Contemporary inhabitants of the Balkan Peninsula belong to several ethnic groups of diverse cultural background. In this study, three ethnic groups from Bosnia and Herzegovina - Bosniacs, Bosnian Croats and Bosnian Serbs - as well as the populations of Serbians, Croatians, Macedonians from the former Yugoslav Republic of Macedonia, Montenegrins and Kosovars have been characterized for the genetic variation of 660 000 genome-wide autosomal single nucleotide polymorphisms and for haploid markers. New autosomal data of the 70 individuals together with previously published data of 20 individuals from the populations of the Western Balkan region in a context of 695 samples of global range have been analysed. Comparison of the variation data of autosomal and haploid lineages of the studied Western Balkan populations reveals a concordance of the data in both sets and the genetic uniformity of the studied populations, especially of Western South-Slavic speakers. The genetic variation of Western Balkan populations reveals the continuity between the Middle East and Europe via the Balkan region and supports the scenario that one of the major routes of ancient gene flows and admixture went through the Balkan Peninsula. It is believed that part of the Illyrians was assimilated and the other part was forced to move south - into the territory of present-day Albania th until 19th century the Peninsula was under the Ottoman control The Balkan Peninsula has been continuously settled by anatomically modern humans (AMH) since the Upper Paleolithic era Today, the Western Balkan territory  is inhabDuring the last two decades the variation of uniparentally inherited markers such as mitochondrial DNA (mtDNA) and the non-recombining part of Y chromosome (NRY) have been exploited in population genetic studies in order to disentangle the problems of the diversity and dispersal of humans both in global and local contexts The study of the variation of mtDNA in the population of Bosnia and Herzegovina has shown - like in case of the variation of NRY - that the majority of detected mtDNA hgs among Bosnians belong to the common West Eurasian gene pool The aim of this study was to characterize, in a larger geographical context, the autosomal gene pool of eight Western Balkan populations from six countries - Bosnia and Herzegovina, Croatia, Serbia, former Yugoslav Republic of Macedonia, Montenegro and Kosovo. All studied samples were characterized also for mtDNA and NRY diversity. One of the main questions we address here is whether the whole genome approach with the accent on the variation of autosomal SNPs is in concordance with the information about genetic affinities of the populations of Western Balkan region, revealed by the studies of uniparental markers.Genome-wide autosomal markers of 70 Western Balkan individuals from Bosnia and Herzegovina, Serbia, Montenegro, Kosovo and former Yugoslav Republic of Macedonia . The genome-wide SNP data generated for this study can be accessed through the data repository of the National Center for Biotechnology Information – Gene Expression Omnibus (NCBI-GEO): dataset nr. GSE59032, 2>0.4) were excluded from the analysis in the window of 200 SNPs (sliding the window by 25 SNPs at a time). The final dataset consisted of 220 727 SNPs and 785 individuals from African, Middle Eastern, Caucasus, European, Central, South and East Asian populations  was reduced with the exclusion of East and South Asians and Africans, in order to increase the resolution level of the populations from the region of interest  for the same dataset used for PCA were estimated between populations, and regional groups for all autosomal SNPs, using the approach of Weir and Cockerham ST values for the populations specified in Pairwise genetic differentiation indices  between different ethno religious groups of Western Balkan region with Middle Eastern populations. The Ottoman rule over the Balkans during 15–19 cc AD led inter alia to the conversion of the local people to Islam, the largest number of whose assumed descendants live in contemporary Bosnia and Kosovo ibd sharing for each of these populations and Middle Eastern populations . The details of the dataset has been characterized in The analysis was designed to compare patterns of shared tracts that are identical by descent (http://faculty.washington.edu/browning/beagle/beagle.html) ibd between pairs of individuals. The fIBD algorithm was applied to the 22 autosomes in 10 iterations and the IBD threshold was set to 1e–10. Since the power of the fIBD algorithm to detect segments shorter than 1 centiMorgan (cM) is low, we considered only ibd segments longer than 1cM. We summarized ibd sharing for six classes of ibd segments . We estimated an average number of ibd segments per pair of individuals for Muslim and non-Muslim populations of Western Balkan vs Middle Eastern populations  algorithm implemented in BEAGLE software package  and non-Muslim  of the Western Balkan populations share around 1.5 ibd segments per pair with the population from the Middle East  patterns of ibd sharing with Middle Eastern populations for shorter classes of ibd