Wide localized excision and reconstruction for minor salivary gland tumours.
Insufficient excision is one of the main factors leading to recurrence of minor salivary gland tumours. Our aim is to perform large resections with immediate reconstruction in order to provide local control, as well as to achieve better cosmetic and functional results. Four consecutive patients suffering from minor salivary gland tumours of the upper aerodigestive tract were treated: two had adenoid cystic carcinomas (one of the lateral oropharyngeal mucosa, one of the retromolar area); one had a polymorphous low-grade carcinoma (at the base of the tongue); and one had mucoepidermoid carcinoma (on the floor of the mouth). Two radial forearm free-flaps, a trapezius myocutaneous pedicled flap (including an adjacent osseous part of the scapula), and a sternocleidomastoid muscular pedicled flap were used, respectively, for reconstruction. Additional external radiation was used for three patients (60 cGy). The postoperative course was uneventful for each patient, and the ability to chew and swallow was rapidly restored. Two patients, locally controlled, with adenoid cystic carcinomas died: one from brain metastases (four years post-surgery), the other with pulmonary metastases (eight-years post-surgery). One patient is alive with a recurrent tumour of the pterygomaxillary fossa (without trismus) sixteen years after the original operation; another patient is free of recurrent disease at the base of the tongue but has recently undergone surgery, with success, for two pulmonary metastases sixty months after the initial surgery. For minor salivary gland carcinoma of the head and neck, large resection with immediate reconstruction affords fast recovery and long-lasting locoregional control. Additional radiotherapy is feasible.


H3K79me2/3 controls enhancer-promoter interactions and activation of the pan-cancer stem cell marker PROM1/CD133 in MLL-AF4 leukemia cells.
MLL gene rearrangements (MLLr) are a common cause of aggressive, incurable acute lymphoblastic leukemias (ALL) in infants and children, most of which originate in utero. The most common MLLr produces an MLL-AF4 fusion protein. MLL-AF4 promotes leukemogenesis by activating key target genes, mainly through recruitment of DOT1L and increased histone H3 lysine-79 methylation (H3K79me2/3). One key MLL-AF4 target gene is PROM1, which encodes CD133 (Prominin-1). CD133 is a pentaspan transmembrane glycoprotein that represents a potential pan-cancer target as it is found on multiple cancer stem cells. Here we demonstrate that aberrant PROM1/CD133 expression is essential for leukemic cell growth, mediated by direct binding of MLL-AF4. Activation is controlled by an intragenic H3K79me2/3 enhancer element (KEE) leading to increased enhancer-promoter interactions between PROM1 and the nearby gene TAPT1. This dual locus regulation is reflected in a strong correlation of expression in leukemia. We find that in PROM1/CD133 non-expressing cells, the PROM1 locus is repressed by polycomb repressive complex 2 (PRC2) binding, associated with reduced expression of TAPT1, partially due to loss of interactions with the PROM1 locus. Together, these results provide the first detailed analysis of PROM1/CD133 regulation that explains CD133 expression in MLLr ALL.


Evaluation of pancreas transplant needle biopsy: reproducibility and revision of histologic grading system.
Tissue samples for the diagnosis of pancreatic allograft rejection are now obtained routinely through the application of the percutaneous needle biopsy technique. The availability of biopsy material (89% adequate for diagnosis in our setting) presents a challenge for pathologists who are asked to provide a fast and accurate diagnosis of rejection and its severity, while at the same time being able to differentiate rejection from other causes of graft dysfunction. To differentiate rejection from other pathologic processes, 26 histologic features were assessed in 92 biopsies performed for confirmation of clinical diagnosis of rejection and the results were compared with 31 protocol biopsies, 12 allograft pancreatectomies with non-rejection pathology, and 30 native pancreas resections with various disease processes. Based on these comparisons, a constellation of findings relating to the vascular, septal, and acinar inflammation was identified for the diagnosis of rejection. Application of these features led us to revise our scheme for grading rejection (ranging from 0-normal to V-severe rejection) to include the categories of "inflammation of undetermined significance" and "minimal rejection." The scheme was used by five pathologist to grade 20 biopsies independently of any clinical data and the interobserver level of agreement was highly significant (kappa=0.83, P<0.0001). This grading scheme was applied blindly to all (183) biopsies from 77 patients with 6-52 months of follow-up. The correlation of the highest degree of rejection on each patient and ultimate graft loss (0% for grades 0-I, 11.5% for grade II, 17.3% for grade III, 37.5% for grade IV, and 100% for grade V) was highly statistically significant (P<0.002). The fraction of grafts lost due to pure immunologic causes increased proportionally to the grade of rejection (0, 50, 66, and 100% for grades II, III, IV, and V, respectively). This study provides strong support for the proposed pancreas rejection grading scheme and confirms its potential for practical use.


Validation of the efficacy of a solar-thermal powered autoclave system for off-grid medical instrument wet sterilization.
This work describes the efficacy of a solar-thermal powered autoclave used for the wet sterilization of medical instruments in off-grid settings where electrical power is not readily available. Twenty-seven trials of the solar-thermal powered system were run using an unmodified non-electric autoclave loaded with a simulated bundle of medical instruments and biological test agents. Results showed that in 100% of the trials the autoclave achieved temperatures in excess of 121°C for 30 minutes, indicator tape displayed visible reactions to steam sterilization, and biological tests showed that microbial agents had been eliminated, in compliance with the Centers for Disease Control and Prevention requirements for efficacious wet sterilization.


Melanin deposition in two Cryptococcus species depends on cell-wall composition and flexibility.
Cryptococcus neoformans and Cryptococcus gattii are two species complexes in the large fungal genus Cryptococcus and are responsible for potentially lethal disseminated infections. These two complexes share several phenotypic traits, such as production of the protective compound melanin. In C. neoformans, the pigment associates with key cellular constituents that are essential for melanin deposition within the cell wall. Consequently, melanization is modulated by changes in cell-wall composition or ultrastructure. However, whether similar factors influence melanization in C. gattii is unknown. Herein, we used transmission EM, biochemical assays, and solid-state NMR spectroscopy of representative isolates and "leaky melanin" mutant strains from each species complex to examine the compositional and structural factors governing cell-wall pigment deposition in C. neoformans and C. gattii. The principal findings were the following. 1) C. gattii R265 had an exceptionally high chitosan content compared with C. neoformans H99; a rich chitosan composition promoted homogeneous melanin distribution throughout the cell wall but did not increase the propensity of pigment deposition. 2) Strains from both species manifesting the leaky melanin phenotype had reduced chitosan content, which was compensated for by the production of lipids and other nonpolysaccharide constituents that depended on the species or mutation. 3) Changes in the relative rigidity of cell-wall chitin were associated with aberrant pigment retention, implicating cell-wall flexibility as an independent variable in cryptococcal melanin assembly. Overall, our results indicate that cell-wall composition and molecular architecture are critical factors for the anchoring and arrangement of melanin pigments in both C. neoformans and C. gattii species complexes.


How severe and prevalent are Ebola and Marburg viruses? A systematic review and meta-analysis of the case fatality rates and seroprevalence.
Ebola and Marburg virus diseases are said to occur at a low prevalence, but are very severe diseases with high lethalities. The fatality rates reported in different outbreaks ranged from 24-100%. In addition, sero-surveys conducted have shown different seropositivity for both Ebola and Marburg viruses. We aimed to use a meta-analysis approach to estimate the case fatality and seroprevalence rates of these filoviruses, providing vital information for epidemic response and preparedness in countries affected by these diseases. Published literature was retrieved through a search of databases. Articles were included if they reported number of deaths, cases, and seropositivity. We further cross-referenced with ministries of health, WHO and CDC databases. The effect size was proportion represented by case fatality rate (CFR) and seroprevalence. Analysis was done using the metaprop command in STATA. The weighted average CFR of Ebola virus disease was estimated to be 65.0% [95% CI (54.0-76.0%), I2 = 97.98%] whereas that of Marburg virus disease was 53.8% (26.5-80.0%, I2 = 88.6%). The overall seroprevalence of Ebola virus was 8.0% (5.0%-11.0%, I2 = 98.7%), whereas that for Marburg virus was 1.2% (0.5-2.0%, I2 = 94.8%). The most severe species of ebolavirus was Zaire ebolavirus while Bundibugyo Ebolavirus was the least severe. The pooled CFR and seroprevalence for Ebola and Marburg viruses were found to be lower than usually reported, with species differences despite high heterogeneity between studies. Countries with an improved health surveillance and epidemic response have lower CFR, thereby indicating need for improving early detection and epidemic response in filovirus outbreaks.


How does pain lead to disability? A systematic review and meta-analysis of mediation studies in people with back and neck pain.
Disability is an important outcome from a clinical and public health perspective. However, it is unclear how disability develops in people with low back pain or neck pain. More specifically, the mechanisms by which pain leads to disability are not well understood. Mediation analysis is a way of investigating these mechanisms by examining the extent to which an intermediate variable explains the effect of an exposure on an outcome. This systematic review and meta-analysis aimed to identify and examine the extent to which putative mediators explain the effect of pain on disability in people with low back pain or neck pain. Five electronic databases were searched. We found 12 studies (N = 2961) that examined how pain leads to disability with mediation analysis. Standardized regression coefficients (β) of the indirect and total paths were pooled. We found evidence to show that self-efficacy (β = 0.23, 95% confidence interval [CI] = 0.10 to 0.34), psychological distress (β = 0.10, 95% CI = 0.01 to 0.18), and fear (β = 0.08, 95% CI = 0.01 to 0.14) mediated the relationship between pain and disability, but catastrophizing did not (β = 0.07, 95% CI = -0.06 to 0.19). The methodological quality of these studies was low, and we highlight potential areas for development. Nonetheless, the results suggest that there are significant mediating effects of self-efficacy, psychological distress, and fear, which underpins the direct targeting of these constructs in treatment.


Reliability of lower limb motor evoked potentials in stroke and healthy populations: how many responses are needed?
To determine the intra- and inter-session reliability of motor evoked potential (MEP) size parameters in the lower limb of patients with stroke, focussing on the number of MEPs collected and the method of measuring MEP size. Transcranial magnetic stimulation was used to elicit MEPs in the soleus muscle of patients with stroke (n=13) and age-matched healthy participants (n=13) during low level muscle activation. Two sets of 10 responses were collected in the first session and a further 10 responses collected in a second session held 7 days later. Four MEP size measurements were made using 4, 6, 8, or all 10 of the MEPs collected. Intra- and inter-session reliability was examined using intraclass correlation coefficients (ICC) and typical percentage error. Intrasession ICC statistics using 6 or more MEPs were >0.85 in the stroke group but intersession values were all <0.5. Reliability was best when measuring parameters from individual MEPs rather than averaged responses. Reliability of intrasession MEP size is excellent in the lower limb of patients with stroke using as few as 6 MEPs but intersession reliability is poor. Comparing MEP size measures across two or more sessions is questionable in the lower limb of patients with stroke.


