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Commensal bacteria play a major role in multiple skin functions by providing the first layer of defense against pathogens and maintaining the skin barrier. Staphylococcus epidermidis is one of the most common skin commensals. In this study, we showed that S. epidermidis ferments glycerol and uses it as a nutrient, while producing short-chain and organic fatty acids, with the most notable being lactic acid. Lactic acid is an alpha-hydroxy acid that inhibits the growth of pathogenic bacteria, without any negative effect on the commensal bacteria itself. Using in vivo experiments, we validated our in vitro results, showing that the skin microbiome is also capable of doing this. Finally, using 2D and 3D skin culture models, we showed that the fermentation of glycerol, mainly lactic acid, as determined by analytical methods, upregulates the expression levels of several key genes that are associated with the barrier properties of the skin. While the hydration effect of glycerol on the skin is well known, our study shows the overall benefits of glycerol on the skin microbiome, while revealing an alternate mode of action of glycerol for multiple skin benefits.

The skin-associated microbiome plays an important role in general well-being and in a variety of treatable skin conditions. In this regard, endogenous antimicrobial peptides have both a direct and indirect role in determining the composition of the microbiota. We demonstrate here that certain small molecular species can amplify the antimicrobial potency of naturally occurring antimicrobial peptides. In this study, we have used niacinamide, a form of vitamin B3 naturally found in foods and widely used in cosmetic skincare products, and two of its structural analogs, to investigate their cooperativity with the human antimicrobial peptide LL37 on the bacterium Staphylococcus aureus. We observed a clear synergistic effect of niacinamide and, to some extent, N-methylnicotinamide, whereas isonicotinamide showed no significant cooperativity with LL37. Adaptively biased molecular dynamics simulations using simplified model membrane substrates and single peptides revealed that these molecules partition into the headgroup region of an anionic bilayer used to mimic the bacterial membrane. The simulated effects on the physical properties of the simulated model membrane are well correlated with experimental activity observed in real biological assays despite the simplicity of the model. In contrast, these molecules have little effect on zwitterionic bilayers that mimic a mammalian membrane. We conclude that niacinamide and N-methylnicotinamide can therefore potentiate the activity of host peptides by modulating the physical properties of the bacterial membrane, and to a lesser extent through direct interactions with the peptide. The level of cooperativity is strongly dependent on the detailed chemistry of the additive, suggesting an opportunity to fine-tune the behavior of host peptides.

Orb2 the Drosophila homolog of Cytoplasmic polyadenylation element binding protein (CPEB) forms prion-like oligomers. These oligomers consist of Orb2A and Orb2B isoforms and their formation are dependent on the oligomerization of the Orb2A isoform. Drosophila with a mutation diminishing Orb2A’s prion-like oligomerization forms long-term memory but fails to maintain it over time. Since, this prion-like oligomerization of Orb2A plays a crucial role in the maintenance of memory, here we aim to find what regulates this oligomerization. In an immunoprecipitation-based screen, we identify interactors of Orb2A in the Hsp40 and Hsp70 families of proteins. Amongst these, we find an Hsp40 family protein Mrj as a regulator of the conversion of Orb2A to its prion-like form. Mrj interacts with Hsp70 proteins and acts as a chaperone by interfering with the aggregation of pathogenic Huntingtin. Unlike its mammalian homolog, we find Drosophila Mrj is neither an essential gene nor causes any gross neurodevelopmental defect. We observe a loss of Mrj results in a reduction in Orb2 oligomers. Further, the knockdown of Mrj in the mushroom body neurons results in a deficit in long-term memory. Our work implicates a chaperone Mrj in mechanisms of memory regulation through controlling the oligomerization of Orb2A and its association with the translating polysomes.

