The aims were to assess: (i) how the instruments were used in practice; (ii) which items of resource use were most frequently measured using patient-recall; and (iii) how estimates compared if more than one method of data collection was used for the same resource itemsWe conducted a review of articles citing trial-related resource use measures catalogued in the Database of Instruments for Resource Use Measurement  used an instrument derived from Beecham and Knapp’s Client Service Receipt Inventory or a variation thereof (81/143). Most measures relied on patient- or proxy-recall (126/146) generally administered during researcher interviews. Use of primary and secondary care was the most widely reported items of resource use (136/146). Twelve studies compared more than one method of data collection for the same resource items and 8/12 showed good correlation between medical records and patient/carer recall or at least indicated one could routinely validate the other. No single administration method was deemed superior.Patient interviews are the most common method of questionnaire administration but studies lack clarity on how they are conducted. Results indicate good precision and accuracy in recall questionnaire use; however, concerns about recall bias still exist.


Pathological and imaging data indicate that amyotrophic lateral sclerosis (ALS) is a multisystem disease involving several cerebral cortical areas. Advanced quantitative magnetic resonance imaging (MRI) techniques enable to explore in vivo the volume and microstructure of the cerebral cortex in ALS. We studied with a combined voxel-based morphometry (VBM) and magnetization transfer (MT) imaging approach the capability of MRI to identify the cortical areas affected by neurodegeneration in ALS patients. Eighteen ALS patients and 18 age-matched healthy controls were examined on a 1.5T scanner using a high-resolution 3D T1 weighted spoiled gradient recalled sequence with and without MT saturation pulse. A voxel-based analysis (VBA) was adopted in order to automatically compute the regional atrophy and MT ratio (MTr) changes of the entire cerebral cortex. By using a multimodal image analysis MTr was adjusted for local gray matter (GM) atrophy to investigate if MTr changes can be independent of atrophy of the cerebral cortex. VBA revealed several clusters of combined GM atrophy and MTr decrease in motor-related areas and extra-motor frontotemporal cortex. The multimodal image analysis identified areas of isolated MTr decrease in premotor and extra-motor frontotemporal areas. VBM and MTr are capable to detect the distribution of neurodegenerative alterations in the cortical GM of ALS patients, supporting the hypothesis of a multi-systemic involvement in ALS. MT imaging changes exist beyond volume loss in frontotemporal cortices. Amyotrophic lateral sclerosis (ALS) is a rapidly progressive neurodegenerative disorder characterized by upper motor neuron (UMN) and lower motor neuron (LMN) degeneration The UMN involvement in ALS cannot be easily established based on the clinical ground because of the confounding effect of LMN involvement. The UMN damage has initially been explored mainly in the corticospinal tract using conventional Magnetization Transfer Imaging (MTI) is a MR technique capable to explore the microstructure of the cerebral cortex in several neurodegenerative diseases In order to explore the topographical distribution of VBM and MTI changes in the cerebral cortex of ALS patients we performed a cross sectional study which included voxel based analysis (VBA) of T1 weighted images and MT ratio as well as a multimodal image analysis aimed to ascertain whether MT changes occur independently of regional volume loss.We examined 18 consecutive right handed ALS patients  and 18 right handed healthy controls . Age and gender between groups were not significantly different (Mann-Withney U test p = 0.07 for age and Pearson Chi-square p = 0.17 for gender). All patients had definitive ALS diagnosis according to the revised El Escorial criteria MR data were acquired on a 1.5 T scanner  with high performance gradients , equipped with an 8-channel head coil with ASSET-technology.A high-resolution 3D spoiled gradient recalled (SPGR) sequence (repetition time (TR)/echo time (TE) = 28 ms/5 ms, flip Angle 40°, number of excitation 0.75, field of view (FOV) 240 mm, 192×192 matrix, 124 oblique-sagittal slices 1.5 mm thick, null gap) was acquired with (Sat) and without (No-Sat) the magnetization transfer (MT) saturation pulse. The MT pulse was a 1200 Hz off-resonance pulse.The VBM analysis was carried out by the FSL-VBM optimized protocol MT image processing was performed as described in a previous work In order to exclude the possibility that differences found in voxel-wise MTr analysis were due to brain atrophy in ALS patients compared to healthy subjects, we performed a multimodal image analysis In order to establish a correlation among clinical variables and VBM or MTr changes, ALSFRS score, MRC for upper limb, MRC for lower limb, disease duration and upper motor neuron sign score was inserted as covariate variables within different GLM design matrix.Statistical maps were superimposed onto Anatomical Automated Labeling (AAL) Atlas The research was conducted according to the principles expressed in the Declaration of Helsinki. The study was approved by the Ethics Commitee of Azienda Ospedaliera Universitaria Pisana, Pisa, Italy. Written informed consent was obtained from all the study partecipants.Results of the between group VBM analysis revealed several clusters of reduced cortical GM in ALS patients compared to healthy controls (p<0.05 FWE corrected for multiple comparisons). They were located in the superior, middle and inferior frontal gyri , in the supplementary motor area (AAL 20) and in the temporal lobe (AAL 85) . These rResults of the between group MTI analysis revealed that the MTr values were significantly lower in ALS patients than in the healthy controls in the following cortical regions: superior, middle and inferior frontal gyrus , gyrus cinguli , supplementary motor area , insula  and temporal lobe  .Results of the between group regional MTI analysis corrected for atrophy revealed that the MTr values were significantly lower in ALS patients than in the healthy controls in the superior, middle and inferior frontal gyri , in the gyrus cinguli , in the supplementary motor area , in the insula  and in the temporal lobe  .No cluster of significant association between atrophy or MTr changes and clinical scales was detected in the respective statistical maps.ALS is emerging as a multi-system disease involving several frontal-temporal structures beside motor structures and functions In order to compare the spatial distribution of atrophy and MTr changes in ALS we adopted in the present study a voxel based approach of MT data Since a region specific MTr decrease in the sensory motor cortex is observed with advancing age Although we do not know the exact meaning of the reduced MTr in the cerebral motor-related and extra-motor cortex of ALS patients, the rate of MT effect depends on the surface chemistry and the biophysical dynamics of the macromolecules, as well on the concentration of macromolecules and water The neuronal loss and degeneration in the V layer of the cortex into the precentral gyrus is considered the pathological hallmark of the UMN involvement in ALS In the present study the voxel-wise analysis of GM volume, MTr and MTr data corrected for regional atrophy failed to show any structural abnormality in the precentral gyrus of ALS patients. This result may reflect both the fact that the pathological changes in primary motor cortex of ALS patients are subtle On the other hand our and other studies showed clusters of atrophy and MTr changes in motor-related area. Clusters of decreased MTr independent of cortical atrophy were observed in the premotor cortex particularly in the superior, middle frontal gyri pertaining to the fronto parietal circuit At multimodal image analysis, the MTr decrease in prefrontal and temporal cortex of our ALS patients survived correction for local atrophy indicating that the microstructural changes exist beyond the areas of cortical atrophy. This interpretation is supported by the observation that MTI can detect microstructural changes in cortical areas unaffected by atrophy also in patients with Alzheimer disease (AD) A longitudinal evaluation of VBM and MTr data could be a further fundamental step in evaluating their spatial and temporal relationship in the progression of motor and cognitive functions in ALS and is currently under way.We recognize as main limitation of our work that we did not obtain a detailed neuropsychological profile of our patients. This hinders investigation of the relationship between behavioural and cognitive functions and regional volume or MTr of the cortical GM in specific brain structures VBM and MTI reveal regional atrophy and microstructural changes of the cerebral cortex in ALS involving not only motor-related areas but also frontotemporal cortices.MTr reduction occurs in non atrophic cortical areas suggesting that microstructural degenerative phenomena can extend beyond the tissue loss in ALS.Table S1Demographic and Clinical data.(DOC)Click here for additional data file.Table S2VBM results: clusters of cortical atrophy.(DOC)Click here for additional data file.Table S3MT Imaging results: clusters of reduced MT ratio.(DOC)Click here for additional data file.Table S4MT Imaging results: clusters of reduced MT ratio adjusted for atrophy.(DOC)Click here for additional data file.