Occupational cancer in developed countries.
Studies of occupational exposures have made major contributions to our understanding of human carcinogenesis. About one third of the factors identified as definite or probable human carcinogens were first investigated in the workplace and these exposures exact a considerable toll on working populations. There are many additional workplace exposures that are suspect carcinogens that require further evaluation to ensure a safe work environment. Information from occupational investigations is also relevant to the general population because many occupational exposures can be found outside the workplace. Much of our understanding about occupational cancer has been obtained from studies largely composed of white men in developed countries. The movement of industry from developed to developing countries underscores the need for future investigations to include more diverse populations.


Progressive disseminated histoplasmosis mimicking a flare of systemic lupus erythematosus: a European case report.
Diagnosing progressive disseminated histoplasmosis (PDH) in patients with systemic lupus erythematosus (SLE) is diagnostically challenging. Since PDH is lethal when untreated, awareness of this infection in patients with SLE is of utmost importance. To the best of our knowledge, this is the first description of a case of PDH in a patient with SLE in Europe. A 56-year-old woman of Surinamese descent with a history of SLE, presented with fever and polyarthritis. Although a flare of SLE was suspected initially, cultures of bone marrow and broncho-alveolar lavage fluid grew Histoplasma capsulatum. This case report highlights that physicians should be aware of progressive disseminated histoplasmosis in patients with SLE treated with immunosuppressive agents. The signs and symptoms can easily mimic a SLE flare, which would then be treated with more aggressive immunosuppression. Failure to recognize the infection will therefore invariably lead to death of the patient. Progressive disseminated histoplasmosis is usually not recognized by doctors in non-endemic areas such as Europe. However, globalisation and more frequent intercontinental traffic of immunocompromised patients currently increases the incidence of histoplasmosis in these areas. It is therefore of life-saving importance that doctors are aware of the features of the infection in areas where H. capsulatum is not endemic.


Oral monoterpene therapy (perillyl alcohol) reduces vein graft intimal hyperplasia.
The development of intimal hyperplasia is recognized as a major impediment to graft patency. D-Limonenes are monoterpenes with a recognized cytostatic effect on cell proliferation by inhibiting posttranslational isoprenylation of p21ras and other small G-proteins. This study examines the effect of perillyl alcohol, an oral hydroxylated D-limonene, on the development of intimal hyperplasia and its associated smooth muscle cell physiological responses in an experimental model of vein bypass grafting. Twenty New Zealand white rabbits had a right carotid interposition bypass graft using the ipsilateral external jugular vein. Ten animals received chronic oral therapy with a perillyl alcohol (200 mg/kg/day; begun 5 days before surgery and continued until harvest) and 10 control animals received vehicle only. All animals were sacrificed on the 28th postoperative day. Vein grafts were harvested either for morphology/videomorphometry (n = 6 per group) or for in vitro isometric tension studies (n = 4; four 5-mm rings per graft). The cell proliferation and incorporation of [3H]thymidine into the cellular DNA of serum-stimulated rabbit aortic smooth muscle cells was also assessed in the presence of increasing concentrations of perillyl alcohol (10(-9)-10(-4) M). Perillyl alcohol treated vein grafts showed a 22% reduction in overall mean intimal thickness (54 +/- 4 microns vs 69 +/- 3 microns; P = 0.006) but a 25% increase in overall mean medial thickness (86 +/- 4 microns vs 61 +/- 3 microns). The intimal ratio of the perillyl alcohol treated vein grafts decreased by 27% compared to controls. Perillyl alcohol induced norepinephrine and serotonin hypersensitivity in vein grafts compared to controls. The IC50 for perillyl alcohol was 176 nM with maximal inhibition at 5 microM. Incubation of smooth muscle cell cultures with increasing concentrations of perillyl alcohol showed a dose-dependent decrease in in vitro cellular proliferation, maximal at 1 microM. Therapy with perillyl alcohol alters the early development of intimal hyperplasia reducing the intimal response but increasing the medial response without significant changes in the physiological responses of the smooth muscle cells. Modulating G-proteins will affect the intimal hyperplastic response in vein grafts.


The relevance of methane emissions from beef production and the challenges of the Argentinean beef production platform.
The livestock sector faces the challenge to respond to the growing demand for animal protein from an expanding population while reducing environmental impact through GHG emissions. Globally about 2.836 million tons of CO2-eq were emitted by the beef production sector equivalent to 46,2 kg CO2-eq per kg carcass weight (CW). From the 1.485 million cattle head spread out over the world, 82% are on extensive grazing systems while only 18% are on high productive intensive systems. Among the top ten beef exporter countries, five are located in Latin America accounting a quarter of the global stock and two of them, Argentina and Uruguay, produce on temperate pastures under grazing systems. In Argentina, the livestock area was reduced in favor of increasing the grain cropping area, which took place in the last two decades. Production systems were intensified to maintain cattle stock. Cattle programs changed from 100% pasture to pasture supplemented with cereal grains and conserved forages, and confinement on grain feeding for fattening was incorporated. Due to land sharing competition with cash crops, no increment of cattle stock is expected therefore improving production efficiency appears as the only way to increase beef production while reducing methane emissions intensity. Beef produced on intensive grazing systems on supplemented pastures maintained organoleptic, nutritional and lipid profile than that of beef produced on pure grazing systems.


Time saved with high speed driving of ambulances.
Transportation with road ambulances is increasing because of a concentration of hospitals to larger units, with high quality in the acute care of the patients. The concentration implies longer distances to receiving units, which increases the transportation time. The purpose of the present study was to investigate the time difference in ambulance transportation with high speed emergency driving, compared to non-emergency driving in normal traffic pace. Data was collected from 30 emergency high speed ambulance transportations in urban and rural areas. These transportations were then repeated experimentally with an ambulance driving at normal traffic pace. The average speed and duration for the emergency transportations were shorter than for the experimental driving, both in urban and rural areas. The mean time saved was 2.9 min (urban areas) and 8.9 min (rural areas). Regardless of the patient's clinical status or need of care the emergency transportations were carried out in higher speed than the experimental driving. However, patients with life threatening conditions were not included in this study. Procedures and methods should be developed to identify the patients for which fast transportation has clinical relevance to the outcomes.


Adverse birth outcomes are associated with circulating matrix metalloproteinases among pregnant women in Puerto Rico.
Matrix metalloproteinases (MMPs) are major extracellular matrix (ECM) remodeling proteinases and regulate uterine remodeling, which is a critical process for healthy pregnancies. The goal of this study was to investigate associations between maternal blood MMPs during pregnancy and birth outcomes among 898 pregnant women in the Puerto Rico PROTECT birth cohort. MMPs (MMP1, MMP2, and MMP9) were quantified using a customized Luminex assay in blood samples collected at two gestational study visits (around 18 and 26 weeks gestation). Linear and logistic regression models were used to regress continuous and binary birth outcomes, respectively, on MMPs at each study visit separately. Sensitivity analyses were conducted to test for effect modification by fetal sex on associations between MMPs and birth outcomes. We observed significant associations between MMP2 at visit 1 and newborn length that were in the opposite direction from the associations between MMP9 at visit 3 and newborn length. MMPs were associated with increased odds of preeclampsia and gestational diabetes mellitus, though case numbers were low. We also observed significant inverse associations with gestational age for MMP9 and MMP2 at visit 1 and visit 3, respectively, and these associations were observed only in mothers carrying male fetuses. Further, MMP2 was associated with heavier female fetuses, whereas MMP9 was associated with lighter female fetuses. We observed significant associations between birth outcomes and MMPs, and the majority of these associations differed by fetal sex. This study highlighted significant MMPs-birth outcomes associations that may provide a basis to explore the impact of MMPs on endometrium health and physiology.


Anti-ulcer effects of antioxidants, quercetin, alpha-tocopherol, nifedipine and tetracycline in rats.
When free radical-scavenging activities of quercetin, alpha-tocopherol, nifedipine and tetracycline were measured by an electron spin resonance technique, all test compounds (10(-5) to 10(-3) M) scavenged both superoxide anions and hydroxyl radicals. The oral administration of quercetin (50 and 100 mg/kg), alpha-tocopherol (8 and 16 mg/kg), nifedipine (20 and 40 mg/kg) or tetracycline (10 and 20 mg/kg) markedly prevented the HCl plus ethanol-induced gastric mucosal injury and the increase in the content of thiobarbituric acid-reactive substances in the injured mucosa in rats. In addition, quercetin (25, 50 and 100 mg/kg), alpha-tocopherol (4, 8 and 16 mg/kg), nifedipine (10, 20 and 40 mg/kg) and tetracycline (5, 10 and 20 mg/kg), given orally, twice daily for 14 consecutive days from the day after acetic acid injection, dose-dependently promoted the ulcer healing and inhibited the increase in the content of thiobarbituric acid-reactive substances in the ulcerated mucosa. These results indicate that quercetin, alpha-tocopherol, nifedipine and tetracycline possess gastric cytoprotective and gastric ulcer healing-promoting actions. In addition, the free radical-scavenging properties of these compounds may be partly related to their anti-ulcer effects.


Beyond Trauma: A Multiple Pathways Approach to Auditory Hallucinations in Clinical and Nonclinical Populations.
That trauma can play a significant role in the onset and maintenance of voice-hearing is one of the most striking and important developments in the recent study of psychosis. Yet the finding that trauma increases the risk for hallucination and for psychosis is quite different from the claim that trauma is necessary for either to occur. Trauma is often but not always associated with voice-hearing in populations with psychosis; voice-hearing is sometimes associated with willful training and cultivation in nonclinical populations. This article uses ethnographic data among other data to explore the possibility of multiple pathways to voice-hearing for clinical and nonclinical individuals whose voices are not due to known etiological factors such as drugs, sensory deprivation, epilepsy, and so forth. We suggest that trauma sometimes plays a major role in hallucinations, sometimes a minor role, and sometimes no role at all. Our work also finds seemingly distinct phenomenological patterns for voice-hearing, which may reflect the different salience of trauma for those who hear voices.


