The hydrokinetic parameters of shunts for hydrocephalus might be inadequate.
Long-term treatment of hydrocephalus continues to be dismal. Shunting is the neurosurgical procedure more frequently associated with complications, which are mostly related with dysfunctions of the shunting device, rather than to mishaps of the rather simple surgical procedure. Overdrainage and underdrainage are the most common dysfunctions; of them, overdrainage is a conspicuous companion of most devices. Even when literally hundreds of different models have been proposed, developed, and tested, overdrainage has plagued all shunts for the last 60 years. Several investigations have demonstrated that changes in the posture of the subject induce unavoidable and drastic differences of intraventricular hydrokinetic pressure and cerebrospinal fluid (CSF) drainage through the shunt. Of all the parameters that participate in the pathophysiology of hydrocephalus, the only invariable one is cerebrospinal fluid production at a constant rate of approximately 0.35 ml/min. However, this feature has not been considered in the design of currently available shunts. Our experimental and clinical studies have shown that a simple shunt, whose drainage capacity complies with this unique parameter, would prevent most complications of shunting for hydrocephalus.


Reduced O-GlcNAcylation and tubular hypoxia contribute to the antifibrotic effect of SGLT2 inhibitor dapagliflozin in the diabetic kidney.
Diabetic kidney disease is a worldwide epidemic, and therapies are incomplete. Clinical data suggest that improved renal outcomes by Na+-glucose cotransporter 2 inhibitor (SGLT2i) are partly beyond their antihyperglycemic effects; however, the mechanisms are still elusive. Here, we investigated the effect of the SGLT2i dapagliflozin (DAPA) in the prevention of elevated O-GlcNAcylation and tubular hypoxia as contributors of renal fibrosis. Type 1 diabetes was induced by streptozotocin in adult male Wistar rats. After the onset of diabetes, rats were treated for 6 wk with DAPA or DAPA combined with losartan (LOS). The effect of hyperglycemia was tested in HK-2 cells kept under normal or high glucose conditions. To test the effect of hypoxia, cells were kept in 1% O2 for 2 h. Cells were treated with DAPA or DAPA combined with LOS. DAPA slowed the loss of renal function, mitigated renal tubular injury markers (kidney injury molecule-1 and neutrophil gelatinase-associated lipocalin), and reduced tubulointerstitial fibrosis. DAPA diminished high glucose-induced protein O-GlcNAcylation and moderated the tubular response to hypoxia through the hypoxia-inducible factor pathway. DAPA alone was as effective as combined treatment with LOS in all outcome parameters. These data highlight the role of ameliorated O-GlcNAcylation and diminished tubular hypoxia as important benefits of SGLT2i treatment. Our results support the link between glucose toxicity, tubular hypoxia, and fibrosis, a vicious trio that could be targeted by SGLT2i in kidney diseases of other origins as well.


Effect of Temperature on Host Preference in Two Lineages of the Brown Dog Tick, Rhipicephalus sanguineus.
Rhipicephalus sanguineus is a species complex of ticks that vector disease worldwide. Feeding primarily on dogs, members of the complex also feed incidentally on humans, potentially transmitting disease agents such as Rickettsia rickettsii, Rickettsia conorii, and Ehrlichia species. There are two genetic Rh. sanguineus lineages in North America, designated as the temperate and tropical lineages, which had occurred in discrete locations, although there is now range overlap in parts of California and Arizona. Rh. sanguineus in Europe are reportedly more aggressive toward humans during hot weather, increasing the risk of pathogen transmission to humans. The aim of this study was to assess the impact of hot weather on choice between humans and dog hosts among tropical and temperate lineage Rh. sanguineus individuals. Ticks in a two-choice olfactometer migrated toward a dog or human in trials at room (23.5°C) or high temperature (38°C). At 38°C, 2.5 times more tropical lineage adults chose humans compared with room temperature, whereas temperate lineage adults demonstrated a 66% reduction in preference for dogs and a slight increase in preference for humans. Fewer nymphs chose either host at 38°C than at room temperature in both lineages. These results demonstrate that risk of disease transmission to humans may be increased during periods of hot weather, where either lineage is present, and that hot weather events associated with climatic change may result in more frequent rickettsial disease outbreaks.


Molecular mechanism of flocculation self-recognition in yeast and its role in mating and survival.
We studied the flocculation mechanism at the molecular level by determining the atomic structures of N-Flo1p and N-Lg-Flo1p in complex with their ligands. We show that they have similar ligand binding mechanisms but distinct carbohydrate specificities and affinities, which are determined by the compactness of the binding site. We characterized the glycans of Flo1p and their role in this binding process and demonstrate that glycan-glycan interactions significantly contribute to the cell-cell adhesion mechanism. Therefore, the extended flocculation mechanism is based on the self-interaction of Flo proteins and this interaction is established in two stages, involving both glycan-glycan and protein-glycan interactions. The crucial role of calcium in both types of interaction was demonstrated: Ca(2+) takes part in the binding of the carbohydrate to the protein, and the glycans aggregate only in the presence of Ca(2+). These results unify the generally accepted lectin hypothesis with the historically first-proposed "Ca(2+)-bridge" hypothesis. Additionally, a new role of cell flocculation is demonstrated; i.e., flocculation is linked to cell conjugation and mating, and survival chances consequently increase significantly by spore formation and by introduction of genetic variability. The role of Flo1p in mating was demonstrated by showing that mating efficiency is increased when cells flocculate and by differential transcriptome analysis of flocculating versus nonflocculating cells in a low-shear environment (microgravity). The results show that a multicellular clump (floc) provides a uniquely organized multicellular ultrastructure that provides a suitable microenvironment to induce and perform cell conjugation and mating. Yeast cells can form multicellular clumps under adverse growth conditions that protect cells from harsh environmental stresses. The floc formation is based on the self-interaction of Flo proteins via an N-terminal PA14 lectin domain. We have focused on the flocculation mechanism and its role. We found that carbohydrate specificity and affinity are determined by the accessibility of the binding site of the Flo proteins where the external loops in the ligand-binding domains are involved in glycan recognition specificity. We demonstrated that, in addition to the Flo lectin-glycan interaction, glycan-glycan interactions also contribute significantly to cell-cell recognition and interaction. Additionally, we show that flocculation provides a uniquely organized multicellular ultrastructure that is suitable to induce and accomplish cell mating. Therefore, flocculation is an important mechanism to enhance long-term yeast survival.


Traditional healers formalised?
Traditional healers are the first to be called for help when illness strikes the majority of South Africans. Their communities have faith in their ability to cure or alleviate conditions managed by doctors, and much more. A visit to such practitioners' websites (they are up with the latest advertising technology!) shows that they promise help with providing more power, love, security or money, protection from evil people and spirits, enhancing one's sex life with penis enlargement and vagina tightening spells, etc. Contemplating such claims, it is easy to be dismissive of traditional healers. But in this issue of the SAMJ Nompumelelo Mbatha and colleagues1 argue that the traditional healers' regulatory council, promised by an Act of Parliament, should be established, followed by (or preferably preceded by) formal recognition by employers of sick certificates issued by traditional healers. Can matters be so simply resolved? What does this mean for doctors and other formally recognised healthcare professionals, and how to respond to such claims and social pressures?


Driving under the influence of non-alcohol drugs--An update. Part II: Experimental studies.
Experimental studies on the impairing effects of drugs of relevance to driving-related performance published between 1998 and 2015 were reviewed. Studies with on-the-road driving, driving simulators, and performance tests were included for benzodiazepines and related drugs, cannabis, opioids, stimulants, GHB, ketamine, antihistamines, and antidepressants. The findings in these experimental studies were briefly discussed in relation to a review of epidemiological studies published recently. The studies mainly concluded that there may be a significant psychomotor impairment after using benzodiazepines or related drugs, cannabis, opioids, GHB, or ketamine. Low doses of central stimulants did not seem to cause impairment of driving behavior.


Conformational Changes in the Endosomal Sorting Complex Required for the Transport III Subunit Ist1 Lead to Distinct Modes of ATPase Vps4 Regulation.
Intralumenal vesicle formation of the multivesicular body is a critical step in the delivery of endocytic cargoes to the lysosome for degradation. Endosomal sorting complex required for transport III (ESCRT-III) subunits polymerize on endosomal membranes to facilitate membrane budding away from the cytoplasm to generate these intralumenal vesicles. The ATPase Vps4 remodels and disassembles ESCRT-III, but the manner in which Vps4 activity is coordinated with ESCRT-III function remains unclear. Ist1 is structurally homologous to ESCRT-III subunits and has been reported to inhibit Vps4 function despite the presence of a microtubule-interacting and trafficking domain-interacting motif (MIM) capable of stimulating Vps4 in the context of other ESCRT-III subunits. Here we report that Ist1 inhibition of Vps4 ATPase activity involves two elements in Ist1: the MIM itself and a surface containing a conserved ELYC sequence. In contrast, the MIM interaction, in concert with a more open conformation of the Ist1 core, resulted in stimulation of Vps4. Addition of the ESCRT-III subunit binding partner of Ist1, Did2, also converted Ist1 from an inhibitor to a stimulator of Vps4 ATPase activity. Finally, distinct regulation of Vps4 by Ist1 corresponded with altered ESCRT-III disassembly in vitro. Together, these data support a model in which Ist1-Did2 interactions during ESCRT-III polymerization coordinate Vps4 activity with the timing of ESCRT-III disassembly.


Bilateral impairments in task-dependent modulation of the long-latency stretch reflex following stroke.
Modulation of the long-latency reflex (LLR) is important for sensorimotor control during interaction with different mechanical loads. Transcortical pathways usually contribute to LLR modulation, but the integrity of pathways projecting to the paretic and non-paretic arms of stroke survivors is compromised. We hypothesize that disruption of transcortical reflex pathways reduces the capacity for stroke survivors to appropriately regulate the LLR bilaterally. Elbow perturbations were applied to the paretic and non-paretic arms of persons with stroke, and the dominant arm of age-matched controls as subjects interacted with Stiff or Compliant environments rendered by a linear actuator. Reflexes were quantified using surface electromyograms, recorded from biceps. LLR amplitude was significantly larger during interaction with the Compliant load compared to the Stiff load in controls. However, there was no significant change in LLR amplitude for the paretic or non-paretic arm of stroke survivors. Modulation of the LLR is altered in the paretic and non-paretic arms after stroke. Our results are indicative of bilateral sensorimotor impairments following stroke. The inability to regulate the LLR may contribute to bilateral deficits in tasks that require precise control of limb mechanics and stability.