The continual emergence of new SARS-CoV-2 variants threatens the effectiveness of worldwide vaccination programs and highlights the need for complementary strategies for a sustainable containment plan. A promising approach is to mobilize the body⍰s own antimicrobial peptides (AMPs), to combat SARS-CoV-2 infection and propagation. We have found that human cathelicidin (LL37), an AMP found at epithelial barriers as well as in various bodily fluids, has the capacity to neutralise multiple strains of SARS-CoV-2. Biophysical and computational studies indicate that LL37⍰s mechanism of action is through the disruption of the viral membrane. This antiviral activity of LL37 is enhanced by the hydrotropic action of niacinamide, which may increase the bioavailability of the AMP. Interestingly, we observed inverse correlation between LL37 levels and disease severity of COVID-19 positive patients, suggesting enhancement of AMP response would be an effective therapeutic avenue to mitigate disease severity and overcome vaccine escape.

Huntington’s disease (HD) is a severe neurodegenerative disorder caused by poly Q repeat expansion in the Huntingtin (Htt) gene. While the Htt amyloid aggregates are known to affect many cellular processes, their role in translation has not been addressed. Here we report that pathogenic Htt expression causes a protein synthesis deficit in cells. We find a functional prion-like protein, the translation regulator Orb2, to be sequestered by Htt aggregates in cells. Co-expression of Orb2 can partially rescue the lethality associated with poly Q expanded Htt. These findings can be relevant for HD as human homologs of Orb2 are also sequestered by pathogenic Htt aggregates. Our work suggests that translation dysfunction is one of the contributors to the pathogenesis of HD and new therapies targeting protein synthesis pathways might help to alleviate disease symptoms.

Real-time PCR is a widely used technique for quantification of gene expression. However, commercially available kits for real-time PCR are very expensive. The ongoing coronavirus pandemic has severely hampered the economy in a number of developing countries, resulting in a reduction in available research funding. The fallout of this will result in limiting educational institutes and small enterprises from using cutting edge biological techniques such as real-time PCR. Here, we report a cost-effective approach for preparing and assembling cDNA synthesis and real-time PCR mastermixes with similar efficiencies as commercially available kits. Our results thus demonstrate an alternative to commercially available kits. Supplementary Information The online version contains supplementary material available at 10.1007/s12038-021-00231-w.

The contact sites between ER and mitochondria regulate several cellular processes including inter-organelle lipid transport, calcium homeostasis and autophagy. However, the mechanisms that regulate the dynamics and functions of these contact sites remain unresolved. We show that annulate lamellae (AL), a relatively unexplored subcellular structure representing subdomains of ER enriched with a subset of nucleoporins, are present at ER-mitochondria contact sites (ERMCS). Depletion of one of the AL-resident nucleoporins, Nup358, results in increased contacts between ER and mitochondria. Mechanistically, Nup358 modulates ERMCS dynamics by restricting mTORC2/Akt signalling. Our results suggest that growth factor-mediated remodelling of ERMCS depends on a reciprocal binding of Nup358 and mTOR to the ERMCS tethering complex consisting of VAPB and PTPIP51. Furthermore, Nup358 also interacts with IP3R, an ERMCS-enriched Ca2+ channel, and controls Ca2+ release from the ER. Consequently, depletion of Nup358 leads to elevated cytoplasmic Ca2+ and autophagy via activation of Ca2+/CaMKK2/AMPK axis. Our study thus uncovers a novel role for AL, particularly for Nup358, in regulating mTORC2-mediated ERMCS remodelling and Ca2+-directed autophagy, possibly via independent mechanisms.

Real-time PCR is a widely used technique for quantification of gene expression. However, commercially available kits for real-time PCR are very expensive. The ongoing coronavirus pandemic has severely hampered the economy in a number of developing countries, resulting in a reduction in available research funding. The fallout of this will result in limiting educational institutes and small enterprises from using cutting edge biological techniques such as real-time PCR. Here, we report a cost-effective approach for preparing and assembling cDNA synthesis and real-time PCR mastermixes with similar efficiencies as commercially available kits. Our results thus demonstrate an alternative to commercially available kits.