Effects of task instructions and target location on reaching kinematics in people with and without cerebrovascular accident: a study of the less-affected limb.
We investigated how verbal instructions and target location interacted to influence reaching movement of the less-affected limb in participants with and without unilateral cerebrovascular accidents (CVAs). Using a counterbalanced repeated-measures design, 26 people with CVA and 24 age-matched healthy people performed the reaching tasks under 4 conditions formed by the crossing of verbal instructions (speed and accuracy emphasis) and target locations (ipsilateral and contralateral to the performing hand). In the control groups, speeded instructions and ipsilateral reaches elicited significantly more preprogrammed movements than did accuracy instruction and contralateral reaches, respectively. Similar patterns of performance in response to task constraints were found in the CVA groups except for movement initiation in the right CVA group. Instruction and locations interacted to constrain reaching movements in both control and CVA groups. The combination of speeded instruction and ipsilateral reach may optimize movement performance of the less-affected limb in stroke patients.


Vitamin E deficiency during chronic childhood cholestasis: presence of sural nerve lesion prior to 2 1/2 years of age.
Vitamin E malabsorption and deficiency during chronic childhood cholestasis has been associated with a progressive ataxic neurologic syndrome. Hyporeflexia, the first sign of neurologic dysfunction, may begin prior to age 2 years, but severe symptoms do not develop until age 5 to 10 years. To establish the age of onset of neuropathologic lesions, we prospectively evaluated four young children with severe cholestasis. Malabsorption and deficiency of vitamin E were documented by low serum vitamin E concentrations, low serum vitamin E to total serum lipids ratios, elevated hydrogen peroxide hemolysis, and impaired absorption of a pharmacologic dose of alpha-tocopherol. Abnormal neurologic findings in two patients were limited to areflexia, ptosis, mild truncal ataxia, and hypotonia; two patients had minimal signs of neurologic dysfunction. Sural nerve histology at age 6 to 25 months revealed a degenerative axonopathy involving large-caliber myelinated fibers, but without quantitative axonal loss. Muscle histology and histochemistry tests yielded normal results. Our study suggests that neurologic injury may occur during the first two years of life in vitamin E-deficient children with cholestatic hepatobiliary disease, obligating aggressive attempts at correcting this deficiency state at a very young age.


HIV and Obesity Comorbidity Increase Interleukin 6 but Not Soluble CD14 or D-Dimer.
Obesity prevalence among people living with HIV (HIV+) is rising. HIV and obesity are proinflammatory states, but their combined effect on inflammation (measured by interleukin 6, IL-6), altered coagulation (D-dimer), and monocyte activation (soluble CD14, sCD14) is unknown. We hypothesized inflammation increases when obesity and HIV infection co-occur. The Veterans Aging Cohort Study survey cohort is a prospective, observational study of predominantly male HIV+ veterans and veterans uninfected with HIV; a subset provided blood samples. Inclusion criteria for this analysis were body mass index ≥ 18.5 kg/m and biomarker measurement. Dependent variables were IL-6, sCD14, and D-dimer quartiles. Obesity/HIV status was the primary predictor. Unadjusted and adjusted logistic regression models were constructed. Data were analyzed for 1477 HIV+ and 823 uninfected participants. Unadjusted median IL-6 levels were significantly higher and sCD14 levels significantly lower in obese/HIV+ compared with nonobese/uninfected (P <0.01 for both). In adjusted analyses, the odds ratio for increased IL-6 in obese/HIV+ patients was 1.76 (95% confidence interval: 1.18 to 2.47) compared with nonobese/uninfected, and obesity/HIV+ remained associated with lower odds of elevated sCD14. We did not detect a synergistic association of co-occurring HIV and obesity on IL-6 or sCD14 elevation. D-dimer levels did not differ significantly between body mass index/HIV status groups. HIV-obesity comorbidity is associated with elevated IL-6, decreases in sCD14, and no significant difference in D-dimer. These findings are clinically significant, as previous studies associated these biomarkers with mortality. Future studies should assess whether other biomarkers show similar trends and potential mechanisms for unanticipated sCD14 and D-dimer findings.


Immortalization of epithelial cells.
The methodologies for isolating cell lines have become very powerful, particularly in terms of retaining differentiated features of the parent cells. Cell lines can be developed from primary or early passage cells as well as from transgenic animals that carry an immortalizing gene. Cell lines from epithelia have been selected for their polar orientation, tight junction formation, and expression of differentiated markers or functions. These cell lines provide useful models for studying cell biology of specific tissues, tumorigenicity, genetic abnormalities, or to help screen for effective methods of gene therapy.


Consensus paper of the WFSBP task force on cannabis, cannabinoids and psychosis.
The liberalisation of cannabis laws, the increasing availability and potency of cannabis has renewed concern about the risk of psychosis with cannabis. The objective of the WFSBP task force was to review the literature about this relationship. Converging lines of evidence suggest that exposure to cannabis increases the risk for psychoses ranging from transient psychotic states to chronic recurrent psychosis. The greater the dose, and the earlier the age of exposure, the greater the risk. For some psychosis outcomes, the evidence supports some of the criteria of causality. However, alternate explanations including reverse causality and confounders cannot be conclusively excluded. Furthermore, cannabis is neither necessary nor sufficient to cause psychosis. More likely it is one of the multiple causal components. In those with established psychosis, cannabis has a negative impact on the course and expression of the illness. Emerging evidence also suggests alterations in the endocannabinoid system in psychotic disorders. Given that exposure to cannabis and cannabinoids is modifiable, delaying or eliminating exposure to cannabis or cannabinoids, could potentially impact the rates of psychosis related to cannabis, especially in those who are at high risk for developing the disorder.


Validation of the IMMIDIET food frequency questionnaire in an adult Belgian population: a report from the Belgian case-control study on bladder cancer risk.
We evaluated the performance of the IMMIDIET food frequency questionnaire (FFQ) used to collect dietary data for the Belgian case-control study on bladder cancer. Thirty-seven men and women aged 50 years and older were recruited from the University Hospital in Leuven, Belgium. Participants completed the IMMIDIET FFQ, a 7-day diet diary and a 24-hour diet recall. Median intakes and inter-quartile ranges were calculated for 27 foods and nutrients from each dietary assessment method. All dietary factors were log-transformed and adjusted for energy using the nutrient density method. Pearson correlation coefficients were used to compare the different dietary assessment methods. Bland-Altman plots were also used to assess levels of agreement between the dietary methods. Energy, fruit and vegetable intake estimates were higher from the IMMIDIET FFQ compared with the two reference methods.The highest deattenuated correlations between the FFQ and 7-day diary were meat (0.58), bread (0.44), fruit (0.38) and fish (0.38). The highest deattenuated correlations between the FFQ and 24-hour recall were for fruit (0.72), fat (0.48), alcohol (0.44), cholesterol (0.42), monounsaturated fatty acid (0.42) and polyunsaturated fatty acid (0.41). Generally, correlation was lower for the micro-nutrients except for phosphorus (0.42), vitamin C (0.41) and calcium (0.40). The IMMIDIET FFQ is an appropriate instrument to measure usual dietary intake for the Belgian case-control study on bladder cancer risk. Further investigation of nutritional assessment methods is necessary.


A Dyadic Analysis of the Associations Between Cumulative Childhood Trauma and Psychological Intimate Partner Violence: The Mediating Roles of Negative Urgency and Communication Patterns.
Prior research has suggested associations between cumulative childhood trauma (CCT), negative urgency, communication patterns, and psychological intimate partner violence (P-IPV), but no study has examined these links using a dyadic approach. This study examined the sequential mediation of negative urgency and communication patterns in the link uniting CCT and P-IPV in a sample of 501 heterosexual couples. Results suggest that more CCT events are associated with higher levels of negative urgency, which in turn are associated with a higher tendency to endorse a demand/withdraw and/or demand/demand communication pattern, and to perpetrate P-IPV. Findings support the need to assess CCT, emotional self-control, dyadic and communication patterns, and P-IPV perpetration and victimization in couples seeking help in order to select interventions that will take into account the individual and dyadic nature of P-IPV.


The effects of long-term administration of oral desmopressin on the baseline secretion of antidiuretic hormone and serum sodium concentration for the treatment of nocturia: a circadian study.
We assessed the effects of long-term oral desmopressin on serum sodium and baseline antidiuretic hormone secretion in elderly patients with nocturia. A total of 15 elderly male patients with severe nocturia (greater than 3 voids nightly) who did not show hyponatremia within 7 days of administration of 0.2 mg desmopressin were enrolled in this study. Desmopressin (0.2 mg) was administered orally nightly for 1 year. Before and 1 month after the 1-year medication 24-hour circadian studies were performed to monitor changes in antidiuretic hormone. Every 3 months during the 1-year medication serum changes and timed urine chemistry were monitored. Desmopressin significantly decreased nocturnal urine output and the number of nocturia episodes (p<0.01). Compared to before treatment desmopressin gradually decreased serum sodium and induced statistically but not clinically significant hyponatremia after 6 months of treatment. After discontinuing desmopressin serum sodium returned to the normal range in all patients. There were no significant differences when baseline and posttreatment endogenous antidiuretic hormone were compared. No serious systemic complications were found during medication. Long-term desmopressin administration gradually decreased the serum concentration and induced significant hyponatremia from 6 months in patients who did not show initial hyponatremia. Long-term administration of desmopressin for 1 year in elderly patients did not affect baseline antidiuretic hormone secretion. For long-term desmopressin administration serum sodium should be assessed regularly, at least every 6 months.


Integration of stochastic models by minimizing alpha-divergence.
When there are a number of stochastic models in the form of probability distributions, one needs to integrate them. Mixtures of distributions are frequently used, but exponential mixtures also provide a good means of integration. This letter proposes a one-parameter family of integration, called alpha-integration, which includes all of these well-known integrations. These are generalizations of various averages of numbers such as arithmetic, geometric, and harmonic averages. There are psychophysical experiments that suggest that alpha-integrations are used in the brain. The alpha-divergence between two distributions is defined, which is a natural generalization of Kullback-Leibler divergence and Hellinger distance, and it is proved that alpha-integration is optimal in the sense of minimizing alpha-divergence. The theory is applied to generalize the mixture of experts and the product of experts to the alpha-mixture of experts. The alpha-predictive distribution is also stated in the Bayesian framework.