Mechanisms regulating IgA class-specific immunoglobulin production in murine gut-associated lymphoid tissues. II. Terminal differentiation of postswitch sIgA-bearing Peyer's patch B cells.
Our previous studies indicated that cloned T cells obtained from Peyer's patches (PP) (Lyt-1+, 2-, Ia+, and H-2K/D+) evoked immunoglobulin (Ig) class switching of PP B cells from sIgM to sIgA cells in vitro; however, these switch T cells could not in themselves provide optimal help for the differentiation of postswitch sIgA-bearing PP B cells to IgA-secreting cells. Thus, in the present report we described studies focused on mechanisms regulating terminal differentiation of the postswitch PP sIgA-bearing B cells. First, to explore the effect of T cell-derived B cell differentiation factor(s) (BCDF) and macrophage factor(s) (MF) on the terminal maturation of PP B cells, LPS-stimulated PP B cells were co-cultured for 7 d with cloned T cells in the presence or absence of the above factors. In the absence of PP cloned T cells the BCDF and MF had only a modest effect on IgA production, whereas in the presence of PP, but not spleen cloned T cells, IgA production was increased. Next, to investigate the effect of T cells derived from a gut-associated lymphoid tissue (GALT), mesenteric lymph nodes (MLN), as well as from spleen on terminal differentiation of postswitch sIgA PP B cells, LPS-driven PP B cells were precultured with the cloned T cells to induce a switch to sIgA, and subsequently cultured with MLN or spleen T cells or a Lyt-2+-depleted T cell subset in the presence of a T-dependent polyclonal mitogen, staphylococcal protein A. Alternatively, in the second culture period BCDF alone was added, instead of T cells and protein A. Here it was found that B cells pre-exposed to switch T cells from PP, but not spleen, were induced to produce greatly increased amounts of IgA in the presence of protein A and T cells or a Lyt-2+-depleted T cell subset as well as in the presence of BCDF alone. Furthermore, in the presence of BCDF alone many B cells expressed cytoplasmic IgA. These observations strongly support the view that the terminal differentiation of postswitch sIgA B cells is governed by helper T cells and macrophages, or factors derived from such cells. Such cells or factors do not affect preswitch B cells.


Pharmacological treatment with mirtazapine rescues cortical atrophy and respiratory deficits in MeCP2 null mice.
Loss of MeCP2 (Methyl CpG binding protein 2) in Rett syndrome (RTT) causes brain weight decrease, shrinkage of the cortex with reduced dendritic arborization, behavioral abnormalities, seizures and cardio-respiratory complications. The observed monoamine neurotransmitters reduction in RTT suggested antidepressants as a possible therapy. We treated MeCP2-null mice from postnatal-day 28 for two weeks with desipramine, already tested in RTT, or mirtazapine, an antidepressant with limited side-effects, known to promote GABA release. Mirtazapine was more effective than desipramine in restoring somatosensory cortex thickness by fully rescuing pyramidal neurons dendritic arborization and spine density. Functionally, mirtazapine treatment normalized heart rate, breath rate, anxiety levels, and eliminated the hopping behavior observed in MeCP2-null mice, leading to improved phenotypic score. These morphological and functional effects of mirtazapine were accompanied by reestablishment of the GABAergic and glutamatergic receptor activity recorded in cortex and brainstem tissues. Thus, mirtazapine can represent a new potential pharmacological treatment for the Rett syndrome.


Characterization of neonatal opioid withdrawal syndrome in Arizona from 2010-2017.
In this paper, we describe a population of mothers who are opioid dependent at the time of giving birth and neonates exposed to opioids in utero who experience withdrawal following birth. While there have been studies of national trends in this population, there remains a gap in studies of regional trends. Using data from the Arizona Department of Health Services Hospital Discharge Database, this study aimed to characterize the population of neonates with neonatal opioid withdrawal syndrome (NOWS) and mothers who were opioid dependent at the time of giving birth, in Arizona. We analyzed approximately 1.2 million electronic medical records from the Arizona Department of Health Services Hospital Discharge Database to identify patterns and disparities across socioeconomic, ethnic, racial, and/or geographic groupings. In addition, we identified comorbid conditions that are differentially associated with NOWS in neonates or opioid dependence in mothers. Our analysis was designed to assess whether indicators such as race/ethnicity, insurance payer, marital status, and comorbidities are related to the use of opioids while pregnant. Our findings suggest that women and neonates who are non-Hispanic White and economically disadvantaged, tend be part of our populations of interest more frequently than expected. Additionally, women who are opioid dependent at the time of giving birth are unmarried more often than expected, and we suggest that marital status could be a proxy for support. Finally, we identified comorbidities associated with neonates who have NOWS and mothers who are opioid dependent not previously reported.


Social disappointment explains chimpanzees' behaviour in the inequity aversion task.
Chimpanzees' refusal of less-preferred food when an experimenter has previously provided preferred food to a conspecific has been taken as evidence for a sense of fairness. Here, we present a novel hypothesis-the social disappointment hypothesis-according to which food refusals express chimpanzees' disappointment in the human experimenter for not rewarding them as well as they could have. We tested this hypothesis using a two-by-two design in which food was either distributed by an experimenter or a machine and with a partner present or absent. We found that chimpanzees were more likely to reject food when it was distributed by an experimenter rather than by a machine and that they were not more likely to do so when a partner was present. These results suggest that chimpanzees' refusal of less-preferred food stems from social disappointment in the experimenter and not from a sense of fairness.


Guillain-Barré Syndrome outbreak associated with Zika virus infection in French Polynesia: a case-control study.
Between October, 2013, and April, 2014, French Polynesia experienced the largest Zika virus outbreak ever described at that time. During the same period, an increase in Guillain-Barré syndrome was reported, suggesting a possible association between Zika virus and Guillain-Barré syndrome. We aimed to assess the role of Zika virus and dengue virus infection in developing Guillain-Barré syndrome. In this case-control study, cases were patients with Guillain-Barré syndrome diagnosed at the Centre Hospitalier de Polynésie Française (Papeete, Tahiti, French Polynesia) during the outbreak period. Controls were age-matched, sex-matched, and residence-matched patients who presented at the hospital with a non-febrile illness (control group 1; n=98) and age-matched patients with acute Zika virus disease and no neurological symptoms (control group 2; n=70). Virological investigations included RT-PCR for Zika virus, and both microsphere immunofluorescent and seroneutralisation assays for Zika virus and dengue virus. Anti-glycolipid reactivity was studied in patients with Guillain-Barré syndrome using both ELISA and combinatorial microarrays. 42 patients were diagnosed with Guillain-Barré syndrome during the study period. 41 (98%) patients with Guillain-Barré syndrome had anti-Zika virus IgM or IgG, and all (100%) had neutralising antibodies against Zika virus compared with 54 (56%) of 98 in control group 1 (p<0.0001). 39 (93%) patients with Guillain-Barré syndrome had Zika virus IgM and 37 (88%) had experienced a transient illness in a median of 6 days (IQR 4-10) before the onset of neurological symptoms, suggesting recent Zika virus infection. Patients with Guillain-Barré syndrome had electrophysiological findings compatible with acute motor axonal neuropathy (AMAN) type, and had rapid evolution of disease (median duration of the installation and plateau phases was 6 [IQR 4-9] and 4 days [3-10], respectively). 12 (29%) patients required respiratory assistance. No patients died. Anti-glycolipid antibody activity was found in 13 (31%) patients, and notably against GA1 in eight (19%) patients, by ELISA and 19 (46%) of 41 by glycoarray at admission. The typical AMAN-associated anti-ganglioside antibodies were rarely present. Past dengue virus history did not differ significantly between patients with Guillain-Barré syndrome and those in the two control groups (95%, 89%, and 83%, respectively). This is the first study providing evidence for Zika virus infection causing Guillain-Barré syndrome. Because Zika virus is spreading rapidly across the Americas, at risk countries need to prepare for adequate intensive care beds capacity to manage patients with Guillain-Barré syndrome. Labex Integrative Biology of Emerging Infectious Diseases, EU 7th framework program PREDEMICS. and Wellcome Trust.


Wheat sprout extract induces changes on 20S proteasomes functionality.
Wheat sprouts contain a very high level of organic phosphates and a powerful cocktail of different molecules such as enzymes, reducing glycosides and polyphenols. The antioxidant properties of wheat sprouts have been widely documented and it has been shown that they are able to protect DNA against free-radicals mediated oxidative damage. Furthermore, we have recently reported on the effects of several polyphenols on 20S proteasomes, underlying the dual role of epigallocatechin-3-gallate as an antioxidant and a proteasome effector in cancer cells. The aim of this study was to investigate the effects of wheat sprout extracts on 20S proteasome functionality. Wheat sprout extracts have been analysed and characterized for their polyphenolic content using the Folin-Ciocalteau reagent and RP-HPLC technique. Comparing our data with a polyphenol standard mixture we identified five different polyphenols: gallic acid, epigallocatechin-3-gallate, epigallocatechin, epicatechin and catechin. The treatment of isolated 20S proteasomes with the extract induced a gradual inhibition of all the tested components, ChT-L, T-L, PGPH and BrAAP, in both the complexes. At low extract concentration a slight activation of the enzyme was evident only for the BrAAP component of the constitutive enzyme and the ChT-L activity of the immunoproteasome. beta-casein degradation rate decreased, particularly with the immunoproteasome. Human Colon adenocarcinoma (Caco) cells, stimulated with 12-O-tetradecanoylphorbol-13-acetate, showed activation of the 20S proteasome activities at short incubation times and an increase in intracellular oxidative proteins. Cells treatment with wheat sprout extract led to proteasome inhibition in unstimulated cells and attenuated the effects mediated by TPA. Finally, exposure to the extract affected the expression levels of pro-apoptotic proteins.