A number of human diseases such as obesity and diabetes are associated with changes or imbalances in the gut microbiota (GM). Laboratory mice are commonly used as experimental models for such disorders. The introduction and dynamic development of next generation sequencing techniques have enabled detailed mapping of the GM of both humans and animal models. Nevertheless there is still a significant knowledge gap regarding the human and mouse common GM core and thus the applicability of the latter as an animal model. The aim of the present study was to identify inter- and intra-individual differences and similarities between the GM composition of particular mouse strains and humans.A total of 1509428 high quality tag-encoded partial 16S rRNA gene sequences determined using 454/FLX Titanium (Roche) pyro-sequencing reflecting the GM composition of 32 human samples from 16 individuals and 88 mouse samples from three laboratory mouse strains commonly used in diabetes research were analyzed using Principal Coordinate Analysis (PCoA), nonparametric multivariate analysis of similarity (ANOSIM) and alpha diversity measures. A reliable cutoff threshold for low abundant taxa estimated on the basis of the present study is recommended for similar trials.Distinctive quantitative differences in the relative abundance of most taxonomic groups between the examined categories were found. All investigated mouse strains clustered separately, but with a range of shared features when compared to the human GM. However, both mouse fecal, caecal and human fecal samples shared to a large extent not only representatives of the same phyla, but also a substantial fraction of common genera, where the number of shared genera increased with sequencing depth. In conclusion, the GM of mice and humans is quantitatively different (in terms of abundance of specific phyla and species) but share a large qualitatively similar core. Shifts in the composition, known as dysbiosis, of the human GM have in several studies been associated with diseases such as allergies Rodents are the mammalian model most extensively used to investigate the relationship between GM and health and disease. The reason for the popularity of mouse models is their well explored genetic and relatively close physiological similarity with humans and the ability to control a wide range of environmental factors which reduces variation in the baseline gut microbiota between individual study objects Lachnospiraceae family have been suggested as possible biomarkers to help predict predispositions to inflammatory bowel disease Many GM related disorders have been linked with bacterial dysbiosis on a higher taxonomic level proving the usefulness of sequencing the GM to e.g. phylum and family level. For example, Turnbaugh et al. showed that a switch from a low fat to a high fat, high sugar diet in mice, which was associated with obesity, lowered the Bacteroidetes/Firmicutes ratio in the gut within one day However, the usefulness of mice models for such studies would to a large extent also depend on similarities in their GM profiles at genus or species level with humans. The aim of the present study was therefore to demonstrate inter- and intra-individual differences and similarities between the GM composition of three laboratory mouse strains commonly used in research in chronic inflammatory diseases with those of humans based on more than 1.5 million high quality sequences of partial 16S rRNA gene verified with tag-encoded 454/FLX Titanium (Roche) pyro-sequencing.A total of 88 mice and 128 human  GM profiles determined using tag-encoded 16S rRNA gene 454/FLX Titanium (Roche) pyro-sequencing were included in the study . All samAll animal experiments were carried out in accordance with the Council of Europe Convention European Treaty Series (ETS) 123 on the Protection of Vertebrate Animals used for Experimental and Other Scientific Purposes, and the Danish Animal Experimentation Act (LBK 1306 from 23/11/2007). The study was approved by the Animal Experiments Inspectorate, Ministry of Justice, Denmark.Human specimens used in this meta-analysis come from the independent study that was approved by The Scientific Ethics Committee of Capital Region, Denmark (reference H-4-2010-137). Written informed consent was obtained from volunteers prior to recruitment.http://www.drive5.com/usearch/). The representative sequences from each cluster were aligned with pyNAST (http://qiime.org/pynast/) and subjected to the Ribosomal Database Project (RDP)-based 16S rRNA gene annotation. For intra-individual assessment all mouse and 16 human samples from the control group were subsampled to an equal number of reads per individual (4500 reads per sample which constitutes to 85% of the second most indigent sample in the dataset). For inter-group comparison the reads were merged according to host (mouse strain/human) and subsampled to an equal number of sequences per category, respectively 80000 reads for mice strains comparisons and 600000 reads for similarity assessment between mice and humans. Both numbers constitute approximately 85% of the least numerous category. Alpha diversity measures such as rarefaction curves based on the estimated species number (97% sequence identity threshold), Chao1 and Shannon indexes were calculated for OTU tables that were unified to 4000 (first most indigent sample) sequences per sample.The dataset was analyzed using the Quantitative Insight Into Microbial Ecology (QIIME) open source software package In order to investigate the influence of the sequencing method into variance between categories, which is caused mostly by the low abundant taxa, a set of 16 samples each sequenced in 4 independent runs were compared within their quadruplicates. A set of 1000 subsampled OTU-tables was generated for each sample (3000 reads per sample). Low abundant taxa were removed until taxa similarity of all replicates within a given sample crossed 99%. An average, minimum cutoff value was therefore calculated based on 16000 subsampled OTU tables using an in-house Matlab (Mathworks) script.Principal Coordinate Analysis (PCoA) plots were generated with the Jackknifed Beta Diversity workflow based on 10 distance metrics calculated using 10 subsampled OTU tables. The -e value (number of sequences taken for each jackknifed subset) was set to 85% of the sequence number within the most indigent sample. Analysis of similarities (ANOSIM) was used to evaluate group differences using weighted and unweighted uniFrac distance metrics that were generated based on rarefied (4500 reads per sample) OTU tables. The relative distribution of the GM genera registered in 88 mouse and 16 human samples was calculated for unified, summarized at the genus level OTU tables.http://metastats.cbcb.umd.edu). From each group 14 samples  were randomly chosen and combinations of group pairs were tested using 1000 permutations . The relationship between sequencing depth and shared GM, classified into phylum and genus level, between mice and humans was plotted based on multiple subsampled OTU tables composed of two categories collecting 794988 human and 714440 mouse GM 16S rRNA gene reads. Simulation of each sequencing depth was repeated 100 times and an average proportion of shared taxonomic groups between the two categories were calculated (abundance threshold for unshared taxa = 0.19%).Differences in taxa abundances at phylum and genus level between categories were verified with Metastats (http://www.cytoscape.org/).A network presenting shared genera between categories was prepared for normalized OTU tables (600000 reads per category) after filtering the low abundant OTUs (abundance threshold for unshared taxa = 0.19%) using the make_otu_network.py script (QIIME). The visualization of the OTU-networks was performed with an open source platform –Cytoscape  yielded 1753858. After removing chimeric sequences a total of 1509428 reads remained, meaning that the ChimeraSlayer algorithm Alpha diversity assessment using rarefaction curves revealed that the fecal GM of the BALB/c mice is the most rich in OTUs (species-level). The average Chao1 index calculated for 4000 reads per sample was 2.5–5 times higher in this group compared with other groups . SpeciesobLep/J groups where age was the only varying factor were only partially separated . The three remaining mouse strains were either devoid of bacteria from this phylum, or the bacteria were below the detection limit.All reads used in this study were classified into 9 phyla after applying the abundance threshold (0.19%), with one phylum noted as unclassified . GeneralAlistipes (12%), while in human specimens – Roseburia (12%) and unclassified genus representing the Clostridiales order (10%) were the dominating genera.The relative abundance of genera depicted for all samples individually shows explicit alterations between studies with the human specimens being the most conspicuous category . In totaMetastats analysis revealed widespread differences in the bacterial relative abundance at both phylum  and genuobV-Lep/J pups  and the NOD group was not detected in the adult B6. obV-Lep/J counterparts (examined in the age of 16 weeks), nor in any of the BALB/c mice. The Akkermansia genus which is the only representative group of the Verrucomicrobia phylum, was therefore the reason of discrepancies on both the phylum . The similarity of the collective mouse and human microbiomes without the abundance threshold was correspondingly 40% and 57% lower at the phylum and the genus level  comparedIn Laboratory mice are commonly used as experimental models for diseases such as diabetes Qualitative and quantitative-based analysis of the three mouse strains fecal GM compared with the mice caecal and human fecal GM (PCoA and ANOSIM) disclosed significant separation of all microbial profiles. The composition of the GM has previously been shown to be influenced by multiple factors including environment and host genetics Diet is an environmental factor known to strongly influence GM composition Early priming of bacterial colonization during early life may also influence GM and immunity later in life Lastly, for adult humans and mice the influence of age on the GM profiles seems to be minor as neither human samples collected from patients at different age (18–50 years) nor NOD mice analyzed at different ages (14–30 weeks) clustered according to age. This supports the high level of GM composition stability in adult individuals which was previously documented among humans and mice Although innovations in high throughput sequencing techniques offer insight into microbial communities at an hitherto up-preceded level of details one need to be aware of constrains that comes with it, for example: sequencing depth, reads error  rate and length or analysis methods In the present study the Firmicutes fraction was more abundant than the Bacterioidetes in most fecal and caecal samples, except for the BALB/c fecal samples, which showed an inverted proportion of these two phyla. Human fecal samples had relatively more Firmicutes and less Bacteriodetes compared to most mouse fecal and caecal samples. This is in accordance with earlier studies reporting that the majority of bacteria in the gut are members of these two phyla and that in mice the Firmicutes fraction seems to be much larger than the Bacteriodetes fraction As seen from Prevotella genus has been found to be inversely correlated with body weight gain, cholesterol accumulation, insulin resistance and diet-induced adiposity Odoribacter genus which relative abundance was recently shown to be increased in the caecum of mice exposed to grid floor induced stress obLep/J group that on the other hand were lacking members of the Akkermansia genus. Akkermansia muciniphila has been suggested to possess anti-inflammatory properties as it was found to be present in lower levels in humans suffering from inflammatory bowel disease compare to the healthy control group For example, the vs. humans) using 0.6 million reads per category disclosed 89% similarity between mouse and human GM genera, with 9 genera being unique for human samples and not detected in any of the three mice strains (abundance threshold 0.19%). Among these 9 genera were Faecalibacterium from the Ruminococcaeae family and Asteroleplasma from phylum Tenericutes that both have been suggested as possible indicators of a healthy human GM since disturbances in the relative distribution of common species from these genera have been linked with the etiology of Crohn’s disease (CD) and ulcerative colitis (UC) Megasphera genus was also unique for humans not being detected in any of the three mouse strains. Megasphera spp. have been imputed to support the growth of colonic mucosa Mistsuokella genus has recently been identified as a GM member of lean as well as obese Indians Faecalibacterium and Dialister but also Sutterella from the Proteobacteria phylum showed poor establishment after transplanting them from human into mice GI tracks Pairwise comparison  and mouse (blue node) categories generated for normalized OTU tables (600000 reads per category) after filtering the low abundant OTUs  using the make_otu_network.py script (QIIME). The visualization of the OTU-networks was performed with an open source platform –Cytoscape .(PDF)Click here for additional data file.Figure S2Number of GM phyla and genera shared between given categories using raw data. Number of taxonomic labels shared between given mice strains using raw data where no abundance threshold was used (A) on the genus level and (B) on the phylum level. 80000 high quality 16s rRNA reads used to represent the GM of each mouse strain were annotated to the Ribosomal Database Project  database. (C) Number of phyla and genera shared between collated categories of humans and mice using raw data (600000 reads per category). Labels “BALB/c (f)”, “BALB/c (c)”, “B6.V-obLep/J (16)” and “B6.V-obLep/J (8)” stand for the gut GM of BALB/c mice determined using fecal and caecal samples and B6.V-obLep/J mice using fecal specimens sampled in 16 and 8 weeks of age respectively.(PDF)Click here for additional data file.Table S1The relative distribution of bacterial genera among categories. The relative distribution of the gut microbial genera among single human  and five mouse groups . The abundance threshold within at least one of the category was set to 0.19%.(PDF)Click here for additional data file.Table S2Differences in the relative abundance of gut microbial phyla between categories. Differences in the gut microbial phyla relative distribution verified using Metastats (http://metastats.cbcb.umd.edu) for all combinations of categories. Each pairwise comparison was performed based on 1000 permutations .(PDF)Click here for additional data file.Table S3Differences in the relative abundance of gut microbial genera between categories. Differences in the gut microbial genera relative distribution verified using Metastats (http://metastats.cbcb.umd.edu) for all combinations of categories. Each pairwise comparison was performed based on 1000 permutations .(PDF)Click here for additional data file.