An analytical model applied to a multicenter pneumococcal enzyme-linked immunosorbent assay study.
Pneumococcal conjugate vaccines will eventually be licensed after favorable results from phase III efficacy trials. After licensure of a conjugate vaccine for invasive pneumococcal disease in infants, new conjugate vaccines will likely be licensed primarily on the basis of immunogenicity data rather than clinical efficacy. Analytical methods must therefore be developed, evaluated, and validated to compare immunogenicity results accurately within and between laboratories for different vaccines. At present no analytical technique is uniformly accepted and used in vaccine evaluation studies to determine the acceptable level of agreement between a laboratory result and the assigned value for a given serum sample. This multicenter study describes the magnitude of agreement among 12 laboratories quantifying an identical series of 48 pneumococcal serum specimens from 24 individuals (quality-control sera) by a consensus immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) developed for this study. After provisional or trial antibody concentrations were assigned to the quality-control serum samples for this study, four methods for comparison of a series of laboratory-determined values with the assigned concentrations were evaluated. The percent error between assigned values and laboratory-determined concentrations proved to be the most informative of the four methods. We present guidelines that a laboratory may follow to analyze a series of quality-control sera to determine if it can reproduce the assigned antibody concentrations within an acceptable level of tolerance. While this study focused on a pneumococcal IgG ELISA, the methods that we describe are easily generalizable to other immunological assays.


Design and molecular dynamics of multifunctional sulfonated poly(dimethylaminoethyl methacrylate)/mica hybrid cryogels through freezing-induced gelation.
This article addresses various strategies that have been explored to design sulfonated poly(dimethylaminoethyl methacrylate)/mica hybrid-gels with optimized network parameters and mechanical/swelling properties. A series of hybrid cryogels and hydrogels containing amino and sulfonic acid groups were prepared from N,N-dimethylaminoethyl methacrylate (DMAEMA) and 2-acrylamido-2-methyl-1-propane sulfonic acid in the presence of inorganic additive mica via a cryogelation process and conventional in situ copolymerization. Cryogelation was used to fine-tune the mechanical properties of the PDMAEMA-based hybrid gels. The effects of pH, temperature and mica content on the network parameters, mechanical properties and swelling behavior were discussed. X-ray diffractometry and Fourier transform infrared spectroscopy confirmed that mica particles had participated in (cryo)polymerization, and the thermal stability and surface morphologies were improved by the addition of mica. The profile of water loss, decomposition of amine groups and breakdown of PDMAEMA chains of the resulting hybrid gels were determined by thermogravimetric analysis. A critical mica concentration was found for the hybrid hydrogels where the degree of swelling attains a maximum value. Below 0.50% (w/v) of mica, the ionic nature of mica dominates its crosslinker effect. The hybrid cryogels were tough and able to recover at room temperature after compression testing. The prepared hybrid-gels showed an enhanced swelling response and on-off switching swelling characteristics in water and in aqueous NaCl solutions. The parameters of equilibrium swelling, the initial swelling rate, the diffusional exponent, and the diffusion coefficient were evaluated and the swelling kinetics of the hybrid hydrogels and cryogels in water followed the pseudo second order model. All the prepared hybrid hydrogel and cryogel materials with tunable mechanical stability and elasticity can be excellent candidates for designing smart materials.


Late onset involuntary movements in chronic schizophrenia: relationship of 'tardive' dyskinesia to intellectual impairment and negative symptoms.
Intellectual impairment, negative symptoms, and medication history were assessed in chronic schizophrenic patients with and without abnormal involuntary movements (tardive dyskinesia). Patients with involuntary movements had received neither longer nor more intensive treatment with neuroleptics or anticholinergics. However, the presence or absence of involuntary movements was prominently associated with the presence or absence of intellectual impairment/negative symptoms; these features are characteristic of the defect state/type II syndrome of schizophrenia, in which structural abnormalities of the brain may be over-represented. The role of subtle organic changes in conferring vulnerability to the emergence of such involuntary movements should be re-evaluated.


The leading methods of suicide in Taiwan, 2002-2008.
Diverse socioeconomic and cultural developments between geographic regions and cities/counties have resulted in different physical availability and socio-cultural acceptability of certain methods of suicide. This study examined the changes in distribution of the leading methods of suicide across cities/counties in Taiwan between 2002-04 and 2006-08. Mortality data for all deaths classified as suicide or as of undetermined intent from 2002 through 2008 were extracted for analysis. The number of deaths and proportion of completed suicides by four main methods were calculated in order to identify the leading lethal methods in each city/county. Hanging was the leading method of suicide in 18 out of 22 cities/counties in 2002-04 but decreased to 10 out of 22 in 2006-08. On the other hand, charcoal burning was not the leading method in any city/county in 2002-04 but increased to 10 out of 22 in 2006-08. The younger the age of the deceased, the more likely the leading method of suicide changed from 2002-04 to 2006-08. Charcoal burning was the most often used method in most cities/counties among those aged 15-44; however, hanging was most frequent for those aged 45 or above. Pesticides were the leading method for the elderly in five counties with a high percentage of agricultural population in 2006-08. The leading method of suicide varied by age group and changed from 2002-04 to 2006-08 in Taiwan. This was due primarily to changes in socio-cultural acceptability of the use of charcoal burning as a method for suicide by younger age groups.


Road casualties and enforcement: distributional assumptions of serially correlated count data.
Road safety data are often in the form of counts and usually temporally correlated. The objective of this research is to investigate the distributional assumptions of road safety data in the presence of temporal correlation. Using the generalized linear model framework, four distributional assumptions are considered: normal, Poisson, quasi-Poisson and negative binomial, and appropriate models are estimated. Monthly casualty and police enforcement data from Greece for a period of six years (January 1998-December 2003) have been used. The developed models include sinusoidal latent terms to capture the temporal serial correlation of observations. Several statistical goodness-of-fit diagnostic tests have been performed for the results of the estimated models, and the predictive capabilities of the models are investigated. The residuals of the quasi-Poisson and negative binomial models do not show any serial correlation. The signs of the estimated coefficients for all models are consistent and intuitive. In particular, a negative coefficient value for the number of breath alcohol controls indicates that the number of persons killed and seriously injured decreases as the intensity of breath alcohol controls increases. The Poisson model fails to capture the overdispersion in the data, thus underestimating the standard errors of the estimated coefficients. The results suggest that the quasi-Poisson and negative binomial outperform the normal and Poisson models in this application. The findings of this research demonstrate a clear link between the intensification of police enforcement and the reduction of traffic accident casualties. In particular, an increase in the number of breath alcohol controls in Greece after 1998 contributed to a reduction in the number of persons killed and seriously injured from traffic accidents.


Proton MR spectroscopy of squamous cell carcinoma of the extracranial head and neck: in vitro and in vivo studies.
To determine the ability of in vitro one-dimensional and two-dimensional proton MR spectroscopy to help differentiate squamous cell carcinoma of the extracranial head and neck from normal tissues and to correlate the in vitro observations with clinical studies. In vitro 1-D and 2-D correlated proton MR spectroscopy (11 T) was performed in tissue specimens of squamous cell carcinoma of the head and neck (n = 19), in normal tissue (n = 13), in metastatic cervical lymph nodes (n = 3), and in a squamous cell carcinoma cell line. In vivo 1-D proton MR spectroscopy (1.5 T) was performed in patients with squamous cell carcinoma (n = 7) and in healthy volunteers (n = 7). The ratio of the areas under the choline (Cho) and creatine (Cr) resonances were calculated for 1-D proton MR spectra for the in vitro tissue studies and correlated with the in vivo studies. Data from in vitro 2-D correlated spectroscopy were analyzed for differences in the presence or absence of various metabolites in samples of tumor and normal tissue. Statistical analysis consisted of 2 x 2 factorial repeated measures analysis of variance (ANOVA), discriminate analysis, and chi2 test. The mean in vitro 1-D proton MR spectroscopic Cho/Cr ratio was significantly higher in tumor than in normal tissue. The difference between the mean ratios appeared to increase with increasing echo time. All in vivo tumor Cho/Cr ratios were greater than the calculated mean in vitro tumor ratio, whereas six of the seven volunteers had no detectable Cho and Cr resonances. Two-dimensional correlated MR spectroscopic data revealed that a variety of amino acids have a significantly greater likelihood of being detected in tumor than in normal tissues. One-dimensional and 2-D proton MR spectroscopy can help differentiate primary squamous cell carcinoma and nodal metastases containing squamous cell carcinoma from normal tissue both in vitro and in vivo. In addition, 2-D spectroscopy can help identify the presence of certain amino acids in squamous cell carcinoma that are not detected in normal tissue.


Formation of silver nanoparticles by human gut microbiota.
Metal nanoparticles have received global attention and one of the most promising nanotechnologies is the use of microorganisms for synthesizing nanoparticles. The oral exposure study has demonstrated the formation of silver (Ag) nanoparticles with in vivo animal models. However, there is limitation in the effect of in vitro cultured human gut microbiota on silver. Here, the phenomenon of biosynthesis of Ag nanoparticles by human gut microbiota has been reported. The size, shape, and morphology of the nanoparticles were characterized using transmission electron microscopy (TEM). Exposure to aqueous AgNO3 solution resulted in the intracellular reduction of Ag+ ions and the formation of approximately spherical Ag nanoparticles with dimensions of 34 ± 10 nm. Electron microscopy analysis of the gut microbial cells by TEM images of thin sections and scanning electron microscopy (SEM), indicated that most of Ag nanoparticles were located on the cytoplasmic membrane and within the cytoplasm, where abundant silver reductases were present. In addition, some nanoparticles were bound to the cell wall, likely via exocytosis. The oral exposure to Ag+ led to in vivo formation of nanoparticles by gut microbiota, and their biological effects and potential applications in human body warrant further investigation.


The water method significantly enhances patient-centered outcomes in sedated and unsedated colonoscopy.
Failure of cecal intubation when using air insufflation during scheduled unsedated colonoscopy in veterans prompted a literature search for a less uncomfortable approach. Water-related maneuvers as "adjuncts" to air insufflation were identified as effective in minimizing discomfort, although medication requirement was not reduced and willingness to repeat unsedated colonoscopy was not addressed. These adjunct maneuvers were combined with turning the air pump off to avoid colon elongation during insertion. Warm water infusion in lieu of air insufflation was evaluated in observational studies. Subsequent refinements evolved into the water method - a combination of air exclusion by aspiration of residual air to minimize angulations at flexures and a dynamic process of water exchange to remove feces in order to clear the view and aid insertion. In subsequent randomized controlled trials, the water method significantly reduced medication requirement, increased the proportion of patients in whom complete unsedated colonoscopy could be achieved, reduced patient recovery time burdens (sedation on demand), decreased abdominal discomfort during and after colonoscopy, enhanced cecal intubation, and increased willingness to repeat the procedure (scheduled unsedated). Supervised education of trainees and self-learning by an experienced colonoscopist were feasible. Lessons learned in developing the water method for optimizing patient-centered outcomes are presented. These proof-of-principle observations merit further research assessment in diverse settings.