Some differences but all at risk: Improving farm safety for young people-An Australian experience.
A significant portion of on-farm deaths and injuries in Australia occur among young people working on the farm. Since most Australian farms are still family owned and operated, young people are an integral part of everyday operations and the farm is a place where these young people live, work and play. This paper describes how the international Gear Up for Ag Health and Safety™ program, originally developed in North America, was further developed for a younger Australian audience (ages 12-19) enrolled in agricultural programs at secondary or vocational schools. In addition, we share insight on demographics, self-reported farm safety behaviours, and the most common farm tasks being performed by program participants utilising a pre-survey originally developed for program customisation. Of particular importance were the most common farming tasks reported by this group. The most common tasks performed on Australian farms included a large variety of vehicle use (farm vehicles, motorbikes, and quadbikes) and handling livestock. Females reported operating vehicles and other farm equipment at the same rates as males. Males were more likely to be working with large heavy machinery and driving trucks, while females were more likely to be working with livestock and using horses for stockwork. Both males and females reported low use of PPE and poor safety habits. In future Australian programs, it will be important to address the conspicuous use of motor vehicles, quadbikes, motor bikes and machinery at early ages, and to target gender-specific tasks to reduce risks on the farm.


Distribution of intermediate-filament proteins in the human enamel organ: unusually complex pattern of coexpression of cytokeratin polypeptides and vimentin.
We applied immunohistochemical techniques and gel electrophoresis to examine the distribution of intermediate filaments in human fetal oral epithelium and the epithelia of the human enamel organ. Both methods demonstrated that human enamel epithelia contain cytokeratins 5, 14, and 17, which are typical of the basal cells of stratified epithelia, as well as smaller quantities of cytokeratins 7, 8, 19, and in trace amounts 18, which are characteristic components of simple epithelial cells. In the external enamel epithelium and stellate-reticulum cells, most of these components appeared to be simultaneously expressed. In contrast, the parental oral epithelium was negative for cytokeratin 7, thus indicating possible "neoexpression" during the course of tooth formation. Immunohistochemical procedures using various monoclonal antibodies against vimentin revealed the transient coexpression of vimentin and cytokeratins in the external enamel epithelium and in stellate-reticulum cells during enamel development. The significance of the coexpression of cytokeratins and vimentin is discussed in relation to previous findings obtained in other normal tissues and in the light of the functional processes characteristic of these epithelia.


Inhalation of activated protein C: A possible new adjunctive intervention in acute respiratory distress syndrome.
Acute respiratory distress syndrome (ARDS) is a potential lethal disease. At present time no evidence based intervention reduces mortality. The pathophysiology of ARDS include intraalveolar fibrin deposition, hyperinflammation and reduced cellular host defense in the airspace. The normal lung activates protein C (PC) to activated protein C (APC), in contrast to the ARDS lung where the PC-APC axis is disrupted. The lungs have targets for inhaled APC as illustrated by a patient case with ARDS, unresponsive to conventional therapy. After inhalation of 190 mug/kg of APC (Drotrecogin alpha activated) three times a day for seven days, a clear reduction in infiltrates on chest X-ray and a 138% increase in oxygenation capacity as reflected by the PaO(2)/FiO(2) ratio was brought about. The patient, however, died later after cardiac arrest after suspected recurrence of the T-cell lymphoma. No local or systemic adverse effects was found related to the iAPC, during, after or at the time of death. It is suggested based on existing studies and the presented case that inhaled APC is a new treatment option in patients with ARDS - a hypothesis which should be substantiated in a larger series of ARDS patients.


Neuropsychiatric lupus or not? Cerebral hypoperfusion by perfusion-weighted MRI in normal-appearing white matter in primary neuropsychiatric lupus erythematosus.
Cerebral perfusion abnormalities have been reported in systemic lupus erythematosus (SLE) but their value in distinguishing lupus from non-lupus-related neuropsychiatric events remains elusive. We examined whether dynamic susceptibility contrast-enhanced perfusion MRI (DSC-MRI), a minimally invasive and widely available method of cerebral perfusion assessment, may assist neuropsychiatric SLE (NPSLE) diagnosis. In total, 76patients with SLE (37 primary NPSLE, 16 secondary NPSLE, 23 non-NPSLE) and 31 healthy controls underwent conventional MRI (cMRI) and DSC-MRI. Attribution of NPSLE to lupus or not was based on multidisciplinary assessment including cMRI results and response to treatment. Cerebral blood volume and flow were estimated in 18 normal-appearing white and deep grey matter areas. The most common manifestations were mood disorder, cognitive disorder and headache. Patients with primary NPSLE had lower cerebral blood flow and volume in several normal-appearing white matter areas compared with controls (P<0.0001) and lower cerebral blood flow in the semioval centre bilaterally, compared with non-NPSLE and patients with secondary NPSLE (P<0.001). A cut-off for cerebral blood flow of 0.77 in the left semioval centre discriminated primary NPSLE from non-NPSLE/secondary NPSLE with 80% sensitivity and 67%-69% specificity. Blood flow values in the left semioval centre showed substantially higher sensitivity than cMRI (81% vs 19%-24%) for diagnosing primary NPSLE with the combination of the two modalities yielding 94%-100% specificity in discriminating primary from secondary NPSLE. Primary NPSLE is characterised by significant hypoperfusion in cerebral white matter that appears normal on cMRI. The combination of DSC-MRI-measured blood flow in the brain semioval centre with conventional MRI may improve NPSLE diagnosis.


RYR1-related congenital myopathy with fatigable weakness, responding to pyridostigimine.
The spectrum of RYR1 mutation associated disease encompasses congenital myopathies, exercise induced rhabdomyolysis, malignant hyperthermia susceptibility and King-Denborough syndrome. We report the clinical phenotype of two siblings who presented in infancy with hypotonia and striking fatigable ptosis. Their response to pyridostigimine was striking, but genetic screening for congenital myasthenic syndromes was negative, prompting further evaluation. Muscle MRI was abnormal with a selective pattern of involvement evocative of RYR1-related myopathy. This directed sequencing of the RYR1 gene, which revealed two heterozygous c.6721C>T (p.Arg2241X) nonsense mutations and novel c.8888T>C (p.Leu2963Pro) mutations in both siblings. These cases broaden the RYR1-related disease spectrum to include a myasthenic-like phenotype, including partial response to pyridostigimine. RYR1-related myopathy should be considered in the presence of fatigable weakness especially if muscle imaging demonstrates structural abnormalities. Single fibre electromyography can also be helpful in cases like this.


Perceptions of living with a device-based treatment: an account of patients treated with deep brain stimulation for Parkinson's disease.
Deep brain stimulation (DBS) is an established treatment for Parkinson's disease. Little is known about patients' own perceptions of living with the implanted hardware. We aimed to explore patients' own perceptions of living with an implanted device. Semistructured interviews with open-ended questions were conducted with 42 patients (11 women) who had been on DBS for a mean of three years. The questions focused on patients' experiences of living with and managing the DBS device. The interviews were transcribed verbatim and analyzed according to the difference and similarity technique in grounded theory. From the patients' narratives concerning living with and managing the DBS device, the following four categories emerged: 1) The device-not a big issue: although the hardware was felt inside the body and also visible from outside, the device as such was not a big issue. 2) Necessary carefulness: Patients expressed the need to be careful when performing certain daily activities in order not to dislocate or harm the device. 3) Continuous need for professional support: Most patients relied solely on professionals for fine-tuning the stimulation rather than using their handheld controller, even if this entailed numerous visits to a remote hospital. 4) Balancing symptom relief and side-effects: Patients expressed difficulties in finding the optimal match between decrease of symptoms and stimulation-induced side-effects. The in-depth interviews of patients on chronic DBS about their perceptions of living with an implanted device provided useful insights that would be difficult to capture by quantitative evaluations.


The good turned ugly: immunopathogenic basis for diabetogenic CD8+ T cells in NOD mice.
Type 1 diabetes (T1D) in both humans and nonobese diabetic (NOD) mice is a T-cell-mediated autoimmune disease in which the insulin-producing pancreatic islet beta-cells are selectively eliminated. As a result, glucose metabolism cannot be regulated unless exogenous insulin is administered. Both the CD4(+) and the CD8(+) T-cell subsets are required for T1D development. Approximately 20 years ago, an association between certain class II major histocompatibility complex (MHC) alleles and susceptibility to T1D was reported. This finding led to enormous interest in the CD4(+) T cells participating in the development of T1D, while the CD8(+) subset was relatively ignored. However, the isolation of beta-cell-autoreactive CD8(+) T-cell clones from the islets of NOD mice helped to generate interest in the pathogenic role of this subset, as has accumulating evidence that certain class I MHC alleles are additional risk factors for T1D development in humans. Three distinct diabetogenic CD8(+) T-cell populations have now been characterized in NOD mice. Here, we review recent investigations exploring their selection, activation, trafficking, and antigenic specificities. As CD8(+) T cells are suspected contributors to beta-cell demise in humans, continued exploration of these critical areas could very possibly lead to tangible benefits for T1D patients and at-risk individuals.


A rare form of Guillain-Barré syndrome: pharyngeal-cervical-brachial variant.
Guillain-Barré syndrome is clinically characterized by acute onset of generalized, symmetrical, and ascending muscle weakness and areflexia from peripheral nerve involvement. In Guillain-Barré syndrome variants, however, some patients have unusual distribution of muscle involvement. Pharyngeal-cervical-brachial variant of Guillain-Barré syndrome is characterized by oropharyngeal, neck, and upper limb muscle involvement. Although Guillain-Barré syndrome is one of several post-infectious diseases that cause limb muscle weakness, the incidence of pharyngeal-cervical-brachial variant is relatively low. Here we report the case of a 16-month-old boy who developed a rare form of Guillain-Barré syndrome, the pharyngeal-cervical-brachial variant of the disease. We concluded that taking all the other etiologic reasons into consideration, pharyngeal-cervical-brachial variant of Guillain-Barré syndrome should be remembered in patients with symptoms of bulbar and upper extremity weakness not only for early diagnosis but also to plan the treatment early and follow up the potential complications.


Monomorphous angiocentric glioma: a distinctive epileptogenic neoplasm with features of infiltrating astrocytoma and ependymoma.
We present 8 examples of a neoplasm with features of both astrocytoma and ependymoma that may represent a distinct clinicopathologic entity. The cerebral hemispheric tumors occurred in patients that were 3, 4, 12, 14, 15, 26, 30, and 37 years of age. All presented with seizures that, with the exception of 2, began in childhood. Magnetic resonance imaging studies showed ill-defined, T2-hyperintense, generally noncontrast-enhancing lesions that, although centered on the cortex or amygdala, extended into the underlying white matter for a short distance. Histologically, the variably infiltrative tumors were distinctively angiocentric with well-developed perivascular pseudorosettes in some cases. Longitudinal and/or circumferential orientations of perivascular cells were common also. The cells were uniform in their cytologic features from case to case and were bipolar in all but one case. A glial nature was inferred from immunoreactivity for GFAP, and ependymal differentiation was suggested by positivity for EMA in three cases and ultrastructural features in one. Overall, the tumors were biologically indolent except for one that recurred and ultimately proved fatal.


