Possible anatomical base of snoring

Variations of the medial and inferior choanal limits may facilitate snoring. These are: 1) short vomer without crista choanalis vomeris and adaptational formations, 2) extremely concave posterior margin of the hard palate leading to disproportion between the muscular and aponeurotic part of m. tensor veli palatini and hence to a flabby soft palate especially during sleep, and 3) neonatal relation of the height and width of the choane resulting in a very large choanal region instead of relative reduction of choanal width compared to its increasing height. All these variations result in a flabby soft palate especially during sleep and may lead to formation of whirles of inspired air and snoring.     

Ricolinostat (ACY‐1215) induced inhibition of aggresome formation accelerates carfilzomib‐induced multiple myeloma cell death

Proteasome inhibition induces the accumulation of aggregated misfolded/ubiquitinated proteins in the aggresome; conversely, histone deacetylase 6 (HDAC6) inhibition blocks aggresome formation. Although this rationale has been the basis of proteasome inhibitor (PI) and HDAC6 inhibitor combination studies, the role of disruption of aggresome formation by HDAC6 inhibition has not yet been studied in multiple myeloma (MM). The present study aimed to evaluate the impact of carfilzomib (CFZ) in combination with a selective HDAC6 inhibitor (ricolinostat) in MM cells with respect to the aggresome‐proteolysis pathway. We observed that combination treatment of CFZ with ricolinostat triggered synergistic anti‐MM effects, even in bortezomib‐resistant cells. Immunofluorescent staining showed that CFZ increased the accumulation of ubiquitinated proteins and protein aggregates in the cytoplasm, as well as the engulfment of aggregated ubiquitinated proteins by autophagosomes, which was blocked by ricolinostat. Electron microscopy imaging showed increased autophagy triggered by CFZ, which was inhibited by the addition of ACY‐1215. Finally, an in vivo mouse xenograft study confirmed a decrease in tumour volume, associated with apoptosis, following treatment with CFZ in combination with ricolinostat. Our results suggest that ricolinostat inhibits aggresome formation, caused by CFZ‐induced inhibition of the proteasome pathway, resulting in enhanced apoptosis in MM cells.     

Needs analysis for developing a virtual-reality NOTES simulator

Introduction and study aim

Natural orifice translumenal endoscopic surgery (NOTES) is an emerging surgical technique that requires a cautious adoption approach to ensure patient safety. High-fidelity virtual-reality-based simulators allow development of new surgical procedures and tools and train medical personnel without risk to human patients. As part of a project funded by the National Institutes of Health, we are developing the virtual transluminal endoscopic surgery trainer (VTEST?) for this purpose. The objective of this study is to conduct a structured needs analysis to identify the design parameters for such a virtual-reality-based simulator for NOTES.

Methods

A 30-point questionnaire was distributed at the 2011 National Orifice Surgery Consortium for Assessment and Research meeting to obtain responses from experts. Ordinal logistic regression and the Wilcoxon rank-sum test were used for analysis.

Results

A total of 22 NOTES experts participated in the study. Cholecystectomy (CE, 68 %) followed by appendectomy (AE, 63 %) (CE vs AE, p = 0.0521) was selected as the first choice for simulation. Flexible (FL, 47 %) and hybrid (HY, 47 %) approaches were equally favorable compared with rigid (RI, 6 %) with p < 0.001 for both FL versus RI and HY versus RI. The transvaginal approach was preferred 3 to 1 to the transgastric. Most participants preferred two-channel (2C) scopes (65 %) compared with single (1C) or three (3C) or more channels with p < 0.001 for both 2C versus 1C and 2C versus 3C. The importance of force feedback and the utility of a virtual NOTES simulator in training and testing new tools for NOTES were rated very high by the participants.

Conclusion

Our study reinforces the importance of developing a virtual NOTES simulator and clearly presents expert preferences. The results of this analysis will direct our initial development of the VTEST? platform.     

Isolation of 10 differentially expressed cDNAs in p53-induced apoptosis: activation of the vertebrate homologue of the drosophila seven in absentia gene.

We report the isolation of 10 differentially expressed cDNAs in the process of apoptosis induced by the p53 tamor suppressor. As a global analytical method, we performed a differential display of mRNA between mouse M1 myeloid leukemia cells and derived clone LTR6 cells, which contain a stably transfected temperature-sensitive mutant of p53. At 32 degrees C wild-type p53 function is activated in LTR6 cells, resulting in programmed cell death. Eight genes are activated (TSAP; tumor suppressor activated pathway), and two are inhibited (TSIP, tumor suppressor inhibited pathway) in their expression. None of the 10 sequences has hitherto been recognized as part of the p53 signaling pathway. Three TSAPs are homologous to known genes. TSAP1 corresponds to phospholipase C beta 4. TSAP2 has a conserved domain homologous to a multiple endocrine neoplasia I (ZFM1) candidate gene. TSAP3 is the mouse homologue of the Drosophila seven in absentia gene. These data provide novel molecules involved in the pathway of wild-type p53 activation. They establish a functional link between a homologue of a conserved developmental Drosophila gene and signal transduction in tumor suppression leading to programmed cell death.     

