Glucose transporter gene expression in rat conceptus during early organogenesis and exposure to insulin-induced hypoglycemic serum

We investigated the glucose transporter gene and protein expression during early organogenesis in the rat and in rat embryos cultured with hypoglycemic serum. Erythrocyte-type glucose transporter (GLUT-1) mRNA was expressed at a high level in embryos; peak levels were reached at days 10.5–11.5 and decreased as gestational age increased. In contrast, the insulin regulaable glucose transporter (GLUT-4) mRNA was not detected. The levels of GLUT-1 protein determined by Western blot analysis increased in parallel with expression of the glucose transporter (GLUT-1) gene and peak levels were observed on days 10.5 and 11.5, which correspond to the main periods of neural tube formation. Immunohistochemical staining of the embryo on day 10.5 showed that GLUT-1 protein was abundantly located in the tissue of neural tube. When embryos were cultured from day 9.5 to day 10.5 with insulin-induced hypoglycemic serum containing 2–3 mM glucose an increased frequency of anterior neural tube defects was observed in association with a significant reduction of the glycolytic flux. Increased levels of GLUT-1 mRNA and protein were not observed during the culture with hypoglycemic serum compared with the levels in embryos cultured in normal serum. Addition of insulin to normal serum (500 U/ml) did not affect the GLUT-1 mRNA and protein levels. GLUT-1 mRNA and protein are strongly expressed in the embryo during early organogenesis, especially in the tissues of the neural tube, and the expression of the glucose transporter did not increase in response to prolonged glycopenia. This may account for the vulnerability of embryogenesis to hypoglycemia during these critical developmental periods.     

Receptor-like cytoplasmic kinase MARIS functions downstream of CrRLK1L-dependent signaling during tip growth