A comparative study of cefamandole versus gentamicin plus clindamycin in the treatment of documented or suspected bacterial peritonitis.
Our data support the premise that antimicrobial therapy for peritonitis must be individualized. In the patient with minimal and moderate contamination who is not suppressed and who will undergo prompt and correct surgical therapy, then a single antimicrobial agent, such as cefamandole or perhaps even a first generation cephalosporin, such as cefazolin, will be adequate therapy even in instances of polymicrobial peritonitis when anticipated resistant organisms are present. In this situation, a single drug will be just as effective and safer when compared with the combination of an aminoglycoside and a specific antianaerobic agent. On the other hand, in the patient with immunosuppression, who is late to come to treatment or who has hospital acquired sepsis with probably a large contamination of resistant organisms, either a third generation cephalosporin with extended coverage or triple drug therapy, including a broad spectrum penicillin, an aminoglycoside and an anaerobic effective agent, should be the treatment of choice. Also, for infections with a culture proved overwhelming anaerobic flora, an antimicrobial specific for these pathogens should be used. The dictum, however, that all instances of peritonitis mandate double or triple drug therapy is a clinical impression based upon experimental models which do not correctly simulate the clinical situation.


Body fat and menstrual resumption in adult females with anorexia nervosa: a 1-year longitudinal study.
The variables predicting the resumption of menses in anorexia nervosa (AN) after weight restoration have not yet been fully established. We therefore aimed to investigate the association between several clinical parameters at inpatient discharge and the resumption of menses at 1-year follow-up in weight-restored adults with AN. Demographic, anthropometric, body composition and eating disorder features were assessed in 54 adult females with AN who had restored normal body weight [body mass index (BMI) ≥ 18.5 kg m(-) ²] at the end of specialist inpatient treatment. These variables were compared between participants who had resumed menses and those who were still amenorrheic 1 year after inpatient discharge. At 1-year follow-up, 35.2% of patients had resumed menstruation. No significant association was found between the resumption of menses and either age, duration of illness or BMI at inpatient admission, nor for BMI, global Eating Disorder Examination score or trunk fat percentage at inpatient discharge. Only total body fat percentage at inpatient discharge was significantly higher in patients who resumed menstruation, as confirmed by combined logistic regression analysis (odds ratio = 1.14, 95% confidence interval = 1.001-1.303, P = 0.049). A higher total body fat percentage at inpatient discharge is associated with the resumption of menses at 1-year follow-up in weight-restored adult females with AN.


Desipramine treatment reduces the long-term behavioural and neurochemical sequelae of early-life maternal separation.
Primate and rodent models of maternal separation have shown that repeated postnatal separation of young from the mother results in long-term changes to neurohormonal systems relevant to depression. To date, however, it remains unclear whether rodents that experience postnatal maternal separation display specific behavioural or biochemical features of depression in adulthood and whether these changes can be prevented by treatment with antidepressant drugs. We report here that maternally separated mice showed significantly shorter swim times on the forced swim test and significantly lower levels of brain-derived neurotrophic factor in dentate gyrus and CA3 regions of the hippocampus compared to control mice when assessed in adulthood. Neither of these differences was apparent in maternally separated mice that received chronic treatment with the antidepressant desipramine after maternal separation. These results suggest that intervention following early stress may eliminate the long-term vulnerability to behavioural and biochemical dysfunction that occurs following this early chronic stress.


Cervical human papillomavirus infection in Tunisian women.
It is well established that certain types of human papillomavirus (HPV) are the sexually transmitted agents etiologically linked to cervical cancer. Sexual habits have been shown to be a major determining factor for HPV infection. A large study was carried out to investigate the prevalence and risk factors associated with cervical infection with HPV in Tunisian women. PCR and restriction enzyme digestion were used to characterize HPV cervical infection in 106 Tunisian married women and 51 legal prostitutes. Epidemiological data were collected and correlated with HPV molecular genotyping. There was a higher relative frequency of HPV-DNA in prostitutes (39%) than in married women (14%) (p = 0.001). Molecular analyses of HPV types showed the most prevalent type in prostitutes to be HPV-16, a high-risk oncogenic type. In married women, the most prevalent type was HPV-6 which is associated with a low risk for cervical cancer. HPV-DNA detection was markedly increased in young adult women and in those having recent sexual experience. Cervical HPV infection in Tunisia is less frequent than in other African countries, but far from uncommon. The decrease of HPV prevalence in older women, regardless of their sexual behavior, may result from an efficient immune response acquired with age.


Biomarkers of oxidative stress study V: ozone exposure of rats and its effect on lipids, proteins, and DNA in plasma and urine.
Ozone exposure effect on free radical-catalyzed oxidation products of lipids, proteins, and DNA in the plasma and urine of rats was studied as a continuation of the international Biomarker of Oxidative Stress Study (BOSS) sponsored by NIEHS/NIH. The goal was to identify a biomarker for ozone-induced oxidative stress and to assess whether inconsistent results often reported in the literature might be due to the limitations of the available methods for measuring the various types of oxidative products. The time- and dose-dependent effects of ozone exposure on rat plasma lipid hydroperoxides, malondialdehyde, F2-isoprostanes, protein carbonyls, methionine oxidation, and tyrosine- and phenylalanine oxidation products, as well as urinary malondialdehyde and F2-isoprostanes were investigated with various techniques. The criterion used to recognize a marker in the model of ozone exposure was that a significant effect could be identified and measured in a biological fluid seen at both doses at more than one time point. No statistically significant differences between the experimental and the control groups at either ozone dose and time point studied could be identified in this study. Tissue samples were not included. Despite all the work accomplished in the BOSS study of ozone, no available product of oxidation in biological fluid has yet met the required criteria of being a biomarker. The current negative findings as a consequence of ozone exposure are of great importance, because they document that in complex systems, as the present in vivo experiment, the assays used may not provide meaningful data of ozone oxidation, especially in human studies.


A simulation study on the efficacy of advanced belt restraints to mitigate the effects of obesity for rear-seat occupant protection in frontal crashes.
Recent field data analyses have shown that the safety advantages of rear seats relative to the front seats have decreased in newer vehicles. Separately, the risks of certain injuries have been found to be higher for obese occupants. The objective of this study is to investigate the effects of advanced belt features on the protection of rear-seat occupants with a range of body mass index (BMI) in frontal crashes. Whole-body finite element human models with 4 BMI levels (25, 30, 35, and 40 kg/m2) developed previously were used in this study. A total of 52 frontal crash simulations were conducted, including 4 simulations with a standard rear-seat, 3-point belt and 48 simulations with advanced belt features. The parameters varied in the simulations included BMI, load limit, anchor pretensioner, and lap belt routing relative to the pelvis. The injury measurements analyzed in this study included head and hip excursions, normalized chest deflection, and torso angle (defined as the angle between the hip-shoulder line and the vertical direction). Analyses of covariance were used to test the significance (P <.05) of the results. Higher BMI was associated with greater head and hip excursions and larger normalized chest deflection. Higher belt routing increased the hip excursion and torso angle, which indicates a higher submarining risk, whereas the anchor pretensioner reduced hip excursion and torso angle. Lower load limits decreased the normalized chest deflection but increased the head excursion. Normalized chest deflection had a positive correlation with maximum torso angle. Occupants with higher BMI have to use higher load limits to reach head excursions similar to those in lower BMI occupants. The simulation results suggest that optimizing load limiter and adding pretensioner(s) can reduce injury risks associated with obesity, but conflicting effects on head and chest injuries were observed. This study demonstrated the feasibility and importance of using human models to investigate protection for occupants with various BMI levels. A seat belt system capable of adapting to occupant size and body shape will improve protection for obese occupants in rear seats.


Modified subaperture tool influence functions of a flat-pitch polisher with reverse-calculated material removal rate.
Numerical simulation of subaperture tool influence functions (TIF) is widely known as a critical procedure in computer-controlled optical surfacing. However, it may lack practicability in engineering because the emulation TIF (e-TIF) has some discrepancy with the practical TIF (p-TIF), and the removal rate could not be predicted by simulations. Prior to the polishing of a formal workpiece, opticians have to conduct TIF spot experiments on another sample to confirm the p-TIF with a quantitative removal rate, which is difficult and time-consuming for sequential polishing runs with different tools. This work is dedicated to applying these e-TIFs into practical engineering by making improvements from two aspects: (1) modifies the pressure distribution model of a flat-pitch polisher by finite element analysis and least square fitting methods to make the removal shape of e-TIFs closer to p-TIFs (less than 5% relative deviation validated by experiments); (2) predicts the removal rate of e-TIFs by reverse calculating the material removal volume of a pre-polishing run to the formal workpiece (relative deviations of peak and volume removal rate were validated to be less than 5%). This can omit TIF spot experiments for the particular flat-pitch tool employed and promote the direct usage of e-TIFs in the optimization of a dwell time map, which can largely save on cost and increase fabrication efficiency.


TUB gene expression in hypothalamus and adipose tissue and its association with obesity in humans.
Mutations in the Tubby gene (TUB) cause late-onset obesity and insulin resistance in mice and syndromic obesity in humans. Although TUB gene function has not yet been fully elucidated, studies in rodents indicate that TUB is involved in the hypothalamic pathways regulating food intake and adiposity. Aside from the function in central nervous system, TUB has also been implicated in energy metabolism in adipose tissue in rodents. We aimed to determine the expression and distribution patterns of TUB in man as well as its potential association with obesity. In situ hybridization was used to localize the hypothalamic regions and cells expressing TUB mRNA. Using RT-PCR, we determined the mRNA expression level of the two TUB gene alternative splicing isoforms, the short and the long transcript variants, in the hypothalami of 12 obese and 12 normal-weight subjects, and in biopsies from visceral (VAT) and subcutaneous (SAT) adipose tissues from 53 severely obese and 24 non-obese control subjects, and correlated TUB expression with parameters of obesity and metabolic health. Expression of both TUB transcripts was detected in the hypothalamus, whereas only the short TUB isoform was found in both VAT and SAT. TUB mRNA was detected in several hypothalamic regions involved in body weight regulation, including the nucleus basalis of Meynert and the paraventricular, supraoptic and tuberomammillary nuclei. We found no difference in the hypothalamic TUB expression between obese and control groups, whereas the level of TUB mRNA was significantly lower in adipose tissue of obese subjects as compared to controls. Also, TUB expression was negatively correlated with indices of body weight and obesity in a fat-depot-specific manner. Our results indicate high expression of TUB in the hypothalamus, especially in areas involved in body weight regulation, and the correlation between TUB expression in adipose tissue and obesity. These findings suggest a role for TUB in human obesity.