Dear Editor,Vagal paragangliomas are uncommon tumors of the parapharyngeal space, approximately 200 cases having been reported. They ariThe other manifestations are hoarseness and less commonly dysphagia, palatal weakness and tongue hemi-atrophy.4] Press Press4] A 37 years old farmer presented with hoarseness and left cervical mass beyond mandible angle. He did not claim of another symptom, on physical examination a firm mobile 2x2 cm mass was moving horizontally. Before treatment planning, we found left vocal cords paralyzed and CT scan demonstrated a heterogenous lesion next to the carotid bifurcation . PlanninThe normal carotid body is a small mass of neurovascular tissue located bilaterally in the medial valley of the carotid artery bifurcation. Although the term "tumor" has historically been assigned to these masses, they are not carcinomas, and the neoplastic process is actually hypertrophy of the carotid body tissue.The four main locations of glomus tissue within the head and neck are: (a) The carotid bifurcation (carotid body tumor), (b) The inferior ganglion region (ganglion nodosum) and cervical portion of the vagus nerve , (c) The jugular bulb region (glomus jugulare) and (d) The middle ear cavity (glomus tympanicum). ParagangChallenging options about surgical versus radiation treatment continues, advocating radiation for small lesions,9] and s and s9]


Nosocomial infection is a constant concern in the obstetric and surgical practice in both developing countries than in developed countries.To assess knowledge, attitudes and practices of the staff of maternity services in the prevention of nosocomial infection in three community health centers in Bujumbura.A qualitative survey was conducted using a standardized questionnaire over 2 months with 76 health workers from three maternity wards of health facilities in the vicinity of all categories. Data collection consisted of two parts: first assessment of knowledge through a questionnaire and also the observation of attitudes and practices of personal hygiene.It was noted: In knowledge: Knowledge of the exact def-inition of nosocomial infection: 75% (doctors), modes of transmission of infection (manu-portage): 50%, hygiene protocols: 25%, and procedures decontamination of contaminated equipment: 16%. At the observation of practices:Compliance with hand washing before and after childbirth: 50% Use of alcohol-based solutions: 20%, antiquated equipment sterilization: 80% Compliance with the sterilization of equipment: 25% Circulation in delivery rooms disorganized: 100%, perceived risk of blood contamination (uterus): 75%, biomedical waste collected and decontaminated evil: 80%, existence of incinerators: 33.3% and participation in training in personal hygiene: 50%.There is a high risk of infectious contamination in maternity services studied for both the staff and for parturient and their newborns.None declared.


This study assessed the feasibility and acceptability of utilising insecticide treated plastic sheeting (ITPS) as a malaria control intervention in Papua New Guinea (PNG).® ITPS was installed in 40 homes across four study sites representing a cross section of malaria transmission risk and housing style. Structured questionnaires were completed at the time of ITPS installation (n=40) and at four weeks post installation (n=40) with the household head. Similarly, focus group discussions (FGDs) with the male and/or female household heads were completed at installation (n=5) and four week follow-up (n=4).Zero Vector® ITPS was successfully installed in a range of homes employing traditional and/or modern building materials in PNG. The ITPS installations remained intact over the course of the four week trial period and were highly acceptable to both male and female household heads. No dissatisfaction with the ITPS product was reported at four week follow-up; however, the installation process was time consuming, participants reported a reduction in mosquito net use following ITPS installation and many participants expressed concern about the longevity of ITPS over the longer term.ZeroVector® ITPS installation is feasible and highly acceptable in a diverse range of PNG contexts and is likely to be favourably received as a vector control intervention if accessible en masse. A longer-term evaluation is required before firm policy or public health decisions can be made regarding the potential application of ITPS in the national malaria control program. The positive study findings suggest a longer-term evaluation of this promising malaria control intervention warrants consideration.ZeroVector


Despite the introduction of more aggressive systemic therapies for the management of rheumatoid arthritis (RA) MTP joint pain & deformity is commonly reported. Previous research has proposed that articular damage in the forefoot of patients with RA is probably caused by synovitis leading to destruction of the bone & soft tissues, which might result in the failure of a complex ligamentous system & the dynamic effect of displacement of the plantar plates (PP).In 24 patients with RA the more symptomatic forefoot was imaged using MRI, high-resolution ultrasound (HRUS) & conventional radiography (CR). The MRI & HRUS images were assessed to determine inflammation & damage at the MTP joints, the presence or absence of the PP & to identify tears of the PP & their location. CR was used to determine radiographic progression at the MTP joints.17 females and 7 males (mean (SD) age of 55.5 (10.5) years, disease duration 10.6 (8.6) years & forefoot pain score was 43.4 (27.9)) took part.Tears were not identified on HRUS. The Larsen score on the index foot correlated with the absence of PP viewed using both MRI  & HRUS . MRI synovitis was not related to PP absence on either MRI  or HRUS  but there was good correlation between MRI bone oedema & the number of absent PP visualised using MRI  & HRUS . There was also a strong correlation between MRI imaged bone erosion & PP absence confirmed using both MRI  & HRUS .In agreement with existing literature, this current study demonstrates that pain & deformity at the MTP joints is highly prevalent despite good disease control. The findings indicate that PP failure is associated more with damage at the lesser MTP joints than active synovitis in patients with RA. Longitudinal follow-up of patients with RA & forefoot pain is required to establish the relationship between local inflammation, disease activity, impairment & structural changes, & identify factors that precede & result in PP pathology.


The endothelial glycocalyx is a recently discovered structure at the luminal side of blood vessels consisting of proteoglycans and glycosaminoglycans, which play an important role in vascular barrier function and cell adhesion. Due to its vulnerability, the endothelial glycocalyx may easily be altered by hypoxia , TNFα 22, oxidizn = 10), severe sepsis (n = 9) and septic shock (n = 18) as defined by the members of the ACCP/SCCM Consensus Conference Committee  by ELISA technique and polymorphonuclear leukocyte (PMN) function by flow cytometry in eight healthy volunteers (HV) and 37 patients who were prospectively enrolled within 24 hours of onset of signs of infection, if they met the criteria for sepsis n = 0, severeP < 0.05; Figure Plasma levels of the glycocalyx components were significantly higher in septic patients than in healthy volunteers and even more pronounced in patients with severe sepsis and septic shock (all Elevated plasma levels of hyaluronan, syndecan and heparan sulfate are suggestive of a glycocalyx shedding from endothelium with increasing sepsis severity. This process might contribute to the vascular dysfunction and development of edema in septic patients.


The aim of the study is identification of correlations between the serum concentrations of iron and the risk of breast cancer among female BRCA1 mutations carriers and unselected breast cancer patients.The first group (99 triplet) selected for the trial were Polish women, positive for at least one of three founder mutations in BRCA1 gene dominating in Poland . Serum was collected at the time of breast cancer diagnosis. Persons with detected tumor were considered as cases and the others were considered as controls. One case and two controls were paired regarding many criteria  to achieve the maximum of similarity between them.The subjects of the second group (28 triplet) selected for the trial were Polish women, positive for at least one of three founder mutations in BRCA1 gene. Serum was collected 1-2 years before breast cancer diagnosis. One case and two controls were matched for year of birth, past history of cancer, adnexectomy status and cigarettes smoking to achieve the maximum of similarity between them.The subjects of the third group were unselected cancer for the trial were Polish women. Serum was collected during breast cancer diagnosis before treatment. One case and one control were matched for year of birth (+/- 3 years), number and location of cancer among Iº relatives, smoking-the number of pack years (+/- 10%), adnexectomy status, CHEK2 mutation.The proportion of cases and control in the first quartile was taken as a reference to calculate the odds ratio, confidence interval and p-value of the multivariate conditional logistic regression.The iron was quantitatively measured by ICP-MS (Inductively Coupled Plasma Mass Spectrometry), .This study shows that concentration levels of iron in blood serum are a strong factors associated with an additionally increased risk of breast cancer among BRCA1 mutation carriers and unselected breast cancer patients.For iron concentration in BRCA1 carriers (serum collected at the time of breast cancer diagnosis) all quartiles above the first one had a decreased risk of breast cancer. The results are shown in Table For iron concentration in BRCA1 carriers (serum collected 1-2 years before breast cancer diagnosis) the last quartile had a decreased risk of breast cancer. The results are shown in Table Individuals classified in the III and IV quartile had lower risk of breast cancer however results were statistically insignificant.The strongest result and lowest cancer risk was for Fe > 1350 μg/l For iron concentration in unselected breast cancer patients (serum was collected during breast cancer diagnosis before treatment) all quartiles were statistically insignificant. The results are shown in Table Individuals classified in the III quartile had lower risk of breast cancer however results were statistically insignificant.The strongest result and lowest cancer risk was for Fe > 1000 μg/l .For iron concentration in unselected breast cancer patients (serum was collected during breast cancer diagnosis before treatment) before 55 years old, third quartile had a decreased risk of breast cancer. The results are shown in Table Individuals classified in the III quartile had lower risk of breast cancer ;The strongest result and lowest cancer risk was for Fe concentration higher than 987 μg/l (III quartile) ; max 1285 μg/l (III quartile)


Inflammation is a vital process induced by microbial infection or tissue injury. The main function of inflammation is to resolve the infection or repair the damage and return the body to a state of homeostasis. Acute inflammation can induce signals that result in two main functions: 1) initiation of an inflammatory cascade that helps limits the infection and 2) activation of the immune response. In this study we determined the impact of the nature and magnitude of acute inflammation on the host anti-tumor activity against Ehrlich Ascites cancer cell (EAC) line as a model system which forms either peritoneal ascites upon intraperitoneal injection.Different inflammatory signals, including agonists for toll-like receptor (TLR) 2 (zymosan) and TLR3 agonist (poly (I:C) or its clinical grade Hiltonol®), complete Freund's adjuvant (CFA), and incomplete Freunds Adjuvant (IFA), and BCG from Bacillus calmatte (TLR9 agonist) were used to induce acute inflammation 1 day after i.p. challenge with EAC.We found that Hiltonol®, poly (I:C), IFA, CFA, and BCG induced inflammatory cells associated with anti-tumor activity that resulted in significant decrease in the tumor growth measured by the total number of tumor cells after 7 days of tumor challenge. Of interest, among the test TLR agonists both poly(I:C) and Hiltonol® (TLR3 agonist) showed the highest anti-tumor effects.Provision of the proper inflammatory signal with optimally defined magnitude and duration during cancer growth will induce inflammatory cells with potent anti-tumor responses leading to significant decreases in tumor growth.


Studies investigating the outcome of conservative scoliosis treatment differ widely with respect to the inclusion criteria used . While tA PubMed and (incomplete) hand search for outcome papers on PT has been performed in order to detect study designs and inclusion criteria used.Real outcome papers (start of treatment in immature samples / end results after the end of growth) have not been found. Some papers investigated mid-term effects of exercises, most were retrospective, few prospective  and someAn agreement of the scientific community on common inclusion criteria for future studies on PT is necessary. We would suggest the following: (1) girls only, (2) age 10 to 13 with the first signs of maturation (Tanner II), (3) Risser 0-2, (4) risk for progression 40 – 60% according to Lonstein and Carlson.A SOSORT consensus paper following a 3-step Delphi process seems necessary in order to establish the inclusion criteria for future studies on PT.