Keratinocytes maintain epidermal integrity through cellular differentiation. This process enhances intraorganelle digestion in keratinocytes to sustain nutritional and calcium-ionic stresses observed in upper skin layers. However, the molecular mechanisms governing keratinocyte differentiation and concomitant increase in lysosomal function is poorly understood. Here, by using primary neonatal human epidermal keratinocytes, we identified the molecular link between signaling pathways and cellular differentiation/lysosome biogenesis. Incubation of keratinocytes with CaCl

Huntington's disease (HD) is a severe neurodegenerative disorder caused by poly Q repeat expansion in the Huntingtin (Htt) gene. While the Htt amyloid aggregates are known to affect many cellular processes, their role in translation has not been addressed. Here we report that pathogenic Htt expression causes a protein synthesis deficit in cells. We find a functional prion-like protein, the translation regulator Orb2, to be sequestered by Htt aggregates in cells. Co-expression of Orb2 can partially rescue the lethality associated with poly Q expanded Htt. These findings can be relevant for HD as human homologs of Orb2 are also sequestered by pathogenic Htt aggregates. Our work suggests that translation dysfunction is one of the contributors to the pathogenesis of HD and new therapies targeting protein synthesis pathways might help to alleviate disease symptoms.

Huntington’s disease (HD) is a severe neurodegenerative disorder caused by poly Q repeat expansion in the Huntingtin (Htt) gene. While the Htt amyloid aggregates are known to affect many cellular processes, its role in translation is not addressed. Here we report pathogenic Htt expression causes protein synthesis deficit in cells. We find a functional prion-like protein, the translation regulator Orb2 to be sequestered by Htt aggregates. Coexpression of Orb2 can partially rescue the lethality associated with poly Q expanded Htt. These findings can be relevant for HD as human homologs of Orb2 also can be sequestered by pathogenic Htt aggregates. Our work suggests that translation dysfunction could be one of the contributors in the pathogenesis of HD and new therapies targeting protein synthesis pathways might help alleviate disease symptoms.

Alzheimer’s disease (AD) is characterized by the accumulation of amyloid plaques surrounded by microglia. In cell culture, microglia internalize fibrillar β-amyloid but do not degrade it efficiently. Unactivated microglia have a relatively high lysosomal pH, which impairs the activity of lysosomal proteases. Previous studies showed that activation of microglia with macrophage colony stimulating factor decreases lysosomal pH and enhances fibrillar β-amyloid degradation. We investigated the role of the lysosomal protease tripeptidyl peptidase 1 (TPP1) in cell culture and in a mouse model of Alzheimer’s disease. Increased levels of TPP1 in unactivated microglia enhanced fibrillar β-amyloid degradation. Conversely, reduction of TPP1 led to decreased fibrillar β-amyloid degradation in activated microglia, macrophages, and other cells that degrade fibrillar β-amyloid efficiently. Reduction of TPP1 in an AD model mouse using a gene-targeted hypomorphic Tpp1 allele increased plaque burden. These results suggest that decreased TPP1 potentiates AD pathogenesis and that strategies to increase TPP1 activity may have therapeutic value.Highlights*In microglia, TPP1 is important for the degradation of fibrillar β-amyloid.*Increased TPP1 in microglia results in enhanced fibrillar β-amyloid degradation.*In an AD mouse model, reduction of TPP1 led to increased amyloid plaque deposition.

Antimicrobial peptides (AMPs) are the body’s natural innate immune defense against a spectrum of pathogenic insults and provide an attractive strategy towards combating the growing problem of antibiotic resistant microorganisms. The prophylactic use of AMPs is contingent upon understanding the regulatory mechanisms governing the release of AMPs from intracellular stores, which occurs via the non-conventional secretory pathway. Analysis of S100A7 (Psoriasin), an abundant AMP in the skin, from its endogenous stores within differentiated keratinocytes has revealed an unexpected biphasic secretory response to bacterial exposure. The core components regulating S100A7 secretion are NFκB/p38MAPK, caspase-1 and IL-1α. The early activation of this core machinery is mediated by toll-like receptor signaling, whereas the chronic response is regulated by the wound-healing machinery mediated by Caspase-8 downregulation. Interestingly, in inflammatory skin diseases in which S100A7 is excessively released, there is a concomitant downregulation of caspase-8, indicating that these pathological scenarios are commandeering the normal wound healing response. These results highlight the potential benefits of targeting intracellular wound healing pathways as a means of exerting control over the release of AMPs from the skin in both homeostatic and disease conditions.