Spinal anesthesia for cesarean section: comparative study between isobaric and hyperbaric bupivacaine associated to morphine.
Bupivacaine preparations, plain or with glucose, are frequently used in the clinical practice. Blockade upper level is determined by local anesthetic spread in the CSF. This study aimed at comparing isobaric and hyperbaric bupivacaine in patients submitted to spinal anesthesia for Cesarean section. In this prospective, randomized and double-blind study 60 patients submitted to spinal anesthesia for Cesarean section were distributed in two groups: IB - (0.5% isobaric bupivacaine, 12.5 mg) and HB - (0.5% hyperbaric bupivacaine, 12.5 mg). After monitoring, venous puncture was performed followed by hydration with lactated Ringers solution. Spinal puncture was paramedially performed at L3-L4 interspace with 27G Quincke needle. Following the CSF dripping, morphine (100 microg) and bupivacaine were separately injected at the speed of 1 ml. 15 s(-1). With the patient back to supine position, two parameters were recorded: onset time (absence of sensitivity in L3) at 1-minute intervals as well as motor and sensory block after 20 minutes. All patients were kept with preanesthetic blood pressure levels until umbilical cord clamping, and if necessary, ephedrine was administered. Neonates were evaluated by Apgars score at 1 and 5 minutes. Sensory and motor blocks were also evaluated at PACU 120 minutes after local anesthetic injection. Groups were homogeneous. Onset time: Group IB (1', 50") and HB (1', 33"), with no statistical difference. Motor and sensory block at twenty minutes showed no significant difference. Ephedrine consumption: IB (11.83 mg) and HB (14.17 mg), showed also no statistical difference. PACU motor block evaluation showed significant differences. We concluded that 12.5 mg isobaric and hyperbaric bupivacaine associated to morphine (100 microg) in spinal anesthesia for Cesarean section in term pregnant women are effective and present similar profiles.


Implementing joint TB and HIV interventions in a rural district of Malawi: is there a role for an international non-governmental organisation?
In a rural district in Malawi, poorly motivated health personnel, shortages of human and financial resources, weak dialogue between existing tuberculosis (TB) and human immunodeficiency virus (HIV) programmes and poor community involvement are constraints to establishing joint TB-HIV interventions. The presence of a non-governmental organisation (NGO), Médecins Sans Frontières (MSF), in the health care delivery system provided an opportunity to bridge some of these gaps. The main inputs provided by MSF included additional staff, supplementary drugs including antiretroviral drugs, technical assistance and infrastructure development. The introduction of a scheme of monthly performance-linked incentives for health personnel proved successful in improving their performance, as judged by attendance rates as well as the quality and quantity of activities. This initiative also provided the district management with a tool for exerting pressure on health staff to improve their performance. The availability of independent NGO funds and a logistics team for construction of new infrastructure allowed the rapid initiation of new interventions at the district level without having to wait for disbursements of funds from the central level. This introduced a new dynamic of decentralised operational flexibility at the district level which improved access to care and support for people with TB-HIV.


Persistence of leukemia-initiating cells in a conditional knockin model of an imatinib-responsive myeloproliferative disorder.
Despite remarkable responses to the tyrosine kinase inhibitor imatinib, CML patients are rarely cured by this therapy perhaps due to imatinib refractoriness of leukemia-initiating cells (LICs). Evidence for this is limited because of poor engraftment of human CML-LICs in NOD-SCID mice and nonphysiologic expression of oncogenes in retroviral transduction mouse models. To address these challenges, we generated mice bearing conditional knockin alleles of two human oncogenes: HIP1/PDGFbetaR (H/P) and AML1-ETO (A/E). Unlike retroviral transduction, physiologic expression of H/P or A/E individually failed to induce disease, but coexpression of both H/P and A/E led to rapid onset of a fully penetrant, myeloproliferative disorder, indicating cooperativity between these two alleles. Although imatinib dramatically decreased disease burden, LICs persisted, demonstrating imatinib refractoriness of LICs.


Electrosurgery and ultrasonics on patients with implantable cardiac devices: Evidence of side effects in the dental practice.
Implantable cardiac pacemakers and cardiac defibrillators (ICDs) have been introduced in the care of patients with cardiac dysrhythmias. Most dental practitioners demonstrate extreme caution when treating patients with ICDs. This paper presents a review of the available literature on these devices and how they interact with dental electrosurgery and ultrasonic device use. Based on the analysis of the literature, this view is not corroborated by the current clinical data, and appears to be misguided. While further in-vivo studies are needed to truly determine the true level of risk, the evidence suggests that there is no contraindication for electrosurgery or ultrasonics use in patients with ICDs. Using the precautions stated in this analysis, the risk of any deleterious effect on ICD function is minimal.


TLR2 promotes Th2/Th17 responses via TLR4 and TLR7/8 by abrogating the type I IFN amplification loop.
TLR2 plays an important role in the removal of Gram-positive bacteria; contrastingly, it also appears to have important protective effects against unrestrained inflammation and subsequent organ injury during infection and autoimmunity. We hypothesized that TLR2 tunes the phenotype of dendritic cells (DCs) activated through other TLRs, thereby fulfilling a crucial role in the modulation of the immune response. TLR2 potently inhibited TLR4- and TLR7/8-induced cytokine production by human DCs. The inhibitory effect of TLR2 on the release of TNF-alpha but not of IL-12p70 was mediated by PI3K. TLR2 inhibits the production of IL-12p70 by dampening the type 1 IFN amplification loop. When DCs were triggered with the potent synergistic combination of LPS (TLR4) and R848 (TLR7/8) in conjunction with a TLR2 ligand, a clear shift to more Th2- and Th17-prone responses in the naive and memory T cell subpopulations was observed. This shift in T cell responses was inherent to the inability of TLR2-stimulated DCs to produce IL-12p70 and was dependent on the production of IL-1 and IL-6.


A nine-dimensional global potential energy surface for NH4(X(2)A(1)) and kinetics studies on the H + NH3↔ H2 + NH2 reaction.
Extensive ab initio calculations of the stationary points in the NH4(X(2)A1) system are reported using both coupled cluster and multi-reference configuration interaction methods. In addition, more than 100 000 points are generated over a large configuration space and energy range (6 eV) using the explicitly correlated unrestricted coupled cluster method with single, double, and perturbative triple excitations with the augmented correlation-consistent polarized triple zeta basis set (UCCSD(T)-F12a/aug-cc-pVTZ). Using the recently proposed permutation-invariant polynomial neural network (PIP-NN) method, these points are accurately fit to an analytical form with a total root mean squared error (RMSE) of 3.4 meV (0.08 kcal mol(-1)). Both the abstraction and exchange channels as well as the metastable ammonium radical (NH4) are included in this potential energy surface. Transition-state theory and quasi-classical trajectory calculations have been performed to obtain the rate constants for the abstraction reaction and its reverse. Comparison with available experimental results is satisfactory, providing supporting evidence for the accuracy of the potential.


Noninvasive monitoring of oxygenation in infants and children: practical considerations and areas of concern.
To review the technical principles of transcutaneous PO2 (tcPO2) monitors and pulse oximeters and to discuss methodological and practical issues related to the use of these devices in infants and children. Literature review and summary of personal experience. tcPO2 measurements are influenced by skin thickness, sensor temperature, amount of contact gel used, and state of peripheral perfusion. The average in vivo response time to a rapid decrease in PaO2 is approximately 16 seconds. Sensitivity to both hypoxemia (PaO2 < 50 mm Hg) and hyperoxemia (PaO2 > 80 to 100 mm Hg) is approximately 85%. Pulse oximeters require careful sensor placement (to avoid optical shunts) and adequate pulse pressures (> 20 mm Hg). They are prone to movement artifact. There are considerable differences in bias and precision between different brands, resulting, for example, in quite different upper alarm limits required to detect hyperoxemia reliably. The normal range of oxygen saturation measured by pulse oximetry, measured with one brand of pulse oximeter (Nellcor) during regular breathing, is 95% to 100% in preterm infants and 97% to 100% in full-term infants and children. Because both devices have their specific shortfalls, they should ideally be used in combination, particularly in critically ill preterm neonates. Where this is not feasible or necessary, clinicians must be aware of the limitations of the device they are using.


Regulation of neural cell adhesion molecule expression on cultured mouse Schwann cells by nerve growth factor.
Schwann cells from early postnatal mouse sciatic nerve were obtained as a homogenous population and shown by indirect immunofluorescence to express the neural cell adhesion molecules L1, N-CAM and J1 and their common carbohydrate epitope L2/HNK-1. L1 and N-CAM are synthesized in molecular forms that are slightly different from those expressed by small cerebellar neurons or astrocytes. As in astrocytes, the J1 antigen is expressed by Schwann cells in multiple forms generally ranging from 160 to 230 kd in the reduced state. J1 is secreted by Schwann cells in a 230-kd mol. wt form. Expression of L1 by Schwann cells can be regulated by nerve growth factor (NGF). L1 expression on the cell surface is increased 1.6-fold in the presence of NGF after 3 days of maintenance in vitro and 3-fold after 16 days. NGF does not change expression of N-CAM. The glia-derived neurite-promoting factor (GdNPF) increases L1 expression by a factor of 1.9 and decreases N-CAM expression by a factor of 0.4 after 3 days in vitro. J1 expression on Schwann cell surfaces remains unchanged in the presence of NGF or GdNPF. Antibodies to NGF abolish the influence of NGF on L1 expression. Addition of NGF antibodies to the Schwann cell cultures without exogenously added NGF decreases L1 expression, indicating that Schwann cells secrete NGF that may influence L1 expression by an autocrine mechanism. Our experiments show for the first time that cell adhesion molecule expression on a non-neuronal cell, the Schwann cell, can be directly regulated by the neurotrophic factor NGF. These observations indicate a considerable degree of 'plasticity' of peripheral glia in regulating cell adhesion molecule expression.


