Transcriptional responses to thermal acclimation in the eurythermal fish Gillichthys mirabilis (Cooper 1864).
Thermal acclimation (acclimatization) capacity may be critical for determining how successfully an ectotherm can respond to temperature change, and adaptive shifts in gene expression may be pivotal for mediating these acclimatory responses. Using a cDNA microarray, we examined transcriptional profiles in gill tissue of a highly eurythermal goby fish, Gillichthys mirabilis, following 4 wk of acclimation to 9 degrees C, 19 degrees C, or 28 degrees C. Overall, gill transcriptomes were not strikingly different among acclimation groups. Of the 1,607 unique annotated genes on the array, only 150 of these genes (9%) were significantly different in expression among the three acclimation groups (ANOVA, false discovery rate < 0.05). Principal component analysis revealed that 59% of the variation in expression among these genes was described by an expression profile that is upregulated with increasing acclimation temperature. Gene ontology analysis of these genes identified protein biosynthesis, transport, and several metabolic categories as processes showing the greatest change in expression. Our results suggest that energetic costs of macromolecular turnover and membrane-localized transport rise with acclimation temperature. The upregulation of several classes of stress-related proteins, e.g., heat shock proteins, seen in the species' response to acute thermal stress was not observed in the long-term 28 degrees C-acclimated fish. The transcriptional differences found among the acclimation groups thus may reflect an acclimation process that has largely remedied the effects of acute thermal stress and established a new steady-state condition involving changes in relative energy costs for different processes. This pattern of transcriptional alteration in steady-state acclimated fish may be a signature of eurythermy.


VirB3 to VirB6 and VirB8 to VirB11, but not VirB7, are essential for mediating persistence of Brucella in the reticuloendothelial system.
The Brucella abortus virB locus contains 12 open reading frames, termed virB1 through virB12, which encode a type IV secretion system. Polar mutations in the virB locus markedly reduce the ability of B. abortus to survive in cultured macrophages or to persist in organs of mice. While a nonpolar deletion of the virB2 gene reduces survival in cultured macrophages and in organs of mice, a nonpolar deletion of virB1 only reduces survival in macrophages, whereas virB12 is dispensable for either virulence trait. Here we investigated the role of the remaining genes in the virB locus during survival in macrophages and virulence in mice. Mutants carrying nonpolar deletions of the virB3, virB4, virB5, virB6, virB7, virB8, virB9, virB10, or virB11 gene were constructed and characterized. All mutations reduced the ability of B. abortus to survive in J774A.1 mouse macrophage-like cells to a degree similar to that caused by a deletion of the entire virB locus. Deletion of virB3, virB4, virB5, virB6, virB8, virB9, virB10, or virB11 markedly reduced the ability of B. abortus to persist in the spleens of mice at 8 weeks after infection. Interestingly, deletion of virB7 did not reduce the ability of B. abortus to persist in spleens of mice. We conclude that virB2, virB3, virB4, virB5, virB6, virB8, virB9, virB10, and virB11 are essential for virulence of B. abortus in mice, while functions encoded by the virB1, virB7, and virB12 genes are not required for persistence in organs with this animal model.


Pediatric coverage of the delivery room: an analysis of manpower utilization.
To determine the frequency and pattern of pediatric calls to the delivery room and the actual type of medical care administered to the newborn in the delivery room. This was a prospective observational study of 2554 births in a university-affiliated tertiary care hospital. Existing protocols required the attendance of a pediatric resident or neonatal fellow at all deliveries other than uncomplicated vaginal term births. The pediatrician's activity in the delivery room was characterized as either "medical care" or "minimal care." Results were analyzed by diagnostic category. Pediatricians attended 646 of the deliveries (25%). Medical care was administered in 204 of the deliveries, representing 31% of the time they were at a delivery and 8% of all deliveries. The three major indications for pediatric delivery room attendance were cesarean sections (n = 253; 39%), presence of meconium in amniotic fluid (n = 117; 18%), and vacuum deliveries (n = 117; 18%). Medical care was required only in 1 of 56 cases of elective repeat cesarean sections, in 1 of 20 cases of a cesarean section for nonprogress of labor, and in 1 of 38 cases when thin meconium was present. In contrast, medical care was needed in 52 of 81 (64%) cases of cesarean sections for fetal distress, in 11 of 11 (100%) of the cesarean sections for multiple births, and in 67 of 89 (85%) cases of thick meconium (p < 0.05). There was a need for medical attendance after the birth in less than 1% of 1908 cases for which the pediatrician was not initially called to delivery room. Because their medical skills were needed only one of three times that pediatricians were called to the delivery room, and then mostly in specific risk situations, more selective use of pediatric manpower for delivery room coverage may lead to a more efficient use of medical resources without any apparent increase in patient morbidity.


Temporal regulation of the Mus81-Mms4 endonuclease ensures cell survival under conditions of DNA damage.
The structure-specific Mus81-Eme1/Mms4 endonuclease contributes importantly to DNA repair and genome integrity maintenance. Here, using budding yeast, we have studied its function and regulation during the cellular response to DNA damage and show that this endonuclease is necessary for successful chromosome replication and cell survival in the presence of DNA lesions that interfere with replication fork progression. On the contrary, Mus81-Mms4 is not required for coping with replicative stress originated by acute treatment with hydroxyurea (HU), which causes fork stalling. Despite its requirement for dealing with DNA lesions that hinder DNA replication, Mus81-Mms4 activation is not induced by DNA damage at replication forks. Full Mus81-Mms4 activity is only acquired when cells finish S-phase and the endonuclease executes its function after the bulk of genome replication is completed. This post-replicative mode of action of Mus81-Mms4 limits its nucleolytic activity during S-phase, thus avoiding the potential cleavage of DNA substrates that could cause genomic instability during DNA replication. At the same time, it constitutes an efficient fail-safe mechanism for processing DNA intermediates that cannot be resolved by other proteins and persist after bulk DNA synthesis, which guarantees the completion of DNA repair and faithful chromosome replication when the DNA is damaged.


Isolation of the liver N-aspartyl-beta-glucosaminidase in aspartylglucosaminuria.
The isolation of liver N-aspartyl-beta-glucosaminidase in human aspartylglucosaminuria, where this enzyme activity is diminished, yields an enzyme molecule with the same molecular weight and pH optimum as the normal enzyme. Its activity is 10% of that of the control preparation. Combination of both enzymes results in the summation of both activities, and the pathological enzyme does not inhibit the control preparation. It is concluded that no change into a totally different isoenzyme has occurred in aspartylglucosaminuria.


Surgical options for left-sided large bowel emergencies.
Current choices of operation for left-sided large bowel emergencies have been established by a questionnaire sent to 218 consultant surgeons asking which operation they would perform under varying circumstances for obstructing sigmoid carcinoma and diverticular disease. A 92% response rate was obtained. Hartmann's procedure (with or without a mucus fistula) is the most popular operation for all conditions. Sigmoid colectomy with primary anastomosis is performed by 40% of surgeons for obstructing carcinomas, but less commonly in other situations. On-table lavage is rarely used, and the majority of anastomoses are not protected by a proximal stoma. Subtotal colectomy is very seldom employed, except when caecal perforation results from an obstructing carcinoma. Some surgeons perform a defunctioning colostomy alone even in the presence of a perforation.


CpG-B oligodeoxynucleotides inhibit TLR-dependent and -independent induction of type I IFN in dendritic cells.
CpG oligodeoxynucleotides (ODNs) signal through TLR9 to induce type I IFN (IFN-alphabeta) in dendritic cells (DCs). CpG-A ODNs are more efficacious than CpG-B ODNs for induction of IFN-alphabeta. Because IFN-alphabeta may contribute to autoimmunity, it is important to identify mechanisms to inhibit induction of IFN-alphabeta. In our studies, CpG-B ODN inhibited induction of IFN-alphabeta by CpG-A ODN, whereas induction of TNF-alpha and IL-12p40 by CpG-A ODN was not affected. CpG-B inhibition of IFN-alphabeta was observed in FLT3 ligand-induced murine DCs, purified murine myeloid DCs, plasmacytoid DCs, and human PBMCs. CpG-B ODN inhibited induction of IFN-alphabeta by agonists of multiple receptors, including MyD88-dependent TLRs (CpG-A ODN signaling via TLR9, or R837 or Sendai virus signaling via TLR7) and MyD88-independent receptors (polyinosinic:polycytidylic acid signaling via TLR3 or ds break-DNA signaling via a cytosolic pathway). CpG-B ODN did not inhibit the IFN-alphabeta positive feedback loop second-wave IFN-alphabeta, because IFN-alphabeta-induced expression of IFN-alphabeta was unaffected, and CpG-B inhibition of IFN-alphabeta was manifested in IFN-alphabetaR(-/-) DCs, which lack the positive feedback mechanism. Rather, CpG-B ODN inhibited early TLR-induced first wave IFN-alpha4 and IFN-beta. Chromatin immunoprecipitation revealed that association of IFN regulatory factor 1 with the IFN-alpha4 and IFN-beta promoters was induced by CpG-A ODN but not CpG-B ODN. Moreover, CpG-A-induced association of IFN regulatory factor 1 with these promoters was inhibited by CpG-B ODN. Our studies demonstrate a novel mechanism of transcriptional regulation of first-wave IFN-alphabeta that selectively inhibits induction of IFN-alphabeta downstream of multiple receptors and may provide targets for future therapeutic inhibition of IFN-alphabeta expression in vivo.


[Forensic medical characteristic of sawed injuries inflicted to the long tubular bones by a power jigsaw].
The main aspects of investigations devoted to forensic medical characteristic of sawed injuries inflicted to the long tubular bones by a power jigsaw are considered. The mathematical model has been developed making it possible to determine the frequency of backward and forward movements of the jigsaw blade from the morphological features of the injuries to long tubular bones of man and to estimate the sawing rate for the further identification of the instrument of crime.


