordance between the results from cytogenetics and flow cytometry was also tested for the first time in oysters, by assessing the ploidy of individuals using both techniques. Results between the two were non-concordant.

Sensory and analytical characterization of the "cool-melting" perception of commercial spreads.

Butters, margarines and table spreads are water-in-oil emulsions. Melting characteristics of these products are important for flavor release and consumer acceptance. One characteristic that is believed to discriminate butters from margarines is a cooling sensation perceived in-mouth while consuming these products. Here, we investigated different methods to characterize sensorically and analytically the "cool-melting" properties of commercial butter and margarines. Our results show that butter indeed can be distinguished from margarines based on their "cool-melting" properties. Furthermore, changes in enthalpy as measured through DSC and solid fat content are good predictors of the "cool-melting" effect of spreads. PRACTICAL APPLICATIONS: By understanding the mechanisms of the "cool-melting" perception of spreads, and linking them to analytical measurements, we can create an in-vitro quantification method of "cool-melting." This method can eventually help directing product development to achieve the desire product profile and increase consumer acceptance and liking of margarines and low-fat spread products. In this study we did not assess the impact of "cool-melting" on consumer perception, which would be the next step in understanding the drivers of liking of spread products.

Genetic engineering of the branched fatty acid metabolic pathway of Bacillus subtilis for the overproduction of surfactin C14 isoform.

Surfactin, a lipopeptide produced by Bacillus subtilis, is one of the most powerful biosurfactants known. This molecule consists of a cyclic heptapeptide linked to a ß-hydroxy fatty acid chain. The isomery and the length of the fatty acid (FA) chain are responsible for the surfactin's activities. In this study, the gene codY, which encode for the global transcriptional regulator and the gene lpdV, located in the bkd operon (lpdV, bkdAA, bkdAB and bkdB genes), which is responsible for the last step of the branched chain amino acid (BCAA) degradation in acyl-CoA were deleted. The influence of these deletions on the quantitative and qualitative surfactin production was analysed. The surfactin production was quantified by RP-HPLC and the surfactin isoforms were characterized using LC-MS-MS and GC-MS analysis. The results obtained in the mutants showed an enhancement of surfactin specific production by a factor of 5.8 for the codY mutant and 1.4 for lpdV mutant. Moreover qualitative analysis of the lpdV mutant reveals that it mainly produced surfactin C14 isoform (2 fold more than the wild type) with linear FA chain. Complete analysis of the extracellular metabolites using 1 H quantitative NMR reveals a reduced production of acetoin in this mutant. This work demonstrates for the first time an original approach to overproduce specifically surfactin with C14 FA chain.

Aneuploid progeny of the American oyster, Crassostrea virginica, produced by tetraploid × diploid crosses: another example of chromosome instability in polyploid oysters.

The commercial production of triploids, and the creation of tetraploid broodstock to support it, has become an important technique in aquaculture of the eastern oyster, Crassostrea virginica. Tetraploids are produced by cytogenetic manipulation of embryos and have been shown to undergo chromosome loss (to become a mosaic) with unknown consequences for breeding. Our objective was to determine the extent of aneuploidy in triploid progeny produced from both mosaic and non-mosaic tetraploids. Six families of triploids were produced using a single diploid female and crossed with three mosaic and non-mosaic tetraploid male oysters. A second set of crosses was performed with the reciprocals. Chromosome counts of the resultant embryos were tallied at 2-4 cell stage and as 6-hour(h)-old embryos. A significant level of aneuploidy was observed in 6-h-old embryos. For crosses using tetraploid males, aneuploidy ranged from 53% to 77% of observed metaphases, compared to 36% in the diploid control. For crosses using tetraploid females, 51%-71% of metaphases were aneuploidy versus 53% in the diploid control. We conclude that somatic chromosome loss may be a regular feature of early development in triploids, and perhaps polyploid oysters in general. Other aspects of chromosome loss in polyploid oysters are also discussed.

Megacystis Microcolon Intestinal Hypoperistalsis Syndrome: Case Reports and Discussion of the Literature.

Megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS) is a rare intestinal dysmotility condition that also involves a dilated urinary bladder. It was believed to be an autosomal recessive condition, but genetic studies have suggested possibly an autosomal dominant inheritance pattern. Prenatal diagnosis can be challenging, but MRI and amniotic fluid/digestive fluid studies may be complementary investigations to improve diagnostic accuracy. Prognosis of MMIHS is generally poor and treatment is mostly supportive. To date, bowel transplantation remains the only viable treatment to restore bowel motility. Here we present two additional cases to contribute towards the scant literature on this condition.

Insights into molecular features of Venerupis decussata oocytes: a microarray-based study.