Summary: Antimicrobial peptides (AMPs) are the body’s natural innate immune defense against a spectrum of pathogens and can also modulate cell proliferation, chemotaxis, angiogenesis, wound healing, and immune cell activity. Harnessing these diverse functions for prophylactic use is contingent upon understanding the regulatory mechanisms governing their unconventional secretion from cells. Analysis of the secretion of S100A7 (Psoriasin), an abundant AMP stored in differentiated keratinocytes of the skin, has revealed an unexpected biphasic secretory response to bacterial exposure. The core components regulating S100A7 secretion are NFκB/p38MAPK, caspase-1, and interleukin (IL)-1α. The initial activation of this core machinery is mediated by Toll-like receptor signaling, whereas the chronic response is mediated by Caspase-8 downregulation. Interestingly, there is a concomitant downregulation of Caspase-8 in inflammatory skin diseases wherein S100A7 is constitutively released. These results highlight the potential of targeting these components to control the release of AMPs from the skin in both homeostatic and disease conditions. : The global explosion of antibiotic-resistant microorganisms has spurred interest in alternative strategies to combat these “superbugs.” Antimicrobial peptides (AMPs) have emerged as a promising solution. Bhatt et al. show downregulation of epidermal caspase-8 can mediate sustained release of AMPs from the skin and provide an effective defense against infection. keywords: caspase, antimicrobial peptides, skin, TLR, IL-1, NFkB, psoriasis, antibiotic resistance

Antimicrobial peptides (AMPs) form a part of the skin's innate immune system. Their primary activity is to provide antimicrobial benefits and hence protect from infections. AMPs that are present on human skin include psoriasin (S100A7), RNase 7, lysozyme, LL-37 and defensins. Niacinamide is a well-known cosmetic ingredient that has been used traditionally for multiple skin benefits. Recent data indicate that niacinamide treatment can boost AMPs in human gut epithelial cells and in neutrophils. Treatment with niacinamide in mice also provided protection from skin infections by enhancing AMPs. In this article, we find that treatment with niacinamide formulation provides long-lasting protection against bacteria, potentially through the activation of an AMP response.

Keratinocytes maintain epidermis integrity and function including physical and antimicrobial barrier through cellular differentiation. This process is predicted to be controlled by calcium ion gradient and nutritional stress. Keratinocytes undergo proteome changes during differentiation, which enhances the intracellular organelle digestion to sustain the stress conditions. However, the molecular mechanism between epidermal differentiation and organelle homeostasis is poorly understood. Here, we used primary neonatal human epidermal keratinocytes to study the link between cellular differentiation, signaling pathways and organelle turnover. Upon addition of calcium chloride (2 mM) to the culture medium, keratinocytes increased their cell size and the expression of differentiation markers. Moreover, differentiated keratinocytes showed enhanced lysosome biogenesis that was dependent on ATF6-arm of UPR signaling but independent of mTOR-MiT/TFE transcription factors. Furthermore, chemical inhibition of mTOR has increased keratinocyte differentiation and relocalized the MiT/TFE TFs to the lysosome membranes, indicating that autophagy activation promotes the epidermal differentiation. Interestingly, differentiation of keratinocytes resulted in dispersal of fragmented Golgi and lysosomes, and the later organelles showed colocalization with Golgi-tethering proteins, suggesting that these lysosomes possibly originated from Golgi, hence named as Golgi-associated lysosomes (GALs). Consistent to this prediction, inhibition of Golgi function using brefeldin A completely abolished the formation of GALs and the keratinocyte differentiation. Thus, ER stress regulates the biogenesis of GALs, which maintains keratinocyte differentiation and epidermal homeostasis.