The role of dose and restriction state on morphine-, cocaine-, and LiCl-induced suppression of saccharin intake: A comprehensive analysis.
Rats avoid intake of a taste cue when paired with a drug of abuse or with the illness-inducing agent, lithium chloride (LiCl). Although progress has been made, it is difficult to compare the suppressive effects of abused agents and LiCl on intake of a gustatory conditioned stimulus (CS) because of the cross-laboratory use of different CSs, different unconditioned stimuli (USs), and different doses of the drugs, different conditioning regimens, and different restriction states. Here we have attempted to unify these variables by comparing the suppressive effects of a range of doses of morphine, cocaine, and LiCl on intake of a saccharin CS using a common regimen in non-restricted, food restricted, or water restricted male Sprague-Dawley rats. The results showed that, while the putatively aversive agent, LiCl, was effective in suppressing intake of the taste cue across nearly all doses, regardless of restriction state, the suppressive effects of both morphine and cocaine were greatly reduced when evaluated in either food or water restricted rats. Greater sensitivity to drug was revealed, at very low doses, when testing occurred in the absence of need (i.e., when the rats were non-restricted). Together, these results provide the first uniform and comprehensive analysis of the suppressive effects of morphine, cocaine, and LiCl as a function of dose and restriction state. In the present case, the suppressive effects of morphine and cocaine are found to differ from those of LiCl and, in some respects, from one another as well.


The effect of two different rehabilitation treatments in cervical dystonia: preliminary results in four patients.
The most widespread approach to rehabilitation of cervical dystonia is electromyographic (EMG) biofeedback. However, consensus is lacking regarding the true effectiveness of this technique. The aim of this study was to evaluate how cervical dystonia was influenced by two rehabilitative treatments, namely a standard biofeedback program and a novel physiotherapy program consisting of postural reeducation exercises and passive elongation of myofascial cervical structures. Both programs were consecutively administered to 4 patients with cervical dystonia. The study design was a behavioral analysis of single cases. The main outcome measures were a head realignment test, a disability questionnaire and a pain visual analogue scale (VAS). Each patient's performance was evaluated before the study and after the first and second program. Furthermore, the disability questionnaire and the pain VAS were administered 3, 6, and 9 months after the end of the treatments. The physiotherapy program showed therapeutic effects comparable to those of EMG biofeedback. Reductions of disability and of pain were still present 3-9 months after the end of the treatments. These preliminary results suggest that the physiotherapy program proposed in the present study may be a promising method for rehabilitation of cervical dystonia.


Characterization of substance P contractile activity on isolated guinea pig lung tissues.
In the presence of the neutral metalloendopeptidase inhibitor, phosphoramidon, substance P (SP) is a highly potent spasmogen for isolated lung parenchymal strips as well as tracheal rings from the guinea pig. We studied the mechanism of action of this peptide, and of the related tachykinin, substance K (SK), on both tissue preparations. The cyclooxygenase inhibitors, indomethacin (1 microM) or aspirin (100 microM), in combination with phosphoramidon (1 microM) effectively block SP-induced contractions in lung parenchymal strips. The lipoxygenase inhibitor, nordihydroguaiaretic acid (10 microM), the H1 antihistamine, pyrilamine (1 microM) and the anticholinergic agent, atropine (1 microM), all had no significant effect on SP-induced contractions. No detectable levels of thromboxane B2, or prostaglandins D2, E2, F2 alpha, or 6-keto-F1 alpha were released into the tissue bathing fluid. These data suggest that the contractile response of guinea pig lung parenchymal strips is mediated by cyclooxygenase metabolites, which are not released in significant concentration from the cells. In the presence of phosphoramidon, SK has a concentration-response curve similar to SP on guinea pig lung parenchymal strips. Its contractile activity is also inhibited by indomethacin but less effectively than SP. In marked contrast, the contractile responses of guinea pig tracheal tissues to the tachykinins were not affected significantly by indomethacin, alone or in combination with phosphoramidon. Additionally, tracheal tissue is 20- to 100-fold more sensitive to SK than SP in the presence or absence, respectively, of the endopeptidase inhibitor.(ABSTRACT TRUNCATED AT 250 WORDS)


Correction of Planovalgus Deformity Through Rotational Reinsertion of the Lateral Layers of the Achilles Tendons in Ambulatory Children With Cerebral Palsy.
Symptomatic planovalgus deformity is a condition commonly seen in patients with cerebral palsy. The authors propose a new procedure for the management of this deformity through rotational reinsertion of the lateral layers of the Achilles tendon, and then they assess its benefit by comparing plantar pressure distribution patterns in children preoperatively and at 6- and 12-month intervals postoperatively. Pedobarographic measurements, range of motion of the ankle, and radiographic indexes were used to assess the outcome of the surgery. The functional abilities of the patients were assessed based on the Gross Motor Function Classification System. A total of 37 feet (22 patients) were included, with a mean ± standard deviation age at surgery of 11.8 ± 2.7 (range 9.1 to 14.5) years. All feet were managed through rotational reinsertion of the lateral layers of the Achilles tendon. Surgical correction of planovalgus has good outcomes. Significant changes were observed with statistical significance at the 5% (p ≤ .05) level in plantar pressure distribution in children preoperatively and at 6- and 12-month intervals postoperatively. The results show that the proposed method of surgery is effective in the correction of planovalgus in ambulatory children with cerebral palsy.


Validation of a tracer technique to determine nonsteady-state ketone body turnover rates in man.
The features of a single-compartment model of total ketone bodies were evaluated using primed constant infusions of [3-14C]acetoacetate (AcAc) and of D-[3-14C]beta-hydroxybutyrate (beta OHB) in 12 postabsorptive subjects. The volume of distribution (VD) of AcAc was 0.18 +/- 0.01 liter/kg (n = 9), and that of beta OHB was similar, 0.18 +/- 0.02 liter/kg (n = 3). The production rate of total ketone bodies was calculated using the combined specific activity of AcAc and of beta OHB. The mean basal total ketone body production rates were similar using either [14C]AcAc (6.5 mumol . kg-1 . min-1) or [14C]beta OHB (6.8 mumol . kg-1 . min-1). To determine the pool fraction that was rapidly mixed during nonsteady state of ketone body inflow, unlabeled AcAc was infused with stepwise increasing and decreasing rates between 5 and 25 mumol . kg-1 . m-1 to mimic nonsteady-state ketone body production rates. The "functional" pool fraction P was determined as the pool fraction that provided the best match between tracer-determined rates of ketone production and rates of AcAc infusion. P of total ketone bodies was almost equal to 1 using either [14C]AcAc (1.05 +/- 0.16) or [14C]beta OHB (1.00 +/- 0.06), suggesting rapid mixing of ketone bodies throughout the entire pool. The described pool model may be used to determine total ketone body kinetics during acute perturbations of the steady state.


Dynamic prefrontal inhibition code mediates reward devaluation.
Repeated reward intake decreases its subjective pleasantness, which is a common phenomenon called reward devaluation. In this issue of Neuron, Yuan et al.1 unravel that blunted inhibitory response of anterior cingulate cortex (ACC) encodes this process, whose hypersensitization leads to anhedonia.


Perceived Walking Speed, Measured Tandem Walk, Incident Stroke, and Mortality in Older Latino Adults: A Prospective Cohort Study.
Walking speed is associated with functional status and all-cause mortality. Yet the relationship between walking speed and stroke, also a leading cause of disability, remains poorly understood, especially in older Latino adults who suffer from a significant burden of stroke. A total of 1,486 stroke-free participants from the Sacramento Area Latino Study on Aging, aged 60 and older at baseline in 1998-1999, were followed annually through 2010. Participants reported their usual walking speed outdoors which was classified into slow, medium, or fast. We also assessed timed tandem walk ability (unable or eight or more errors vs less than eight errors). We ascertained three incident stroke endpoints: total stroke, nonfatal stroke, and fatal stroke. Using Cox proportional hazards models, we estimated hazard ratios (HRs) for stroke at different walking speed and timed tandem walk categories. Over an average of 6 years of follow-up (SD = 2.8), the incidence rate of total strokes was 23.2/1,000 person-years for slow walkers compared to 15.6/1,000 person-years for medium walkers, and 7.6/1,000 person-years for fast walkers. In Cox models adjusted for sociodemographics, cardiovascular risk, cognition and functional status, and self-rated health, the hazard of total stroke was 31% lower for medium walkers (HR: 0.69, 95% confidence interval [CI]: 0.47, 1.02) and 56% lower for fast walkers (HR: 0.44, 95% CI: 0.24, 0.82) compared with slow walkers. We found similar associations with timed tandem walk ability (fully adjusted HR: 0.66, 95% CI: 0.45, 0.98). Our findings suggest perceived walking speed captures more than self-rated health alone and is a strong risk factor for stroke risk in Latino older adults.


Tuberculosis in Somalia.
This is a description of a tuberculosis treatment programme in a country at war where security and the absence of order pose problems to health care delivery. It is also a description of an epidemic of tuberculosis where treatment and diagnosis are difficult and the methods used have changed little in many years. More international pressure is needed.


Variants in the 15q24/25 locus associate with lung function decline in active smokers.
Genetic variation in nicotinic acetylcholine receptor subunit genes (nAChRs) is associated with lung function level and chronic obstructive pulmonary disease (COPD). It is unknown whether these variants also predispose to an accelerated lung function decline. We investigated the association of nAChR susceptibility variants with lung function decline and COPD severity. The rs1051730 and rs8034191 variants were genotyped in a population-based cohort of 1,226 heavy smokers (COPACETIC) and in an independent cohort of 883 heavy smokers, of which 653 with COPD of varying severity (LEUVEN). Participants underwent pulmonary function tests at baseline. Lung function decline was assessed over a median follow-up of 3 years in COPACETIC. Current smokers homozygous for the rs1051730 A-allele or rs8034191 G-allele had significantly greater FEV(1)/FVC decline than homozygous carriers of wild-type alleles (3.3% and 4.3%, p = 0.026 and p = 0.009, respectively). In the LEUVEN cohort, rs1051730 AA-carriers and rs8034191 GG-carriers had a two-fold increased risk to suffer from COPD GOLD IV (OR 2.29, 95% confidence interval [CI] = 1.11-4.75; p = 0.025 and OR = 2.42, 95% [CI] = 1.18-4.95; p = 0.016, respectively). The same risk alleles conferred, respectively, a five- and four-fold increased risk to be referred for lung transplantation because of end-stage COPD (OR = 5.0, 95% [CI] = 1.68-14.89; p = 0.004 and OR = 4.06, 95% [CI] = 1.39-11.88; p = 0.010). In Europeans, variants in nAChRs associate with an accelerated lung function decline in current smokers and with clinically relevant COPD.