Growing plant cells need to rigorously coordinate external signals with internal processes. For instance, the maintenance of cell wall (CW) integrity requires the coordination of CW sensing with CW remodeling and biosynthesis to avoid growth arrest or integrity loss. Despite the involvement of receptor-like kinases (RLKs) of the Catharanthus roseus RLK1-like (CrRLK1L) subfamily and the reactive oxygen species-producing NADPH oxidases, it remains largely unknown how this coordination is achieved. ANXUR1 (ANX1) and ANX2, two redundant members of the CrRLK1L subfamily, are required for tip growth of the pollen tube (PT), and their closest homolog, FERONIA, controls root-hair tip growth. Previously, we showed that ANX1 overexpression mildly inhibits PT growth by oversecretion of CW material, whereas pollen tubes of anx1 anx2 double mutants burst spontaneously after germination. Here, we report the identification of suppressor mutants with improved fertility caused by the rescue of anx1 anx2 pollen tube bursting. Mapping of one these mutants revealed an R240C nonsynonymous substitution in the activation loop of a receptor-like cytoplasmic kinase (RLCK), which we named MARIS (MRI). We show that MRI is a plasma membrane-localized member of the RLCK-VIII subfamily and is preferentially expressed in both PTs and root hairs. Interestingly, mri-knockout mutants display spontaneous PT and root-hair bursting. Moreover, expression of the MRI^R240C mutant, but not its wild-type form, partially rescues the bursting phenotypes of anx1 anx2 PTs and fer root hairs but strongly inhibits wild-type tip growth. Thus, our findings identify a novel positive component of the CrRLK1L-dependent signaling cascade that coordinates CW integrity and tip growth.Growing plant cells are in constant communication with their environment, monitoring external signals that lead to internal reactions. Because cellular growth depends on a tight coordination of external and internal processes, signaling between the extracellular matrix, i.e., the primary cell wall (CW), and the internal growth machinery plays a central role in its regulation. The turgor pressure-resisting CW that shields plant cells from a changing environment exhibits remarkable but seemingly contradictory properties: rigidity and extensibility. Growing cells must find a balance between loosening their CWs sufficiently to allow expansion but not so much that they lose CW integrity. Because any environmental perturbation affecting the properties of the CW can upset this fragile balance, plant cells have developed complex sensing mechanisms to coordinate the state of the CW with the internal growth machinery (1, 2). These CW sensing mechanisms must be particularly robust in fast-growing cells, such as the tip-growing root hairs and pollen tubes (PTs), the male gametophytes of flowering plants. PTs elongate rapidly and grow over long distances within female tissues to deliver the sperm cells to the female gametophytes, which are deeply embedded in the ovules. Considering the vast excess of pollen grains that germinate on a receptive stigma, there is strong competition, and PTs must grow as fast as possible to reach unfertilized ovules; otherwise they will not contribute to the next generation. However, PTs must ensure that, while elongating extremely rapidly, they do not lose their integrity. In PTs of the model plant Arabidopsis thaliana, the plasma membrane-localized ANXUR (ANX1 and ANX2) receptor-like kinases (RLKs) play a major role in controlling CW integrity and growth. On one hand, loss of function of the redundant RLKs ANX1 and ANX2 leads to precocious PT rupture shortly after germination, resulting in male sterility (3, 4). This phenotype also is seen when the ANX-RLKs are degraded by the endoplasmic reticulum-associated degradation pathway in the turan mutant affecting N-glycosylation (5). On the other hand, overexpression of ANX-RLKs inhibits pollen germination and PT elongation, most likely because of the oversecretion of CW material (6). The ANX-RLKs and their closest homolog FERONIA (FER) belong to the Catharanthus roseus RLK1-like (CrRLK1L) subfamily that has been investigated extensively as putative CW sensors involved in coordinating cell growth, cell–cell communication, defense against pathogens, and hormone signaling as well as CW remodeling and integrity (reviewed in refs. 7–9). FER, the most thoroughly studied member of this subfamily, controls many developmental processes, such as intercellular communication during fertilization (10–12), cell elongation (13), calcium and hormone signaling (10, 14, 15), mechanosensing (16), plant defense (17, 18), and growth control of root hairs (19). Despite the many reports describing the role of CrRLK1Ls in various signaling processes, the mechanistic basis of their function and their relationship with other pathways remain poorly understood. FER was recently found to mediate the inhibition of primary root growth by forming a receptor–ligand pair with the rapid alkalinization factor 1 (RALF1) peptide (20). Moreover, it was shown that the intracellular kinase domains of three CrRLK1Ls members are interchangeable and that the various CrRLK1Ls share downstream signaling components (21). This finding has been confirmed for at least one signaling component so far, namely the reactive oxygen species (ROS)-producing NADPH oxidases, also called “respiratory burst oxidase homologs” (Rboh). It was reported that RbohD/RbohF for THESEUS1 (THE1), RbohC for FER, and RbohH/RbohJ for ANX1/2 act downstream of the CrRLK1Ls in primary roots, root hairs, and PTs, respectively (6, 19, 22).Here, we report, based on an anx1 anx2 male sterility suppressor screen, the identification of the receptor-like cytoplasmic kinase (RLCK) MARIS (MRI), a novel component of the CrRLK1L-mediated signaling pathway in tip-growing cells. MRI is expressed preferentially in PTs and root hairs, and its disruption triggers spontaneous bursting of PTs and root hairs. The suppressor mutant carries a R240C amino acid substitution in the activation loop of MRI. Expression of MRI^R240C partially rescues the bursting phenotypes of anx1 anx2 and rbohH rbohJ PTs as well as fer root hairs while strongly inhibiting WT tip growth, indicating that this mutant form of MRI overactivates the CrRLK1L-dependent pathway downstream of the NADPH oxidases.     

Abridged republication of FIGO's staging classification for cancer of the ovary,fallopian tube,and peritoneum

Ovarian, fallopian tube, and peritoneal cancers have a similar clinical presentation and are treated similarly, and current evidence supports staging all 3 cancers in a single system. The primary site (i.e. ovary, fallopian tube, or peritoneum) should be designated where possible. The histologic type should be recorded. Intraoperative rupture (“surgical spill”) is IC1; capsule ruptured before surgery or tumor on ovarian or fallopian tube surface is IC2; and positive peritoneal cytology with or without rupture is IC3. The new staging includes a revision of stage III patients; assignment to stage IIIA1 is based on spread to the retroperitoneal lymph nodes without intraperitoneal dissemination. Extension of tumor from omentum to spleen or liver (stage IIIC) should be differentiated from isolated parenchymal metastases (stage IVB). Cancer 2015;121:3435–43. © 2015 American Cancer Society.     