Disseminated intravascular coagulation (DIC) is observed after not only sepsis but also trauma. DIC is associated with concomitant activation of coagulofibrinolytic disorder and systemic inflammation with endothelial dysfunction and microvascular permeability. The angiogenic factors, including vascular endothelial growth factor (VEGF), angiopoietin (Ang), and their receptors, play crucial roles in angiogenesis and microvascular permeability. The aim of the study was to assess: the relationship between angiogenic factors, their soluble receptors and organ dysfunction associated with DIC precipitated by severe trauma; and the effects of DIC-induced platelet consumption, thrombin generation and tissue hypoxia on the expression of these factors and receptors.Fifty-seven patients with severe trauma were divided into two subgroups: 30 DIC patients and 27 non-DIC patients. The serum levels of angiogenic factors were measured on admission (day 1), day 3, and day 5. We compared serum levels of these angiogenic factors between with and without DIC groups and evaluated their predictive value for organ dysfunction and outcome.DIC patients showed higher Sequential Organ Failure Assessment (SOFA) scores, soluble fibrin and lactate levels. The serum levels of VEGF, Ang1, and the sTie2 levels were lower in the DIC patients than the non-DIC patients. The serum levels of sVEGFR1, Ang2 and the Ang2/Ang1 ratio in the DIC patients were higher than in those without DIC. The sVEGFR2 levels showed no statistically significant difference between the patients with and without DIC. The levels of sVEGFR1, Ang2 and the Ang2/Ang1 ratio correlated with the SOFA score. In particular, sVEGFR1 and Ang2 were independent predictors of an increase in the SOFA score. The lactate levels independently predicted increases in the levels of sVEGFR1 and Ang2 and platelet consumption also independently predicted the increase in Ang2 levels in severe trauma patients with DIC.Angiogenic factors and their soluble receptors, particularly sVEGFR1, play pivotal roles in the development of organ dysfunction in DIC associated with severe trauma. The DIC-induced tissue hypoxia and platelet consumption plays crucial roles in inducing sVEGFR1 and Ang2, and in determining the prognosis of the severity of organ dysfunction.


Molecular epidemiologic findings suggest an ancient focus of TB. Mycobacterium tuberculosis in India is largely unknown. A collection of 91 isolates originating from 12 different regions spread across the country were analyzed by genotyping using 21 loci with variable-number tandem repeats (VNTRs), by spoligotyping, by principal genetic grouping (PGG), and by deletion analysis of M. tuberculosis–specific deletion region 1. The isolates showed highly diverse VNTR genotypes. Nevertheless, highly congruent groupings identified by using the 4 independent sets of markers permitted a clear definition of 3 prevalent PGG1 lineages, which corresponded to the "ancestral" East African–Indian, the Delhi, and the Beijing/W genogroups. A few isolates from PGG2 lineages and a single representative of the presumably most recent PGG3 were identified. These observations suggest a predominance of ancestral M. tuberculosis genotypes in the Indian subcontinent, which supports the hypothesis that India is an ancient endemic focus of TB.Although India has the highest prevalence of tuberculosis (TB) worldwide, the genetic diversity of  Mycobacterium tuberculosis, its main causative agent, is widely disseminated and is one of the most successful human pathogens today, with 2 billion persons infected. Most of the disease's effects are now concentrated in countries with few resources; India has the highest number of cases (Tuberculosis (TB) in humans has been described since ancient times. M. tuberculosis  of genetic elements named mycobacterial interspersed repetitive units (MIRUs)  were used to assess consistency of the genetic relationships obtained by VNTR typing and spoligotyping at a broader evolutionary level. SNPs in the katG and gyrA genes classify M. tuberculosis isolates into 3 principal genetic groups (PGGs) thought to have evolved sequentially from group 1 to group 3  according to the previously described method . MIRU-VNTR profiles were compared to each other by using the neighbor-joining algorithm. For spoligotypes, the Jaccard index was calculated to allow for the construction of a dendrogram by using the unweighted pair-group method with arithmetic averages. The spoligotypes were compared to fingerprints in an international database , X (1 case), and T (8 cases) spoligotype families were detected among the isolates of the PGG2/3group. The single PGG3 isolate (ICC399) had a T spoligotype, which includes both PGG2 and PGG3 strains   groups. This correlation, also seen in previous studies on isolates from Singapore  has been observed in southern India (,,The few previously identified TbD1+ strains were isolated from patients from East Africa and South Asia. These strains have low copy numbers of IS India ,,. Also co India ,,.6110 distribution (6110 low copy-number fingerprints and of known spoligotypes (,6110 (The prevalence of these low-copy-number strains in regions of such high endemicity has raised the question of the true extent of genetic variation beyond their restricted ISgotypes ,. Moreove,,In addition to the TbD1+ isolates, 2 other major PGG1 families were well represented in this Indian collection. They were qualified as modern groups by their TbD1– status. The recently identified Delhi type (M. tuberculosis ancestral strains and the relatively poor representation of the most recent lineages in this Indian collection lend support to the hypothesis that India is a relatively ancient endemic focus of TB (M. tuberculosis complex, subsequent to the recently proposed emergence of tubercle bacilli in eastern Africa millions of years ago (The predominance of


We reviewed neuromedin U (NMU) and neuromedin S (NMS) involvement in the regulation of the hypothalamo–pituitary–adrenal (HPA) axis function. NMU and NMS are structurally related and highly conserved neuropeptides. They exert biological effects via two GPCR receptors designated as NMUR1 and NMUR2 which show differential expression. NMUR1 is expressed predominantly at the periphery, while NMUR2 in the central nervous system. Elements of the NMU/NMS and their receptors network are also expressed in the HPA axis and progress in molecular biology techniques provided new information on their actions within this system. Several lines of evidence suggest that within the HPA axis NMU and NMS act at both hypothalamic and adrenal levels. Moreover, new data suggest that NMU and NMS are involved in central and peripheral control of the stress response. Bombesin-like neuromedins—neuromedin B (NMB) and neuromedin C (NMC)  and neuromedin L (NML)  , for which no substantial homology with other known neuropeptides was found  and its anorexigenic action have enhanced interest in physiological role of NMU and NMS  and the other 8 (NMU8) amino acid residues   gene, on the other hand, was identified based on its sequence similarity with NMUR1  NMU-like immunoreactivity was observed in perikaria of the dorsal nucleus of the hypothalamus and the caudal part of the infundibulum . In the rat hypothalamus expression of NMS gene was nearly 3-fold higher than that of NMU gene  and protein (dot blot method) were identified in 2000  injection of NMU demonstrated a strong increase in Fos-immunoreactive nuclei in the PVN and supraoptic nucleus (SON) of the rat hypothalamus  release by rat hypothalamic explants NMU also regulates HPA axis in birds. Icv administration of NMU in chicks significantly upregulated mRNA expression of CRH in the hypothalamus  blood ACTH level remained unchanged  into hemiadrenalectomized rats did not significantly change corticotropin levels  resulted in a dose-dependent increase of plasma ACTH concentrations, an effect significantly reduced by pretreatment with anti-NMU IgG  and a sustained (24 h) elevation of plasma corticosterone concentration  NMU activates CRH containing neurons and stimulates CRH secretion, which in turn triggers pituitary ACTH and adrenal corticosterone/cortisol secretion. In this regard it is not astonishing that NMU and NMS are involved in central and peripheral control of the stress response. These studies were initiated by Hanada et al.  who obseStress related behavior induced by icv NMU or NMS administration was also observed in cattle  glycol (PEG) (“PEGylation”) may be helpful in these attempts (Ingallinella et al., The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Aortic valve stenosis (AVS) is the most common valvular disease. Its evaluation is of growing interest because of its increasing incidence with the aging population. Previous studies demonstrated the usefulness of Phase-Contrast Magnetic Resonance (PCMR) images in the evaluation of AVS. However, because of the lack of automated methods for PCMR data analysis, this technique remains time-consuming and operator-dependent.Therefore, the aims of this study were 1) to develop a semi-automated method for aortic flow analysis from PCMR images, and 2) to evaluate several approaches of aortic valve area (AVA) estimation.2) and 12 healthy subjects  who had the same day a trans-thoracic echocardiography (TTE) and PCMR acquisitions at the levels of the aortic valve and the left ventricle outflow tract (LVOT). PCMR data analysis included a semi-automated segmentation, based on pixels connectivity in terms of velocity sign, to delineate the aortic flow on all systolic frames, as well as a functional parameters extraction from aortic velocity and flow rate curves such as aortic maximal velocity (VmaxAo) and AVA. AVA was calculated using: 1) Hakki’s formula which is a simplification of Gorlin’s formula, resulting in AVA1= cardiac output divided by √systolic pressure gradient, 2) the continuity equation with the most pertinent method found in previous studies in PCMR resulting in AVA2 = LVOT stroke volume (LVOT SV) divided by aortic valve velocity time integral (VTIAo), and 3) the continuity equation with another approach previously described in echocardiography but never used in PCMR. It resulted in AVA3= LVOT peak Flow rate (QLVOT) divided by VmaxAo . Hakki’s formula underestimated the AVA with regard to TTE in the absence of severe stenosis. AVA3 provided the best results .The LVOT and aortic flows were successfully delineated on all phases for all subjects. Comparison of PCMR and echocardiographic Vmaxts Table  in termsOur semi-automated approach for AVS evaluation from PCMR provided reproducible velocity measurements and AVA estimates in good agreement with echocardiographic values, and were able to characterize the severity of AVS.