A 28-year-old gravida 2 para 1 booked under consultant care for her second pregnancy. The patient had had a previous traumatic delivery four years earlier at a different hospital, with a difficult ...

Key content Urinary incontinence (UI) affects up to 69% of the female population at some point in their lives but remains under-reported. There are three main types of UI: stress, urgency and mixed. It is recommended to try conservative approaches as first-line measures in the management of all types of UI. These include lifestyle interventions such as adjustment of fluid intake and weight loss, physical and behavioural therapies (pelvic floor muscle training, electrical stimulation, vaginal cones and bladder training programmes) and occasionally containment devices. Initial management of UI, in most cases, renders itself to primary care settings: this involves appropriate assessment of women's symptoms, including quality of life (QOL) assessment, appropriate simple investigations (such as urine analysis) and conservative treatment. Appropriate referral pathways to secondary and tertiary levels of care are necessary. Regular audit should take place to assess the efficacy of management options and referral pathways. UI has significant adverse affects on women's QOL. Hence patient reported outcome measures (PROMs) have been highlighted in recent NHS reports as a means of assessing effectiveness of care from the patient's perspective by gauging patient health status or health-related QOL. Learning objectives To gain an understanding of the assessment and various conservative management options for common types of UI in women. To explore the existing evidence base for such conservative management modalities and to analyse the effect of the current economic climate and reorganisation of services on the conservative management approach. To recognise the important role of patient reported outcome measures as well as service and user involvement, in the assessment of clinical and cost-effectiveness of various treatment options of UI in women. Ethical issues Should conservative measures such as weight loss and smoking cessation be a pre-requisite for offering further management options? In the present climate of scarce health resources should more emphasis be placed on the active role of patients in their own management? Will reorganisation of the NHS pose any threat to the conservative measures for treating UI in women? Is it acceptable to take a ‘one size fits all’ approach to management of common types of UI?

Microglial lysosomes lack ClC-7, which leads to incomplete lysosomal acidification. In quiescent microglia, ClC-7 is targeted for proteasomal degradation apparently by an endoplasmic reticulum-associated degradation (ERAD) pathway. Microglial activation recruits ClC-7 to lysosomes and causes lysosomal acidification, which leads to efficient amyloid degradation., Incomplete lysosomal acidification in microglia inhibits the degradation of fibrillar forms of Alzheimer's amyloid β peptide (fAβ). Here we show that in primary microglia a chloride transporter, ClC-7, is not delivered efficiently to lysosomes, causing incomplete lysosomal acidification. ClC-7 protein is synthesized by microglia but it is mistargeted and appears to be degraded by an endoplasmic reticulum–associated degradation pathway. Activation of microglia with macrophage colony-stimulating factor induces trafficking of ClC-7 to lysosomes, leading to lysosomal acidification and increased fAβ degradation. ClC-7 associates with another protein, Ostm1, which plays an important role in its correct lysosomal targeting. Expression of both ClC-7 and Ostm1 is increased in activated microglia, which can account for the increased delivery of ClC-7 to lysosomes. Our findings suggest a novel mechanism of lysosomal pH regulation in activated microglia that is required for fAβ degradation.

SummaryPrions are proteins that can assume at least two distinct conformational states, one of which is dominant and self-perpetuating. Previously we found that a translation regulator CPEB from Aplysia, ApCPEB, that stabilizes activity-dependent changes in synaptic efficacy can display prion-like properties in yeast. Here we find that, when exogenously expressed in sensory neurons, ApCPEB can form an amyloidogenic self-sustaining multimer, consistent with it being a prion-like protein. In addition, we find that conversion of both the exogenous and the endogenous ApCPEB to the multimeric state is enhanced by the neurotransmitter serotonin and that an antibody that recognizes preferentially the multimeric ApCPEB blocks persistence of synaptic facilitation. These results are consistent with the idea that ApCPEB can act as a self-sustaining prion-like protein in the nervous system and thereby might allow the activity-dependent change in synaptic efficacy to persist for long periods of time.PaperClip