Catechin-rich oil palm leaf extract enhances bone calcium content of estrogen-deficient rats.
Postmenopausal estrogen deficiency often causes bone density loss and osteoporosis. This study evaluated the effects of an oral administration of oil palm leaf extract (OPL) on bone calcium content and structure, bone density, ash weights, and serum total alkaline phosphatase (T-ALP) of estrogen-deficient ovariectomized (OVX) rats. Female Sprague-Dawley rats were divided into five experimental groups: 1) intact (normal control); 2) ovariectomized (OVX control), and OVX rats supplemented with 3) 2% (w/v) green tea (OVX + GT), 4) OPL 150 mg/kg of body weight, or 5) OPL 300 mg/kg of body weight in the drinking water. After 3 mo, the OVX control rats had significantly decreased femur and tibia masses (-5% and -3%, respectively), ash (-15% and -10%), calcium content (-0.5% and -2.7%), and bone density and T-ALP concentrations (-40%) compared with intact rats. The catechin-rich OPL dose dependently increased the OVX bone density and structure, femur and tibia masses (by +8% and +12% respectively), ash (by +30% and +20% respectively), calcium (by +3% and +5%), and T-ALP concentrations (by +76%) compared with the OVX rats. The increases by OPL were higher than that in OVX + GT and control intact rats. The catechin-rich OPL increased the bone mass in estrogen-deficient rats by increasing osteoblast activities to higher levels than in normal rats and those supplemented with GT. This was shown by the modulation of serum T-ALP levels, bone calcium content, total mineral content, and bone histologic structure. The OPL is a potential inexpensive ingredient for protection against osteoporosis and influences bone metabolism by encouraging bone formation.


Sampling of the diffraction field.
When optical signals, like diffraction patterns, are processed by digital means the choice of sampling density and geometry is important during analog-to-digital conversion. Continuous band-limited signals can be sampled and recovered from their samples in accord with the Nyquist sampling criteria. The specific form of the convolution kernel that describes the Fresnel diffraction allows another, alternative, full-reconstruction procedure of an object from the samples of its diffraction pattern when the object is space limited. This alternative procedure is applicable and yields full reconstruction even when the diffraction pattern is undersampled and the Nyquist criteria are severely violated. Application of the new procedure to practical diffraction-related phenomena, like in-line holography, improves the processing efficiency without creating any associated artifacts on the reconstructed-object pattern.


Molecular evidence that most but not all carcinosarcomas of the uterus are combination tumors.
The pathogenesis of carcinosarcoma is still a subject of controversy. In the present study, molecular techniques were applied to determine the pathogenesis of uterine carcinosarcomas. The patterns of chromosome X inactivation were analyzed, targeting a portion of exon 1 of the human androgen receptor (HUMARA) in malignant epithelial and mesenchymal components. The presence of p53 and K-ras mutations were also analyzed. H&E-stained sections of paraffin-embedded, formalin-fixed tissues were microdissected to obtain both epithelial and nonepithelial lesions from 25 carcinosarcomas, and DNAs were extracted by proteinase K digestion. Following treatment with methylation-sensitive restriction endonuclease (HhaI or HpaII), PCR amplification was performed using nested primers targeted to the HUMARA locus. Mutations in the p53 gene and K-ras gene were found in eight (32%) and six (24%) tumors, respectively. The patterns of chromosome X inactivation were different between the carcinomatous and sarcomatous components of three carcinosarcomas, indicating that these three tumors represent collision tumors. By contrast, the patterns of chromosome X inactivation, K-ras sequence, and p53 sequence were identical in both carcinomatous and sarcomatous components in 21 carcinosarcomas, indicating that these 21 tumors represent combination tumors. One case produced equivocal results that precluded determination of whether it represented a collision or combination tumor. These observations show that although most carcinosarcomas are combination tumors, some develop as collision tumors. The determination of histogenesis in individual cases of carcinosarcoma using molecular markers may be worthwhile, because the result could help predict the prognosis of individual cases and help guide clinical management.


Influence of fermentation conditions on the surface properties and adhesion of Lactobacillus rhamnosus GG.
The surface properties of probiotic bacteria influence to a large extent their interactions within the gut ecosystem. There is limited amount of information on the effect of the production process on the surface properties of probiotic lactobacilli in relation to the mechanisms of their adhesion to the gastrointestinal mucosa. The aim of this work was to investigate the effect of the fermentation pH and temperature on the surface properties and adhesion ability to Caco-2 cells of the probiotic strain Lactobacillus rhamnosus GG. The cells were grown at pH 5, 5.5, 6 (temperature 37°C) and at pH 6.5 (temperature 25°C, 30°C and 37°C), and their surfaces analysed by X-ray photoelectron spectrometry (XPS), Fourier transform infrared spectroscopy (FT-IR) and gel-based proteomics. The results indicated that for all the fermentation conditions, with the exception of pH 5, a higher nitrogen to carbon ratio and a lower phosphate content was observed at the surface of the bacteria, which resulted in a lower surface hydrophobicity and reduced adhesion levels to Caco-2 cells as compared to the control fermentation (pH 6.5, 37°C). A number of adhesive proteins, which have been suggested in previous published works to take part in the adhesion of bacteria to the human gastrointestinal tract, were identified by proteomic analysis, with no significant differences between samples however. The temperature and the pH of the fermentation influenced the surface composition, hydrophobicity and the levels of adhesion of L. rhamnosus GG to Caco-2 cells. It was deduced from the data that a protein rich surface reduced the adhesion ability of the cells.


Genetic polymorphism of GSTM1, CYP2E1 and CYP2D6 in Egyptian bladder cancer patients.
Polymorphic changes in the GSTM1, CYP2E1 and the CYP2D6 genes have been reported to be individually associated with increased susceptibility to certain cancers. In the present study, the relationship between genetic polymorphism for these genes and development of urinary bladder cancer among Egyptian patients was investigated. Our results indicate that the frequency of bladder cancer patients with the GSTM1 null genotype is significantly higher than that of the normal controls (86.3 and 47.6%, respectively) with an odds ratio (OR) of 6.97 (95% CL -1.59-30.57, Fisher's exact P = 0.008). In contrast, our investigation failed to demonstrate any difference in the distribution of CYP2E1 polymorphism between bladder cancer patients and controls as detected by PstI restriction fragment length polymorphism (RFLP) analysis. RFLP analysis of the CYP2D6 gene revealed a non-significant increase in the number of extensive metabolizers (EM) among the patients compared to the controls (68 versus 48%). However, the EM genotypes enhances the risk further for individuals harboring the GSTM1 null genotype as individuals harboring both the EM and the GSTM1 null genotypes have an odds ratio of 14.0 (95% CL = 1.3- 151.4, Fisher's exact P = 0.02) compared to individuals harboring the EM and the GSTM1 +/+ genotypes. In conclusion, our results indicate that genetic polymorphism, especially in GSTM1 and CYP2D6 could play an important role as host risk factors for development of urinary bladder cancer among Egyptians.


Wetting morphologies on an array of fibers of different radii.
We investigate the equilibrium morphology of a finite volume of liquid placed on two parallel rigid fibers of different radii. As observed for identical radii fibers, the liquid is either in a column morphology or adopts a drop shape depending on the inter-fiber distance. However the cross-sectional area and the critical inter-fiber distance at which the transition occurs are both modified by the polydispersity of the fibers. Using energy considerations, we analytically predict the critical inter-fiber distance corresponding to the transition between the column and the drop morphologies. This distance depends both on the radii of the fibers and on the contact angle of the liquid. We perform experiments using a perfectly wetting liquid on two parallel nylon fibers: the results are in good agreement with our analytical model. The morphology of the capillary bridges between fibers of different radii is relevant to the modeling of large arrays of polydisperse fibers.


Glycosylated insulin complexed to Concanavalin A. Biochemical basis for a closed-loop insulin delivery system.
The oligosaccharides maltose, maltotriose, mannotriose, and mannotetrose have been chemically attached to insulin molecules. Incubation of oligosaccharide and insulin at different molar ratios, with or without addition of cyanoborohydride, showed a nearly linear increase in carbohydrate attachment over time. The intravenous t1/2 of 125I-labeled sugar-insulin derivatives was identical to that of unmodified insulin (3.0 min). Biologic activity of these derivatives, assessed in rats by use of a blood glucose depression assay, did not differ significantly from control. These glycosylated insulin molecules are reversibly bound to the glucose-binding lectin Concanavalin A (Con A). Such sugar-insulin/lectin complexes serve as an insulin reservoir from which sugar-insulin molecules are displaced by glucose. Release of sugar-insulin molecules is a function of the particular sugar-insulin and of the ambient glucose concentration. Glucose displacement of glycosylated insulin complexed to Con A is in direct proportion to the amount of glucose present in the surrounding fluid. At each glucose concentration, the relative binding affinity of the maltotriose derivative is less than that of the mannotriose derivative, while the relative binding affinity of both maltotriose and mannotriose are less than that of the mannotetrose derivative. Prolonged incubation at 37 degrees C causes sugar-insulin, like unmodified insulin, to spontaneously aggregate into high-mol-wt, nondiffusable complexes. This aggregation phenomenon was found to be markedly inhibited when glycosylated insulins were synthesized utilizing partially sulfated insulin. Results from the studies described in this report provide the biochemical basis for a closed-loop, glucose-controlled insulin delivery system, utilizing glycosylated insulin complexed to Con A.


Recurrent meningococcal meningitis. A case report.
A 17-year-old Coloured youth presented at Tygerberg Hospital on six occasions between 1978 and 1981. He was admitted three times for meningococcal meningitis and three times for a meningitic illness clinically suggestive thereof. He had been admitted twice before to the City Hospital, Cape Town, in 1966 and 1968, for presumed meningococcal meningitis. He was fully evaluated in the outpatient department and found to have no detectable total haemolytic complement activity. There was no evidence of classic or alternative pathway activation. He has been tentatively designated as having a deficiency in the C6 or C8 components of the terminal membrane attack mechanism of the complement cascade.