Rheumatoid arthritis (RA) is a prototypic autoimmune disease characterized by a prominent humoral autoimmunity. Of particular relevance is the local production of autoantibodies such as rheumatoid factor and anti-citrullinated protein antibodies in the inflamed synovial tissue. The mechanisms underlying break of B cell tolerance and local autoantibody production remains poorly understood. This study was conducted in order to identify cellular and molecular pathways implicated in RA-specific humoral autoimmunity.Synovial tissue samples were obtained by arthroscopy from untreated individuals with RA (n=33) and inflammation matched SpA controls (n=58). Gene expression profiling was performed on tissue samples of patients with established arthritis using 44K Whole Genome Human microarrays (Agilent). Top differentially expressed genes were validated on three independent cohorts by Taqman based RT-qPCR and immunohistochemistry. Collagen-induced arthritis (CIA) and Experimental autoimmune encephalomyelitis (EAE) experiments were conducted using Bob1 knockout mice and their littermate controls.−/− mice were fully resistant to CIA induction compared to their wild-type littermates. This remarkable protection from CIA is explained by failure to produce pathogenic anti-collagen autoantibodies in the absence of Bob1. In contrast, Bob1−/−mice were susceptible to MOG protein induced EAE and incidence and severity of clinical disease were not altered in these mice comparing to wild-type littermates, suggesting that absence of Bob1 does not impact on antigen-presentation/costimulatory capacity of B cells.Microarray screening for genes differentially expressed in the inflamed synovium, the key target of the disease process in RA, revealed a prominent and disease-specific B cell/plasma cell signature with the B cell-specific transcriptional co-activator Bob1 and its transcriptional target BCMA among the most upregulated genes. Validation by RT-qPCR on two independent cohorts representing early and established arthritis confirmed microarray data and demonstrated elevated expression of Bob1 and BCMA not only in established RA, but also at the early phase of the disease. Quantitative evaluation of immunohistochemical stainings of synovial tissue with monoclonal antibody for Bob1 revealed significant increase in Bob1 positive cells in RA synovium (p<0.01). Next we determined whether lack of functional Bob1 modifies disease onset or severity in CIA. Interestingly, the results showed that Bob1The specific increase in Bob1 expressing cells in RA synovitis and the resistance of Bob1-defecient mice to development of CIA indicate that Bob1/BCMA axis may contribute to humoral autoimmunity in RA. The relationship between an aberrant Bob1 expression and the break of peripheral tolerance in RA is currently under investigation.


We investigated the effects of dorsal root stimulation (flexor related dL2 and extensor related dL5) on fictive locomotion evoked pharmacologically (by 5-HT+NMDA+DA) in the isolated neonatal mouse spinal cord. In our experiments, electrical stimulation produced a wide variety of effects depending on stimulation frequency, intensity, and the drug concentrations used. At stimulation intensities near threshold we were able to produce phase advances and delays that resolved within a single step cycle. During these single-cycle alterations we observed different effects after stimulating dL2 and dL5. Stimulation of dL2 during ipsilateral extension typically produced an early onset of the next flexion and termination of current extension in the ipsilateral activity with or without rhythm disturbances on the contralateral side of the cord (subject to drug concentrations and stimulus intensity). These disturbances represented a true phase resetting characterized by a full flexor phase expression independent of ending the stimulation producing the disturbance. In contrast, stimulation of dL5 often produced a complex bilateral effect starting from a brief activation of ipsilateral extension (for the stimulus duration) with a corresponding reduction in the flexor activity followed by a reactivation of ipsilateral flexor activity (a full second burst that could represent a rebound evoked by end of stimulation) and then by an enhanced next extensor burst. Our results were consistent with the previously proposed two-level architecture of the spinal central pattern generator (CPG) consisting of a top-level rhythm generator (RG) and pattern formation (PF) circuits ,3. The a


ERBB2. In this review, we will focus on the functional role of these somatic mutations that cause ERBB2 receptor activation. We will additionally discuss the current preclinical and clinical therapeutic strategies for targeting mutationally activated ERBB2.Despite the ongoing “war on cancer,” cancer remains one of the major causes of human morbidity and mortality. A new paradigm of targeted therapies holds the most promise for the future, making identification of tumor-specific therapeutic targets of prime importance. ERBB2/HER2, best known for its role in breast cancer tumorigenesis, can be targeted by two types of pharmacological manipulation: antibody therapy against the extracellular receptor domain and small molecule compounds against the intracellular tyrosine kinase domain. Aberrant activation of ERBB2 by gene amplification has been shown to participate in the pathophysiology of breast, ovarian, gastric, colorectal, lung, brain, and head and neck tumors. However, the advent of next-generation sequencing technologies has enabled efficient identification of activating molecular alterations of  Rising incidences of neoplasia worldwide are estimated to translate into 13 million cancer deaths by 2030 , HER3/ERBB3, and HER4/ERBB4  amplification and overexpression, (ii) molecular alterations of the receptor, and (iii) inhibition of phosphatase activity  revealed more rapid and acute activation of different signaling pathways compared with the full-length receptor and the 648-CTF fragment   was demonstrated to be active in patients with breast cancer who had progressed on trastuzumab therapy  were found to respond to neratinib  in lung epithelium revealed that the combination of afatinib and an mTOR inhibitor (rapamycin) were effective in mediating tumor shrinkage , a second irreversible inhibitor of EGFR and ERBB2, was demonstrated in Ba/F3 cells expressing an ERBB2-mutant with an insertional mutation at codon 776 and in transgenic lung cancer models , a third irreversible pan-ERBB inhibitor, is currently under clinical investigation due to promising preclinical studies  or with other targeted therapies  present valid options to combat ERBB2-driven oncogenesis.in vitro and in vivo experiments (Garcia-Garcia et al., Recent clinical data showed a significant overall survival benefit of patients with heavily pretreated metastatic ERBB2-positive breast cancer upon dual ERBB2 blockade through trastuzumab and lapatinib (Blackwell et al., Although activating mutations of ERBB2 were identified in various tumor types and several potential therapeutic options are at hand, specific screening for these lesions has not been translated into clinical routine yet.Despite the plethora of ERBB2 targeted compounds, we currently lack a sound understanding why tumor shrinkage is short-lived and only a relatively small percentage of patients benefit from these therapies. Major mechanisms of primary and acquired resistance to anti-ERBB therapeutics include (reviewed in Tortora, Retrospective studies on tumors expressing truncated p95HER2 fragments revealed that these tend to be resistant to any current therapeutic antibody approach as the required epitopes are missing (Scaltriti et al., In vitro analyses identified three point mutations, L755S, L755P, and T798M to confer resistance to lapatinib (Kancha et al., in vitro and in vivo (Kobayashi et al., Analogous to a commonly observed event during TKI treatment of EGFR-driven lung adenocarcinomas, lapatinib application was shown to induce secondary mutations within the ERRB2 kinase domain consequently leading to TKI resistance. During the past decades, the ERBB2 signaling cascade gained significant importance in the oncogenesis of many tumor types. The discovery of primary activating mutations and the emergence of acquired secondary mutations represent sophisticated challenges for effective treatment approaches. Our next steps in evaluating potential ERBB2-directed therapeutics clearly rely on: adequate diagnostic properties for specific patient selection and identification of tissue-specific mechanisms of resistance to initiate well-designed clinical trials of combinational treatment strategies.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Hypoxia/ischemia induces the neural stem cells (NSCs) proliferation in mammalian brain; but the mechanisms remain unknown.In this study, we investigated the effects of metabotropic glutamate receptor 5 (mGluR5) on NSC proliferation under hypoxia by 3--2,5-diphenyltetrazolium bromide (MTT) assay, diameter measurement of neurospheres, bromodeoxyuridine (BrdU) incorporation assay and cell cycle analysis. The cell death of NSCs was evaluated by terminal dUTP nick-end labeling (TUNEL) assay and Hoechst staining. The expression of cyclin D1 and the activation of mitogen-activated protein kinases (MAPKs) signaling pathway were analyzed by immunoblotting assay.The results showed that hypoxia promoted the mGluR5 expression on NSCs. Under hypoxia, mGluR5 agonist DHPG and CHPG significantly increased NSC proliferation in cell activity, diameter of neurospheres, bromodeoxyuridine (BrdU) incorporation and cell division, and expression of cyclin D1 with decreasing of cell death. mGluR5 siRNA and antagonist MPEP decreased the NSC proliferation and expression of cyclin D1 with increasing of cell death. Phosphorylated JNK and ERK increased with the proliferation of NSCs after mGluR5 agonist DHPG and CHPG treatment under hypoxia, while p-p38 level decreased.These results demonstrated that the expression of mGluR5 was upregulated during the proliferation of NSCs stimulated by hypoxia in vitro. The activation of ERK and JNK signaling pathway and the expression of cyclin D1 were increased in the process. These finding suggesting the involvement of mGluR5 in NSC proliferation and providing a target molecule in neural repair after ischemia/hypoxia injury of CNS.


Grid cells in the medial entorhinal cortex (MEC) encode location through firing fields that form grid-like maps of the environment. At the same time network activity in the MEC is dominated by oscillations in the theta 4-12 Hz) and gamma (30-100 Hz) bands. Our recent experimental data established that feedback inhibition between excitatory stellate cells and inhibitory fast spiking interneurons dominates the synaptic connectivity in layer II of the MEC, and that continuous attractor models derived from these properties are sufficient to explain both the network oscillations and grid firing fields [2 Hz and


To the Editor: I read with interest the article by Jones-Engel et al. (Because current evidence shows that SV40 infections are identified in some humans and that the virus is associated with selected human malignancies ( In response: Dr. Vilchez (Specificity of SV40 seroreactivity in macaques has recently been demonstrated by competitive inhibition assays (


To the Editor: Since 1987, morbillivirus  outbreaks among pinnipeds and cetaceans in the Northern Hemisphere have caused high rates of death (Phocine distemper virus (PDV) and Cetacean morbillivirus (CeMV). PDV has been isolated from pinnipeds, and 3 strains of CeMV  have been isolated from dolphins and whales (Tursiops truncatus) has been confirmed in the Southern Hemisphere (in the southwestern Pacific Ocean) (Lagenodelphis hosei) stranded off Brazil and Argentina in 1999 indicate the exposure of South Atlantic cetaceans to morbillivirus , a coastal marine and estuarine species that occurs off the Atlantic Coast of South and Central America. Serologic surveys indicate that morbilliviruses infect marine mammals worldwide  , a monoclonal antibody against the nucleoprotein antigen of canine distemper virus that cross-reacts with cetacean morbilliviruses (Morbillivirus spp.–specific primers were used for PCR: 5′-ATGTTTATGATCACAGCGGT-3′ (forward) and 5′-ATTGGGTTGCACCACTTGTC-3′ (reverse) (http://megasoftware.net/) was used to construct a neighbor-joining phylogenic tree based on the sequenced amplicon from this study (GenBank accession no. KF711855) and 12 other GenBank sequences that represent the 6 morbillivirus species already described in the literature. The analysis placed the Guiana dolphin strain at the CeMV clade, but segregated it from the already described dolphin morbillivirus strains PMV, DMV, and PWMV . We extracted RNA from frozen lung samples by using TRIzol Reagent  according to the manufacturer’s instructions and amplified a 374-bp conserved fragment of the phosphoprotein (P) gene by reverse transcription PCR. The following and PWMV . The samIn summary, sequence analysis of the morbillivirus from the dead Guiana dolphin suggests that the virus is a novel strain of the CeMV species; this conclusion is supported by phylogenic analysis and geographic distribution of the virus and by its distinct host. Emaciation, marked lymphoid depletion, interstitial pneumonia, and meningoencephalitis are common findings in morbillivirus-infected animals (Morbillivirus outbreaks have caused a high number of deaths among pinnipeds and cetaceans and are a major risk to previously unexposed nonimmune populations of aquatic mammals (