Aims To evaluate whether the treatment based on urodynamics (UDS) leads to better treatment response compared to where the treatment is based on symptoms alone. Study design, materials and methods All patients referred with lower urinary tract symptoms (LUTS) were offered the opportunity to participate in this patient preference trial. Patients were asked to complete a Kings Health Questionnaire (KHQ) and a 3-day bladder diary. The patients were then offered treatment based on their preference for conservative therapy based on either symptoms alone or with additional UDS or randomization to the same two alternatives, if they had no preference. The primary outcomes studied were improvement in KHQ and reduction in Incontinence Episode Frequency (IEF) at 6 months. Logistic regression analysis was done to evaluate the effect of choice of treatment and effect of incontinence on KHQ. Results There was no statistically significant difference between the KHQ scores pre and post UDS in patient preference or randomized groups. Follow up attendance rates were significantly better in those who chose to undergo urodynamics when compared to those who chose conservative treatment. Conclusions This study suggests although UDS does not improve IEF, it may have utility in patient choice and subsequent compliance. Another relevant finding was the previously unreported high uptake of UDS by women when given the choice of whether to undergo UDS first or have treatment based on symptoms alone. Neurourol. Urodynam. 29:522–526, 2010. © 2009 Wiley-Liss, Inc.

Three Canadian coals, namely, Bienfait lignite, Montgomery sub-bituminous C, and Coal Valley high volatile bituminous C were activated in a fixed-bed reactor. For each coal, two different sizes of ...

Cultured microglia internalize fibrillar amyloid Abeta (fAbeta) and deliver it to lysosomes. Degradation of fAbeta by microglia is incomplete, but macrophages degrade fAbeta efficiently. When mannose-6 phosphorylated lysosomal enzymes were added to the culture medium of microglia, degradation of fAbeta was increased, and the increased degradation was inhibited by excess mannose-6-phosphate, which competes for binding and endocytic uptake. This suggests that low activity of one or more lysosomal enzymes in the microglia was responsible for the poor degradation of fAbeta. To further characterize the degradation of fAbeta in late endosomes and lysosomes, we analyzed fAbeta-derived intracellular degradation products in macrophages and microglia by mass spectrometry. Fragments with truncations in the first 12 N-terminal residues were observed in extracts from both cell types. We also analyzed material released by the cells. Microglia released mainly intact Abeta1-42, whereas macrophages released a variety of N-terminal truncated fragments. These results indicate that initial proteolysis near the N-terminus is similar in both cell types, but microglia are limited in their ability to make further cuts in the fAbeta.

Microglia are the main immune cells of the brain, and under some circumstances they can play an important role in removal of fibrillar Alzheimer amyloid beta peptide (fAbeta). Primary mouse microglia can internalize fAbeta, but they do not degrade it efficiently. We compared the level of lysosomal proteases in microglia and J774 macrophages, which can degrade fAbeta efficiently, and we found that microglia actually contain higher levels of many lysosomal proteases than macrophages. However, the microglial lysosomes are less acidic (average pH of approximately 6), reducing the activity of lysosomal enzymes in the cells. Proinflammatory treatments with macrophage colony-stimulating factor (MCSF) or interleukin-6 acidify the lysosomes of microglia and enable them to degrade fAbeta. After treatment with MCSF, the pH of microglial lysosomes is similar to J774 macrophages (pH of approximately 5), and the MCSF-induced acidification can be partially reversed upon treatment with an inhibitor of protein kinase A or with an anion transport inhibitor. Microglia also degrade fAbeta if lysosomes are acidified by an ammonia pulse-wash or by treatment with forskolin, which activates protein kinase A. Our results indicate that regulated lysosomal acidification can potentiate fAbeta degradation by microglia.