Activation of the human homologue of the Drosophila sina gene in apoptosis and tumor suppression.

Developmentally regulated genes in Drosophila, which are conserved through evolution, are potential candidates for key functions in biological processes such as cell cycle, programmed cell death, and cancer. We report cloning and characterization of the human homologue of the Drosophila seven in absentia gene (HUMSIAH), which codes for a 282 amino acids putative zinc finger protein. HUMSIAH is localized on human chromosome 16q12-q13. This gene is activated during the physiological program of cell death in the intestinal epithelium. Moreover, human cancer-derived cells selected for suppression of their tumorigenic phenotype exhibit constitutively elevated levels of HUMSIAH mRNA. A similar pattern of expression is also displayed by the p21waf1. These results suggest that mammalian seven in absentia gene, which is a target for activation by p53, may play a role in apoptosis and tumor suppression.     

KIF1A‐related disorders in children: A wide spectrum of central and peripheral nervous system involvement

KIF1A‐related disorders (KRD) were first described in 2011 and the phenotypic spectrum has subsequently expanded to encompass a range of central and peripheral nervous system involvement. Here we present a case series demonstrating the range of clinical, neurophysiological, and radiological features which may occur in childhood‐onset KRD. We report on all the children and young people seen at a single large tertiary centre. Data were collected through a retrospective case‐notes review. Twelve individuals from 10 families were identified. Eight different mutations were present, including four novel mutations. Two patients displayed a very severe phenotype including congenital contractures, severe spasticity and/or dystonia, dysautonomia, severe sensorimotor polyneuropathy and optic atrophy, significant white matter changes on brain MRI, respiratory insufficiency, and complete lack of neurodevelopmental progress. The remaining 10 patients represented a spectrum of severity with common features including a movement disorder with spasticity and/or dystonia, subtle features of dysautonomia, sensory axonal neuropathy, varying degrees of optic atrophy and of learning and/or behavioural difficulties, and subtle or absent—but sometimes progressive—changes in white matter on MRI. Epilepsy was common among the more severely affected children. This case series demonstrates that KRD comprise a range of neurological disorders, with both the milder and the more severe forms combining central and peripheral (including autonomic) nervous system deficits.     

Cell encapsulation and oxygenation in nanoporous microcontainers

With strides in stem cell biology, cell engineering and molecular therapy, the transplantation of cells to produce therapeutic molecules endogenously is an attractive and achievable alternative to the use of exogenous drugs. The encapsulation of such cell transplants in semi-permeable, nanoporous constructs is often required to protect them from immune attack and to prevent their proliferation in the host. However, effective graft immunoisolation has been mostly elusive owing to the absence of a high-throughput method to create precisely controlled, high-aspect-ratio nanopores. To address the clinical need for effective cell encapsulation and immunoisolation, we devised a biocompatible cell-encapsulating microcontainer and a method to create highly anisotropic nanopores in the microcontainer’s surface. To evaluate the efficacy of these nanopores in oxygenating the encapsulated cells, we engineered 9L rat glioma cells to bioluminesce under hypoxic conditions. The methods described above should aid in evaluating the long term survival and efficacy of cellular grafts.     

Postnatal changes in the styloid process,vagina processus styloidei,and stylomastoid foramen in relation to the function of muscles originating from the styloid process

We investigated age-related changes in the styloid process in 88 skulls, aged from 5 months to 85 years of age. The osseous styloid process was not well developed in children. Its length increased significantly with age (from 2.3 mm in 11–20 age group to 16.3 mm in 61–85 group). In adolescent specimens (11–20 years of age), the median distance from the styloid process to the stylomastoid foramen was 0.7–0.8 mm, whereas in adult and old age specimens the two structures were completely adjacent or very close, usually less than 0.2 mm. The process was missing in 5% of the adult specimens. There was a statistically significant positive association between the length of the styloid process with age (r = 0.3210, 95% CI 0.0817–0.5254; P = 0.0097), whereas the distance from the styloid process to the stylomastoid foramen significantly decreased with age (r = −0.4518, 95% CI −0.6167 to −0.2490; P = 0.0001). Changes in the length and shape of the styloid process reflected altered function of the three muscles originating from the styloid process—m. stylopharyngeus, m. stylohyoideus and m. styloglossus. They have a common function of lifting the aerodigestive elements upward and backward, after the descent of the aerodigestive tract and final morphological differentiation of the vocal system during puberty. Relationship between altered muscle function and the morphology of the styloid process are important for understanding the clinical syndromes related to the styloid process, such as Eagle’s syndrome. Presented as a poster at the 78 Jahresversammlung 2007 der Deutschen Gesellschaft für Hals-Nasen-Ohren-Heilkunde, Kopf- und Hals-Chirurgie e.V., Munich, 17–20 May 2007.     