Estimating the implicit component of visuomotor rotation learning by constraining movement preparation time.
When sensory feedback is perturbed, accurate movement is restored by a combination of implicit processes and deliberate reaiming to strategically compensate for errors. Here, we directly compare two methods used previously to dissociate implicit from explicit learning on a trial-by-trial basis: 1) asking participants to report the direction that they aim their movements, and contrasting this with the directions of the target and the movement that they actually produce, and 2) manipulating movement preparation time. By instructing participants to reaim without a sensory perturbation, we show that reaiming is possible even with the shortest possible preparation times, particularly when targets are narrowly distributed. Nonetheless, reaiming is effortful and comes at the cost of increased variability, so we tested whether constraining preparation time is sufficient to suppress strategic reaiming during adaptation to visuomotor rotation with a broad target distribution. The rate and extent of error reduction under preparation time constraints were similar to estimates of implicit learning obtained from self-report without time pressure, suggesting that participants chose not to apply a reaiming strategy to correct visual errors under time pressure. Surprisingly, participants who reported aiming directions showed less implicit learning according to an alternative measure, obtained during trials performed without visual feedback. This suggests that the process of reporting can affect the extent or persistence of implicit learning. The data extend existing evidence that restricting preparation time can suppress explicit reaiming and provide an estimate of implicit visuomotor rotation learning that does not require participants to report their aiming directions.NEW & NOTEWORTHY During sensorimotor adaptation, implicit error-driven learning can be isolated from explicit strategy-driven reaiming by subtracting self-reported aiming directions from movement directions, or by restricting movement preparation time. Here, we compared the two methods. Restricting preparation times did not eliminate reaiming but was sufficient to suppress reaiming during adaptation with widely distributed targets. The self-report method produced a discrepancy in implicit learning estimated by subtracting aiming directions and implicit learning measured in no-feedback trials.


[Ca2+]i oscillations in sympathetic neurons: an experimental test of a theoretical model.
[Ca2+]i oscillations have been described in a variety of cells. This study focuses on caffeine-induced [Ca2+]i oscillations in sympathetic neurons. Previous work has shown that these oscillations require Ca2+ entry from the extracellular medium and Ca(2+)-induced Ca2+ release from a caffeine- and ryanodine-sensitive store. The aim of the study was to understand the mechanism responsible for the oscillations. As a starting point, [Ca2+]i relaxations were examined after membrane depolarization and exposure to caffeine. For both stimuli, post-stimulus relaxations could be described by the sum of two decaying exponential functions, consistent with a one-pool system in which Ca2+ transport between compartments is regulated by linear Ca2+ pumps and leaks. After modifying the store to include a [Ca2+]i-sensitive leak, the model also exhibits oscillations such as those observed experimentally. The model was tested by comparing measured and predicted net Ca2+ fluxes during the oscillatory cycle. Three independent fluxes were measured, describing the rates of 1) Ca2+ entry across the plasma membrane, 2) Ca2+ release by the internal store, and 3) Ca2+ extrusion across the plasma membrane and uptake by the internal store. Starting with estimates of the model parameters deduced from post-stimulus relaxations and the rapid upstroke, a set of parameter values was found that provides a good description of [Ca2+]i throughout the oscillatory cycle. With the same parameter values, there was also good agreement between the measured and simulated net fluxes. Thus, a one-pool model with a single [Ca2+]i-sensitive Ca2+ permeability is adequate to account for many of the quantitative properties of steady-state [Ca2+]i oscillations in sympathetic neurons. Inactivation of the intracellular Ca2+ permeability, cooperative nonlinear Ca2+ uptake and extrusion mechanisms, and functional links between plasma membrane Ca2+ transport and the internal store are not required.


Association between mast cells of different phenotypes and angiogenesis in colorectal cancer.
It is known that mast cells proliferate in solid tumours and increase tumour angiogenesis. Nevertheless, there is no consensus regarding their role in colorectal cancer (CRC). In this study, we aimed to clarify the relationship of mast cells positive for tryptase (MCts) and tryptase-chymase (MCtcs) with microvessel density (MVD) in the intratumoral zone and the invasive edge of 80 CRC patient tumours. We evaluated these parameters and associated their expression with clinicopathological parameters, including survival rate. Tumour sections from each patient were immunostained for tryptase to evaluate MCts, chymase to evaluate MCtcs, and CD34 to evaluate microvessel counts under x100 microscopy. The number of MCs of both phenotypes and the MVD counts were higher in the invasive edge than in the intratumoral zone (p<0.001). MCt numbers were higher than those of MCtcs in all Astler-Coller stages in both regions. A positive correlation between MVD and MCts or MCtcs was observed (Pearson's test p<0.001). Neither the number of MCs nor MVD was associated with overall survival (log rank test). However, only 8.3% of patients with low numbers of MCtcs in the invasive edge succumbed to the disease, compared to 32% with high numbers of MCtcs. Our results indicate that angiogenesis and MC hyperplasia are events which appear early during CRC development. The correlation of MC phenotypes with MVD is in agreement with the role attributed to MCs, that of angiogenesis enhancement. Collectively, these findings suggest that screening during the early malignization of CRC can provide valuable clinical information.


Diagnostic performance of using one- or two-HbA1c cut-point strategies to detect undiagnosed type 2 diabetes and impaired glucose regulation within a multi-ethnic population.
We compared test performance and cost per case for strategies detecting diabetes on the oral glucose tolerance test (OGTT) using either (a) glycated haemoglobin (HbA1c) ≥ 6.5% (48 mmol/mol) or (b) two HbA1c thresholds where the first cut-point 'rules out' and the second 'rules in' diabetes. HbA1c values in between the thresholds require confirmatory glucose testing for diagnosis. We conducted an analysis of adults aged 40-75 years from the Leicester Ethnic Atherosclerosis and Diabetes Risk (LEADER) cohort (Leicester, UK), from 2002 to 2008, who underwent oral glucose tolerance testing (OGTT) and HbA1c testing. From 8696 individuals (mean age 57.3 years, 73% white Europeans (WE) and 27% South Asians (SA)), HbA1c ≥ 6.5% produced sensitivity of 62.1% for detecting diabetes in WE and 78.9% in SA. Using two selected thresholds, HbA1c ≤ 5.8% (rule-in, 40 mmol/mol) and HbA1c ≥ 6.8% (rule-out, 51 mmol/mol) produced high sensitivity/specificity (> 91.0%) for detecting diabetes, however, 28.8% of the cohort with HbA1c 5.9%-6.7% required a subsequent glucose test. The two cut-point threshold produced a lower cost per case of diabetes detected in WE, compared to HbA1c ≥ 6.5% of £38.53 (1.89 to 86.81) per case, but was more expensive in SA by £84.50 (69.72 to 100.92) per case. Using a risk score to determine HbA1c testing, the same costs per case became £63.33 (23.33 to 113.26) in WE and £69.21 (55.60 to 82.41) in SA. Using a two-threshold strategy may have some benefits over a single cut-point; however, 28.8% of individuals required two blood tests.


Impact of glucose-lowering agents on the risk of cancer in type 2 diabetic patients. The Barcelona case-control study.
The aim of the present study is to evaluate the impact of glucose-lowering agents in the risk of cancer in a large type 2 diabetic population. A nested case-control study was conducted within a defined cohort (275,164 type 2 diabetic patients attending 16 Primary Health Care Centers of Barcelona). Cases (n = 1,040) comprised those subjects with any cancer diagnosed between 2008 and 2010, registered at the Cancer Registry of Hospital Vall d'Hebron (Barcelona). Three control subjects for each case (n = 3,120) were matched by age, sex, diabetes duration, and geographical area. The treatments analyzed (within 3 years prior to cancer diagnosis) were: insulin glargine, insulin detemir, human insulin, fast-acting insulin and analogues, metformin, sulfonylureas, repaglinide, thiazolidinediones, dipeptidyl peptidase-4 inhibitors, and alpha glucosidase inhibitors. Conditional logistic regressions were used to calculate the risk of cancer associated with the use of each drug adjusted by age, BMI, dose and duration of treatment, alcohol use, smoking habit, and diabetes duration. No differences were observed between case and control subjects for the proportion, dose or duration of exposure to each treatment. None of the types of insulin and oral agents analyzed showed a significant increase in the risk of cancer. Moreover, no cancer risk was observed when glargine was used alone or in combination with metformin. Our results suggest that diabetes treatment does not influence the risk of cancer associated with type 2 diabetes. Therefore, an eventual increase of cancer should not be a reason for biasing the selection of any glucose-lowering treatment in type 2 diabetic population.


Deficiency of brain structural sub-network underlying post-ischaemic stroke apathy.
This study aimed to reveal the structural basis of post-ischaemic stroke apathy, especially in relation to disruptions in structural connectivity. Eighty-eight participants were included. The Apathy Evaluation Scale, clinician version, was used to characterize the severity of apathy. Diffusion tensor imaging tractography was used to examine white matter integrity and to reconstruct white matter networks using 90 nodes based on the automated anatomical labeling atlas. The degree for each node was extracted to determine the relationship to the severity of apathy. Apathy was not significantly associated with damage to any single brain region. The degrees of 24 nodes (limbic system, three nodes; frontal lobe, six; basal ganglia, two; temporal lobe, three; parietal lobe, three; insula, two; occipital lobe, five) were significantly correlated to the Apathy Evaluation Scale scores. These 24 nodes constituted an apathy-related sub-network and its global and local efficiencies were negatively correlated with apathy levels (global, r = -0.54, P < 0.01; local, r = -0.64, P < 0.01). Multivariate logistic regression indicated that decreased global efficiency of this sub-network was an independent risk factor for apathy (odds ratio 0.03, 95% confidence interval 0.01-0.04, P = 0.007). Efficiencies of the non-apathy-related sub-network (the remaining 66 nodes) did not correlate or predict the presence of apathy. Post-stroke apathy is not due to the dysfunction of a single region or circuit. Rather, it results from disconnection of a complex sub-network of brain regions. This provides new insights into the neuroanatomical basis of post-stroke apathy.