An experimental model of postnatal jaundice in the suckling rat. Suppression of induced hyperbilirubinemia by Sn-protoporphyrin.
A model of experimental postnatal hyperbilirubinemia in the rat has been developed utilizing the heme precursor delta-aminolevulinic acid (ALA) to produce jaundice during a selective time period after birth. This time period is defined as that between 7 d postnatally, when the initial postpartum alterations of serum bilirubin and heme metabolism in the neonate have subsided, and 21 d, when the hepatic conjugation mechanism for the bile pigment appears fully developed. Administration of ALA in this time period led to a rapid, consistent, and significant dose-dependent increase in serum bilirubin levels in the newborn animals. Heme administration produced a qualitatively similar but enhanced effect. Both compounds, in addition, induced a dose-dependent increase in hepatic heme oxygenase activity concomitant with the increase in serum bilirubin levels. Neither compound increased serum bilirubin levels significantly when administered at or after 21 d postnatally. Administration of the synthetic metalloporphyrin, Sn-protoporphyrin, to ALA-treated neonates resulted in a dose-dependent decrease in serum bilirubin levels and hepatic heme oxygenase activity. Mn- and Zn-protoporphyrin in comparable doses did not significantly inhibit ALA-induced hyperbilirubinemia. Sn-protoporphyrin also inhibited the hyperbilirubinemia produced by heme in the suckling animals. ALA administration to newborn rats during the specific postnatal period described provides a simple and convenient model of experimental jaundice in the developing neonate which permits an examination of the potential ability of synthetic metalloporphyrins or other compounds to suppress induced hyperbilirubinemia in the newborn animal. The ability to induce a consistent and significant degree of jaundice in the postnatal rat by the method described may also be useful for other types of studies concerned with the biological disposition and effects of endogenously formed bilirubin in the neonate. The results of this study confirm in another model system the potent ability of Sn-protoporphyrin to suppress jaundice in the neonate, and suggest that suppression of heme oxidation by synthetic heme analogues may represent a useful therapeutic approach to the problem of severe hyperbilirubinemia in human premature newborn.


Induction of the Keap1/Nrf2/ARE pathway by oxidizable diphenols.
Inducible Keap1/Nrf2/ARE pathway determines the ability of multicellular organisms to adapt to conditions of stress caused by oxidants and electrophiles through upregulating proteins with versatile cytoprotective functions. Para- and ortho-hydroquinones were among the first identified small-molecule inducers of this pathway. Their oxidative lability strongly suggested that the electrophilic quinone metabolites, and not the hydroquinones themselves, were the ultimate inducers. Molecular orbital calculations re-enforced this notion by showing linear correlations between inducer potency and: (i) the ability of diphenols to release electrons, and (ii) the electron affinity of their corresponding quinones. Consequently, a two-step mechanism was proposed which involves oxidation of the diphenols to their corresponding quinone derivatives, followed by modification of specific cysteine residues of the sensor protein Keap1. Our finding that Cu(2+), as well as other transition metals, enhanced induction by oxidizable diphenols provided a rationale to test this hypothesis. We found that hypoxia inhibits the potentiation of diphenolic inducer activity afforded by copper as oxygen is required to oxidize Cu(+) and regenerate Cu(2+). In the stably transfected AREc32 reporter cell line, exposure to 2-tert-butyl-1,4-hydroquinone (tBHQ) for 30min induced ARE-luciferase (measured 24h later) only in the presence of copper (Cu(2+) or Cu(+)), whereas induction by tert-butyl-1,4-quinone (tBQ) was copper-independent. tBQ, but not tBHQ, reacts with cysteine residues of Keap1. Other para- and ortho-hydroquinones, such as catechol estrogens, dopamine, and l-DOPA, also induce ARE-driven transcription in a Cu(2+)-dependent manner. Thus, based on theoretical and experimental evidence, the oxidation of para- and ortho-hydroquinones to their corresponding electrophilic quinones is a requisite step for the activation of the Keap1/Nrf2/ARE pathway.


Detection and subtyping of human herpesvirus-8 in renal transplant patients before and after remission of Kaposi's sarcoma.
Kaposi's sarcoma (KS) is a complication of renal transplantation. If the human herpesvirus-8 (HHV-8) causes KS, the virus should be present in all KS lesions and be drastically reduced or cleared from involved tissue on remission of the KS. Fourteen renal transplant patients with cutaneous KS, including autopsy material from two cases, were investigated for the presence of HHV-8. A second skin biopsy was taken from 11 survivors, after remission of KS, from normal skin in the same anatomical region as the first biopsy. Remission was induced by reduction or cessation of immunosuppression. A peripheral blood sample was collected simultaneously with the repeat biopsy. A nested polymerase chain reaction assay was used to detect HHV-8 DNA in the biopsy tissue and peripheral blood mononuclear cells followed by direct sequencing of polymerase chain reaction product to detect any nucleotide changes. HHV-8 DNA was detected in all the cutaneous KS and all the visceral KS samples, as well as a number of KS-free organs including the thyroid, salivary gland, and myocardium that have not been described before. Mutations in the viral DNA could be demonstrated in all patients. The mutations found were related more to that seen in AIDS-KS cases than that found in African endemic KS cases. HHV-8 sequences could be detected in follow-up frozen skin biopsies of five patients but were negative in the equivalent formalin-fixed specimens. Viral DNA was also detected in 2 of 11 peripheral blood mononuclear cell samples collected at the time of the follow-up skin biopsies. Reduction or withdrawal of immunosuppression allows the immune system to recover sufficiently to reduce viral replication with subsequent viral persistence and low grade viral replication that coincides with clinical remission of the KS lesions. This provides further evidence for the important etiological role played by HHV-8 in the pathogenesis of posttransplant KS.


Metabolic surgery profoundly influences gut microbial-host metabolic cross-talk.
Bariatric surgery is increasingly performed worldwide to treat morbid obesity and is also known as metabolic surgery to reflect its beneficial metabolic effects especially with respect to improvement in type 2 diabetes. Understanding surgical weight loss mechanisms and metabolic modulation is required to enhance patient benefits and operative outcomes. The authors applied a parallel and statistically integrated bacterial profiling and metabonomic approach to characterise Roux-en-Y gastric bypass (RYGB) effects in a non-obese rat model. Substantial shifts of the main gut phyla towards higher concentrations of Proteobacteria (52-fold), specifically Enterobacter hormaechei, are shown. Low concentrations of Firmicutes (4.5-fold) and Bacteroidetes (twofold) in comparison with sham-operated rats were also found. Faecal extraction studies revealed a decrease in faecal bile acids and a shift from protein degradation to putrefaction through decreased faecal tyrosine with concomitant increases in faecal putrescine and diaminoethane. Decreased urinary amines and cresols were found and indices of modulated energy metabolism were demonstrated after RYGB, including decreased urinary succinate, 2-oxoglutarate, citrate and fumarate. These changes could also indicate renal tubular acidosis, which is associated with increased flux of mitochondrial tricarboxylic acid cycle intermediates. A surgically induced effect on the gut-brain-liver metabolic axis is inferred from modulated faecal γ-aminobutyric acid and glutamate. This profound co-dependence of mammalian and microbial metabolism, which is systematically altered after RYGB surgery, suggests that RYGB exerts local and global metabolic effects. The effect of RYGB surgery on the host metabolic-microbial cross-talk augments our understanding of the metabolic phenotype of bariatric procedures and can facilitate enhanced treatments for obesity-related diseases.


High-efficiency stable transformation of the model fern species Ceratopteris richardii via microparticle bombardment.
Ferns represent the most closely related extant lineage to seed plants. The aquatic fern Ceratopteris richardii has been subject to research for a considerable period of time, but analyses of the genetic programs underpinning developmental processes have been hampered by a large genome size, a lack of available mutants, and an inability to create stable transgenic lines. In this paper, we report a protocol for efficient stable genetic transformation of C. richardii and a closely related species Ceratopteris thalictroides using microparticle bombardment. Indeterminate callus was generated and maintained from the sporophytes of both species using cytokinin treatment. In proof-of-principle experiments, a 35S::β-glucuronidase (GUS) expression cassette was introduced into callus cells via tungsten microparticles, and stable transformants were selected via a linked hygromycin B resistance marker. The presence of the transgene in regenerated plants and in subsequent generations was validated using DNA-blot analysis, reverse transcription-polymerase chain reaction, and GUS staining. GUS staining patterns in most vegetative tissues corresponded with constitutive gene expression. The protocol described in this paper yields transformation efficiencies far greater than those previously published and represents a significant step toward the establishment of a tractable fern genetic model.


Maturation state of dendritic cells during the extracorporeal photopheresis and its relevance for the treatment of chronic graft-versus-host disease.
Chronic graft-versus-host disease (cGVHD) represents a major cause of morbidity and mortality after allogeneic bone marrow transplantation (BMT). Extracorporeal photochemotherapy (ECP), a technique used in the treatment of cutaneous T-cell lymphoma, has also shown clinical efficacy in the treatment of refractory cGVHD. In this study, the dynamics of dendritic cell (DC) activation were investigated during the process of photopheresis in patients treated for refractory cGVHD. It is reported that myeloid DCs can be isolated from the photopheretic products before retransfusion to the patient. It is shown that DCs in ECP product are in the immature state with respect to the phenotypic and functional characteristics. In contrast to the in vitro-generated monocyte-derived DCs and DCs not treated by 8-methoxypsoralen and UVA, they produce significant amounts of interleukin-10 (IL-10). They efficiently capture apoptotic lymphocytes and do not induce proliferation of T lymphocytes. They preserve the capacity to be activated by polyriboinosinic polyribocytidylic acid and lipopolysaccharide, however. ECP also induces rapid and massive apoptosis of alloreactive lymphocytes. A model of the potential implication of IL-10-producing DCs in the down regulation of harmful alloreactive immune reaction is presented. It is believed that this study provides a novel insight into the mechanisms of action of ECP in the control of cGVHD.