Lactobacillus bulgaricus Prevents Intestinal Epithelial Cell Injury Caused by Enterobacter sakazakii-Induced Nitric Oxide both In Vitro and in the Newborn Rat Model of Necrotizing Enterocolitis

Enterobacter sakazakii is an emerging pathogen that has been associated with outbreaks of necrotizing enterocolitis (NEC) as well as infant sepsis and meningitis. Our previous studies demonstrated that E. sakazakii induces NEC in a newborn rat model by inducing enterocyte apoptosis. However, the mechanisms responsible for enterocyte apoptosis are not known. Here we demonstrate that E. sakazakii induces significant production of nitric oxide (NO) in rat intestinal epithelial cells (IEC-6) upon infection. The elevated production of NO, which is due to increased expression of inducible NO synthase, is responsible for apoptosis of IEC-6 cells. Notably, pretreatment of IEC-6 cells with Lactobacillus bulgaricus (ATCC 12278) attenuated the upregulation of NO production and thereby protected the cells from E. sakazakii-induced apoptosis. Furthermore, pretreatment with L. bulgaricus promoted the integrity of enterocytes both in vitro and in the infant rat model of NEC, even after challenge with E. sakazakii. Infection of IEC-6 cells with E. sakazakii upregulated several genes related to apoptosis, cytokine production, and various signaling pathways, as demonstrated by rat gene array analysis, and this upregulation was subdued by pretreatment with L. bulgaricus. In agreement with these data, L. bulgaricus pretreatment protected newborn rats infected with E. sakazakii from developing NEC, resulting in improved survival.Necrotizing enterocolitis (NEC) is a worldwide problem in very-low-birth-weight infants, with a highly variable incidence, affecting 2.6% to 28% of these infants. The precise etiology of NEC is unknown but is widely considered multifactorial. Three major factors have been proposed, including the presence of a pathogenic organism, the challenge of enteral feeding, and altered enteric mucosal integrity. Although mortality rates among infants with NEC may have decreased as a result of improved supportive and surgical care, effective preventive strategies are lacking. The initial management of infants who are suspected of having NEC relies upon aggressive fluid resuscitation and the prompt initiation of broad-spectrum antibiotics (30). Thus, it would be of extreme value to develop a preventive or therapeutic strategy in the management of this disease. Prevention offers benefits over reactive intervention because despite successful treatment, once infants are affected with NEC, they continue to be at risk for multiple morbidities, including short gut syndrome, stricture formation, and even poor neurologic outcomes.Several pathogens have been associated with NEC. However, thus far, none fulfills Koch''s postulates (17). Enterobacter sakazakii is a bacterium that has been implicated in outbreaks of sepsis, meningitis, and NEC in newborns. Such outbreaks have been attributed to E. sakazakii contamination of powdered human infant formula (43). A taxonomic reclassification of E. sakazakii to consist of five species within a new genus, Cronobacter, was recently proposed (20). However, due to the familiarity of the name Enterobacter sakazakii in the literature, we continue to use the same in this study. Additionally, E. sakazakii infection confers a high mortality rate in affected infants (40). Our recent studies have shown that oral feeding of E. sakazakii induces NEC in an experimental animal model (16). We further demonstrated that E. sakazakii binding to rat intestinal epithelial cells (IEC-6) in vitro and in vivo causes enterocyte apoptosis. However, the mechanisms involved in E. sakazakii-induced apoptosis of IEC-6 cells have not yet been identified.Although E. sakazakii may be an important pathogenic trigger in the development of NEC, it has been recognized that there are both quantitative and qualitative changes in the fecal flora before the onset of NEC. A decline in the variety of species and a shift to a predominance of members of the Enterobacteriaceae before the onset of NEC were identified. Gewolb et al. reported that Bifidobacterium and Lactobacillus are found in the stools of <5% of extremely-low-birth-weight infants within the first month of life (11). These data suggest that low levels of colonization of Bifidobacterium and Lactobacillus in low-birth-weight infants may serve as a predisposing factor in microbial infection. Recently, there have been several randomized prospective human clinical trials that have shown a decreased incidence of NEC after prophylactic oral administration of certain probiotic species (2, 4, 15, 29). Although the results for humans are promising, very little is understood regarding the mechanisms by which probiotics may alter disease susceptibility, and an appreciation of the role of specific probiotics is absent. However, there are several theoretical mechanisms through which probiotics may protect against the development of NEC, including blocking enterocyte-binding sites used by pathogenic species through modulation of the intestinal immune system or creating a locally hostile environment (8).NO is a short-lived, highly reactive molecule that plays a key role in the pathogenesis of intestinal barrier failure in NEC (3). NO is produced by three isoforms of NO synthase (NOS). Two of the isoforms, endothelial NOS (eNOS) and neuronal NOS, are expressed at constitutively low levels. The third isoform, inducible NOS (iNOS), is not expressed under normal conditions but is dramatically increased during inflammation, resulting in high levels of NO production (34). Elevated levels of iNOS have been demonstrated in infants with NEC as well as in adults with inflammatory bowel disorders (14, 37). The role of iNOS in E. sakazakii-induced NEC is unknown, as is the role of probiotics in iNOS induction. In this study, we report the following two important phenomena: (i) E. sakazakii stimulates the production of NO, leading to apoptosis of IEC-6 cells; and (ii) oral feeding of Lactobacillus bulgaricus prior to infection with E. sakazakii significantly reduces mortality rates in an experimental animal model of NEC.