segments . This is slightly higher with Turks, and lower with Saudis, Syrians, Iranians and Palestinians  and only Bosniacs have higher ibd sharing with Turks (p = 0.0097 for 5–6 cM ibd segments)  we first analyzed the number of dle East . This isulations . Next, wstinians . For lonegments) . Howeveribd segments reveals similar patterns of ibd sharing for Muslim and non-Muslim Western Balkan populations with populations of Middle East, providing thereby little support to a gene flow scenario during the conversion to Islam (15–19 cc AD) in the Balkans. Our analysis of ibd sharing agrees with other analyses  of the Western Balkan populations, the influence of the clan structure to the present genetic variation should be seen the most in the Y chromosomal gene pool of the studied populations – this, however, is not the case. To conclude: the linguistic or religious differences seem to have had no impact on the present variation of uniparentally inherited or autosomal markers in a region.Compared to the rest of the Western Balkan populations, the Kosovars have a somewhat different cultural and demographic background. All studied Western Balkan populations, except Kosovars, belong to the South Slavic branch of the Indo-European (IE) language family n Balkan . Howevern Balkan , suggestn Balkan , Text S1We have analyzed and present here the new data of genome-wide autosomal diversity of five Western Balkan populations. The variation analysis of 660K autosomal SNPs of 70 individuals from Western Balkan populations revealed that the genetic uniformity that has been shown by studies of uniparentally inherited markers of these populations can be seen also at the whole-genome level. Thus, culturally diverse Western Balkan populations are genetically very similar to each other. These results, together with the high-resolution analysis of the variation of mtDNA and NRY, let us to affirm that the genetic profiles of Western Balkan populations resemble that of their closest geographical neighbors, and in the global context are in concordance with the geographical distribution of the studied population.The major variants of the gene pool of present-day Western Balkan populations have developed from a common source without being influenced by major population-specific bottlenecks. In a more general perspective, our results reflect clear genetic continuity between the Middle Eastern and European populations. It has been suggested recently that the Neolithic migrants from Anatolia took mainly the maritime coastal route and island hopping to reach Europe The autosomal analysis as well as mtDNA and NRY data presented in this study contribute to an existing database and for understanding the origins of the peopling of this part of Europe.Figure S1ADMIXTURE plots of autosomal SNPs of Western Balkan region in a global context on the resolution level of 3 to 15 assumed ancestral populations (K). A. Box and whiskers plot of the cross validation (CV) indexes of all 15×100 runs of ADMIXTURE; B. Log-likelihood (LL) scores of all 15×100 runs of ADMIXTURE. Inset shows the variation in the fractions  of runs that reached the highest LL values; C. Bar plot displaying individual ancestry estimates for studied populations.(TIF)Click here for additional data file.Figure S2Principal component (PC) analysis (PC1 versus PC2) of the variation of autosomal SNPs in Western Balkan populations (highlighted) in Eurasian context (see (TIF)Click here for additional data file.Figure S3Principal component (PC) analysis (PC1 versus PC3) of the variation of autosomal SNPs in Western Balkan populations (highlighted) in Eurasian context (see (TIF)Click here for additional data file.Figure S4Principal component (PC) analysis (PC1 versus PC4) of the variation of autosomal SNPs in Western Balkan populations (highlighted) in Eurasian context (see (TIFF)Click here for additional data file.Figure S5Principal component (PC) analysis (PC1 versus PC5) of the variation of autosomal SNPs in Western Balkan populations (highlighted) in Eurasian context (see (TIF)Click here for additional data file.Figure S6Principal component (PC) analysis (PC2 versus PC3) of the variation of autosomal SNPs in Western Balkan populations (highlighted) in Eurasian context (see (TIF)Click here for additional data file.Figure S7Principal component (PC) analysis (PC2 versus PC4) of the variation of autosomal SNPs in Western Balkan populations (highlighted) in Eurasian context (see (TIF)Click here for additional data file.Figure S8Principal component (PC) analysis (PC2 versus PC5) of the variation of autosomal SNPs in Western Balkan populations (highlighted) in Eurasian context (see (TIF)Click here for additional data file.Figure S9Principal component (PC) analysis (PC3 versus PC4) of the variation of autosomal SNPs in Western Balkan populations (highlighted) in Eurasian context (see (TIF)Click here for additional data file.Figure S10Principal component (PC) analysis (PC3 versus PC5) of the variation of autosomal SNPs in Western Balkan populations (highlighted) in Eurasian context (see (TIF)Click here for additional data file.Figure S11Principal component (PC) analysis (PC4 versus PC5) of the variation of autosomal SNPs in Western Balkan populations (highlighted) in Eurasian context (see (TIF)Click here for additional data file.Figure S12Principal component (PC) analysis based on the frequencies of mtDNA (panels A and C) and NRY haplogroups (panels B and D) of Western Balkan (WB) populations in a context of selected Central and South Europeans and Iranians from the Middle East Click here for additional data file.Figure S13Average total length of genome shared identical by descent between Bosniacs, Kosovars and Near Eastern populations. Panels A-E indicate five length classes of ibd segments: 1–2, 2–3, 3–4, 4–5, 5–6 cM, respectively. Bosniacs and Kosovars are tested Muslim populations from Western Balkans; Macedonians, Montenegrins, Bosnian Croats and Serbs, Croatians, Serbians are non-Muslim populations from Western Balkans, used as a background. Red color of the Western Balkan population name and red circle around the symbol of Middle Eastern population indicates significantly higher ibd sharing between these populations as compared to non-Muslim background.(TIF)Click here for additional data file.Figure S14Median-joining network of mtDNA hg H lineages of Western Balkan populations: A: subhg H1 and its sub-branches; B: other subhgs of hg H. A total number of 19 H1 and 49 other H haplotypes are reported. Numbers on links indicate the mutations: blue color indicates HVS1 and HVS2 mutations, black color coding region mutations. Polymorphic nucleotide sites are numbered according to Reconstructed Sapiens Reference Sequence. Node size is proportional to absolute haplotype frequency, as it is reported in figure legend.(TIFF)Click here for additional data file.Figure S15Median joining network of mtDNA hg U lineages. A total number of 34 U haplotypes are presented. A diagnostic mutation A1811G has been added into the network, but not genotyped in the sample. For further details, see the legend of (TIF)Click here for additional data file.Figure S16Median joining network of mtDNA lineages for hgs J and T in the Western Balkan region. A total number of 24 J and 10 T haplotypes are presented. For further details, see the legend of (TIF)Click here for additional data file.Figure S17Median joining network of mtDNA lineages of hgs HV, V and R0a. A total number of 11 V, 8 HV and 2 R0a haplotypes are reported.For further details, see the legend of (TIF)Click here for additional data file.Figure S18Median joining network of mtDNA lineages from hgs W, I and N1b. A total number of 6 W, 1 I and 3 N1b of haplotypes are reported. For further details, see the legend of (TIF)Click here for additional data file.Figure S19Phylogenetic tree of mtDNA K1a13a complete sequences. Two samples of Bosnian Croats (BHCB15 and BHCHZ20), and two from Croatia [Croatia.m.(S)199 and Croatia.m.(S)34] are sequenced in this study, the others are from Phylotree mtDNA Build 15. The mutations are given relative to the Reconstructed Sapiens Reference Sequence.(TIF)Click here for additional data file.Figure S20Phylogenetic tree of mtDNA N1a complete sequences. One sequence of Croatian from Croatia is sequenced in this study, the others are from Phylotree mtDNA Build 15. The mutations are given relative to the Reconstructed Sapiens Reference Sequence.(TIF)Click here for additional data file.Figure S21Phylogenetic tree of mtDNA R0a2 complete sequences. Macedonian (MAC16) and Croat from Bosnia and Herzegovina (BHCCB19) are sequenced in this study, the others are from Phylotree mtDNA Build 15. The mutations are given relative to the Reconstructed Sapiens Reference Sequence.(TIF)Click here for additional data file.Table S1Sample of populations used for autosomal analyses.(XLS)Click here for additional data file.Table S2The ethnolinguistic characteristics of studied Western Balkan populations.(XLS)Click here for additional data file.Table S3F3-statistic calculated for all possible triplets of f3 of TreeMix dataset.(XLSX)Click here for additional data file.Table S4Average number of IBD segment per pair of individuals.(XLSX)Click here for additional data file.Table S5MtDNA HVS-1 and HVS-2 haplotypes of analyzed Western Balkan populations relative to Reconstructed Sapiens Reference Sequence.(XLSX)Click here for additional data file.Table S6Y chromosome variation in Western Balkan populations.(XLSX)Click here for additional data file.Table S7MtDNA gene and nucleotide diversity of analyzed Western Balkan populations.(XLSX)Click here for additional data file.Table S8st-distances of mtDNA HVS1 variation between Western Balkan populations.F(XLSX)Click here for additional data file.Table S9Results of AMOVA and Mantel test based on mtDNA HVS-1 haplotype or haplogroup frequencies.(XLSX)Click here for additional data file.Table S10Estimated coalescense time for the most frequent mtDNA haplogroups in studied Western Balkan populations.(XLSX)Click here for additional data file.Text S1Description of the sample and methods of the analyses of mtDNA and NRY.(DOCX)Click here for additional data file.Text S2Results of the analyses of mtDNA and NRY variation.(DOC)Click here for additional data file.