The ascorbic acid redox state controls guard cell signaling and stomatal movement.
H(2)O(2) serves an important stress signaling function and promotes stomatal closure, whereas ascorbic acid (Asc) is the major antioxidant that scavenges H(2)O(2). Dehydroascorbate reductase (DHAR) catalyzes the reduction of dehydroascorbate (oxidized ascorbate) to Asc and thus contributes to the regulation of the Asc redox state. In this study, we observed that the level of H(2)O(2) and the Asc redox state in guard cells and whole leaves are diurnally regulated such that the former increases during the afternoon, whereas the latter decreases. Plants with an increased guard cell Asc redox state were generated by increasing DHAR expression, and these exhibited a reduction in the level of guard cell H(2)O(2). In addition, a higher percentage of open stomata, an increase in total open stomatal area, increased stomatal conductance, and increased transpiration were observed. Guard cells with an increase in Asc redox state were less responsive to H(2)O(2) or abscisic acid signaling, and the plants exhibited greater water loss under drought conditions, whereas suppressing DHAR expression conferred increased drought tolerance. Our analyses suggest that DHAR serves to maintain a basal level of Asc recycling in guard cells that is insufficient to scavenge the high rate of H(2)O(2) produced in the afternoon, thus resulting in stomatal closure.


Exercise and Muscle Lipid Content, Composition, and Localization: Influence on Muscle Insulin Sensitivity.
Accumulation of lipid in skeletal muscle is thought to be related to the development of insulin resistance and type 2 diabetes. Initial work in this area focused on accumulation of intramuscular triglyceride; however, bioactive lipids such as diacylglycerols and sphingolipids are now thought to play an important role. Specific species of these lipids appear to be more negative toward insulin sensitivity than others. Adding another layer of complexity, localization of lipids within the cell appears to influence the relationship between these lipids and insulin sensitivity. This article summarizes how accumulation of total lipids, specific lipid species, and localization of lipids influence insulin sensitivity in humans. We then focus on how these aspects of muscle lipids are impacted by acute and chronic aerobic and resistance exercise training. By understanding how exercise alters specific species and localization of lipids, it may be possible to uncover specific lipids that most heavily impact insulin sensitivity.


Hearing testing in autistic spectrum disorder: is it unnecessary in low and middle-income countries?
In low and middle-income countries where the health care is in a weakened state, spending money on unaffordable and probably unnecessary investigations might be substituted by a reliable, simple and more informative tool that can deal with the problem. To examine current medical practice of measuring auditory brainstem response for all children with autistic spectrum disorder, and assess the value of this test in these children and its applicability in low and middle-income countries such as Libya. We reviewed the medical records of all children with autistic spectrum disorder who presented to neurodevelopment clinics of Al-Khadra Teaching Hospital, Tripoli, Libya between January 2010 and December 2014. In 71 of 2368 children with autistic spectrum disorder, the family were concerned about their children's hearing and reaction to loud noises. Auditory brainstem response confirmed that 26 of these 71 children had sensorineural hearing loss. We consider auditory brainstem response measurement to be unnecessary in children with autistic spectrum disorder without clinical signs suggesting hearing impairment and without any parental concerns about hearing.


Language MEG predicts postoperative verbal memory change in left mesial temporal lobe epilepsy.
To clarify whether preoperative language magnetoencephalography (MEG) predicts postoperative verbal memory (VM) changes in left mesial temporal lobe epilepsy (LMTLE). We reviewed 18 right-handed patients with LMTLE who underwent anterior temporal lobectomy or selective amygdala hippocampectomy, 12 with (HS+) and 6 without hippocampal sclerosis (HS-). Patients underwent neuropsychological assessment before and after surgery. MEG was measured with an auditory verbal learning task in patients preoperatively and in 15 right-handed controls. Dynamic statistical parametric mapping (dSPM) was used for source imaging of task-related activity. Language laterality index (LI) was calculated by z-score of dSPM in language-related regions. LI in the region of HS+ and HS- was compared to controls. The correlation between LI and postoperative VM change was assessed in HS+ and HS-. Preoperative LI in supramarginal gyrus showed greater right-shifted lateralization in both HS+ and HS- than in controls. Right-shifted LI in supramarginal gyrus was correlated with postoperative VM increase in HS+ (p = 0.019), but not in HS-. Right-shifted language lateralization in dSPM of MEG signals may predict favorable VM outcome in HS+ of LMTLE. Findings warrant further investigation of the relation between regional language laterality index and postoperative verbal memory changes.


Identification of Mycoplasmatales: characterization procedures.
A large collection of avian Mycoplasma cultures was used in studies to improve and develop biological and biochemical characterization techniques. Differential patterns among 11 avian serotypes were shown by carbohydrate fermentation, tetrazolium- and methylene blue-reduction reactions, breakdown of arginine, and the formation of film on egg yolk-agar. Some cultures fermented as many as 14 carbohydrates. Polyhydric alcohols and pentoses were among the compounds fermented. An improved procedure for determining methylene blue reduction by Mycoplasma was developed. These simple, rapid procedures are reproducible and should be useful in grouping Mycoplasma isolates prior to definitive identification by serological or other means.


Phenotypic variability in patients with osteogenesis imperfecta caused by BMP1 mutations.
Osteogenesis Imperfecta (OI) is an inherited bone fragility disorder most commonly associated with autosomal dominant mutations in the type I collagen genes. Autosomal recessive mutations in a number of genes have also been described, including the BMP1 gene that encodes the mammalian Tolloid (mTLD) and its shorter isoform bone morphogenic protein-1 (BMP1). To date, less than 20 individuals with OI have been identified with BMP1 mutations, with skeletal phenotypes ranging from mild to severe and progressively deforming. In the majority of patients, bone fragility was associated with increased bone mineral density (BMD); however, the full range of phenotypes associated with BMP1 remains unclear. Here, we describe three children with mutations in BMP1 associated with a highly variable phenotype: a sibship homozygous for the c.2188delC mutation that affects only the shorter BMP1 isoform and a further patient who is compound heterozygous for a c.1293C>G nonsense mutation and a c.1148G>A missense mutation in the CUB1 domain. These individuals had recurrent fractures from early childhood, are hypermobile and have no evidence of dentinogenesis imperfecta. The homozygous siblings with OI had normal areal BMD by dual energy X-ray absorptiometry whereas the third patient presented with a high bone mass phenotype. Intravenous bisphosphonate therapy was started in all patients, but discontinued in two patients and reduced in another due to concerns about increasing bone stiffness leading to chalk-stick fractures. Given the association of BMP1-related OI with very high bone material density, concerns remain whether anti-resorptive therapy is indicated in this ultra-rare form of OI.© 2016 Wiley Periodicals, Inc.


A heteroleptic cyclometalated iridium(III) fluorophenylpyridine complex from partial defluorohydrogenation reaction: synthesis, photophysical properties and mechanistic insights.
In this paper, a C-F bond activation reaction of a chloro-bridged iridium(III) dimer (dfppy)(2)Ir(μ-Cl)(2)Ir(dfppy)(2) (1) (dfppy denotes 2-(4,6-difluorophenyl)pyridyl) in the presence of sodium methoxide has been reported, leading to the formation of a heteroleptic cyclometalated iridium(III) fluorophenylpyridine complex 2. HPLC-mass analysis confirmed the release of formaldehyde in the product mixture. When sodium benzyloxide was used as the base, complex 2 was also generated with the release of a benzaldehyde derivative. Complex 2 has been fully characterized by (1)H-NMR, (19)F-NMR and X-ray crystallographic methods, confirming the partial loss of one of the fluorine atoms on one of the cyclometalated phenylpyridyl ligands. Photophysical studies of complex 2 show that it has a similar absorption spectrum to that of Ir(III)(dfppy)(3). However, the emission spectrum shows a red shift maximum emission band at 478 nm due to the loss of a single fluorine atom, highlighting the critical effect of fluorine on the photoluminescence of these Ir(III) complexes. Finally, intensive mechanistic studies including HPLC-mass analysis, (1)H-NMR, and (19)F-NMR studies demonstrate that the formation of complex 2 should involve a critical β-hydride elimination of Ir(III)-alkoxide intermediate and the participation of Ir-hydride and/or Ir-fluoride intermediates.


Clinical and genetic features of familial Mediterranean fever in Japan.
Familial Mediterranean fever (FMF) is thought to be a rare disorder in Japan, and the clinical features of Japanese patients with FMF remain unclear. Our aim was to elucidate the clinical characteristics of FMF in Japanese patients. We analyzed clinical and genetic data of 80 patients based on the results of a nationwide questionnaire survey and review of the literature. From clinical findings of 80 Japanese patients, high-grade fever was observed in 98.8%, chest attacks (pleuritis symptoms) in 61.2%, abdominal attacks (peritonitis symptoms) in 55.0%, and arthritis in 27.5%. Twenty-four percent of patients experienced their first attacks before 10 years of age, 40% in their teens, and 36% after age 20 years. Colchicine was effective in many patients at a relatively low dose (< 1.0 mg/day). AA amyloidosis was seen in only 1 patient. Common MEFV mutation patterns were E148Q/M694I (25.0%), M694I alone (17.5%), and L110P/E148Q/M694I (17.5%), and no patient carried the M694V mutation, the most common mutation in Mediterranean patients with FMF. A larger than expected number of patients with FMF exist in Japan, and the clinical presentation of Japanese FMF patients seems to be relatively milder than those of Mediterranean FMF patients. AA amyloidosis rarely occurs in Japanese patients, probably due to difference in patterns of the MEFV genotype between Japanese and Mediterranean patients.


Methamphetamine Exposure Combined with HIV-1 Disease or gp120 Expression: Comparison of Learning and Executive Functions in Humans and Mice.
Methamphetamine dependence is a common comorbid condition among people living with HIV, and may exacerbate HIV-associated neurocognitive disorders. Animal models of neuroAIDS suggest that the gp120 protein may also cause cognitive impairment. The present work evaluated the separate and combined effects of HIV/gp120 and methamphetamine on learning and executive functions in both humans and transgenic mice. Human participants were grouped by HIV serostatus (HIV+ or HIV-) and lifetime methamphetamine dependence (METH+ or METH-). A neurocognitive test battery included domain-specific assessments of learning and executive functions. Mice (gp120+ and gp120-) were exposed to either a methamphetamine binge (METH+) or saline (METH-), then tested in the attentional-set-shifting task to assess learning and executive functions. In humans, HIV status was associated with significant impairments in learning, but less so for executive functions. The frequency of learning impairments varied between groups, with the greatest impairment observed in the HIV+/METH+ group. In mice, gp120 expression was associated with impairments in learning but not reversal learning (executive component). The greatest proportion of mice that failed to complete the task was observed in the gp120+/METH+ group, suggesting greater learning impairments. Our cross-species study demonstrated that HIV in humans and gp120 in mice impaired learning, and that a history of methamphetamine exposure increased the susceptibility to HIV-associated neurocognitive deficits in both species. Finally, the similar pattern of results in both species suggest that the gp120 protein may contribute to HIV-associated learning deficits in humans.