Atrial fibrillation (AF) is the commonest sustained arrhythmia afflicting the adult population. Identifying and ablating triggers in the left atrium (LA) or more often the pulmonary veins (PV) with radiofrequency energy to restore sinus rhythm is fast becoming the mainstay of treatment. The success of the procedure often requires a combination of imaging techniques with an electroanatomical mapping system. We describe the use of three dimensional (3D) delayed enhancement cardiovascular magnetic resonance imaging (DE-MRI) to provide a road map of the anatomy as well as regions of scar tissues in the LA and PV for AF ablation.Images were acquired with a Siemens Avanto 1.5 tesla scanner. Standard acquisition of cardiac anatomy and function were performed as well as the atrial short-tau inversion recovery (STIR) sequences in the axial plane. In addition, 3D steady-state free precession (SSFP) axial plane, 3D magnetic resonance angiography (MRA) coronal plane and 3D DE-MRI axial plane of both LA and PV were acquired.The 3D SSFP axial plane, 3D MRA coronal plane and 3D DE-MRI axial plane of both LA and PV obtained were then incorporated into an electroanatomical mapping system which generated a 3D road map of the LA and PV together with regions of scar tissues as shown by DE-MRI. The merging of this road map with the electroanatomical map enables AF ablation to be done effectively.Together with the electroanatomical mapping system, 3D DE-MRI provides an invaluable non-invasive assessment of the anatomy and scar tissues of the LA and PV for AF ablation.No external funding.


Idiopathic toe walking (ITW) has been associated with ankle equinus, and while equinus has been linked with foot deformity in adults, there has been limited investigation on its impact on structural foot change in children. This study used the weight bearing lunge (WBL) test and Foot Posture Index-6 (FPI-6) to evaluate the foot and ankle measures of children with an ITW gait.Sixty children between the ages of four and eight years were grouped into an ITW (N=30) and a non-toe walking (NTW) (N=30) cohort. The ankle range of movement and FPI-6 was calculated during appropriate weight–bearing test and stance.There was a highly significant difference in the WBL test measures between the ITW cohort and the NTW cohort. The FPI-6 comparison was not significant. The WBL test was also not predictive of an abnormal FPI-6 in the ITW cohort.These results demonstrate that ITW gait style impacts only on the available dorsiflexion at the ankle. The WBL measure may be utilised within the clinical setting to guide and monitor treatment interventions.


Staphylococcus aureus. In an adaptive response, bacteria have evolved systems to acquire the metals in the face of this competition while effluxing excess or toxic metals to maintain a bioavailability of transition metals that is consistent with a particular inorganic “fingerprint” under the prevailing conditions. This review highlights recent biological, chemical and structural studies focused on manganese (Mn) acquisition and homeostasis and connects this process to oxidative stress resistance and iron (Fe) availability that operates at the human host-pathogen interface.Pathogenic bacteria acquire transition metals for cell viability and persistence of infection in competition with host nutritional defenses. The human host employs a variety of mechanisms to stress the invading pathogen with both cytotoxic metal ions and oxidative and nitrosative insults while withholding essential transition metals from the bacterium. For example, the S100 family protein calprotectin (CP) found in neutrophils is a calcium-activated chelator of extracellular Mn and Zn and is found in tissue abscesses at sites of infection by  The most abundant transition metals in humans are iron (Fe) and zinc (Zn) , Mn(IV) and Mn(V)] and is most strongly linked to oxidative stress resistance mediated by Mn superoxide dismutase (Mn-SOD) . This is generally expressed in nmol or ng of each metal per mg total protein and is readily measured by inductively coupled plasma mass spectrometry (ICP-MS) of acid-solubilized cells. A remarkable aspect of the inorganic fingerprint relevant to this discussion is the fact that the total Mn:Fe ratio varies by over seven orders of magnitude when various single-celled organisms are compared with one another  when grown in the presence of oxygen  and Fe(II), if left unregulated  metal ions to the exclusion of all others  or hydrogen peroxide (H2O2)  atoms from enzymes harboring mononuclear Fe and 4Fe-4S cluster cofactors  dissociation, essentially analogous to that which occurs in the peroxide sensor Fe-PerR  H2O2 induces the OxyR regulon which includes genes encoding a manganese import pump MntH and the DNA binding iron-scavenging protein Dps .Manganese as a micronutrient is critical to the viability and virulence of many Gram-positive and Gram-negative bacterial pathogens . First, increased Mn(II) availability allows for increased metallation of Mn-SOD which efficiently catalyzes the dismutation of the superoxide anion radical to Hi) and Mn-carbonate complexes are efficient catalysts of superoxide disproportionation and the chemical mechanism of this reaction has recently been investigated in detail  to one based on Mn(II) in order to protect key enzymes from inactivation by ROS. Another example of this type of cofactor replacement is the iron sequestration protein, Dps, in Kineococcus radiotolerans  via cofactor substitution in the oxidative stress response  functions as an antioxidant through a combination of enzymatic degradation of oxidants by Mn-SOD and other Mn(II)-containing enzymes, nonenzymatic degradation of oxidants by LMW-Mn complexes, and metalloenzyme cofactor substitution to prevent Fe-induced peroxide chemistry and subsequent enzyme inactivation Figure . The exto Figure , may repo Figure .The ability of a bacterial pathogen to obtain sufficient Mn(II) is critically important for pathogenesis and as such, Mn(II)-dependent metal sensor proteins control the expression of operons that encode additional virulence factors unrelated to the acquisition of Mn(II)  are known to modest resolution  reveal essentially closed, metal bound-like structures, with BtuF indicative of a slightly more open and conformationally dynamic structure that collapses around the Co(II)-ligand complex -specific cluster A-I SBPs have been reported including those from distant bacterial phyla such as cyanobacteria  depending on the transporter  is often too tight to measure at the protein concentrations required to make the measurement . As can be seen, determined KdMn values range from the low nM to several hundred nM, with some indication that Zn(II) may bind more weakly than Mn(II)  is robust since this value was extracted from a nonstoichiometric binding isotherm acquired in the presence of 50 mM citrate as a Mn(II) competitor chelator  from the SBP into the transport cavity  can only enter the cytoplasm efficiently through Mn(II)-specific transporters, elucidation of the affinity of each for Mn(II) vs. noncognate Zn(II) and the rates at which Mn(II) is transported across the plasma membrane takes on added significance when considered in the context of the discovery of calprotectin (see below). This is also true from the perspective of fundamental inorganic chemistry since Mn(II) complexes will tend to be far less thermodynamically stable than “isostructural” Zn(II) complexes Figure  Waldron. Metal tt Figure , despiteE. coli, Mycobacterium tuberculosis, or Bacillus anthracis in this case) that secretes these siderophores to capture bioavailable Fe from the host  and Zn(II) to sequester these metals from the invading bacterium. For example, the S100A7 homodimer limits growth and invasion at epithelial surfaces through chelation of Zn(II) . One simple explanation is that more resistant bacteria express Mn(II) uptake systems that possess a higher affinity than CP for Mn(II) and thus will compete more effectively with CP for extracellular Mn(II). This requires knowledge of the structure and Mn(II) and Zn(II) binding affinities of CP. Initial studies established that a single S100A8/S100A9 heterodimer is capable of binding two molar equivalents of transition metal, designated S1 and S2; however, while Zn(II) could fill both sites, Mn(II) could fill only one with high affinity (S1)  which is well-modeled in metal binding experiments as two functionally independent heterodimers  as a competitor ligand for Zn(II) revealed KdZn1 = 0.13 nM and KdZn2 = 185 nM (–Ca) and KdZn1 ≤ 0.01 nM and KdZn2 ≤ 0.24 nM (+Ca)  Mn(II) site conforming to octahedral coordination geometry  site, thus likely connecting Mn(II) sequestration to the biological activity of CP. Although CP may have a lower affinity for Mn(II) than bacterially encoded SBPs  transport into the cytoplasm  with significantly higher affinity (≥100-fold), a finding consistent with the Irving-Williams series  and Fe(II) can influence Mn(II) acquisition and intracellular Mn(II)-dependent metalloregulation of transcription. For example, Zn(II) has been shown to inhibit Mn(II) uptake by binding irreversibly to s Figure  - or Mn(II)-specific metal-sensing repressor -MntR and two related Fur family members, the peroxide sensing, Fe(II)-binding PerR  levels to rise to a level sufficient level to fill the mutant PerR regulatory site with cognate Fe(II)  and Mn(II) homeostasis systems may well be more relevant to bacterial cell physiology and pathogenesis than is Zn(II)-Mn(II) crosstalk. These two metals lie at the same weakly competitive end of the Irving-Williams series and the Fe:Mn ratio might be considered a reporter of microbial lifestyle, capable of altering the altering the resistance of an organism to ROS -specific in vivo. Remarkably, this Fe(II)-specificity is lost in a perR mutant strain. Here, the combined impact of increasing Fur concentrations and intracellular Mn(II) levels relative to Fe(II), leads to conditions where Fur binds Mn(II), which in turn, leads to inappropriate Mn(II)-mediated repression of the fur regulon, including genes responsible for Fe(II) uptake. Thus, PerR may directly impact Fe(II) homeostasis by modulating Fur levels in response to a change in the Mn:Fe ratio, i.e., that which might occur under conditions of high Mn(II) and Fe(II)-deplete conditions :Fe(II) ratio :Fe(II) ratios under conditions of low iron is also critical to virulence in sepsis models  effluxer, MntE, that is required for virulence -Mn(II) crosstalk influences bacterial pathogenesis is not firmly established, a number of recent studies suggest that maintenance of an optimal Mn(II):Fe(II) ratio can impact the virulence of pathogenic bacteria. For example, the ability of Recent studies of bacterial transition metal physiology and crosstalk places manganese acquisition by human microbial pathogens on center stage of the host-pathogen “arms race”  proteins attached on the cell surface known to bind zinc -specific uptake systems compete for the same metal on the basis of their respective affinities, there is much more to be learned about this process. This includes elucidation of the rates and rate-limiting steps of Mn(II) transport, and structural studies of intact bacterial ABC transporters positioned at the “front line” of Mn(II) acquisition. This is particularly interesting since Mn(II) is generally handicapped relative to other divalent metal ions, notably Zn(II), in chelate stability, and as a result, other factors including formation of kinetically trapped Mn(II) metalloenzyme complexes in the cell (Whittaker, Further insights into molecular mechanisms of host nutritional immunity against bacterial pathogens will continue to rely on concerted and collaborative efforts of microbiologists, coordination chemists and structural biologists in an effort to win the “tug-of-war” over transition metals at the host-pathogen interface through the development of intervention strategies based on metals in biology of infectious disease.John P. Lisher and David P. Giedroc wrote the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Acinetobacter baumannii(CRAB) is an emerging pathogen of healthcare associated infection and little is known about the effectiveness of the active surveillance culture of CRAB.Carbapenem resistant This study aims at evaluating the effect of active surveillance culture of CRAB upon intensive care unit(ICU) admission on the occurrence of new CRAB bacteremia in ICU.Since February 2011, the active surveillance culture of CRAB was performed in all patients admitted to medical ICU in Seoul national university hospital. Contact precaution was applied for the patients who had positive surveillance culture results. Respiratory specimen were obtained and the isolates were cultured overnight in blood agar plates which contain imipenem. To assess the effectiveness of active surveillance culture of CRAB and contact precaution, the rate of new CRAB bacteremia was compared between baseline period(February 2012-January 2011) and intervention period(June 2011-February 2012). The new CRAB bacteremia case was defined as the CRAB bacteremia occurred later than 48 hours of ICU admission in patients without positive CRAB clinical culture in the prior 12 months and within the 48 hours of ICU admission.P=0.019).In the baseline period there were 242 total admissions to the ICU(7355 patients-days) and in the intervention period there were 266 total admissions to the ICU(5432 patients-days). During the intervention period, 21(7.9%) patients showed positive CRAB surveillance culture results. The rate of new CRAB bacteremia is 2.72 cases per 1000 patients-days in the baseline period and 0.92 case per 1000 patients-days in the intervention period(When active surveillance culture of CRAB and contact precaution for the patients of positive results were applied in a medical ICU, the rate of new CRAB bacteremia was lowered and the time betweennew CRAB bacteremia and ICU admission was lengthened.None declared.