The production of Venerupis decussata relies on wild seed collection, which has been recently compromised due to recruitment failure and severe mortalities. To address this issue and provide an alternative source of seed, artificial spawning and larval rearing programs were developed. However, hatchery-based seed production is a relatively new industry and it is still underdeveloped. A major hurdle in the European clam seed production is the control of spawning and reproduction, which is further hindered by the impossibility of obtaining fertile gametes by gonadal "stripping", as meiosis re-initiation is constrained to a maturation process along the genital ducts. In the present study, oocytes were collected from 15 females and microarray analyses was performed to investigate gene expression profiles characterizing released and stripped ovarian oocytes. A total of 198 differentially expressed transcripts between stripped and spawned oocytes were detected. Functional analysis carried out on these transcripts highlighted the importance of a few biological processes, which are most probably implicated in the control of oocyte competence. Significant differences were observed for transcripts encoding proteins involved in meiosis progression (e.g. dual specificity phosphatase CDC25), WNT signalling (e.g. frizzled class receptor 8, wingless-type MMTV integration site family member 4), steroid synthesis (e.g. progestin and adipoQ receptor family member 3, cytochrome P450-C17), mRNA processing (e.g. zinc finger protein XlCOF28), calcium regulation (e.g. regucalcin, calmodulin) and ceramide metabolism (ceramidase B, sphingomyelinase). This study provides new information on transcriptional profiles putatively associated with ovarian egg infertility, and suggests potential mechanisms regulating early oocyte development in clams. Genes which were differentially expressed between stripped and spawned oocytes might have a pivotal role during maturation process in the gonadal duct and could be interesting targets for further functional studies aiming to make ovarian oocytes fertilizable.

A microarray-based analysis of gametogenesis in two Portuguese populations of the European clam Ruditapes decussatus.

The European clam, Ruditapes decussatus is a species with a high commercial importance in Portugal and other Southern European countries. Its production is almost exclusively based on natural recruitment, which is subject to high annual fluctuations. Increased knowledge of the natural reproductive cycle of R. decussatus and its molecular mechanisms would be particularly important in providing new highly valuable genomic information for better understanding the regulation of reproduction in this economically important aquaculture species. In this study, the transcriptomic bases of R. decussatus reproduction have been analysed using a custom oligonucleotide microarray representing 51,678 assembled contigs. Microarray analyses were performed in four gonadal maturation stages from two different Portuguese wild populations, characterized by different responses to spawning induction when used as progenitors in hatchery. A comparison between the two populations elucidated a specific pathway involved in the recognition signals and binding between the oocyte and components of the sperm plasma membrane. We suggest that this pathway can explain part of the differences in terms of spawning induction success between the two populations. In addition, sexes and reproductive stages were compared and a correlation between mRNA levels and gonadal area was investigated. The lists of differentially expressed genes revealed that sex explains most of the variance in gonadal gene expression. Additionally, genes like Foxl2, vitellogenin, condensing 2, mitotic apparatus protein p62, Cep57, sperm associated antigens 6, 16 and 17, motile sperm domain containing protein 2, sperm surface protein Sp17, sperm flagellar proteins 1 and 2 and dpy-30, were identified as being correlated with the gonad area and therefore supposedly with the number and/or the size of the gametes produced.

Evaluation of chronic hypertension in pregnant young women.

Women with chronic hypertension are at higher risk of adverse obstetric outcomes. It is essential that the condition is identified and evaluated appropriately in early pregnancy. Therefore, an audit has been carried out to assess how well young pregnant women with chronic hypertension were investigated for secondary cause in South Auckland, compared with the recommendations of the Australasian Society for the Study of Hypertension in Pregnancy. The evaluation of chronic hypertension by history taking, physical examination, laboratory assessment and radiology tests was highly variable. Only 76% of women had appropriate follow-up for their hypertension. Screening for secondary causes was not consistent, and the majority had incomplete investigation.

Diastereoselective synthesis of cularine alkaloids via enium ions and an easy entry to isoquinolines by aza-Wittig electrocyclic ring closure.

In preliminary communications, we reported the diastereoselective synthesis of cularine and sarcocapnine via the intramolecular ring closure of nitrenium and oxenium ions, a new highly diastereoselective reductive methylation with (+)-8-phenylmenthyl chloroacetate followed by reduction with sodium borohydride, and a facile entry to the isoquinoline precursors by aza-Wittig electrocyclic ring closure. We now report the full details of the syntheses of (+)-O-demethylcularine, (+)-cularine, (+)-sarcocapnidine, (+)-sarcocapnine, and (+)-crassifoline and describe different methods of synthesis of their precursors.