Some aspects of the nursing care for patients on hemodialysis.
Currently the majority of patients receiving hemodialysis for chronic renal failure are treated as outpatients in hemodialysis units of large hospitals. In most of these units it is the nurses that handle the entire dialysis procedure with little if any direct supervision from physicians. The nurses are also the ones on the health team that have the most contact with the patients. Thus the nurse must have a thorough knowledge of the pathophysiology of renal failure, of the mechanics and technical aspects of the dialyzer, of the expected outcomes and complications of hemodialysis and, particularly, the needs of the hemodialysis patient. The nurses that choose to work with chronic hemodialysis patients can enjoy much satisfaction in assisting in maintaining both a productive life and a long-term relationship with chronic renal failure patients. The nurse makes the difference.


Fully endoscopic transnasal approach to the jugular foramen: anatomic study and clinical considerations.
To describe a transnasal endoscopic route to the jugular foramen and the endoscopic anatomy of the infratemporal fossa. Endoscopic transnasal dissection of the infratemporal fossa was performed in 3 injected fresh heads (1 head only in arteries and 2 heads in arteries and veins). Two other double-injected specimens were dissected externally (2 of them side laterally and 1 anteriorly) to compare the different views and better understand the 3-dimensionality of the region.Detailed endoscopic anatomy of the infratemporal fossa was clearly observed. The realization of a septal and posterior maxillary window allows surgeons to gain space to the jugular foramen. The ability to manage the vessels, especially the veins, and identify the muscles is mandatory. The fundamental role of the vidian canal in targeting the anterior genu of the internal carotid artery is confirmed. The role of the maxillary and mandibular branches of the trigeminal nerve and the eustachian tube in this kind of approach is critical. A fully transnasal endoscopic route to the jugular foramen is feasible. The most important landmark for this kind of approach is the eustachian tube.


[Atheroembolism and cardiac surgery: importance of preoperative management].
Atheroembolization is a recognized complication of cardiac surgical procedures, and has been implicated in postoperative stroke, renal failure, multiorgan failure, and death. Preoperative identification of patients at risk for developing atheroemboli is essential. The aim of this study was to determine preoperative risk factors for atheroemboli and to assess the postoperative course of the patients who developed atheroembolic syndrome. A retrospective record review was conducted. From 1/1990 to 12/1994, 5.486 patients underwent coronary artery bypass grafting (CABG), valve operations, or other cardiac surgical procedures at the Massachusetts General Hospital. Among this population, 107 patients (1.9%) developed atheroembolic syndrome. There were compared to a concurrent population of 925 patients operated during the same period. Patients who developed atheroemboli were older, with an increased incidence (p < 0.01) of hypertension, cerebrovascular disease, and aortoiliac disease. Many had a complicated course after catheterization, with renal insufficiency (35%) and evidence of peripheral emboli (12%). Average Intensive Care Unit stay, hospital stay, and hospital cost of these patients were 16.8 days, 48.4 days and $88,000 respectively, compared to 1.5 days, 9.6 days and $23,000 for the concurrent population. Of these 107 patients only two were discharged home, the others either died (48 patients, or 25% of all cardiac surgical deaths during this period), or went to rehabilitation or chronic hospital facilities. Twenty-seven autopsies were performed and invariably showed a diffusely diseased aorta, with calcification, mural thrombus, and ulceration. Atheroembolization during cardiac surgical procedures has profound medical and economic consequences. Because of the diffuse nature of aortic disease, measures approaching the disease as a local process are likely to be unsuccessful. Appropriate evaluation would ideally identify patients with extensive aortic atheromatous disease, prior to rather than during surgery.


A comparison of dyadic interactions and coping with still-face in healthy pre-term and full-term infants.
Pre-term birth has a significant impact on infants' social and emotional competence, however, little is known about regulatory processes in pre-term mother-infant dyads during normal or stressful interactions. The primary goals of this study were to investigate the differences in infant and caregiver interactive behaviour and dyadic coordination of clinically healthy pre-term compared to full-term infant-mother dyads and to examine pre-term infants' capacity for coping with stress using the face-to-face still-face paradigm (FFSF). Fifty mother-infant dyads, including 25 pre-term infants and 25 full-term infants were videotaped during the FFSF. All infants were 6-9 months of age (corrected for gestational age in the pre-term group). Infant and maternal socio-emotional expressivity and self-regulatory behaviours were coded and measures of dyadic coordination (Matching, Reparation Rate, and Synchrony) were calculated. There were no significant differences in infant and caregiver socio-emotional behaviours between the two groups and both groups demonstrated the still-face (SF) effect and the reunion effect. There was a difference in self-regulatory behaviour. Pre-term infants were more likely than full-term infants to use distancing (e.g., by turning away, twisting, or arching) from their mothers during the FFSF. Additionally, during the Reunion episode of the FFSF pre-term infants showed more social monitoring compared to full-term infants. Regardless of the birth status, the dyads showed less coordination and a slower rate of reparation during the Reunion episode than during the Play episode. The higher proportion of distancing in the pre-term group and the increase in social monitoring suggest that even in normal interactions pre-term infants may experience a higher level of stress and have less capacity for self-regulation compared to the full-terms and that pre-term infants appear to use a compensatory strategy of increased social monitoring to cope with the stress of renegotiating the interaction during Reunion. The findings suggest that pre-term infants have different regulatory and interactive capacities than full-term infants.


Identification of rice genes induced in a rice blast-resistant mutant.
To clarify mechanisms of rice blast resistance in rice plants we used suppression subtractive hybridization (SSH) to isolate genes induced upon rice blast inoculation in a rice blast-resistant mutant. A total of 26 rice cDNAs were isolated and found to have elevated expression upon rice blast infection in a rice blast-resistant derivative, SHM-11, of the rice cultivar, Sanghaehyanghyella. Sequencing of the cDNAs revealed that many of the proteins they encoded had been previously described as involved in plant responses against pathogen attack. Two interesting groups of the defense-related proteins consisted of three different PR5 homologues and four different protease inhibitors, all highly expressed in the rice blast mutant. Genes encoding proteins involved in signal transduction and regulation were also identified, including translation initiation factor eIF5A, C2 domain DNA binding protein, putative rice EDS and putative receptor like kinase. Most of the identified cDNAs were highly expressed 24 h after blast inoculation. Our results suggest that a pathway regulating defense gene expression may be altered in the mutant, resulting in early induction of the defense genes upon fungal infection.


Genetic admixture supports an ancient hybrid origin of the endangered Hawaiian duck.
Speciation is regarded primarily as a bifurcation from an ancestral species into two distinct taxonomic units, but gene flow can create complex signals of phylogenetic relationships, especially among different loci. We evaluated several hypotheses that could account for phylogenetic discord between mitochondrial DNA (mtDNA) and nuclear DNA (nuDNA) within Hawaiian duck (Anas wyvilliana), including stochastic lineage sorting, mtDNA capture and widespread genomic introgression. Our results best support the hypothesis that the contemporary Hawaiian duck is descended from an ancient hybridization event between the mallard (Anas platyrhynchos) and Laysan duck (Anas laysanensis). Whereas mtDNA clearly shows a sister relationship between Hawaiian duck and mallard, nuDNA is consistent with a genetic mosaic with nearly equal contributions from Laysan duck and mallard. In addition, coalescent analyses suggest that gene flow from either mallard or Laysan duck, depending on the predefined tree topology, is necessary to explain contemporary genetic diversity in Hawaiian ducks, and these estimates are more consistent with ancient, rather than contemporary, hybridization. Time since divergence estimates suggest that the genetic admixture event occurred around the Pleistocene-Holocene boundary, which is further supported by circumstantial evidence from the Hawaiian subfossil record. Although the extent of reproductive isolation from either putative parental taxon is not currently known, these species are phenotypically, genetically and ecologically different, and they meet primary criteria used in avian taxonomy for species designation. Thus, the available data are consistent with an admixed origin and support the hypothesis that the Hawaiian duck may represent a young hybrid species.


Method of bacterial killing differentially affects the human innate immune response to Staphylococcus epidermidis.
In vitro investigations of human innate immune responses to extracellular bacteria commonly utilise killed preparations in preference to live organisms. The effects of the bacterial preparation method on the activation of innate signalling pathways by the common opportunistic pathogen Staphylococcus epidermidis (SE) are unknown. Mononuclear cell cytokine expression patterns induced by live (LSE), heat-killed (HKSE) and ethanol-killed SE (EKSE) were characterized at the transcriptional and translational level. Toll-like receptor (TLR)-activating capacity of the preparations was analysed using TLR-transfected human embryonic kidney cells. Live SE activated NF-κB, STAT1, type I interferon, and inflammasome pathways. Killed preparations engaged the NF-κB pathway, but had significantly lower capacity to activate other innate immune pathways. Killing of extracellular bacteria has significant qualitative and quantitative effects on key aspects of innate responses in vitro. Interpretation of in vitro data and extrapolation of findings should take into account the potential effects of bacterial preparation and should not assume that responses to killed bacteria are predictive of responses to live organisms.


Immunosuppressive effect of mesenchymal stem cells favors tumor growth in allogeneic animals.
Mesenchymal stem cells (MSCs) are largely studied for their potential clinical use. Recently, they have gained further interest after demonstration of an immunosuppressive role. In this study, we investigated whether in vivo injection of MSCs could display side effects related to systemic immunosuppression favoring tumor growth. We first showed in vitro that the murine C3H10T1/2 (C3) MSC line and primary MSCs exhibit immunosuppressive properties in mixed lymphocyte reaction. We demonstrated that this effect is mediated by soluble factors, secreted only on "activation" of MSCs in the presence of splenocytes. Moreover, the immunosuppression is mediated by CD8+ regulatory cells responsible for the inhibition of allogeneic lymphocyte proliferation. We then demonstrated that the C3 MSCs expressing the human bone morphogenetic protein 2 (hBMP-2) differentiation factor were not rejected when implanted in various allogeneic immunocompetent mice and were still able to differentiate into bone. Importantly, using a murine melanoma tumor model, we showed that the subcutaneous injection of B16 melanoma cells led to tumor growth in allogeneic recipients only when MSCs were coinjected. Although the potential side effects of immunosuppression induced by MSCs have to be considered in further clinical studies, the usefulness of MSCs for various therapeutic applications still remains of great interest.