In this work we present a large-scale three stage model of the early mammalian olfactory system, including the olfactory epithelium (OE), the olfactory bulb (OB) and the olfactory (piriform) cortex (OC). All neurons in the network are modeled with a single or few compartments using the Hodgkin-Huxley formalism and are implemented in the NEURON simulator for parallel execution ,2. We inThe OE model comprises families of olfactory receptor neurons (ORNs) with different sensitivities, each family expressing one type of olfactory receptor (OR) with a vector of affinity values for each ligand . These dResults from runs with network sizes comprising thousands of model neurons show that this biophysically plausible network model generates response patterns of cells reminding of their real counterparts (see Figure


Loss of phosphatase and tensin homologue (PTEN) function evaluated by loss of PTEN protein expression on immunohistochemistry (IHC) has been reported as both prognostic in metastatic colorectal cancer and predictive of response to anti-EGFR monoclonal antibodies although results remain uncertain. Difficulties in the methodological assessment of PTEN are likely to be a major contributor to recent conflicting results.We assessed loss of PTEN function in 51 colorectal cancer specimens using Taqman® copy number variation (CNV) and IHC. Two blinded pathologists performed independent IHC assessment on each specimen and inter-observer variability of IHC assessment and concordance of IHC versus Taqman® CNV was assessed.Concordance between pathologists (PTEN loss vs no loss) on IHC assessment was 37/51 (73%). In specimens with concordant IHC assessment, concordance between IHC and Taqman® copy number in PTEN loss assessment was 25/37 (68%).Assessment PTEN loss in colorectal cancer is limited by the inter-observer variability of IHC, and discordance of CNV with loss of protein expression. An understanding of the genetic mechanisms of PTEN loss and implementation of improved and standardized methodologies of PTEN assessment are required to clarify the role of PTEN as a biomarker in colorectal cancer. Survival for patients with metastatic colorectal cancer (mCRC) has improved significantly over the past 15 years, largely due to improved systemic treatment options-6. In adPTEN is an important negative regulator of PI3K/AKT pathway and controls cell proliferation, survival and angiogenesis. Loss of PTEN function leads to persistent activation of the PI3K pathway and has been observed in breast, prostate, glioblastoma, endometrial and colon cancers,13. LossSeveral crucial factors make testing and interpretation of PTEN difficult. Loss of PTEN function results from several genetic mechanisms including small scale PTEN gene mutations , allelic loss at chromosome 10 and epigenetic silencing via hypermethylation of the PTEN promoter region. PTEN geFurther complicating the situation, the frequency of loss of PTEN expression increases from progression from normal colonic mucosa to adenoma, primary CRC and ultimately metastasis. The resClearly the role of PTEN is more complex than KRAS gene mutation where a single identifiable mechanism (activating mutation), largely concordant between primary and secondary tumours, confers near complete resistance to anti-EGFR MoAbs. Understanding this complexity is central to interpreting the current literature relating to PTEN and its potential role as a predictive biomarker. Recently reported cohorts of mCRC patients receiving anti-EGFR MoAbs have used PTEN loss of IHC expression to report loss of PTEN function. While this represents the functional outcome of several genetic mechanisms of PTEN loss, IHC relies on subjective interpretation and has the potential for inter reporter variation. Furthermore there is variability over the definition of 'loss of PTEN’ based on IHC scoring. In the largest cohort of mCRC patients, PTEN loss was defined as no staining in any cells at any intensity, while oOur group undertook an analysis of PTEN status in the AGITG MAX study of mCRC patients to identify the rate of inter-observer variability in IHC assessment and the rate of discordance between IHC and PCR assessment of PTEN status.The MAX study design and eligibility criteria have been reported previously.The primary objective of this Phase III randomized trial was to evaluate the effect of adding bevacizumab to capecitabine (with or without mitomycin C) on progression free survival (PFS) among patients receiving first line chemotherapy for mCRC. Four hundred and seventy-one patients were enrolled between July 2005 and June 2007. We have used the TaqMan® Copy Number Assay  to assess for PTEN allelic loss and have previously reported that loss of PTEN copy number was not prognostic nor predictive of outcome in the MAX trial cohort. In thisFormalin-fixed, paraffin-embedded (FFPE) samples of tumor tissue from archival specimens collected at the time of diagnosis were retrieved from storage at hospital pathology departments. For Copy Number PCR, genomic DNA was extracted from FFPE tissue sections with the use of the QIAamp DNA FFPE tissue kit . Manual micro-dissection was performed on samples with less than 80% malignant cells when visualized by microscopy. The same tissue blocks were used to make tissue microarrays (TMAs) and were assessed for PTEN expression by IHC. Researchers who assessed PTEN IHC expression were blinded to the PCR results.Immunohistochemical staining was carried out on TMAs using the PTEN monoclonal antibody 6H2.1  that has been used previously-36. EssePTEN, at cytoband 10q23.31a, location Chr.10:89727445 on NCBI build 37 (Life Technologies). The assay is a duplex PCR for the PTEN gene and the reference gene, RNaseP , set up according to the supplier’s protocol and run on the Rotorgene 6000 real time PCR instrument . The results are calculated as a ratio relative to a 2-copy control using the 2-∆∆Ct method (Rotorgene software), and multiplied by 2 to give the copy number. We tested DNA from colon cancer cell lines to determine the reproducibility of the assay and to select cell lines to use as copy number controls. HT29 (ATCC) is known to have 3 copies of chromosome 10 as determined by spectral karyotyping and comparative genomic hybridization[The PTEN TaqMan® copy number assay  was performed using 10 ng DNA in quadruplicate PCR. The primers provided in the assay were entirely within exon 9 of dization. For theFifty-nine tumor specimens were analysed for loss of copy number by Taqman® and for loss of protein expression by IHC. Eight samples were found to contain no tumor tissue and were excluded from further analysis..Two blinded pathologists assessed 51 specimens independently for PTEN protein expression with IHC. Pathologist JC assessed 29/51 (57%) as having PTEN expression loss, while pathologist AR assessed 17/51 (33%) as having loss of PTEN expression. Concordance between pathologists on final IHC assessment (PTEN loss/no loss) was 37/51 (73%), indicating in 14/51 (27%) of specimens there was discordance in the final assessment of IHC PTEN loss  had ≤1.5 copy number and were thus classified as PTEN loss.® PCR concordance analysis. Fifteen specimens had PTEN loss on IHC of which 10 (67%) also had PTEN allelic loss on Taqman® PCR. Seventeen specimens had PTEN allelic loss on Taqman® PCR of which 10 (58%) had PTEN loss on IHC. Fifteen specimens had preserved PTEN on both IHC and Taqman® PCR analysis. Overall concordance between IHC and Taqman® copy number in PTEN loss assessment was 25/37 (68%)  whether protein expression is reduced and whether such reduction confers a growth advantage is unknown. Sood et al. also demonstrated monoallelic PTEN dysfunction (by mutation or promoter methylation) resulted in loss of protein expression in only 38% of samples, while biallelic inactivation resulted in loss of PTEN expression in 80% of cases. Ali et In our cohort 25% of cases without PTEN allelic loss demonstrated complete absence of PTEN expression on IHC. These findings confirm alternative genetic mechanisms, beyond allelic loss, are responsible for loss of PTEN protein expression. Several authors have undertaken more comprehensive analysis of PTEN status on CRC specimens and provide an important insight into the often coexisting genetic mechanisms of PTEN dysfunction. Goal et al. demonstrated hypermethylation of the PTEN promoter region occurred in 10/132 (7.6%) sporadic CRC specimens, with a higher rate (19.1%) in microsatellite unstable CRCs. PTEN mutations coexisted in 4/10 (40%) of hypermethylated PTEN specimens. Eighty percent of patients with promoter hypermethylation had reduced (+1) or loss of PTEN protein expression and in the 3 cases of complete loss of PTEN staining, promoter hypermethylation coexisted with PTEN mutation or allelic loss. PerroneAlternatively, focusing on loss of protein expression at least represents the functional outcome of any such genetic insult. We have demonstrated the current limitations of IHC for this purpose. In our cohort, IHC assessment of PTEN loss by two pathologists was 33% and 57%, with overall concordance of 73%. As this was designed as a validation subset we did not ask the two pathologists to discuss the results that were not concordant, nor seek a further opinion, methods commonly described in papers reporting PTEN IHC aimed at reducing the apparent discordance rate. In all Overall the current literature highlight the difficulty in accurately measuring PTEN function to date; measurement of a single genetic insult, while minimizing inter-observer variability, does not capture the often coexisting mechanisms required for biallelic inactivation. The use of IHC, while potentially a better measure of PTEN function, is observer dependent and there remains a lack of consensus on optimal methodology and scoring.Given the limitations of PTEN assessment discussed here, it is not surprising reports of the predictive value of PTEN as a biomarker in CRC remain conflicting,17,27,31The lack of standardization in assessing loss of PTEN function appears to have contributed significantly to the conflicting results from retrospective cohort studies. Further elucidation of PTEN as a potential biomarker for colorectal cancer relies on defining PTEN loss of function and standardizing analytical methods and scoring systems. Future studies assessing PTEN function may be better served by obtaining a more comprehensive analysis of PTEN function by assessing PTEN mutation, hypermethylation of PTEN promoter, PTEN allelic loss and protein expression on each specimen. An alternative approach may be to explore improved methods of measuring reduced protein expression beyond IHC, given reduced or absent protein expression should reflect the functional outcome of PTEN loss irrespective of the genetic mechanism. Immuno-PCR may provide an option of combining the protein-specific capability of antibodies with the objective quantification of real-time PCR. This wiCNV: Copy number variation; CV: Coefficient of variability; EGFR: Epidermal growth factor receptor; FFPE: Formalin-fixed, paraffin-embedded; FISH: Fluorescent in situ hybridization; IHC: Immunohistochemistry; mCRC: Metastatic colorectal cancer; MoAbs: Monocloncal antibodies; PTEN: Phosphatase and tensin homologue; TMAs: Tissue microarrays; VEGF: Vascular endothelial growth factor.The authors declare that they have no competing interest.CH drafted the manuscript. JH conceived the study and coordinated all laboratory aspect of the study. VB participated in the design of the study and assisted with writing of manuscript. JW was involved in performing the DNA isolation and copy number PCR. NT contributed to trial conception and design as well as assisting with manuscript preparation. JC undertook the immunohistochemistry of the specimens and contributed to study design. AR undertook the immunohistochemistry of the specimens and contributed to study design. CL undertook data analysis and statistical analysis. TP conceived the study and participated in its design and coordination of all aspects of the study including drafting of the manuscript. ART designed study and assisted with manuscript. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2407/14/128/prepub