Urinary incontinence is a common problem which increases in prevalence in association with advancing age and has a significant adverse effect upon well-being and quality of life. It is not the "benign" condition that many take it for. Overactive bladder (frequency-urgency syndrome) is the commonest bladder problem in late life, affecting up to 41% of over-75-year-old individuals, and the elderly experience more severe disease. This small series should provide the reader with a comprehensive overview of the current thinking in the assessment and management of patients with overactive bladder syndrome, explores the history of the condition and current approaches to its medical and surgical management and explores where management may change in more complex populations. The current state and future developments in pharmacological therapy are also outlined.

The EMQ element constitutes 60% of the new MRCOG Part 2 examination. Candidates therefore need to be well-versed in approaching this type of question and extensive practice in answering questions is recommended. This new book contains 150 sample EMQs, together with answers that have full detailed explanations and references for further reading – including RCOG resources such as Green Top Guidelines - to enhance knowledge and understanding. The book also contains a section of handy hints and tips on passing the examination to improve candidates' chances of success. EMQs for the MRCOG Part 2 is an invaluable study aid to help pass the EMQ component of the MRCOG Part 2.

Pelvic organ prolapse is a common gynaecological problem and the mechanisms underlying prolapse development are not yet clear but it is thought that increases in abdominal pressure, such as those routinely involved in heavy lifting and long periods of standing, may cause progressive pelvic floor damage over time. The aim of this study was to investigate the effects of strenuous physical activity on the development of prolapse. A narrative literature review was carried out to investigate the effects of occupation and recreational activity on the pathogenesis of pelvic organ prolapse. A marked paucity of literature relevant to the research question makes it difficult to draw firm conclusions. Further research is greatly needed to explore potentially preventable factors in this frequently occurring condition. The review reveals some evidence linking strenuous physical activity with pelvic organ prolapse but this is neither consistent nor adequately powered to reach any firm conclusions.

Amyloids are ordered protein aggregates that are typically associated with neurodegenerative diseases and cognitive impairment. By contrast, the amyloid-like state of the neuronal RNA binding protein Orb2 in Drosophila was recently implicated in memory consolidation, but it remains unclear what features of this functional amyloid-like protein give rise to such diametrically opposed behaviour. Here, using an array of biophysical, cell biological and behavioural assays we have characterized the structural features of Orb2 from the monomer to the amyloid state. Surprisingly, we find that Orb2 shares many structural traits with pathological amyloids, including the intermediate toxic oligomeric species, which can be sequestered in vivo in hetero-oligomers by pathological amyloids. However, unlike pathological amyloids, Orb2 rapidly forms amyloids and its toxic intermediates are extremely transient, indicating that kinetic parameters differentiate this functional amyloid from pathological amyloids. We also observed that a well-known anti-amyloidogenic peptide interferes with long-term memory in Drosophila. These results provide structural insights into how the amyloid-like state of the Orb2 protein can stabilize memory and be nontoxic. They also provide insight into how amyloid-based diseases may affect memory processes., This research was supported by funds from SAF2013-49179-C2-1-R JPND_CD_FP-688- 059 (AC14/00037 ISCIII) to MCV, SIMR to KS, SAF2013-49179-C2-2-R JPND_CD_FP-688-059 (AC14/00037 ISCIII) to DVL, BFU2012-36825, S2011/BMD-2457 (Comunidad de Madrid)

Hysterectomy, removal of the uterus, has traditionally been regarded as the definitive surgical treatment for heavy menstrual bleeding. And is one of the most commonly performed operations in the UK. (Marjoribanks, 2006) Whilst menstrual disorders are the most frequent reasons for performing hysterectomy other common indications include chronic pelvic pain, fibroids, malignancy of the uterus, cervix or ovaries and genital prolapse. Hysterectomy is a major surgical procedure with potential for significant physical and emotional complications. It also carries additional social and economic costs. (Lethaby, 1999)

Hysterectomy is one of the commonest gynaecological procedures undertaken in the UK and in the USA. (Gupta & Manyonda, 2011) It is carried out both for benign indications (namely abnormal uterine bleeding and pelvic pain) and for malignancies of the female genital tract. In some cases there are multiple indications for hysterectomy, for example, menorrhagia and dysmenorrhoea (Williams & Clark, 2000) and chronic pelvic pain can accompany a variety of other gynaecological diagnoses. Bilateral salpingo-oophrectomy is often carried out concurrently and this may be followed by postoperative commencement of hormone replacement therapy (HRT).