Relationship between plasma level of vitamin D and post operative atrial fibrillation in patients undergoing CABG.
Atrial fibrillation is the most common arrhythmia after cardiac surgery. Several studies have shown the impact of vitamin D on heart disease; however, there have been few studies for the incidence of AF and its relationship with vitamin D levels. According to the different results of these studies, we decided to evaluate the relation of plasma levels of vitamin D and postoperative atrial fibrillation in patients undergoing coronary artery bypass surgery (CABG). This cross-sectional study was performed on 50 patients after CABG surgery. Simple random sampling was done. Twenty five patients who developed AF within 48 hours after CABG with Cardiopulmonary bypass (CPB) were enrolled in the case group and 25 patients who did not develop AF within 48 hours after CABG with CPB were enrolled in the control group. Plasma levels of vitamin D in both groups of patients were recorded. Collected data were analyzed by the SPSS software version 17. There was no significant difference in terms of demographic characteristics, comorbidities, lipid profile and kidney function between two groups. The mean plasma level of vitamin D was 27.4 ± 2.22 ng/ml in the case group and was 28.2 ± 1.18 ng/ml in the control group it (p= 0.803). Plasma levels of vitamin D were almost the same in both groups and there was no statistically significant difference between the groups with and without atrial fibrillation following CABG.


First in human randomised trial of J2H-1702: A novel 11β-hydroxysteroid dehydrogenase type 1 inhibitor for non-alcoholic steatohepatitis treatment.
11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), which is an enzyme that converts cortisone to cortisol, plays a role in the regulation of glucose metabolism and inflammation. J2H-1702 is a novel 11β-HSD1 inhibitor, and the inhibition of 11β-HSD1 has been shown to improve insulin sensitivity, reduce inflammation, and prevent the development of nonalcoholic steatohepatitis (NASH) in preclinical models. We aimed to assess the pharmacokinetics (PKs), pharmacodynamics (PDs), safety, and tolerability of J2H-1702 after a single-dose oral administration. A randomised, double-blinded, placebo-controlled, single-dose, dose-escalation study was conducted on 50 healthy volunteers. Blood and urine samples were collected to assess the PK and PD of J2H-1702. The peak plasma concentration of J2H-1702 was observed at 2-2.9 h after a single-dose oral administration. J2H-1702 reduced 11β-HSD1 activity compared to the placebo at all dose levels. The drug reached its maximal inhibitory effect within 12-24 h post-dose administration, and the inhibitory effect was maintained till 1 day after administration of the study drug. The drug showed typical first-order elimination kinetics, with a mean elimination half-life of 9.8-14.7 h. Systemic exposure to J2H-1702 increased in a dose-dependent manner. J2H-1702 was well tolerated after a single oral administration of up to 300 mg. A total of 11 treatment-emergent adverse events (TEAEs) occurred in seven (14%) participants, all of which were mild and resolved spontaneously. The most common TEAE was diarrhoea (8%), followed by dizziness (4%). The results of this study suggest that J2H-1702 could be developed as an effective therapeutic option for NASH.


CIM Journal Club: Gene therapy for spinal muscular atrophy Comment on Mendell et al. N Engl J Med 2017;377:1713-22.
In their landmark paper, Mendell et al. show that infants with spinal muscular atrophy (SMA) reached important motor milestones and survived longer when treated with AVXS-101 (AveXis), a viral vector containing DNA encoding the survival of motor neuron protein (SMN). Patients not only crawled, stood and walked independently, but learned to speak. These results are very encouraging for patients with SMA and offer hope for pediatric and adult patients with other types of motor neuron diseases.


Standard of care for cancer-related anemia: improving hemoglobin levels and quality of life.
The introduction of recombinant human erythropoietin (rHuEPO) has proven to be a major advance in the therapeutic options available for managing anemia in cancer patients. The results of placebo-controlled clinical trials and large, community-based, open-label studies have confirmed that epoetin alfa, a recombinant human erythropoietin, significantly reduces transfusion requirements, and reliably increases hemoglobin (Hb) levels in anemic (Hb level <12 g/dl) cancer patients undergoing chemotherapy. Increased Hb improves patients' energy level and their ability to perform the activities of daily living, as well as their overall quality of life (QOL). These findings are independent of tumor type and disease status and are comparable in patients receiving nonplatinum- and platinum-based chemotherapeutic regimens. Furthermore, more than a decade of use in clinical trials and by physicians in routine clinical practice has demonstrated that epoetin alfa is safe and well tolerated when used to treat cancer patients with anemia. The availability of epoetin alfa as an alternative to transfusion has changed practices in anemia management; physicians can now treat anemia with the goal of achieving adequate Hb levels to relieve anemia-related fatigue, a major symptom contributing to decreased QOL in cancer patients. Incremental benefit analysis has shown that increasing Hb level from 11 g/dl to 12 g/dl yields the greatest improvement in QOL per 1 g/dl increase in Hb. The demonstrated efficacy of epoetin alfa for increasing Hb levels and improving patient QOL have made this agent a rationale choice for management of cancer-related anemia. Ongoing research will continue to provide new insights into best management of anemia with epoetin alfa in cancer patients.


A median redshift of 2.4 for galaxies bright at submillimetre wavelengths.
A significant fraction of the energy emitted in the early Universe came from very luminous galaxies that are largely hidden at optical wavelengths (because of interstellar dust grains); this energy now forms part of the cosmic background radiation at wavelengths near 1 mm (ref. 1). Some submillimetre (submm) galaxies have been resolved from the background radiation, but they have been difficult to study because of instrumental limitations. This has impeded the determination of their redshifts (z), which is a crucial element in understanding their nature and evolution. Here we report spectroscopic redshifts for ten submm galaxies that were identified using high-resolution radio observations. The median redshift for our sample is 2.4, with a quartile range of 1.9-2.8. This population therefore coexists with the peak activity of quasars, suggesting a close relationship between the growth of massive black holes and luminous dusty galaxies. The space density of submm galaxies at redshifts over 2 is about 1,000 times greater than that of similarly luminous galaxies in the present-day Universe, so they represent an important component of star formation at high redshifts.


Sensitization of trigeminal brainstem pathways in a model for tear deficient dry eye.
Chronic dry eye disease (DE) is associated with an unstable tear film and symptoms of ocular discomfort. The characteristics of symptoms suggest a key role for central neural processing; however, little is known about central neuroplasticity and DE. We used a model for tear deficient DE and assessed effects on eye blink behavior, orbicularis oculi muscle activity (OOemg), and trigeminal brainstem neural activity in male rats. Ocular-responsive neurons were recorded at the interpolaris/caudalis transition (Vi/Vc) and Vc/upper cervical cord (Vc/C1) regions under isoflurane, whereas OOemg activity was recorded under urethane. Spontaneous tear volume was reduced by ∼50% at 14 days after exorbital gland removal. Hypertonic saline-evoked eye blink behavior in awake rats was enhanced throughout the 14 days after surgery. Saline-evoked neural activity at the Vi/Vc transition and in superficial and deep laminae at the Vc/C1 region was greatly enhanced in DE rats. Neurons from DE rats classified as wide dynamic range displayed enlarged convergent periorbital receptive fields consistent with central sensitization. Saline-evoked OOemg activity was markedly enhanced in DE rats compared with controls. Synaptic blockade at the Vi/Vc transition or the Vc/C1 region greatly reduced hypertonic saline-evoked OOemg activity in DE and sham rats. These results indicated that persistent tear deficiency caused sensitization of ocular-responsive neurons at multiple regions of the caudal trigeminal brainstem and enhanced OOemg activity. Central sensitization of ocular-related brainstem circuits is a significant factor in DE and likely contributes to the apparent weak correlation between peripheral signs of tear dysfunction and symptoms of irritation.


Technical improvement of human pancreatic islet isolation.
A key factor for successful islet isolation is to place the optimal amount of enzyme into the pancreatic ducts prior to starting digestion of pancreatic glands. To improve this procedure, we introduced novel techniques to identify and repair tissue damage resulting in leakage of collagenase solution. One hundred twelve standardized consecutive islet isolations were for the effects of dye and glue on islet yield, islet function using a perifusion assay, and the possibility of clinical transplantation. One group of pancreata (n = 26) obtained en bloc together with duodenum were carefully detached with ligation of accessory ducts in an isolation unit (WPD group), whereas the pancreata were dissected from the duodenum in the operating room in the other 86 isolations. In 28 of 86 isolations, whole glands were used (WP group), while only the body and tail area were applied in the remaining 58 isolations (PP group). Both dye and glue effectively prevented leakage of collagenase from the gland. Both islet yield and success rate were higher with these tools without adverse effects on islet function or collagenase activity. The success rate of isolations and islet yield were significantly higher in the WPD group (P = .02 and .001, respectively). Dye and glue may be useful tools to improve human islet isolation procedures. In addition, the use of the whole pancreas further improves the outcome.


Column chromatography analysis of brain tissue: an advanced laboratory exercise for neuroscience majors.
Neurochemical analysis of discrete brain structures in experimental animals provides important information on synthesis, release, and metabolism changes following behavioral or pharmacological experimental manipulations. Quantitation of neurotransmitters and their metabolites following unilateral drug injections can be carried out using standard chromatographic equipment typically found in most undergraduate analytical laboratories. This article describes an experiment done in a six session (four hours each) component of a neuroscience research methods course. The experiment provides advanced neuroscience students experience in brain structure dissection, sample preparation, and quantitation of catecholamines using high performance liquid chromatography (HPLC) and protein analysis using ultraviolet spectroscopic methods. The students are exposed to useful laboratory techniques such as standard solution preparation and calibration curve construction, centrifugation, quantitative pipetting, and data evaluation and graphical presentation. Typically, only students that participate in independent neuroscience research are familiar with these types of quantitative skills. The usefulness of this type of experimental design for understanding behavioral or pharmacological effects on neurotransmitter systems is emphasized through a final report requiring a comprehensive literature search.