Patient and practitioner compliance with silicone hydrogel and daily disposable lens replacement in the United States.
The objectives of this study were to assess current recommendations for replacement frequency (RF) of silicone hydrogel (SH) and daily disposable (DD) lenses, to determine compliance with these recommendations, and to investigate the reasons given for noncompliance. A package containing 20 patient surveys was sent to 309 eye care practitioners (ECPs) in the United States who had agreed to participate in the study. One thousand eight hundred fifty-nine completed surveys were received from 158 ECPs and 1,654 surveys were eligible for analysis. Questions related to patient demographics, lens type, lens wearing patterns, the ECP instructions for RF, and the actual patient reported RF. ECPs were asked to provide lens information and their recommendation for RF after the surveys had been completed and sealed in envelopes. All responses were anonymous. Sixty-six percent of patients were women and their mean age was 34 +/- 12 years. Eighty-eight percent of lenses were worn for daily wear, 12.8 +/- 3.2 hours a day, 6.2 +/- 1.5 days a week. Lens type distribution was 16% DD, 45% 2 week (2W) SH, and 39% 1 month (1M) SH. ECP recommendations for RF varied according to the lens type; 1% of 1M (95% CI 0.2-1.7), 4% of DD (95% CI 2.1-7.2), and 18% of 2W (95% CI 15.1-20.7) patients were given instructions that did not conform to the manufacturers' recommended RF (MRRF). When considering only those patients who were given the correct instructions for RF, 38% were not compliant with the MRRF; noncompliance rates varied according to the lens type and were 12% for DD (95% CI 8.6-17.2), 28% for 1M (95% CI 24.9-32.1), and 52% for 2W (95% CI 47.8-55.8). The most frequent reasons for over wearing lenses were "forgetting which day to replace lenses" (51%) and "to save money" (26%). Fifty-three percent believed that a reminder system would aid compliance; the most popular methods being a cell phone reminder or text message (29%) and a nominated day each week or month (26%). Discussions between the ECPs and the patients were more extensive for patients who were compliant with the MRRF. ECPs recommended RFs more frequently with DD and 1M SH lenses than with 2W SH lenses, consistent with manufacturers' recommendations. Patients were less compliant with RF than ECPs for all lens types investigated. Patients were most compliant with RF when wearing DD lenses and least compliant when wearing 2W SH lenses. Better communication facilitated greater compliance with RF. More than half of those not replacing lenses, when recommended, reported that this was because they forgot which day to replace their lenses.


Protective effects of combined micronutrients on islet beta-cells of streptozotocin-induced diabetic mice.
There is a tendency for the incidence of diabetes in a population to increase with an improvement in living standards. This would imply the involvement of nutritional factors in the development of diabetes, and so nutritional considerations could be a key aspect in the research and development of an effective remedy for diabetes. In this study, combined micronutrients (selenium, vitamin E, vanadium, and chromium) were orally supplemented to streptozotocin-induced diabetic mice. Results showed that combined micronutrients could decrease the high blood glucose levels (p<0.05 or p<0.01) of diabetic mice. The protective effects of combined micronutrients on structures of beta-cells in pancreatic islets of diabetic mice were observed histopathologically and ultrastructurally. In addition, the supplementation of combined micronutrients increased insulin expression by beta-cells in pancreatic islets of diabetic mice at both translational and transcriptional levels. The immune molecular mechanisms involved were preliminarily regarded as downregulation of the expression of pathogenic T-helper 1 lymphocyte (Th1) cytokine tumor necrosis factor-alpha (TNF-alpha) (p<0.01) along with upregulation of the expression of protective T-helper 2 lymphocyte Th2 cytokine interleukin 10 (IL-10) (p<0.01) which ameliorates the Th1/Th2 imbalance in diabetes. In conclusion, supplementation of combined micronutrients to diabetic mice could effectively improve disordered glucose metabolism, protect islet structures, and improve the function of beta-cells in pancreatic islets, which are affected by differential regulation of the expression of Th1/Th2 cytokines involved in the pathogenesis of diabetes.


Contribution of low-molecular-weight antioxidants to the antioxidant capacity of raw and processed lentil seeds.
In this study four cultivars of lentil originating from Spain were examined: cv Paula, cv Agueda, cv Almar and cv Alcor. Since consumption of these seeds after heat treatment and as sprouts has been popularised, the impact of cooking (up to 30 min) and germination process (in dark, at 25 degrees C, for up to 4 days) on peroxyl radical-trapping capacity (PRTC) and Trolox-equivalent antioxidant capacity (TEAC) of the processed seeds was addressed. Also, changes in the content of low-molecular-weight antioxidants (LMWA) and soluble proteins in the course of cooking and germination were studied. The analyzed LMWAwere: total phenolics, tocopherols (alpha-T, beta-T, gamma-T, delta-T), reduced glutathione, and L-ascorbic acid. On the basis of the results obtained, the contribution of LMWA and soluble proteins to the PRTC and TEAC of raw, cooked, and germinated lentil seeds was calculated by multiple mean values for the content of investigated compounds and their relative potential with respect to Trolox. The results showed avery high molar percentage contribution of phenolic compounds and low contribution of tocopherols, glutathione, soluble proteins, and ascorbate (only in germinated seeds) to the total TEAC and total PRTC calculated as a sum of data provided for phosphate-buffered and 80% methanolic extracts of raw and processed lentil seeds.


Optimal strategies for learning multi-ancestry polygenic scores vary across traits.
Polygenic scores (PGSs) are individual-level measures that aggregate the genome-wide genetic predisposition to a given trait. As PGS have predominantly been developed using European-ancestry samples, trait prediction using such European ancestry-derived PGS is less accurate in non-European ancestry individuals. Although there has been recent progress in combining multiple PGS trained on distinct populations, the problem of how to maximize performance given a multiple-ancestry cohort is largely unexplored. Here, we investigate the effect of sample size and ancestry composition on PGS performance for fifteen traits in UK Biobank. For some traits, PGS estimated using a relatively small African-ancestry training set outperformed, on an African-ancestry test set, PGS estimated using a much larger European-ancestry only training set. We observe similar, but not identical, results when considering other minority-ancestry groups within UK Biobank. Our results emphasise the importance of targeted data collection from underrepresented groups in order to address existing disparities in PGS performance.


Return to play criteria after hamstring muscle injury in professional football: a Delphi consensus study.
Hamstring muscle injury (HMI) is the most common injury in professional football and has a high re-injury rate. Despite this, there are no validated criteria to support return to play (RTP) decisions. To use the Delphi method to reach expert consensus on RTP criteria after HMI in professional football. All professional football clubs in England (n=92) were invited to participate in a 3-round Delphi study. Round 1 requested a list of criteria used for RTP decisions after HMI. Responses were independently collated by 2 researchers under univocal definitions of RTP criteria. In round 2 participants rated their agreement for each RTP criterion on a 1-5 Likert Scale. In round 3 participants re-rated the criteria that had reached consensus in round 2. Descriptive statistics and Kendall's coefficient of concordance enabled interpretation of consensus. Participation rate was limited at 21.7% (n=20), while retention rate was high throughout the 3 rounds (90.0%, 85.0%, 90.0%). Round 1 identified 108 entries with varying definitions that were collated into a list of 14 RTP criteria. Rounds 2 and 3 identified 13 and 12 criteria reaching consensus, respectively. Five domains of RTP assessment were identified: functional performance, strength, flexibility, pain and player's confidence. The highest-rated criteria were in the functional performance domain, with particular importance given to sprint ability. This study defined a list of consensually agreed RTP criteria for HMI in professional football. Further work is now required to determine the validity of the identified criteria.


Comparative genomics of hepatitis A virus, hepatitis C virus, and hepatitis E virus provides insights into the evolutionary history of Hepatovirus species.
The intraspecies genomic diversity of the single-strand RNA (+) virus species hepatitis A virus (Hepatovirus), hepatitis C virus (Hepacivirus), and hepatitis E virus (Orthohepevirus) was compared. These viral species all can cause liver inflammation (hepatitis), but share no gene similarity. The codon usage of human hepatitis A virus (HAV) is suboptimal for replication in its host, a characteristic it shares with taxonomically related rodent, simian, and bat hepatitis A virus species. We found this codon usage to be strikingly similar to that of Triatoma virus that infects blood-sucking kissing bugs. The codon usage of that virus is well adapted to its insect host. The codon usage of HAV is also similar to other invertebrate viruses of various taxonomic families. An evolutionary ancestor of HAV and related virus species is hypothesized to be an insect virus that underwent a host jump to infect mammals. The similarity between HAV and invertebrate viruses goes beyond codon usage, as they also share amino acid composition characteristics, while not sharing direct sequence homology. In contrast, hepatitis C virus and hepatitis E virus are highly similar in codon usage preference, nucleotide composition, and amino acid composition, and share these characteristics with Human pegivirus A, West Nile virus, and Zika virus. We present evidence that these observations are only partly explained by differences in nucleotide composition of the complete viral codon regions. We consider the combination of nucleotide composition, amino acid composition, and codon usage preference suitable to provide information on possible evolutionary similarities between distant virus species that cannot be investigated by phylogeny.


Phagosomal signalling of the C-type lectin receptor Dectin-1 is terminated by intramembrane proteolysis.
Sensing of pathogens by pattern recognition receptors (PRR) is critical to initiate protective host defence reactions. However, activation of the immune system has to be carefully titrated to avoid tissue damage necessitating mechanisms to control and terminate PRR signalling. Dectin-1 is a PRR for fungal β-glucans on immune cells that is rapidly internalised after ligand-binding. Here, we demonstrate that pathogen recognition by the Dectin-1a isoform results in the formation of a stable receptor fragment devoid of the ligand binding domain. This fragment persists in phagosomal membranes and contributes to signal transduction which is terminated by the intramembrane proteases Signal Peptide Peptidase-like (SPPL) 2a and 2b. Consequently, immune cells lacking SPPL2b demonstrate increased anti-fungal ROS production, killing capacity and cytokine responses. The identified mechanism allows to uncouple the PRR signalling response from delivery of the pathogen to degradative compartments and identifies intramembrane proteases as part of a regulatory circuit to control anti-fungal immune responses.