In recent years, emerging evidence has shown the potential risks of arterial hyperoxia, but observational studies failed to indisputably demonstrate its impact on clinical outcome of critically ill patients. Importantly, the lack of a clinical definition of hyperoxia and methodological limitations hamper the interpretation and clinical relevance of these studies.We aimed to systematically evaluate previously used metrics for defining hyperoxia and associations with outcome of intensive care unit (ICU) admission.2). Worst PaO2 was defined as the PaO2 associated with the lowest PaO2/FiO2 ratio. In logistic regression models, we analysed the associations between hospital mortality and severe hyperoxia (PaO2> 200 mmHg) defined by either identified metric, calculated over the total ICU admission.Arterial blood gas (ABG) analyses between July 2011 and July 2014 were extracted from the patient data management system (PDMS) database of three large tertiary care ICUs in the Netherlands. Data from all admissions where more than one ABG was available were supplemented with anonymous demographic and admission and discharge data from the Dutch National Intensive Care Evaluation (NICE) registry. Identified oxygen metrics from a systematic review of the literature included the first, highest, worst, and mean arterial oxygen tension  died before hospital discharge. All identified metrics showed statistically significant associations between severe hyperoxia and hospital mortality. The risk estimates differed substantially between the metrics used for defining hyperoxia (P < 0.001). The strongest effects were found for the mean and the highest PaOIn this multicenter cohort study, severe arterial hyperoxia was associated with hospital mortality and this association was independent of the used metric for hyperoxia. However, the metric choice greatly influences the effect size for hospital outcome.This work was financially supported by an unrestricted grant issued by the Netherlands Organization for Health Research and Development (ZonMw).ADDIN EN.REFLIST


Patent foramen ovale (PFO) occurs in a quarter of the population but half of those with cryptogenic stroke (CS). Difficulty in identifying the pathogenic PFO versus the innocent bystander has contributed to controversy surrounding outcomes following PFO closure. We aimed to investigate whether right atrial flow patterns could help define the mechanism for CS in the setting of PFO.3, temporal resolution:50-55ms, 20 phases, SENSE, factor 2) was used. Analysis was performed with GTflow v2.0 (Gyrotools). Contours were placed manually in the superior vena cava (SVC) and inferior vena cava (IVC) in the axial plane at the junction with the right atrium. Net flow was assessed at these points. The relative position of the SVC and IVC at these points was measured. A pathline analysis of atrial flow was performed .4D flow cardiac magnetic resonance (CMR) was performed in 10 subjects  with proven PFO and CS and in 10 controls  with normal trans-thoracic echocardiograms. CMR was performed at 3T  with a 6-channel cardiac array. A retrospectively ECG-gated and respiratory-gated TFE sequence x240mm(FH)x142mm(LR), spatial resolution:3mmBody mass index (BMI) and heart rate (HR) were comparable but blood pressure was higher in the PFO/CS group with three patients taking anti-hypertensives (Table). Four variations of right atrial flow were observed Figure :a) vortical b) helico-vortical c) helical [SVC and IVC flow twist together in a helix]d) multiple vortices [arising variably from SVC and IVC flow]Vortical flow was most prevalent in the control group whilst the PFO/CS subjects were more likely to show one of the other flow patterns (Graph). Peak systolic and diastolic flow in both SVC and IVC were comparable between groups. However, peak diastolic flow in the IVC occurred earlier in the cardiac cycle when corrected for heart rate in the PFO/CS group than controls . The antero-posterior distance between the SVC and IVC was comparable between groups. The right-left distance between the SVC and IVC was greater in the PFO/CS group than the controls A spectrum of right atrial flow patterns can be seen in the normal heart. The relative position of the SVC and IVC may influence the generation of these. Our findings suggest that variant flow patterns in the right atrium may contribute to the pathogenesis of CS in the presence of PFO, we hypothesise, by promoting a passage for paradoxical embolism. Further work is needed to better define the subsets of CS patients in whom the PFO should be regarded as pathological.Academy of Medical Sciences and Newcastle Hospital Charities.


The frequency of contact allergens is often regionally different and hence it regional identification is important for the prevention of allergic contact dermatitis (ACD).Aim of this study was evaluation of the frequency of particular positive results of patch test (APT).A total number of 127 patients from Outpatient Allergic Clinic MED-ALL Medical Centre (age 2-76 years old) with suspected ACD were examined using the most common allergens from the patch tests from the European Standard Series including nickel, pallatium, chrome, cobalt, fragrance-mix I and II, balsam of Peru, formaldehyde and propolis and one extra allergen -- house dust mite (HDM). The data obtained were subjected to statistical analysis.The highest positivity of APT was seen against HDM (67%), then nickel (51%), chrome (40%), cobalt (39%), pallatium (16%), propolis (16%), fragrance-mix I , balsam of Peru , fragrance-mix II  formaldehyde . In the group of patients with positive results with HDM higher percentage is in younger group<18 years old (70%) with almost 68% of cases with strong positive results (++ or +++).HDM allergens play an important role in determining the clinical severity of allergic dermatitis and should be included in basic patch test series.