Growth of the two layers of the chick sclera is modulated reciprocally by visual conditions.
Although visual deprivation causes increased ocular elongation and myopia in both birds and mammals, changes in sclera appear to be in opposite directions. Because avian sclera has a cartilaginous layer as well as the fibrous layer found in mammals, we examined whether the scleral responses to various visual manipulations differ between the two layers. To produce increases in ocular elongation and myopia, monocular diffusers or negative lenses were fitted to eyes. Conversely, to produce decreases in ocular elongation, diffusers were removed (restoring normal vision) or monocular positive lenses were fitted. Scleral layers were then dissected apart, and incorporation of labeled precursors into glycosaminoglycans (GAGs), DNA, and protein was assessed. Tissue coculture experiments were used to assess humoral interactions between scleral layers and with the choroid. In the cartilaginous layers, the incorporation of label into proteoglycans and DNA was significantly higher in eyes elongating faster than normal because of wearing diffusers or negative lenses and significantly lower than normal in eyes elongating slower than normal because of removal of the diffuser or wearing positive lenses. In the fibrous layers, the reverse was the case. Coculturing cartilaginous sclera from normal eyes with fibrous sclera from myopic or recovering eyes produced the same increase or decrease in sulfate incorporation into GAGs in the cartilaginous layer as though the tissue measured was from the animal providing the conditioning tissue. Coculturing with choroid, especially from recovering eyes, also inhibited cartilaginous sclera. The fibrous layer of the avian sclera shows changes in sulfate incorporation into GAGs during deprivation and recovery from deprivation in the same direction as does the mammalian sclera, whereas the cartilaginous layer changes in the opposite direction. The responses of the cartilaginous layer may be controlled by the fibrous layer, although they are influenced by the choroid as well.


Measles viruses with altered envelope protein cytoplasmic tails gain cell fusion competence.
The cytoplasmic tail of the measles virus (MV) fusion (F) protein is often altered in viruses which spread through the brain of patients suffering from subacute sclerosing panencephalitis (SSPE). We transferred the coding regions of F tails from SSPE viruses in an MV genomic cDNA. Similarly, we constructed and transferred mutated tail-encoding regions of the other viral glycoprotein hemagglutinin (H) gene. From the mutated genomic cDNAs, we achieved rescue of viruses that harbor different alterations of the F tail, deletions in the membrane-distal half of the H tail, and combinations of these mutations. Viruses with alterations in any of the tails spread rapidly through the monolayer via enhanced cell-cell fusion. Double-tail mutants had even higher fusion competence but slightly decreased infectivity. Analysis of the protein composition of released mutant viral particles indicated that the tails are necessary for accurate virus envelope assembly and suggested a direct F tail-matrix (M) protein interaction. Since even tail-altered glycoproteins colocalized with M protein in intracellular patches, additional interactions may exist. We conclude that in MV infections, including SSPE, the glycoprotein tails are involved not only in virus envelope assembly but also in the control of virus-induced cell fusion.


Interrelationships of ion transport in rat submaxillary duct epithelium.
The transport of Na+, K+, Cl-, and HCO3(-) across the epithelium of the rat submaxillary salivary duct is postulated to be due to the coupling of the basolateral Na+-K+-ATPase with various ion transport systems in the luminal and basolateral membranes. Na+ reabsorption depends on the presence of a rheogenic (Na+ conductance) and an electroneutral (Na+:H exchange) pathway, both of which are sensitive to amiloride. K+ secretion is postulated to be mediated by a K+: H+ antiport, coupling between Na+ reabsorption and K+ secretion, thus depending on local H+ ion concentration. The ratio between electroneutral Na+ influx and K+ efflux, therefore, determines the rate of HCO3(-) secretion. In the absence of Na+ influx, although K+ efflux falls, HCO3(-) secretion rises to a value equal to that of K+ secretion. The maintenance of K+ secretion in the absence of luminal Na+ requires an additional Na+-entry step across the basolateral membrane, also postulated to be due to Na+:H+ exchange.


Acute stroke intervention.
Ischemic strokes will make up most (>80%) of the three-quarters of a million strokes that will occur in Americans this year. Reperfusion therapy is the fundamental strategy for the treatment of acute ischemic stroke. Reperfusion therapy may be accomplished noninvasively (intravenous thrombolysis) or invasively with catheter-based treatments (intra-arterial thrombolysis, thrombectomy, or angioplasty). Currently, a large majority of patients with acute ischemic stroke do not receive any form of reperfusion therapy owing to their delayed presentation (>3 hours) and lack of skilled man power for on-demand endovascular treatment. Paradoxically, improved success rates for reperfusion have been reported with the newer thrombectomy catheters, called "stentreivers." An option for broadening access for patients who need endovascular therapy would be to use interventional cardiologists with carotid stent experience who can help to provide 24 × 7 × 365 coverage.


Using cyclosporine Neoral immediately after liver transplantation. United Kingdom Neoral Pilot Study Group.
Cyclosporine Neoral was used as an alternative to intravenous Sandimmun in an open-label pilot study performed at five British liver transplant centres. Twenty primary orthotopic liver graft recipients received Neoral (5 mg/kg body weight bid via a nasogastric tube) together with azathioprine and glucocorticoids from the day of transplantation. None of the twenty patients required supplemental intravenous cyclosporine during follow-up (minimum 44 days). Cyclosporine blood levels (analyzed at a central laboratory using a specific fluorescence polarisation immunoassay) reached a mean trough value of 205 micrograms/L after the second Neoral dose. During the first postoperative week there was no significant difference between trough cyclosporine levels in patients with duct-to-duct (n = 14) or Roux-en-Y (n = 5) biliary drainage. Blood profiles showed a progressive increase in the rate and extent of cyclosporine absorption from days 1 to 3 to 5 (mean tmax 5.4, 2.7, and 2.3 h, respectively; mean area under the curve 1092, 1716, and 1758 micrograms/L.h per 100-mg dose, respectively). Acute rejection episodes occurred in 37% of cases within the first postoperative month and the early incidence of nephrotoxicity (16%) and neurotoxicity (37%) was unexceptional. Our preliminary data demonstrate that the 20 liver transplant recipients in this pilot study were safely and effectively managed without intravenous cyclosporine by administering Neoral from the day of transplantation.


Shear stress induces preimplantation embryo death that is delayed by the zona pellucida and associated with stress-activated protein kinase-mediated apoptosis.
In this study, we discovered that embryos sense shear stress and sought to characterize the kinetics and the enzymatic mechanisms underlying induction of embryonic lethality by shear stress. Using a rotating wall vessel programmed to produce 1.2 dynes/cm2 shear stress, it was found that shear stress caused lethality within 12 h for E3.5 blastocysts. Embryos developed an approximate 100% increase in mitogen-activated protein kinase 8/9 (formerly known as stress-activated protein kinase/junC kinase 1/2) phosphorylation by 6 h of shear stress that further increased to approximately 350% by 12 h. Terminal deoxynucleotidyltransferase dUTP nick end labeling/apoptosis was at baseline levels at 6 h and increased to approximately 500% of baseline at 12 h, when irreversible commitment to death occurred. A mitogen-activated protein kinase 8/9 phosphorylation inhibitor, D-JNKI1, was able to inhibit over 50% of the apoptosis, suggesting a causal role for mitogen-activated protein kinase 8/9 phosphorylation in the shear stress-induced lethality. The E2.5 (compacted eight-cell/early morula stage) embryo was more sensitive to shear stress than the E3.5 (early blastocyst stage) embryo. Additionally, zona pellucida removal significantly accelerated shear stress-induced lethality while having no lethal effect on embryos in the static control. In conclusion, preimplantation embryos sense shear stress, chronic shear stress is lethal, and the zona pellucida lessens the lethal and sublethal effects of shear stress. Embryos in vivo would not experience as high a sustained velocity or shear stress as induced experimentally here. Lower shear stresses might induce sufficient mitogen-activated protein kinase 8/9 phosphorylation that would slow growth or cause premature differentiation if the zona pellucida were not intact.


Comorbidity of Physical Disorders Among Patients With Severe Mental Illness With and Without Substance Use Disorders: A Systematic Review and Meta-Analysis.
Objective: Physical disorders in patients with severe mental illness (SMI) are common and they tend to be underdiagnosed by clinicians, which might lead to negative treatment outcomes. The presence of substance use disorders could further aggravate the situation. There are existing systematic reviews on physical disorders among individuals with SMI in general but none of these previous reviews stratified their findings by substance use disorder status. This study aimed to synthesize the evidence on the frequency of comorbid physical disorders among patients with SMI with or without substance use disorders. Methods: We searched for studies published in English between 1988 and 2017 in MEDLINE, Embase, CINAHL, PsycINFO, Global Health, Web of Science, Scopus, WHO Global Health Library (Global Index Medicus), Google Scholar, OpenGrey, the Grey Literature Report, Cochrane Library, International Standardized Randomized Controlled Trial Number Registry, WHO International Clinical Trials Registry Platform, ClinicalTrials.gov, Australian and New Zealand Clinical Trials Registry, and PROSPERO. There was no geographical restriction and the target population was adults (≥18 years) with diagnosed SMI including schizophrenia, bipolar disorder, and other psychotic illnesses. The outcome of interest was physical disorder. Results: A total of 6,994 records were retrieved. Only 30 papers (representing 24 studies) met our inclusion criteria and 13 studies were included in the meta-analysis. The prevalence of most of the reported physical disorders was higher in SMI patients with substance use disorders than in those without substance use disorders. When ranked according to pooled prevalence level, hypertension (35.6%), tardive dyskinesia (35.4%), and hepatitis C (26.9%) were the most prevalent physical disorders among SMI patients with substance use disorders. For SMI patients without substance use disorders, hypertension (32.5%), tardive dyskinesia (25.1%), and endocrine disease (19.0%) were more common. Estimates for diabetes (7.5% vs. 7.5%) and cardiovascular diseases (11.8% vs. 11.3%) were similar across groups. Conclusions: Physical disorders among SMI patients vary by substance use disorder status. Clinicians managing SMI in patients should screen for physical disorders and substance use disorders and provide treatment or referral. Registration: International Prospective Register of Systematic Reviews (PROSPERO) registration number CRD42017072286.


Clinical utility of the 70-gene MammaPrint profile in a Japanese population.
van't Veer and colleagues developed a 70-gene prognosis profile known as MammaPrint to identify breast cancer patients who were at low risk of developing metastases. We evaluated the prognostic value of the 70-gene MammaPrint profile in Japanese women with node-negative breast cancer. Frozen tumour samples from 102 eligible node-negative breast cancer patients aged 70 or younger were characterized with the MammaPrint array. The patients were treated with breast-conserving therapy or mastectomy with axillary lymph node dissection between December 1998 and August 2001. About 73 percent received adjuvant hormonal therapy and 28 percent received adjuvant chemotherapy. The gene expression profiles obtained by MammaPrint classified the patients as high- or low-genomic risk. The median follow-up was 7.1 years. Among the 102 patients, 20 (20%) were classified as low-genomic risk and 82 (80%) were classified as high-genomic risk. The probability of distant metastasis-free survival at five years was 100% for the low-risk group and 94% for the high-risk group. The 70-gene MammaPrint prognosis profile accurately identified Japanese breast cancer patients at low risk of developing recurrences. In fact, 100% of the individuals in the low-risk category remained metastasis-free for the duration of the observation period.