Noninvasive Fetal Electrocardiography Part I: Pan-Tompkins' Algorithm Adaptation to Fetal R-peak Identification.
Indirect fetal electrocardiography is preferable to direct fetal electrocardiography because of being noninvasive and is applicable also during the end of pregnancy, besides labor. Still, the former is strongly affected by noise so that even R-peak detection (which is essential for fetal heart-rate evaluations and subsequent processing procedures) is challenging. Some fetal studies have applied the Pan-Tompkins' algorithm that, however, was originally designed for adult applications. Thus, this work evaluated the Pan-Tompkins' algorithm suitability for fetal applications, and proposed fetal adjustments and optimizations to improve it. Both Pan-Tompkins' algorithm and its improved version were applied to the "Abdominal and Direct Fetal Electrocardiogram Database" and to the "Noninvasive Fetal Electrocardiography Database" of Physionet. R-peak detection accuracy was quantified by computation of positive-predictive value, sensitivity and F1 score. When applied to "Abdominal and Direct Fetal Electrocardiogram Database", the accuracy of the improved fetal Pan-Tompkins' algorithm was significantly higher than the standard (positive-predictive value: 0.94 vs. 0.79; sensitivity: 0.95 vs. 0.80; F1 score: 0.94 vs. 0.79; P<0.05 in all cases) on indirect fetal electrocardiograms, whereas both methods performed similarly on direct fetal electrocardiograms (positive-predictive value, sensitivity and F1 score all close to 1). Improved fetal Pan-Tompkins' algorithm was found to be superior to the standard also when applied to "Noninvasive Fetal Electrocardiography Database" (positive-predictive value: 0.68 vs. 0.55, P<0.05; sensitivity: 0.56 vs. 0.46, P=0.23; F1 score: 0.60 vs. 0.47, P=0.11). In indirect fetal electrocardiographic applications, improved fetal Pan-Tompkins' algorithm is to be preferred over the standard, since it provides higher R-peak detection accuracy for heart-rate evaluations and subsequent processing.


Risk factors for surgery due to rotator cuff disease in a population-based cohort.
Few risk factors for rotator cuff disease (RCD) and corresponding treatment have been firmly established. The aim of this study was to evaluate the relationship between numerous risk factors and the incidence of surgery for RCD in a large cohort. A population-based cohort of people aged between 40 and 69 years in the UK (the UK Biobank) was studied. People who underwent surgery for RCD were identified through a link with NHS inpatient records covering a mean of eight years after enrolment. Multivariate Cox proportional hazards regression was used to calculate hazard ratios (HRs) as estimates of associations with surgery for RCD accounting for confounders. The risk factors which were considered included age, sex, race, education, Townsend deprivation index, body mass index (BMI), occupational demands, and exposure to smoking. Of the 421,894 people who were included, 47% were male. The mean age at the time of enrolment was 56 years (40 to 69). A total of 2,156 people were identified who underwent surgery for RCD. Each decade increase in age was associated with a 55% increase in the incidence of RCD surgery (95% confidence interval (CI) 46% to 64%). Male sex, non-white race, lower deprivation score, and higher BMI were significantly associated with a higher risk of surgery for RCD (all p < 0.050). Greater occupational physical demands were significantly associated with higher rates of RCD surgery (HR = 2.1, 1.8, and 1.4 for 'always', 'usually', and 'sometimes' doing heavy manual labour vs 'never', all p < 0.001). Former smokers had significantly higher rates of RCD surgery than those who had never smoked (HR 1.23 (95% CI 1.12 to 1.35), p < 0.001), while current smokers had similar rates to those who had never smoked (HR 0.94 (95% CI 0.80 to 1.11)). Among those who had never smoked, the risk of surgery was higher among those with more than one household member who smoked (HR 1.78 (95% CI 1.08 to 2.92)). The risk of RCD surgery was not significantly related to other measurements of secondhand smoking. Many factors were independently associated with surgery for RCD, including older age, male sex, higher BMI, lower deprivation score, and higher occupational physical demands. Several of the risk factors which were identified are modifiable, suggesting that the healthcare burden of RCD might be reduced through the pursuit of public health goals, such as reducing obesity and modifying occupational demands. Cite this article: Bone Joint J 2020;102-B(3):352-359.


The ecological relationship of tobacco smoking to the prevalence of symptoms of asthma and other atopic diseases in children: the International Study of Asthma and Allergies in Childhood (ISAAC).
This study examined the relationship between parental smoking and asthma and other atopic diseases at the ecological level. The prevalence of atopic symptoms in 6-7- and 13-14-year old children was assessed in 91 centres (from 38 countries) and 155 centres (from 56 countries) respectively in the International Study of Asthma and Allergy in Childhood (ISAAC). These symptoms were related to prevalence of tobacco smoking for each country by gender as reported by the World Health Organisation. There was a significant negative association between the prevalence of smoking by men and the prevalence of symptoms of asthma and rhinitis, but not eczema in the 13-14-year age group. There was a significant positive relationship between prevalence of smoking by women and prevalence of wheeze in the last 12 months, but not for any other symptoms in the 13-14-year age group. In the 6-7-year age group, there was a significant negative correlation between prevalence of smoking by men and the prevalence of wheeze causing sleep disturbance and a close to significant negative association at the 5% level with the prevalence of wheeze in the last 12 months and rhinitis. Thus, for the countries that are included in this analysis, countries that have high adult male smoking rates have a lower risk of asthma and rhinitis symptoms in children. It should be stressed that this analysis does not involve information on individual exposures and therefore does not contradict the well-established association of active and passive smoking in individuals with the occurrence of asthma symptoms in the same individuals. Rather, it indicates that this well-established individual-level association does not account for the international differences in asthma prevalence, and that other risk factors for asthma must be responsible for the observed international patterns.


Staphylococcus aureus protein A binds to osteoblasts and triggers signals that weaken bone in osteomyelitis.
Osteomyelitis is a debilitating infectious disease of the bone. It is predominantly caused by S. aureus and is associated with significant morbidity and mortality. It is characterised by weakened bones associated with progressive bone loss. Currently the mechanism through which either bone loss or bone destruction occurs in osteomyelitis patients is poorly understood. We describe here for the first time that the major virulence factor of S. aureus, protein A (SpA) binds directly to osteoblasts. This interaction prevents proliferation, induces apoptosis and inhibits mineralisation of cultured osteoblasts. Infected osteoblasts also increase the expression of RANKL, a key protein involved in initiating bone resorption. None of these effects was seen in a mutant of S. aureus lacking SpA. Complementing the SpA-defective mutant with a plasmid expressing spa or using purified protein A resulted in attachment to osteoblasts, inhibited proliferation and induced apoptosis to a similar extent as wildtype S. aureus. These events demonstrate mechanisms through which loss of bone formation and bone weakening may occur in osteomyelitis patients. This new information may pave the way for the development of new and improved therapeutic agents to treat this disease.


Constructed wetland integrated microbial fuel cell system: looking back, moving forward.
In the last 10 years, the microbial fuel cell (MFC) has been extensively studied worldwide to extract energy from wastewater via electricity generation. More recently, a merged technique of embedding MFC into a constructed wetland (CW) has been developed and appears to be increasingly investigated. The driving force to integrate these two technologies lies in the fact that CWs naturally possess a redox gradient (depending on flow direction and wetland depth), which is required by MFCs as anaerobic anode and aerobic cathode chambers. No doubt, the integration of MFC with a CW will upgrade the CW to allow it to be used for wastewater treatment and, simultaneously, electricity generation, making CWs more sustainable and environmentally friendly. Currently, published work shows that India, China, Ireland, Spain, Germany and Malaysia are involved in the development of this technology although it is in its infant stage and many technical issues are faced on system configuration, operation and maximisation of electricity production. This paper aims to provide an updated review and analysis of the CW-MFC development. Focuses are placed on the experience gained so far from different researchers in the literature and further research directions and proposals are discussed in great detail.


Unified biogenesis of ambiguine, fischerindole, hapalindole and welwitindolinone: identification of a monogeranylated indolenine as a cryptic common biosynthetic intermediate by an unusual magnesium-dependent aromatic prenyltransferase.
Biochemical characterization of aromatic prenyltransferase AmbP1 and its close homologs WelP1/FidP1 in hapalindole-type alkaloid biosynthetic pathways is reported. These enzymes mediate the magnesium-dependent selective formation of 3-geranyl 3-isocyanovinyl indolenine (2) from cis-indolyl vinyl isonitrile and geranyl pyrophosphate. The role of the magnesium cofactor in AmbP1/WelP1/FidP1 catalysis is unusual for a microbial aromatic prenyltransferase, as it not only facilitates the formation of 2 but also prevents its rearrangement to an isomeric 2-geranyl 3-isocyanovinyl indole (3). The discovery of 2 as a cryptically conserved common biosynthetic intermediate to all hapalindole-type alkaloids suggests an enzyme-mediated Cope rearrangement and aza-Prins-type cyclization cascade is required to transform 2 to a polycyclic hapalindole-like scaffold.


Prevention and management of intraoperative crisis in VATS and open chest surgery: how to avoid emergency conversion.
Video assisted thoracic surgery (VATS) has become a routinely utilized approach to complex procedures of the chest, such as pulmonary resection. It has been associated with decreased postoperative pain, shorter length of stay and lower incidence of complications such as pneumonia. Limitations to this modality may include limited exposure, lack of tactile feedback, and a two-dimensional view of the surgical field. Furthermore, the lack of an open incision may incur technical challenges in preventing and controlling operative misadventures leading to major hemorrhage or other intraoperative emergencies. While these events may occur in the best of circumstances, prevention strategies are the primary means of avoiding these injuries. Unplanned conversions for major intraoperative bleeding or airway injury during general thoracic surgical procedures are relatively rare and often can be avoided with careful preoperative planning, review of relevant imaging, and meticulous surgical technique. When these events occur, a pre-planned, methodical response with initial control of bleeding, assessment of injury, and appropriate repair and/or salvage procedures are necessary to maximize outcomes. The surgeon should be well versed in injury-specific incisions and approaches to maximize adequate exposure and when feasible, allow completion of the index operation. Decisions to continue with a minimally invasive approach should consider the comfort and experience level of the surgeon with these techniques, and the relative benefit gained against the risk incurred to the patient. These algorithms may be expected to shift in the future with increasing sophistication and capabilities of minimally invasive technologies and approaches.


Predicting Amino Acid Substitution Probabilities Using Single Nucleotide Polymorphisms.
Fast genome sequencing offers invaluable opportunities for building updated and improved models of protein sequence evolution. We here show that Single Nucleotide Polymorphisms (SNPs) can be used to build a model capable of predicting the probability of substitution between amino acids in variants of the same protein in different species. The model is based on a substitution matrix inferred from the frequency of codon interchanges observed in a suitably selected subset of human SNPs, and predicts the substitution probabilities observed in alignments between Homo sapiens and related species at 85-100% of sequence identity better than any other approach we are aware of. The model gradually loses its predictive power at lower sequence identity. Our results suggest that SNPs can be employed, together with multiple sequence alignment data, to model protein sequence evolution. The SNP-based substitution matrix developed in this work can be exploited to better align protein sequences of related organisms, to refine the estimate of the evolutionary distance between protein variants from related species in phylogenetic trees and, in perspective, might become a useful tool for population analysis.