At present there are two methods to determine the chromosome health of unborn fetus in pregnant women. One is through maternal serum screening that is considered as simple but less sensitive method and the second options involves invasive methods like amniocentesis or chorionic villus sampling (CVS) for women who are found to be positive in maternal serum screening. Recent studies have demonstrated that non-invasive prenatal testing (NIPT) using cell-free fetal DNA (cfDNA) present in circulating maternal blood is considered as one the most effective method of screening for trisomy 21.The single nucleotide polymorphism (SNP) based Non-invasive Pre-natal test, being offered by Quest Diagnostics in technical collaboration with Natera Inc to determine chromosomal copy number by looking for specific patterns of (SNPs). This second generation technology analyzes cfDNA in a single reaction targeting 19,500 single nucleotide polymorphisms which are the most informative portions of an individual’s DNA selected across all the analyzed chromosomes.et al., 2013 paper on total of 242 cases that include thirty two cases aneuploidy included trisomy 21 (n=25) which were correctly identified with 100% sensitivity and 100% specificity with no false positive and false negative. Another study by Zimmerman et al., 2012 study on 166 samples from pregnant women, including 11 trisomy 21, three trisomy 18, two trisomy 13, two 45, X, and two 47,XXY samples including 146 low risk pregnancies. The study also correctly reported the chromosome copy number of all five chromosomes in 145 samples that passed a DNA quality test.Two published study demonstrated in the women undergone both invasive chorionic villus samplings followed by karyotyping in conjunction with non-invasive prenatal testing the efficacy of test. Data from Nicolaides The studies had demonstrated that SNP based NIPT is considered as an effective method of screening for common chromosomal abnormalities with detection rate of more than 99% and false positive rate of less than 0.1%. The initial finding shows NIPT approach has potential to avoid invasive procedure and can be provided an option to high risk pregnancies i.e. those with advanced maternal age, screens positive for biochemical screening and history of pregnancies with previous aneuploidies.


Addiction is associated with medical consequences, due to direct drug effects, poor adherence to health-care recommendations, and poor quality of care. But little is known about how the spectrum of illicit drug use in primary care patients affects quality of medical care measures for chronic conditions. The objectives of this study were to determine the association between drug use  and quality measures indicating failure to meet criteria for good blood pressure (BP) and blood glucose (BG) control among primary care patients identified by screening as using illicit drugs who also have hypertension and/or diabetes.The study population was adult patients presenting for primary care visits in an urban, safety-net hospital with recent illicit drug use or prescription drug misuse identified by screening and either hypertension or diabetes mellitus. The outcomes were failure to control: a blood pressure (primary), defined as systolic BP of 140 or higher or diastolic BP of 90 or higher, and b) blood glucose (secondary), defined as hemoglobin A1c of 8 percent or higher  and Healthcare Effectiveness Data and Information Set [HEDIS] measures). Main independent variables were: days of drug use in the past month; drug type ; and severity of use . We fit separate longitudinal logistic regression models for each measure of drug use and each outcome.Overall, 40 percent (66/164) of the sample with hypertension failed to meet criteria for BP control and 42 percent (26/62) of those with diabetes failed to meet criteria for BG control. No significant associations were detected for any measure of drug use and BP or BG control, except those reporting cocaine use had higher odds of failing to meet criteria for BG control compared to those reporting marijuna use (adjusted odds ratio (AOR) 8.8, 95% CI: 1.9, 41.9). Higher severity of drug use was associated with higher odds of failing to meet criteria for BG control in an unadjusted model ; however, this association was not significant after controlling for age, gender, and race/ethnicity.In this cohort of primary care patients with drug use, type of drug but not frequency of use or severity was significantly associated with failure to meet quality measure criteria for control of hypertension and diabetes.Trial registrationidentifying number NCT00876941


In nerve transfer, we transfer a physiologically active nerve to a distal, more important but irreparably paralyzed nerve. This surgical procedure is best done early within 6 months of the injury. Nerve transfer can be broadly classified into four categories: (1) Extraplexal; (2) Intraplexal and (3) Targeted nerve transfer.Prime extraplexal donors are the spinal accessory, phrenic and intercostal nerves. There are many combination of nerve transfer, depending upon the loss of action and available motors.For shoulder abduction: 1) Spinal accessory to supra-scapular: this transfer can be done from a dorsal approach; 2) Triceps branch of radial nerve to axillary from an anterior approachFor elbow flexion: 1) Spinal accessory to biceps and brachialis split sural nerve transfer: this extraplexal transfer involves using a reversed sural nerve graft and the two ends are used for the biceps and brachialis separately; 2) Ulnar to brachialis and median to bicepsFor Elbow extension: 1) Ulnar nerve to triceps; 2)Intercostal to TricepsFor wrist extension: Median nerve to ECRB branch of Radial nerveFor Finger extension: Transferring the FDS branch of median to posterior interosseous nerveFor distal Ulnar nerve neurotisation to prevent Claw deformity and sensation:Pronator quadratus branch of median to deep motor branch of Ulnar nerve