The influence of the Rashba spin-orbit coupling on the two-dimensional magnetoexcitons.
The influence of the Rashba spin-orbit coupling (RSOC) on the two-dimensional (2D) electrons and holes in a strong perpendicular magnetic field leads to different results for the Landau quantization in different spin projections. In the Landau gauge the unidimensional wave vector describing the free motion in one in-plane direction is the same for both spin projections, whereas the numbers of Landau quantization levels are different. For an electron in an s-type conduction band they differ by one, as was established earlier by Rashba (1960 Fiz. Tverd. Tela 2 1224), whereas for heavy holes in a p-type valence band influenced by the 2D symmetry of the layer they differ by three. The shifts and the rearrangements of the 2D hole Landau quantization levels on the energy scale are much larger in comparison with the case of conduction electron Landau levels. This is due to the strong influence of the magnetic field on the RSOC parameter. At sufficiently large values of this parameter the shifts and rearrangements are comparable with the hole cyclotron energy. There are two lowest spin-split Landau levels for electrons as well as four lowest ones for holes in the case of small RSOC parameters. They give rise to eight lowest energy bands of the 2D magnetoexcitons, as well as of the band-to-band quantum transitions. It is shown that three of them are dipole-active, three are quadrupole-active and two are forbidden. The optical orientation under the influence of circularly polarized light leads to optical alignment of the magnetoexcitons with different orbital momentum projections in the direction of the external magnetic field.


Who's stealing your money?
Is there a gap between your productivity and profitability? Look inside your organization and you may find hundreds of thousands of dollars that were stolen by someone associated with your practice. This article offers examples of stealing that occurred in hundreds of medical practices and hospitals nationwide and outlines six components that you can install this year to protect your bottom line.


Fishing for leadership: The role diversification plays in facilitating change agents.
Leadership is often viewed as being critical to successful natural resource management. This research focuses on a set of leaders identified through a social network analysis of fishers in a rural coastal region. Leaders' connections to different fisheries are evaluated, and these actors are found to be significantly more diversified than other fishers in the area. Drawing on theory related to institutional entrepreneurship and a series of in-depth interviews with these actors, this paper puts forward several hypotheses to explain how diverse social-ecological connections facilitate leadership. Three mechanisms are identified. Being diversified facilitates: (1) production of alternative visions; (2) framing of tractable strategies to sustain local marine resource; and (3) participation in the management process. While more research is needed to understand the relationship between diversification and leadership, these exploratory results suggest that leadership is, in part, a manifestation of ecological circumstance, supporting recent assertions that scholarship on leadership in natural resource management settings could benefit from being more attentive to the processes that shape leadership rather than fixating on individuals and their personal attributes. Given that fisheries policies increasingly constrain diversification, policymakers and managers should consider how specialization of fishers might change the form and function of leaders in the future.


Compilation of a standardised international folate database for EPIC.
This paper describes the methodology applied for compiling an "international end-user" folate database. This work benefits from the unique dataset offered by the European Prospective Investigation into Cancer and Nutrition (EPIC) (N=520,000 subjects in 23 centres). Compilation was done in four steps: (1) identify folate-free foods then find folate values for (2) folate-rich foods common across EPIC countries, (3) the remaining "common" foods, and (4) "country-specific" foods. Compiled folate values were concurrently standardised in terms of unit, mode of expression and chemical analysis, using information in national food composition tables (FCT). 43-70% total folate values were documented as measured by microbiological assay. Foods reported in EPIC were either matched directly to FCT foods, treated as recipes or weighted averages. This work has produced the first standardised folate dataset in Europe, which was used to calculate folate intakes in EPIC; a prerequisite to study the relation between folate intake and diseases.


Crystal structure of human macrophage elastase (MMP-12) in complex with a hydroxamic acid inhibitor.
Human macrophage elastase (MMP-12) is a member of the family of matrix metalloproteinases (MMPs) that plays, like other members of the family, an important role in inflammatory processes contributing to tissue remodelling and destruction. In particular, a prominent role of MMP-12 in the destruction of elastin in the lung alveolar wall and the pathogenesis of emphysema has been suggested. It is therefore an attractive therapeutic target. We describe here the crystal structure of the catalytic domain of MMP-12 in complex with a hydroxamic acid inhibitor, CGS27023A. MMP-12 adopts the typical MMP fold and binds a structural zinc ion and three calcium ions in addition to the catalytic zinc ion. The enzyme structure shows an ordered N terminus close to the active site that is identical in conformation with the superactivated form of MMP-8. The S1'-specificity pocket is large and extends into a channel through the protein, which puts MMP-12 into the class of MMPs 3, 8 and 13 with large and open specificity pockets. The two crystallographically independent molecules adopt different conformations of the S1'-loop and its neighbouring loop due to differing crystal packing environments, suggesting that flexibility or the possibility of structural adjustments of these loop segments are intrinsic features of the MMP-12 structure and probably a common feature for all MMPs. The inhibitor binds in a bidentate fashion to the catalytic zinc ion. Its polar groups form hydrogen bonds in a substrate-like manner with beta-strand sIV of the enzyme, while the hydrophobic substituents are either positioned on the protein surface and are solvent-exposed or fill the upper part of the specificity pocket. The present structure enables us to aid the design of potent and selective inhibitors for MMP-12.


Mycobiota and aflatoxin B1 in feed for farmed sea bass (Dicentrarchus labrax).
Thesafety characteristics of feed used in fish and crustacean aquaculture systems are an essential tool to assure the productivity of those animal exploitations. Safety of feed may be affected by different hazards, including biological and chemical groups. The aim of this preliminary study was to evaluate fungi contamination and the presence of aflatoxins in 87 samples of feed for sea bass, collected in Portugal. Molds were found in 35 samples (40.2%) in levels ranging from 1 to 3.3 log10 CFU∙g(-1). Six genera of molds were found. Aspergillus flavus was the most frequent, found in all positive samples, with a range from 2 to 3.2 log(10) CFU∙g(-1). Aspergillus niger was found in 34 samples (39.1%), ranging from 1 to 2.7 log(10) CFU∙g(-1). Aspergillus glaucus was found in 26 samples (29.9%) with levels between 1 and 2.4 log(10) CFU∙g(-1). Penicillium spp. and Cladosporium spp. were both found in 25 samples (28.7%). Fusarium spp. was found in 22 samples (25.3%), ranging from 1 to 2.3 log(10) CFU∙g(-1). All feed samples were screened for aflatoxins using a HPLC technique, with a detection limit of 1.0 μg∙kg(-1). All samples were aflatoxin negative.


The role of uric acid in the pathogenesis of human cardiovascular disease.
Hyperuricaemia is common in subjects with cardiovascular disease, but is not commonly considered a true risk factor. Recent studies suggest that uric acid is biologically active and can stimulate oxidative stress, endothelial dysfunction, inflammation and vasoconstriction. Epidemiological studies have found that uric acid can independently predict the development of hypertension, as well as stroke and heart failure. Experimentally raising uric acid in animals increases blood pressure, and pilot studies suggest that lowering uric acid in humans can reduce blood pressure in hypertensive individuals. Uric acid may also have emerging roles in the pathogenesis of kidney disease, metabolic syndrome and diabetes. More studies need to be performed on the pathophysiology and clinical consequences of hyperuricaemia in cardiovascular disease.


Urine testing for cocaine abuse: metabolic and excretion patterns following different routes of administration and methods for detection of false-negative results.
Although cocaine is typically the second-most identified drug of abuse in drug-testing programs, there is surprisingly little quantitative information on excretion patterns following different routes of administration. This report details the urinary excretion and terminal elimination kinetics for cocaine and eight metabolites [benzoylecgonine (BZE), ecgonine methylester (EME), norcocaine (NCOC), benzoylnorecgonine (BNE), m-hydroxy-BZE (m-HO-BZE), p-hydroxy-BZE (p-HO-BZE), m-hydroxy-COC (m-HO-COC), and p-hydroxy-COC (p-HO-COC)]. Six healthy males were administered approximately equipotent doses of cocaine by the intravenous (IV), smoking (SM), and inhalation (IN) routes of administration. Urine specimens were collected for a minimum of three days after drug administration, screened by immunoassay (EMIT and TDX, 300 ng/mL), and analyzed by GC-MS. Mean Cmax values were generally as follows: BZE > EME > COC > BNE approximately p-HO-BZE > m-HO-BZE > m-HO-COC > NCOC > p-HO-COC. Elimination half-lives for cocaine and metabolites were generally shorter following s.m., intermediate after i.v., and longest following i.n. administration. m-HO-BZE demonstrated the longest half-life (mean range 7.0-8.9 h), and cocaine displayed the shortest (2.4-4.0 h). Mean detection times were extended progressively by lowering cutoff concentrations. The maximum increases were approximately 55% at 50 ng/mL for the TDx assay (e.g., the detection time for the last consecutive positive changed from 32.8 h to 50.6 h for i.v. cocaine) and up to 39% for GC-MS at a cutoff concentration of 40 ng/mL (e.g., the detection time for the last consecutive positive changed from 34.8 h to 48.1 h for i.v. cocaine). Sensitivity, specificity, and predictive values for EMIT and TDx were comparable at the 300-ng/mL cutoff concentration; but at lower cutoff concentrations, predictive values of positive results for TDx were diminished indicating a higher risk of false-positive results, that is, positive results that failed to meet administrative cutoff criteria. Detection of positive results was significantly enhanced through the use of the "Zero Threshold Criteria Method", a method developed by the authors to differentiate false-negatives from true-negatives. The method was based on establishing mean immunoassay response (MIR) baselines and variance (SD) in assays of drug-free specimens. Arbitrary thresholds (MIR + 0.5 SD, MIR + 1 SD, MIR + 2 SD) were utilized to evaluate all negative specimens. Apparent true positives were identified by the presence of BZE at or above 40% GC-MS cutoff concentrations. With these criteria, up to 111 false-negative specimens were confirmed as true-positive specimens; this was in addition to the 208 true positives detected at recommended cutoff concentrations. This represents a 50% increase in positive detection rates through the use of this methodology. Such methodology is recommended for further evaluation by drug-testing programs for enhancement of positive detection rates and as an alternative to creatinine testing for dealing with dilute specimens that test negative by initial tests, but contain quantifiable concentrations of drugs of abuse.