The study is aimed at i) optimization of techniques for preparing microcapsules; ii) generation of multicellular tumor spheroids (MTS) by culturing tumor cells in the microcapsules; iii) study of anticancer treatment effects for both photodynamic therapy (PDT) and anti-cancer drug screening. The model allows to estimate drug doses or parameters for PDT To form MTS, tumor cell lines  were encapsulated in polyelectrolyte microcapsules (200-600 μm), and cultivated for 3-4 weeks . MicrocaMTS based MCF-7 cells were prepared and used to study effects of PDT. To study the effect of irradiation parameters on cell viability, 2 photosensitizers (PS), namely photosense and chlorine e6 were used. Phototoxicity of PS depended on PS concentration and light energy density in both monolayer culture (MLC) and MTS. Study of cell morphology in MLC and MTS before and after PDT revealed that light energy density increase within the range of 30-70 J/cm2 resulted in cell apoptosis. However, cell survival in MTS was much higher than this in the MLC. MTS were also used to test some antitumor therapeutics . An enhanced cell resistance in MTS compared to MLC both for normal and Dox-resistant cells  were observed. MTS were also proposed to evaluate cytotoxicity not only of novel therapeutics but also nanosized drug delivery systems .


NCC is considered as potential therapeutic target for several pathological disorders like neuropathic pain and stroke disease. For understanding mechanism of action at the atomic level crystal structure provide valuable inside but lack of crystal structure of ion channel lead sequence analysis of different types of voltage dependent Ca2+ channel (VDCC) and we found identical/similar active site residues, which was confirmed by site-directed mutagenesis analysis of L-type Ca2+ channel (LCC). Based on these observations, we have developed for the first time the homology model of the closed state of NCC receptor at the ligand-sensing region by using bacterial K+ channel receptor. Further, molecular docking using different dihydropyridine (DHP) blockers identified NCC receptor hot spot binding residues, which is in consonance with that of the LCC. These residues are potential for further biochemical investigations. To understand binding and stability behavior of NCC with the DHP (amlodipine) in closed state, 50 nano second molecular dynamics simulation in lipid bilayer membrane environment were carried out. This analysis revealed the closed state stabilizing by binding of ligand into inner part of S6 region.The voltage dependent N-type Ca


Post-zygotic aneuploidy is the prominent genetic feature of the human brain (1). Genomically mosaic brain results from that the excess of aneuploid neurons due to early developmental disturbances (somatic genome variations), abnormal cell cycle regulation and altered programmed cell death. As the result, aneuploidization of the brain is a likely susceptibility factor (mechanism) for brain disorders including Alzheimer’s disease.The proportion of aneuploid cells was determined in brain areas differentially affected by neurodegeneration  by molecular-cytogenetic and immunohistochemical techniques  in brain tissues of individuals with AD and controls as described ealier (2).Increased levels of aneuploidy (monosomy and trisomy) involving chromosome 21 and chromosome X was observed in AD brain. The high level of aneuploidy involving chromosome 21 was observed in the AD cerebrum and hippocampus. In total, the incidence of abnormal (aneuploid) neural cells was significantly higher in degenerating brain areas  comparing to the less degenerating area (cerebellum).Our data indicates that AD brain areas subjected to neurodegeneration are more significantly affected by aneuploidy . We propose that widespread postzygotic aneuploidization of selected brain areas is a mechanism for AD neurodegeneration. Such area-specific distribution of aneuploidy can be explained by the accumulation of aneuploid cells during postnatal life or abnormal selective pressure against non-aneuploid cells (3). Finally, these data provide for the speculation that acquired neural aneuploidy could be generated during both developing and adult neurogenesis/gliogenesis.


To the Editor: Diversity of genotypes among Pneumocystis jirovecii (human-specific Pneumocystis species) isolates mainly involves internal transcribed spacer (ITS) loci (Pneumocystis pneumonia (PCP). Studies of the DHPS locus have found mutations at positions 165 and 171, which confer potentially lower sensitivity to sulfonamides to mutant P. jirovecii organisms  from 15 PCP patients and in 15 air samples collected 1 meter from each patient’s head  of the 15 pulmonary samples; none of the15 patients had received sulfonamide treatment at the time of PCP diagnosis. These results were not unexpected because frequency of finding DHPS mutants in PCP patients in Paris who had no prior sulfonamide treatment is high (Several lines of evidence suggest that P. jirovecii genotypes in pairs of pulmonary and room air samples argue in favor of P. jirovecii exhalation by infected patients. The exhalation of P. jirovecii organisms emphasizes the risk for their nosocomial transmission. Our data provide additional arguments in favor of the application of measures to prevent the airborne transmission of P. jirovecii in hospitals.Matches of


Results of a recent National Insitute of Drug Abuse-funded pilot randomized controlled trial of Mindful Awareness in Body-oriented Therapy (MABT) for women in substance use disorder treatment showed improved substance use outcomes for MABT compared to treatment as usual. In addition 80% of the MABT participants maintained a daily mindful body awareness practice at nine month follow-up. The purpose of this subsequent study was to explore the perceived role of MABT and mindful body awareness practice in the recovery process.A qualitative design utilizing a focus group method was employed. Participants from the larger RCT that had completed the MABT intervention (n =18) were recruited for the focus group, and five attended. Semi-structured questions were used to explore participants’ utilization of mindful body awareness practice in substance use disorder recovery, and the usefulness of various aspects of MABT delivery. Content analysis was used. Two researchers individually coded for themes across participant responses and together verified the coded themes.The primary themes were: (1) motivation to maintain body awareness practices one year post-intervention was due to improved ability to access and process emotions when using these practices; (2) exploration of uncomfortable feelings was perceived as critical for relapse prevention, as substance use was associated with escalated emotions and an inability to attend to feelings; (3) MABT facilitated awareness of inner experience and the connection between emotions and the body; they had not been able to achieve this awareness with prior therapeutic approaches. Also, three aspects of MABT were considered critical for learning mindful body awareness practice: (1) individual delivery to allow for safe exploration of emotional experiences, (2) manual/touch-based processes to learn interoceptive skills, and (3) homework to integrate body awareness practice into daily life.These findings point to the importance of studying emotional awareness in mind-body interventions, and in women’s substance use disorder prevention and treatment specifically.


Treatment of chronic urticaria consists of antihistamines as the first-line treatment. For more severe symptoms, combinations can be necessary as well as dose augmentations. The recent guidelines suggest the possibility of using omalizumab in resistant cases. We treated 2 patients with cold-urticaria (CO), 1 with cholinergic urticaria(CH) and 11 with chronic spontaneous urticaria (CSU) with omalizumab, who had not benefited from the recommended first-line, second-line and third-line treatments.Patients were required to document their CU symptoms once daily with urticaria activity scores for 7 days (7UAS). Briefly, the symptoms were monitored in terms of numbers of wheals , and the intensity of their pruritus , for a total of 42 points. To evaluate the efficacy of the omalizumab treatment, 7UAS obtained at baseline was compared to that at the third and sixth month of the therapy. Omalizumab was given at 150 μg/month irrespective of IgE levels and increased to 300 mg if needed (no response). The concomitant medication was slowly reduced according to clinical response.The 7UAS improved significantly in all severe urticaria patients with omalizumab as early as one month (not shown) after initiation of therapy and was sustained for the 6 month observation Figure . 7UAS inOur results show that omalizumab improves significantly recommended treatment-resistant urticaria patients (13/14) in terms of clinical symptomatology (7UAS) and drug reduction in a real life setting. None of the patients reported any adverse effect.