We found that non-functional alleles of white, unable to produce a coding transcript, could reactivate in trans the expression of a wild type copy of the same gene silenced by heterochromatin. This new epigenetic phenomenon of transcriptional trans-reactivation is heritable, relies on the presence of homologous RNA’s and is affected by mutations in genes involved in post-transcriptional gene silencing. Our data suggest a general new unexpected level of gene expression control mediated by homologous RNA molecules in the context of heterochromatic genes.In order to study the role played by cellular RNA pools produced by homologous genomic loci in defining the transcriptional state of a silenced gene, we tested the effect of non-functional alleles of the  trans-reactivation. We found that genes, unable to produce a functional coding transcript, but with the potential of transcribing other RNA’s within their gene body, strongly reactivate the transcription of a wildtype copy of the same gene silenced by heterochomatin. This new epigenetic phenomenon is heritable, relies on the presence of diffusible RNAs able to carry and transfer epigenetic information and is affected by mutations in genes involved in Post-Transcriptional Gene Silencing. Our data strongly suggest that homologous non-coding RNA can reactivate the expression of genes silenced by heterochromatin, thus defining a new unpredicted level of gene expression control in the context of heterochromatic genes.We discovered a new epigenetic phenomenon we called  In recent years it has become increasingly evident that the expression of eukaryotic genomes is far more complex than it had been previously explored. An emerging body of evidence, coming from next generation sequencing approaches, is showing that the genomes of all studied eukaryotes are almost entirely transcribed, generating an enormous number of non-coding RNAs (ncRNAs) . The ENCparamutation v) (C). All autosomal lines tested carried an X chromosome with a w1118 allele. In order to clean the lines from the w1118 allele, we back crossed the original lines with deletions of the w gene (Df(1)w), before testing them for their interaction with the w* suppressing alleles. To help follow the transmission of individual chromosomes, female chromosomes are represented in pink while male chromosomes are shown in blue.Detailed description of cross schemes and fly genotypes tested as experimental and control classes to measure (JPG)Click here for additional data file.S5 Fig(A) Loss of function mutations in smaug (smg1) and pumilio (pum13), two RNA binding protein involved in mRNA destabilization in flies, are embryonic lethal. Thus, the ability of smg1 and pum13 loss of function alleles to modify the levels of eye pigmentation (trans-reactivation) we scored in wm4h/w* females (+) was tested in heterozygosis (pum and smg). Polytene chromosome FISH using genomic probes covering the entire w gene (with the exception of the first intron) and coding sequences for the hsp70 gene (mapping chromosome 3R) on homozygous wwt/wwt(B), and transheterozygous wm4h/Δw(C) combinations. FISH signals for white (red) and hsp70 (green) genomic sequences are indicated by arrowheads. The asterisk indicate the region of pericentric heterochromatin. The X indicates the chromosome where the w gene maps.(JPG)Click here for additional data file.S6 Figtrans-generational inheritance of wm4htrans-reactivation from (F1) trans-heterozygous wm4h/w* females when crossed with parental w*/Y (A) or Δw /Y (B) males. To help follow the transmission of individual chromosomes, female chromosomes are represented in pink while male chromosomes are shown in blue. (C) Trans-generational inability of a wild type w+ allele (coming from an OreR stock) to trans-reactivate in F2 the wm4h locus. Eye pigment quantification and representative eye pictures of the parental (P), F1 and F2 progenies for each genotype tested are shown.Detailed description of cross schemes and fly genotypes tested as experimental and control classes to measure (JPG)Click here for additional data file.S7 Fig(A) and control (B) classes to measure trans-reactivation of wm4h during the process of gynogenesis. (C) Cross scheme used to test in heterozygosis the ability of loss of function alleles (Mut) to modify trans-reactivation. (D) Eye pigment quantification was conducted on adult flies of the indicated genotype and the Log2 ratio of wm4h/w* trans-reactivation in the mutant background over the normal levels of wm4h/w* trans-reactivation  was used as a measure of the influence of the mutation tested in the onset of trans-reactivation. To help follow the transmission of individual chromosomes, female chromosomes are represented in pink while male chromosomes are shown in blue.Detailed description of cross schemes and fly genotypes tested as experimental (JPG)Click here for additional data file.S1 Tablew* alleles tested for their ability to modify in trans-heterozygosity the wm4h eye color variegation. Stocks that increased eye color variegation are highlighted in orange (Suppressors), the ones that did not have any effect are highlighted in light red (-). Tested white genomic deletions are highlighted in light grey. Finally, the mutation class, the mutagen employed and the Nature of the lesion, for each w* allele tested is reported when known.Genotypes of stocks carrying the (DOCX)Click here for additional data file.