The cotton rat provides a novel model to study genital herpes infection and to evaluate preventive strategies.
Prevention of genital herpes and other sexually transmitted infections (STI) is a critical health priority because of the overwhelming impact on women and infants and the epidemiological association with human immunodeficiency virus (HIV)/AIDS. Small animal models are essential for evaluating strategies for prevention or treatment of STI. Neither the murine nor the guinea pig model of genital herpes fully recapitulates human disease. We demonstrate that herpes simplex virus type 2 (HSV-2) readily infects inbred cotton rats (Sigmodon hispidus). Consistent infection does not require pretreatment with medroxyprogesterone, and primary disease resembles that observed in humans. The animals develop genital lesions and fully recover. During primary infection, viral DNA is also detected in liver, lungs, brain, and kidneys. Clinical self-limited recurrences occur spontaneously but may also be induced by dexamethasone. Pretreatment of cotton rats with PRO 2000 gel, a candidate vaginal microbicide being evaluated in clinical trials to prevent HSV and HIV, protects cotton rats from HSV. Together, these studies suggest that the cotton rat may provide an excellent model to study genital herpes and to evaluate preventive strategies.


Evaluation of dipstick analysis among elderly residents to detect bacteriuria: a cross-sectional study in 32 nursing homes.
Few studies have evaluated dipstick urinalysis for elderly and practically none present confidence intervals. Furthermore, most previous studies combine all bacteria species in a "positive culture". Thus, their evaluation may be inappropriate due to Yule-Simpson's paradox. The aim of this study was to evaluate diagnostic accuracy of dipstick urinalysis for the elderly in nursing homes. In this cross-sectional study voided urine specimens were collected from 651 elderly individuals in nursing homes. Dipstick urinalysis for nitrite, leukocyte esterase and urine culture were performed. Sensitivity, specificity, positive and negative predictive values with 95% confidence intervals were calculated. Visual readings were compared to readings with a urine chemistry analyzer. 207/651 (32%) of urine cultures showed growth of a potentially pathogenic bacterium. Combining the two dipsticks improved test characteristics slightly compared to using only one of the dipsticks. When both dipsticks are negative, presence of potentially pathogenic bacteria can be ruled out with a negative predictive value of 88 (84-92)%. Visual and analyzer readings had acceptable agreement. When investigating for bacteriuria in elderly people at nursing homes we suggest nitrite and leukocyte esterase dipstick be combined. There are no clinically relevant differences between visual and analyzer dipstick readings. When dipstick urinalysis for nitrite and leukocyte esterase are both negative it is unlikely that the urine culture will show growth of potentially pathogenic bacteria and in a patient with an uncomplicated illness further testing is unnecessary.


Discrete states of a protein interaction network govern interphase and mitotic microtubule dynamics.
The cytoplasm of eukaryotic cells is thought to adopt discrete "states" corresponding to different steady states of protein networks that govern changes in subcellular organization. For example, in Xenopus eggs, the interphase to mitosis transition is induced solely by activation of cyclin-dependent kinase 1 (CDK1) that phosphorylates many proteins leading to a reorganization of the nucleus and assembly of the mitotic spindle. Among these changes, the large array of stable microtubules that exists in interphase is replaced by short, highly dynamic microtubules in metaphase. Using a new visual immunoprecipitation assay that quantifies pairwise protein interactions in a non-perturbing manner in Xenopus egg extracts, we reveal the existence of a network of interactions between a series of microtubule-associated proteins (MAPs). In interphase, tubulin interacts with XMAP215, which is itself interacting with XKCM1, which connects to APC, EB1, and CLIP170. In mitosis, tubulin interacts with XMAP215, which is connected to EB1. We show that in interphase, microtubules are stable because the catastrophe-promoting activity of XKCM1 is inhibited by its interactions with the other MAPs. In mitosis, microtubules are short and dynamic because XKCM1 is free and has a strong destabilizing activity. In this case, the interaction of XMAP215 with EB1 is required to counteract the strong activity of XKCM1. This provides the beginning of a biochemical description of the notion of "cytoplasmic states" regarding the microtubule system.


A clinical pilot study of lignin--ascorbic acid combination treatment of herpes simplex virus.
Antiviral drugs as well as natural remedies have been used to reduce symptoms and the rate of recurrences of herpes simplex virus type 1 (HSV-1) infection, a common disease. To evaluate anti-HSV-1 activity of a pine cone lignin and ascorbic acid treatment, a clinical pilot study was carried out. Forty-eight healthy patients of both genders between 4 and 61 years old (mean: 31+/-16 years), with active lesions of HSV-1, took part in the study. According to the HSV-1 stage at the presentation, the patients were classified into the prodromic (16 patients), erythema (11 patients), papule edema (1 patient), vesicle/pustule (13 patients) and ulcer stages (7 patients). One mg of lignin-ascorbic acid tablet or solution was orally administered three times daily for a month. Clinical evaluations were made daily the first week and at least three times a week during the second week after the onset and every six months during the subsequent year to identify recurrence episodes. The patients who began the lignin-ascorbic acid treatment within the first 48 hours of symptom onset did not develop HSV-1 characteristic lesions, whereas those patients who began the treatment later experienced a shorter duration of cold sore lesions and a decrease in the symptoms compared with previous episodes. The majority of the patients reported the reduction in the severity of symptoms and the reduction in the recurrence episodes after the lignin-ascorbic acid treatment compared with previous episodes, suggesting its possible applicability for the prevention and treatment of HSV-1 infection.


Risk Factors for Hemorrhagic and Ischemic Stroke in Sub-Saharan Africa.
In sub-Saharan Africa (SSA), there is a significant burden of ischemic stroke (IS) and hemorrhagic stroke (HS), although data on risk factors for each type are sparse. In this systematic review we attempt to characterize the risk factors. We systematically reviewed (PubMed, EMBASE, WHOLIS, Google Scholar, Wiley online, and the Cochrane Central Register of Controlled Trials (CENTRAL)) case-control studies and case series from 1980 to 2016 that reported risk factors for IS and/or HS in SSA. For each risk factor we calculated random-effects pooled odds ratios (ORs) for case-control studies and pooled prevalence estimates for case series. Results. We identified 12 studies, including 4,387 stroke patients. Pooled analysis showed that patients who had diabetes (OR = 2.39; 95% CI: 1.14-5.03) and HIV (OR = 2.46 (95% CI: 1.59-3.81) were at a significantly greater risk of suffering from all stroke types. There were insufficient data to examine these factors by stroke type. Among case series, the pooled prevalence of hypertension was higher for HS than for IS (73.5% versus 62.8%), while diabetes mellitus (DM) and atrial fibrillation (AF) were more prevalent among IS compared to HS (15.9% versus 10.6% and 9.6% versus 2.3%, respectively). There remain too few data from SSA to reliably estimate the effect of various factors on the risk of IS and HS. Furthermore, the vast majority of cases were identified in hospital and so are unlikely to be representative of the totality of stroke cases in the community.


Carbon Monoxide-Releasing Molecule-401 Suppresses Polymorphonuclear Leukocyte Migratory Potential by Modulating F-Actin Dynamics.
Carbon monoxide-releasing molecules (CORMs) suppress inflammation by reducing polymorphonuclear leukocyte (PMN) recruitment to the affected organs. We investigated modulation of PMN-endothelial cell adhesive interactions by water-soluble CORM-401 using an experimental model of endotoxemia in vitro. Human umbilical vein endothelial cells (HUVEC) grown on laminar-flow perfusion channels were stimulated with 1 μg/mL lipopolysaccharide for 6 hours and perfused with 100 μmol/L CORM-401 (or inactive compound iCORM-401)-pretreated PMN for 5 minutes in the presence of 1.0 dyn/cm2 shear stress. PMN co-cultures were perfused for additional 15 minutes with PMN-free medium containing CORM-401/inactive CORM-401. The experiments were videorecorded (phase-contrast microscopy), and PMN adhesion/migration were assessed off-line. In parallel, CORM-401-dependent modulation of PMN chemotaxis, F-actin expression/distribution, and actin-regulating pathways [eg, p21-activated protein kinases (PAK1/2) and extracellular signal-regulated kinase (ERK)/C-Jun N-terminal kinase (JNK) mitogen-activated protein kinases (MAPK)] were assessed in response to N-formyl-methionyl-leucyl-phenylalanine (fMLP) stimulation. Pretreating PMN with CORM-401 did not suppress PMN adhesion to HUVEC, but significantly reduced PMN transendothelial migration (P < 0.0001) and fMLP-induced PMN chemotaxis (ie, migration directionality and velocity). These changes were associated with CORM-401-dependent suppression of F-actin levels/cellular distribution and fMLP-induced phosphorylation of PAK1/2 and ERK/JNK MAPK (P < 0.05). CORM-401 had no effect on p38 MAPK activation. In summary, this study demonstrates, for the first time, CORM-401-dependent suppression of neutrophil migratory potential associated with modulation of PAK1/2 and ERK/JNK MAPK signaling and F-actin dynamics.


Increased small bowel epithelial turnover in interleukin-1 receptor knockout mice.
To determine whether interleukin-1 (IL-1) affects the cellular homeostasis of small bowel mucosa, the authors studied apoptosis and proliferation in small bowel epithelium in two groups of C57 mice: an IL-1 receptor knockout group, and a control wild-type group. Gut mucosal integrity is maintained by a balance of cell proliferation and cell death. Recent reports suggest that IL-1, a proinflammatory cytokine, increases cell death by apoptosis in some epithelial cells. Twenty-four male C57BL6 IL-1 receptor (type I) knockout mice were killed, and small bowel was removed for study. Twenty-four wild-type mice (C57-BL6) served as controls. Body weights, bowel length, and mucosal morphology were examined for phenotypic differences. Apoptosis was quantified by terminal deoxyuridine nick-end labeling (TUNEL) immunohistochemical staining and cellular proliferation by proliferation cell nuclear antigen staining. Whole mucosal protein was analyzed for nuclear factor-kappaB expression. Groups were analyzed by t test. The absence of IL-1 type I receptor in knockout mice was verified by reverse transcriptase-polymerase chain reaction. IL-1 receptor null mice were larger than wild-type controls, with a longer small bowel; however, the index of small bowel length to total body weight did not differ between groups. The percentage of apoptotic cells was higher in IL-1 receptor null mice than in wild-type mice; the proliferation index also increased. Mucosal height and other measures of mucosal morphology were not different. Genotypic absence of IL-1 receptors was associated with decreased expression of nuclear factor-kappaB in whole mucosal protein extracts. Both apoptosis and proliferation increased in gut epithelial cells of mice without IL-1 receptors, suggesting increased cell turnover with no change in net balance. This model represents an opportunity to examine potential mechanisms of gut epithelial turnover in vivo, under both normal conditions and in conditions of mucosal proliferation and atrophy.