To Assess the Efficacy of Modality Slow Motion Dynamic Video Endoscopy of Eustachian Tube in Chronic Middle Ear Pathologies

Eustachian tube dysfunction (ETD) has a role in chronic otitis media (COM) is a well known fact and number of modalities have come up for its assessment—slow motion dynamic videoendoscopy (SMDV) being the latest. (i) To find prevalence of ETD in COM and (ii) to check the efficacy of SMDV in detecting dysfunction as compared to gold standard tympaometry (TYM). It’s a prospective analytical study with total of 100 patients of chronic otitis media, hence 200 ears were tested for ETD by TYM and SMDV. Prevalence of ETD was noted as 35.5 % in COM whereas the two testing modalities TYM versus SMDV showed agreement in 161 cases out of the 200 ears giving a moderate agreement of statistical significance (Kappa test) suggesting SMDV to be included in standard of protocol when assessing ETD along with TYM.     

Feeding difficulties in craniofacial microsomia: A multicenter retrospective analysis of 755 patients

A retrospective cohort study was initiated to analyse the prevalence, risk factors and treatment modalities of feeding difficulties in patients with craniofacial microsomia. This study included 755 subjects with craniofacial microsomia from three craniofacial centres. Medical charts were reviewed for severity of the deformity, documented feeding difficulties, age at which feeding difficulties first presented and treatment, presence of cleft lip/palate, extracraniofacial anomalies, and obstructive sleep apnoea. In total, 199 patients (26.4%) had documented feeding difficulties. Patients with bilateral involvement, Pruzansky-Kaban III classification, cleft lip/palate, or obstructive sleep apnoea were significantly more at risk for developing feeding difficulties and significantly more often needed additional feeding via a nasogastric tube than patients without these risk factors.     

术前置胃管时间对全麻胃肠道手术患者皮质醇的影响

目的观察术前置胃管时间对全麻胃肠道手术患者皮质醇的影响。方法将40例胃肠道手术需要置胃管的患者随机分为两组,各20例。对照组于术晨病房常规置胃管,观察组于全麻诱导后置胃管。每组分别监测术晨,麻醉诱导前,气管插管后2min及5min时平均动脉压、心率及血清皮质醇。结果两组不同时段各观察指标比较,干预主效应均P<0.05。结论将手术患者置胃管时间放在麻醉诱导后进行能减少患者的不适感,避免血压、心率及皮质醇急剧升高,减轻应激反应。     

Safety and Efficacy of Video Laparoscopic Surgical Debulking of Recurrent Ovarian,Fallopian Tube,and Primary Peritoneal Cancers

Background and Objective:

Studies on the role of laparoscopy in secondary or tertiary cytoreduction for recurrent ovarian cancer are limited. Our objective is to describe our preliminary experience with laparoscopic secondary/tertiary cytoreduction in patients with recurrent ovarian, fallopian, and primary peritoneal cancers.

Methods:

This is a retrospective analysis of a prospective case series. Women with recurrent ovarian, fallopian tube, or primary peritoneal cancers deemed appropriate candidates for laparoscopic debulking by the primary surgeon(s) were recruited. The patients underwent exploratory video laparoscopy, biopsy, and laparoscopic secondary/tertiary cytoreduction between June 1999 and October 2009. Variables analyzed include stage, site of disease, extent of cytoreduction, operative time, blood loss, length of hospital stay, complications, and survival time.

Results:

Twenty-three patients were recruited. Only one surgery involved conversion to laparotomy. Seventeen (77.3%) of the patients had stage IIIC disease at the time of their initial diagnosis, and 20 (90.9%) had laparotomy for primary debulking. Median blood loss was 75 mL, median operative time 200 min, and median hospital stay 2 d. No intraoperative complications occurred. One patient (4.5%) had postoperative ileus. Eighteen (81.8%) of the patients with recurrent disease were optimally cytoreduced to < 1cm. Overall, 12 patients have no evidence of disease (NED), 6 are alive with disease (AWD), and 4 have died of disease (DOD), over a median follow-up of 14 mo. Median disease-free survival was 71.9 mo.

Conclusions:

In a well-selected population, laparoscopy is technically feasible and can be utilized to optimally cytoreduce patients with recurrent ovarian, fallopian, or primary peritoneal cancers.