The prevention and treatment of caries and non-carious lesions by means of remineralization has been studied for many decades. The theoretical basis for remineralization as a means of caries prevention and treatment is based on the retention of protein matrix in dental enamel at early stage of caries (white spot) and on the possibility of its remineralization.The detection of new structural formations and emerging information about morph-chemical properties of human and animal enamel have facilitated identification of a new approach not only to for caries prevention but also the creation of new, more effective formulations for oral health hygiene as well as, structural and metabolic reparation of tooth enamel.For ultrastructure and morph-chemical tooth enamel reconstruction, we applied dentifrice gel ROCS Medical Minerals which contains calcium glycerophosphate, magnesium chloride and xylitol. Participants were 10–30 years old, without any significant medical conditions. Each patient received 15 applications of the gel. Total of 57 subjects were divided to 4 groups depending on the diagnosis. All the subjects had indications for remineralizing therapy.st group consisted of 15 patients with early stage of caries (white spot).• The 1nd group was made up of 19 patients with systemic hypoplasia (spotted form).• The 2rd group consisted of 10 young subjects with significant hypersensitivity without any visible lesions .• The 3th group consisted of 13 subjects with hypersensitivity due to wedge-shaped lesions, abrasion or enamel erosion.• The 4The application of gel containing the new mineral complex of magnesium and calcium resulted in a positive remineralizing effect which was obtained in patients with early stages of caries as well as those with non-caries lesions. Further, white spots (early stage of caries) completely disappeared in 80% cases after 15 applications of the dentifrice. Thus, it appears that 15 procedures of remineralizing therapy with ROCS Medical Minerals are enough for mineralizing of white hypoplastic spots. Most patients with hypersensitivity (82% and 93% in two groups) noticed significant reduction of painful reactions after two 15-minutes applications. Finally, all subjects noticed a significant improvement of teeth appearance .In conclusion, the results suggest high efficacy of the mineral magnesium-calcium remineralizing complex in patients with early stages of caries as well as those with non-carious lesions.


Recent studies in the United Kingdom ,2] have have2] hWe surveyed Scottish intensive care trainees to ascertain whether such deficits are widespread and persistent.Trainees were invited to complete a web-based questionnaire on three occasions in June 2014. Responses in the form of ticked boxes or free text were collated and analysed for trends.Ninety-nine trainees replied, of whom 96 were based in anaesthesia, one in emergency medicine and two in allied medical specialties. Alarmingly, 70 respondents (71%) had been involved in managing an airway emergency in a patient with a tracheostomy or laryngectomy, 63 (90%) on a critical care unit and 40 (57%) on a general ward. Deficits in staff training and equipment were identified as contributory by 28 (40%) and 10 (14%) respondents, respectively. Most trainees (68%) felt quite confident in caring for neck breathers, with confidence increasing as training progressed. However, the majority of trainees accrued such confidence through exposure to these patients on theatre lists (80%). Only a minority had participated in simulated emergencies (33%) or a formal training course (14%). Almost half (49%) rated their training as deficient or absent and the vast majority (96%) felt there was a place for more formal tuition in caring for patients with a tracheostomy or laryngectomy.Our findings suggest that we are failing to prepare trainees for the challenges of managing airway emergencies in neck breathers. Divergence of anaesthesia and intensive care training in the UK may further compound this problem by reducing exposure to controlled subglottic airway management in theatre. The Scottish Intensive Care Society Trainee Committee has sought to address these deficits in training and reduce the burden of avoidable harm by incorporating sessions on tracheostomy management into its educational programme.


High numbers of tumour infiltrating lymphocytes has been associated with a significantly improved prognosis. Some of the key cell types involved are Cork Cancer Research Centre (CCRC) has long utilised an ablative cancer treatment that involves delivering a pulse of electrical energy (electroporation) directly to the tumour. This treatment results in the formation of reversible pores in the cell membrane, allowing up to a 1000 fold increase in chemotherapy drug absorption (Electrochemotherapy). A significant benefit of this non thermal ablation is the release of tumour associated antigens into the microenvironment, leading to activation of APCs and subsequent cross presentation of the antigens to circulating T and B cells. While ECT facilitates the priming of a favourable immune response, the combination of iCOS antibody, capable of stimulating T cell proliferation, causes a very robust mediated regression of treated tumours. In addition, ICOS is involved in humoral immune responses  and increases the production of IL-4.+ and CD8+ T cells, CD45R+ B cells and CD11c+ DCs, post treatment.Preclinical work in aggressive murine models has yielded very encouraging results. The combinational therapy has achieved cures in both colorectal (CT26) and metastatic lung (LLC) cancer cell lines. Follow up immunohistochemical staining confirmed the presence of tumour infiltrating CD3


Sedation management during weaning from mechanical ventilation is an important task for ICU nurses. Nurse-led sedation protocols have a favorable impact on weaning outcome , 2.To identify sedation practices among ICU nurses and to determine barriers towards use of sedation protocols.A cross-sectional, self-administrated survey was developed. Consensus on content validity was achieved through a Delphi procedure among experts. The survey was distributed and collected during the annual congress of the Flemish Society of Critical Care Nurses (Dec. 2014).423 attendants completed the survey (response rate: 66%) of which 342 ICU nurses were included for further analysis. These are employed in general (73%) or university hospitals (27%) and are working in mixed (66%), surgical (18%) or medical (13%) ICUs.Short working agents are most frequently chosen for analgo-sedation. A majority of respondents administer sedatives in continuous infusion with bolus doses (81%). Less than half of nurses have a sedation protocol (44%) but only minority consistently uses it (8%). Nurses in university hospitals reported higher availability of sedation protocols compared to general hospitals . Patient-targeted protocols are most available (53%). They are mostly developed particularly by MDs (79%) and nurses (52%). Level of sedation is generally evaluated per 2 hrs (56%) and with use of a RASS scale (59%). Daily interruption of sedation (DIS) is applied variably  and usually to evaluate the neurological status (86%) and to shorten the duration of ventilation (44%). 78% of nurses report not to apply DIS at night. Patients' discomfort is the most important barrier to execute DIS (49%). Nearly half of respondents pointed out respiratory deterioration is the greatest concern during DIS (47%). For 40% of nurses it is risk of self-extubation and removal of the catheters. Agitation/confusion (66%) and lack of comfort and pain (46%) are the most frequently reported problems associated with sedation reduction.There appears to be a considerable discrepancy between recommendations and sedation practice. This data demonstrate room for quality improvement initiatives.