Sacrocolpopexy, be it laparoscopic or abdominal, is associated with a risk of mesh extrusion. We report an interesting case of mesh extrusion with subsequent removal of the mesh from the vagina by the patient. We take this opportunity to review the literature regarding incidence, predisposing factors and complications of sacrocolpopexy mesh extrusion with specific reference to this case.

On a etudie l'effet de la pression jusqu'a 5600 kPa et de la temperature jusqu'a 175°C sur l'oxydation de faibles concentrations de H 2 S dans du gaz naturel dans un reacteur a lit fixe sur un catalyseur de charbon active. Ce systeme a la pression de 5600 kPa montre une plus grande activite catalytique (conversion du H 2 S de pratiquement 100%) sur une longue periode et une moins grande selectivite vers SO 2 qu'a la pression atmospherique. La desorption du soufre d'un catalyseur charge se produit d'abord dans les macropores (>100 nm) du catalyseur qui contiennent une part substantielle de la charge de soufre, puis dans les micropores (

Urinary incontinence is a common problem with serious effect on the quality of life. Bladder training aims to increase the interval between voids, either by a mandatory or self-adjustable schedule, so that incontinence is avoided. This study aimed to assess the effectiveness of inpatient bladder retraining.A retrospective case-note analysis was conducted over a period of 24 months. Outcome measures were decrease in incontinence episode frequency (IEF) and nocturia and increase in interval between voids. Subjective improvement was assessed on a four-point scale.The study revealed statistically significant decrease in IEF and nocturia and increase in the interval between voids. Twenty-three percent was cured of their symptoms, 36% reported improvement, 27% did not find any change, whereas 14% reported that they were worse off their after bladder retraining.The study confirms the usefulness of inpatient bladder retraining as a treatment option, especially in people refractory to outpatient management.

To evaluate whether the treatment based on urodynamics (UDS) leads to better treatment response compared to where the treatment is based on symptoms alone.All patients referred with lower urinary tract symptoms (LUTS) were offered the opportunity to participate in this patient preference trial. Patients were asked to complete a Kings Health Questionnaire (KHQ) and a 3-day bladder diary. The patients were then offered treatment based on their preference for conservative therapy based on either symptoms alone or with additional UDS or randomization to the same two alternatives, if they had no preference. The primary outcomes studied were improvement in KHQ and reduction in Incontinence Episode Frequency (IEF) at 6 months. Logistic regression analysis was done to evaluate the effect of choice of treatment and effect of incontinence on KHQ.There was no statistically significant difference between the KHQ scores pre and post UDS in patient preference or randomized groups. Follow up attendance rates were significantly better in those who chose to undergo urodynamics when compared to those who chose conservative treatment.This study suggests although UDS does not improve IEF, it may have utility in patient choice and subsequent compliance. Another relevant finding was the previously unreported high uptake of UDS by women when given the choice of whether to undergo UDS first or have treatment based on symptoms alone.

The mammalian protein Eps15 is phosphorylated by EGF receptor tyrosine kinase and has been shown to interact with several components of the endocytic machinery. We have identified a hypomorphic Eps15 mutant in Drosophila which shows reversible paralysis and an altered physiology at restrictive temperatures. In addition, the temperature-sensitive paralytic defect of shibire mutant is enhanced by this mutant. Eps15 is enriched in the larval neuromuscular junction in endocytic 'hot spots' in a pattern similar to Dynamin. Eps15 mutants show a decrease in the alpha-Adaptin levels at the larval neuromuscular junction synapse. Genetic and biochemical studies of interactions with components of the endocytic machinery suggest that Eps15 has an important role in synaptic vesicle recycling and regulates recruitment of alpha-Adaptin.