Purification of a 100 kDa phospholipase A2 from spleen, lung and kidney: antiserum raised to pig spleen phospholipase A2 recognizes a similar form in bovine lung, kidney and platelets, and immunoprecipitates phospholipase A2 activity.
Phospholipase A2 (PLA2) plays a key role in the production of intracellular and extracellular chemical mediators such as arachidonic acid, eicosanoids and platelet-activating factor, which modulate membrane channel activity, signal transduction, are vasoactive and chemotactic, and are implicated in many pathophysiological mechanisms of inflammation and tissue injury. We previously identified, purified and characterized an arachidonic acid-selective cytosolic 100-110 kDa PLA2 from bovine platelets and rat kidney that is activated during cell stimulation. The purification schemes previously published resulted in low yields of enzyme, insufficient for extensive biochemical characterization. We report the purification of a large-molecular-mass (100 kDa) PLA2 from pig spleen, bovine kidney and bovine lung, using a novel large-scale purification scheme. The enzyme was purified to near homogeneity from an acidified extract obtained from 4.8 kg of pig spleen by sequential use of DEAE-cellulose anionic exchange, Butyl-Toyopearl hydrophobic chromatography and DEAE-5PW h.p.l.c., and further purified by non-denaturing PAGE. This purification scheme will permit the preparation of quantities of purified native enzyme sufficient to study its properties and regulation. To generate antiserum against the PLA2 enzyme, the 100 kDa protein was excised and electroeluted from SDS/PAGE gels of the active fractions after DEAE-5PW h.p.l.c., and this was used as antigen. This polyclonal antibody against pig spleen 100 kDa PLA2 protein reacted with 100 kDa bands in preparations partially purified from bovine platelets, kidney and lung as well as pig spleen, and immunoprecipitated PLA2 activity from these sources. The antibody also immunoprecipitated a 100 kDa protein from cytosolic fractions of cultured renal mesangial cells, human erythroleukaemia cells and human monocytic U937 cells. Considerable PLA2 activity was present in the immunoprecipitates. To our knowledge this antibody is unique in its ability to permit measurement of PLA2 activity in the immunoprecipitate itself, and will be a useful tool for the study of the regulation and the activation mechanisms of the native PLA2 enzyme.


Dissimilation of tryptophan and related indolic compounds by ruminal microorganisms in vitro.
Intraruminal doses of L-tryptophan cause acute pulmonary edema and emphysema in cattle. The D and L isomers of tryptophan and 22 related indolic compounds were incubated with ruminal microorganisms in vitro. Incubation of L-[U-benzene ring-(14)C]tryptophan with ruminal microorganisms for 24 h resulted in 39% of the added radioactivity being incorporated into skatole, 7% into indole, and 4% into indoleacetate (IAA). D-Tryptophan was not degraded to any of these metabolites. The major pathway of skatole formation from L-tryptophan appeared to be by the decarboxylation of IAA. Incubation of [2-(14)C]IAA with ruminal microorganisms for 24 h resulted in 38% incorporation into skatole. L-[5-Hydroxy]tryptophan was degraded to 5-hydroxyskatole and 5-hydroxyindole, whereas 5-hydroxyindoleacetate was degraded to only 5-hydroxyskatole. Incubation of indolepyruvate, indolelactate, and indolealdehyde with ruminal microorganisms resulted in the formation of both skatole and indole. Under similar conditions, indoleacetaldehyde was converted to IAA and tryptophol. The addition of increasing concentrations of glucose (0 to 110 mM) reduced the formation of both skatole and indole from L-tryptophan and resulted in the accumulation of IAA. Antibiotics reduced the degradation of L-tryptophan to skatole and indole, with kanamycin and neomycin particularly effective in reducing the decarboxylation of IAA to skatole.


Using reflection to influence practice: student perceptions of daily reflection in clinical education.
Reflection is a key element in learning from experience, but the impact of most programmes of reflection on daily practice remains unclear. We investigated students' perceptions of adding a daily written reflection assignment to a clinical rotation. Third-year medical students on a single two-week rotation completed daily reflections analyzing their performance. Programme evaluation used a 33-question anonymized survey. Quantitative data were summarized and qualitative responses coded for recurring themes. Twenty-six students completed the survey (90 % response rate). Eighty-five percent of students felt that the daily reflections had a positive impact on their learning from clinical experience. Seventy-seven percent of students reported that the programme changed their awareness of their thoughts and actions, and 80 % felt that it improved their recall of experiences. A greater sense of mindfulness and focus on self-improvement were major themes that emerge from students' descriptions of the role of daily reflections in their learning. Overall, daily reflections demonstrated a positive learning influence. This exploratory study suggests students may benefit from more frequent, short reflections as opposed to more typically spaced reflective assignments.


Reasons Not to Turf a Patient Whose "Belonging" in a Hospital Is Unclear.
Through the lens of metaphor and the arts, this article aims to illuminate how persons who are ill tarry through uncertainty to receive care, and, in response, clinicians must resist turfing such patients in a health system that often confers upon patients unclear criteria for belonging. In addition, this article considers relationships among clinicians, patients, and their loved ones through the perspectives of Maris and Ludlow, characters in the book, A Hospital Odyssey, by Gwyneth Lewis. The article suggests that engaged curiosity and empathy are helpful responses to clinical detachment, distraction, and disengagement.


Prevalence of age-related macular degeneration in the Republic of Ireland.
Age-related macular degeneration (AMD) remains the most common cause of visual loss among subjects over 50 years of age in the developed world. The Irish Longitudinal study on Ageing (TILDA) is a population-based study of subjects aged 50 years or older, designed to investigate factors that influence ageing, and has enabled this investigation of the prevalence of AMD in the Republic of Ireland (ROI). Data collected from a nationally representative sample of community-living older adults aged 50 years and over in ROI over the period November 2009 to July 2011. 5035 participants attended the TILDA health centre for assessment. Retinal photographs were obtained in 4859 of these participants. Retinal grading was performed in a masked fashion using a modified version of the International Classification and Grading System for AMD. Adjusting for lower response rates among older subjects, the estimated overall prevalence of any AMD was 7.2% (95% CI 6.5% to 7.9%) in the population aged 50 years or older. The estimated prevalence of early AMD was 6.6% (95% CI 5.9% to 7.3%), and the estimated prevalence of late AMD was 0.6% (95% CI 0.4% to 0.8%). Statistically significant associations with AMD included increasing age and family history of the condition. This is the first study to provide prevalence estimates of AMD in ROI and will inform eye care professionals and policymakers involved in the delivery and planning of care for those afflicted with this condition.


An amplification unit in human melanoma cells showing partial homology with sequences of human papillomavirus type 9 and with nuclear antigen 1 of the Epstein-Barr virus.
By partial homology with the DNA of human papillomavirus type 9 a cellular amplification unit was detected which is amplified in melanoma cells but not in Epstein-Barr virus-transformed B cells of two melanoma patients. A 2.4-kilobase EcoRI fragment of this amplification unit was cloned and designated mel/HPV9. At the chromosomal level we detected mel/HPV9 in homogeneously staining regions or in abnormally banded regions containing different marker chromosomes of both melanoma cell lines. DNA sequence analysis of a part of mel/HPV 9 revealed homology with the third internal repeat array of Epstein-Barr virus nuclear antigen 1.


Infection of cultured early mouse embryos with Semliki Forest and rubella viruses.
Early mouse embryos at the four- to eight-cell stage or the blastocyst stage could be infected with the A7 strain of Semliki Forest virus (SFV) after the removal of the zona pellucida, either by Pronase treatment or following hatching of blastocysts. With SFV, rapid virus production and eventual cytolysis resulted from infection at either stage. For four- to eight-cell embryos the cytopathic effect was delayed and a proportion of embryos developed to the blastocyst stage. Four- and eight-cell embryos could not be infected with rubella virus (RV), even after removal of the zona pellucida. RV infection of zona-free blastocysts resulted in a productive but non-cytolytic infection which did not affect embryonic development to the early egg-cylinder stage. RV did not multiply in inner cell mass cells isolated from embryos at the blastocyst stage, although SFV did multiply in such cells.


Sunburn in Grapes: A Review.
Sunburn is a physiological disorder that affects the visual and organoleptic properties of grapes. The appearance of brown and necrotic spots severely affects the commercial value of the fruit, and in extreme cases, significantly decreases yield. Depending on the severity of the damage and the driving factors, sunburn on grapes can be classified as sunburn browning (SB) or as sunburn necrosis (SN). Sunburn results from a combination of excessive photosynthetically active radiation (PAR) and UV radiation and temperature that can be exacerbated by other stress factors such as water deficit. Fruit respond to these by activating antioxidant defense mechanisms, de novo synthesis of optical screening compounds and heat-shock proteins as well as through morphological adaptation. This review summarizes the current knowledge on sunburn in grapes and compares it with relevant literature on other fruits. It also discusses the different factors affecting the appearance and degree of sunburn, as well as the biochemical response of grapes to this phenomenon and different potential mitigation strategies. This review proposes further directions for research into sunburn in grapes.


The C-terminal dilysine motif confers endoplasmic reticulum localization to type I membrane proteins in plants.
The tomato Cf-9 disease resistance gene encodes a type I membrane protein carrying a cytosolic dilysine motif. In mammals and yeast, this motif promotes the retrieval of type I membrane proteins from the Golgi apparatus to the endoplasmic reticulum (ER). To test whether the C-terminal KKXX signal of Cf-9 is functional as a retrieval motif and to investigate its role in plants, green fluorescent protein (GFP) was fused to the transmembrane domain of Cf-9 and expressed in yeast, Arabidopsis, and tobacco cells. The fusion protein was targeted to the ER in each of these expression systems, and mutation of the KKXX motif to NNXX led to secretion of the fusion protein. In yeast, the mutant protein reached the vacuole, but plants secreted it as a soluble protein after proteolytic removal of the transmembrane domain. Triple hemagglutinin (HA)-tagged full-length Cf-9 was also targeted to the ER in tobacco cells, and cleavage was also observed for the NNXX mutant protein, suggesting an endoprotease recognition site located within the Cf-9 lumenal sequence common to both the GFP- and the HA-tagged fusions. Our results indicate that the KKXX motif confers ER localization in plants as well as mammals and yeast and that Cf-9 is a resident protein of the ER.


First-line icotinib versus cisplatin/pemetrexed plus pemetrexed maintenance therapy for patients with advanced EGFR mutation-positive lung adenocarcinoma (CONVINCE): a phase 3, open-label, randomized study.
Icotinib has been previously shown to be non-inferior to gefitinib in non-selected advanced non-small-cell lung cancer patients when given as second- or further-line treatment. In this open-label, randomized, phase 3 CONVINCE trial, we assessed the efficacy and safety of first-line icotinib versus cisplatin/pemetrexed plus pemetrexed maintenance in lung adenocarcinoma patients with epidermal growth factor receptor (EGFR) mutation. Eligible participants were adults with stage IIIB/IV lung adenocarcinoma and exon 19/21 EGFR mutations. Participants were randomly allocated (1 : 1) to receive oral icotinib or 3-week cycle of cisplatin plus pemetrexed for up to four cycles; non-progressive patients after four cycles were maintained with pemetrexed until disease progression or intolerable toxicity. The primary end point was progression-free survival (PFS) assessed by independent response evaluation committee. Other end points included overall survival (OS) and safety. Between January 2013 and August 2014, 296 patients were randomized, and 285 patients were treated (148 to icotinib, 137 to chemotherapy). Independent response evaluation committee-assessed PFS was significantly longer in the icotinib group (11.2 versus 7.9 months; hazard ratio, 0.61, 95% confidence interval 0.43-0.87; P = 0.006). No significant difference for OS was observed between treatments in the overall population or in EGFR-mutated subgroups (exon 19 Del/21 L858R). The most common grade 3 or 4 adverse events (AEs) in the icotinib group were rash (14.8%) and diarrhea (7.4%), compared with nausea (45.9%), vomiting (29.2%), and neutropenia (10.9%) in the chemotherapy group. AEs (79.1% versus 94.2%; P < 0.001) and treatment-related AEs (54.1% versus 90.5%; P < 0.001) were significantly fewer in the icotinib group than in the chemotherapy group. First-line icotinib significantly improves PFS of advanced lung adenocarcinoma patients with EGFR mutation with a tolerable and manageable safety profile. Icotinib should be considered as a first-line treatment for this patient population.


Medical undergraduate primary care teaching across the UK: what is being taught?
All UK medical schools use primary care settings to deliver their undergraduate courses. However there is no national undergraduate curriculum for primary care and it is thought that the learning objectives of primary care teaching vary considerably between medical schools. The overall aim was to establish what is being taught within and by primary care across UK medical schools. We did this by collating learning objectives from the primary care department at each school. In order to categorise and compare the list of learning objectives from each school we mapped the learning objectives to the postgraduate curriculum of the Royal College of General Practitioners (RCGP). Cross sectional survey sent to heads of teaching of primary care at all 32 UK medical schools. GP teacher handbooks for primary care modules at each medical school were requested. Information was extracted based on key headings from the RCGP postgraduate curriculum. Topics taught by primary care at all medical schools include: consulting and communication skills, leading and working in teams, and developing yourself and others. Novel topics, taught at a few medical schools include: learning disability, genetics and multi-morbidity. The majority of medical schools address aspects of over half of the RCGP postgraduate curriculum headings in their learning objectives for primary care. This project provides valuable information about primary care teaching at an undergraduate level across the UK. Although it confirms widespread variation in learning objectives, it also highlights considerable common ground and opportunities for sharing teaching resources between schools.


Food fight: the battle over redefining competitive foods.
Environmental and policy influences are potentially the most powerful-and yet the least understood-strategies for reversing the current childhood obesity epidemic. This essay focuses on the school lunch environment and examines the key legal and policy factors that affect competitive foods or foods of minimal nutritional value (FMNV) in schools. The essay also analyzes the latest, proposed federal legislation on redefining competitive foods. FMNV compete for children's coins and calories in the school food environment. The emerging scientific record on the negative impact competitive foods have on children's diet and health is significantly stronger than when Congress and the United States Department of Agriculture (USDA) secretary first deemed the connection sufficient to take action. The Child Nutrition Promotion and School Lunch Protection Act of 2006, if enacted, will provide additional authority to the USDA and supply science-based federal guidance to local schools. Opposition to this legislation may come from the local school districts, the food industry, parents, and children. Defining competitive foods in a federally consistent manner and eliminating competitive foods from our nation's schools are food fight that will evidently be fought by legislative efforts at the local, state, and federal level. The food industry, as history illustrates, will likely work hard to weaken any regulatory efforts and could potentially demand that this fight end up requiring judicial review.


Complement C3 serum levels in anorexia nervosa: a potential biomarker for the severity of disease?
Anorexia nervosa carries the highest mortality rate of any psychiatric disorder. Even the most critically ill anorexic patients may present with normal 'standard' laboratory values, underscoring the need for a new sensitive biomarker. The complement cascade, a major component of innate immunity, represents a driving force in the pathophysiology of multiple inflammatory disorders. The role of complement in anorexia nervosa remains poorly understood. The present study was designed to evaluate the role of complement C3 levels, the extent of complement activation and of complement hemolytic activity in serum, as potential new biomarkers for the severity of anorexia nervosa. This was a prospective cohort study on 14 patients with severe anorexia nervosa, as defined by a body mass index (BMI) <14 kg/m2. Serum samples were obtained in a biweekly manner until hospital discharge. A total of 17 healthy subjects with normal BMI values served as controls. The serum levels of complement C3, C3a, C5a, sC5b-9, and of the 50% hemolytic complement activity (CH50) were quantified and correlated with the BMIs of patients and control subjects. Serum C3 levels were significantly lower in patients with anorexia nervosa than in controls (median 3.7 (interquartile range (IQR) 2.5-4.9) vs 11.4 (IQR 8.9-13.7, P <0.001). In contrast, complement activation fragments and CH50 levels were not significantly different between the two groups. There was a strong correlation between index C3 levels and BMI (Spearman correlation coefficient = 0.71, P <0.001). Complement C3 serum levels may represent a sensitive new biomarker for monitoring the severity of disease in anorexia nervosa. The finding from this preliminary pilot study will require further investigation in future prospective large-scale multicenter trials.


Dietary and lifestyle indices for hyperinsulinemia and colorectal cancer risk: a case-control study.
The incidence of colorectal cancer (CRC) has increased in Iran, and determining the dietary patterns that can contribute to reducing or increasing the risk of CRC will help better control this disease. Therefore, in the current study, we assessed the association between the empirical lifestyle index for hyperinsulinemia (ELIH) and the empirical dietary index for hyperinsulinemia (EDIH) with the CRC odds. The present case (n = 71)-control (n = 142) study was carried out in several CRC surgical units of hospitals in Tehran, Iran. A semi-quantitative food frequency questionnaire containing 168 items was used to assess participants' dietary intakes. The EDIH and ELIH scores were calculated by food groups and some variables such as body mass index and physical activity. Logistic regression models were applied to evaluate the association between the EDIH and ELIH scores with CRC odds. According to baseline features of the study participants, there were significant differences between the controls and cases in ELIH score, fiber intake, taking aspirin, and family history of CRC in first- and second-degree relatives. Also, we found that the odds of CRC increased significantly in the last tertile compared to the first tertile in EDIH and ELIH in the adjusted model (odds ratio (OR) = 3.12; 95% confidence interval (CI): 1.30-7.48 and OR = 4.72; 95% CI: 1.15-19.39, respectively). In conclusion, the result of this study indicated that CRC odds was significantly greater in subjects with higher EDIH and ELIH scores. Also, according to the results of this study, lifestyle and diet with insulinemic potential can influence the CRC risk.


Frameworks for evaluating health research capacity strengthening: a qualitative study.
Health research capacity strengthening (RCS) projects are often complex and hard to evaluate. In order to inform health RCS evaluation efforts, we aimed to describe and compare key characteristics of existing health RCS evaluation frameworks: their process of development, purpose, target users, structure, content and coverage of important evaluation issues. A secondary objective was to explore what use had been made of the ESSENCE framework, which attempts to address one such issue: harmonising the evaluation requirements of different funders. We identified and analysed health RCS evaluation frameworks published by seven funding agencies between 2004 and 2012, using a mixed methods approach involving structured qualitative analyses of documents, a stakeholder survey and consultations with key contacts in health RCS funding agencies. The frameworks were intended for use predominantly by the organisations themselves, and most were oriented primarily towards funders' internal organisational performance requirements. The frameworks made limited reference to theories that specifically concern RCS. Generic devices, such as logical frameworks, were typically used to document activities, outputs and outcomes, but with little emphasis on exploring underlying assumptions or contextual constraints. Usage of the ESSENCE framework appeared limited. We believe that there is scope for improving frameworks through the incorporation of more accessible information about how to do evaluation in practice; greater involvement of stakeholders, following evaluation capacity building principles; greater emphasis on explaining underlying rationales of frameworks; and structuring frameworks so that they separate generic and project-specific aspects of health RCS evaluation. The third and fourth of these improvements might assist harmonisation.


Pathology of Minamata disease.
Minamata disease, or methylmercury poisoning, was first discovered in 1956 around Minamata Bay, Kumamoto Prefecture, Japan. A similar epidemic occurred in 1965 along the Agano River, Niigata Prefecture, Japan. The neuropathology of Minamata disease has been well studied; this review focuses on human cases of Minamata disease in Kumamoto Prefecture. Nervous system lesions associated with Minamata disease have a characteristic distribution. In the cerebral cortex, the calcarine cortex was found to be involved in all cases of Minamata disease, particularly along the calcarine fissure. The destruction of nerve tissue was prominent in the anterior portions of the calcarine cortex. Occasionally, the centrifugal route from the visual and visual association areas (internal sagittal stratum) showed secondary degeneration in prolonged cases after acute onset. Postcentral, precentral, and temporal transverse cortices showed similar changes, though they were less severe. Intense lesions in the precentral cortex caused the development of secondary bilateral degeneration of the pyramidal tracts. In the cerebellum, the lesions occurred deeper in the hemisphere. The granule cell population was most affected. In the peripheral nerves, sensory nerves were more affected than motor nerves. Secondary degeneration of Goll's tracts was occasionally seen in prolonged or chronic cases.