Few studies have investigated the use of viscoelastic devices for monitoring of treatment with LMWHs and to our knowledge there are no studies comparing different LMWHs or different visocoelastic methods.Enoxaparin (Klexane) and tinzaparin (Innohep) were added to 2 ml citrated blood from 10 intensive care patients to obtain plasma concentrations of 0, 0.5, 1.0 and 1.5 IU/ml enoxaparin and tinzaparin, respectively. The study was approved by the local ethics committee and with written consent (relatives). Clot formation and clot retraction was studied using ROTEM and ReoRox.ROTEM analysis showed prolonged clot formation (CT) with increasing concentrations of enoxaparin and tinzaparin (more so). ReoRox analysis showed that the initiation of clot formation COT1) increased with increasing doses of enoxaparin and tinzaparin (more so), as did the progression of clot formation (COT2 - COT1), thus resulting in a prolongation until complete clot formation (COT2). See Table  increaseClot initiation was prolonged with both drugs and detected by both ROTEM and ReoRox. Clot formation was more decreased with tinzaparin than enozaparin and only detected by ReoRox.


Impaired right ventricle (RV) function is associated with worse outcome in ARDS. Pulmonary artery pressure waveform analysis provides information about phenomena that affect RV performance. In particular, pulmonary vascular wave reflection (WR) is directly related with RV stress. We hypothesised that open lung ventilation (OLV), compared with conventional ARDS-net ventilation (CV), would improve conditions for RV performance in an ARDS model. This hypothesis was tested by measuring pulmonary vascular wave reflection (WR).To evaluate the effect of two mechanical ventilation (MV) strategies on WR in an experimental model of ARDS.8 anesthetized and muscle relaxed pigs were submitted to a two-hit lung injury model combining repeated lung lavages with injurious MV. After lung injury was induced, animals were randomized (4 pigs in each group) to one of two strategies of MV: OLV, PEEP 2cmH2O above the PEEP corresponding with the maximal dynamic compliance in a decreasing PEEP trial after a recruitment manoeuvre; or CV, PEEP adjusted according to the ARDSnetwork table. In both groups tidal volume was 6ml/kg, respiratory rate to maintain PaCO2 between 55-65 mmHg and FIO2 to maintain PaO2 55-80 mmHg.1 and quantify WR. The following indexes of WR were calculated: Backward wave amplitude (APbw); Reflection index (RI) which is the ratio between the backward wave amplitude and the sum of backward and forward wave amplitude. Evaluation was done before (BL) and after lung injury (ARDS) and after 4 hours of management in the respective MV strategy.Pulmonary artery (PA) flow and pressure waveforms (1000Hz sampling rate) were acquired by a high-fidelity microtip manometer and an instantaneous transonic pulmonary flow probe placed in the main PA by a small lateral thoracotomy. These signals were used to separate the forward and backward components of the pressure waveformWe did not find any significant changes by induction of ARDS but both APbw  and RI  were lower in OLV comparing with CV.In this experimental ARDS-model OLV decreased WR in the pulmonary vascular system comparing with CV, indicated that OLV could reduce the stress on the RV and improve conditions for RV performance.


Traumatic injury of the central nervous system (CNS) has severe impact on the patients’ quality of life and initiates many molecular and cellular changes at the site of insult. Traumatic CNS injury results in direct damage of the axons of CNS neurons, loss of myelin sheaths, destruction of the surrounding vascular architecture and initiation of an immune response. Class III semaphorins (SEMA3s) are present in the neural scar and influence a wide range of molecules and cell types in and surrounding the injured tissue. SEMA3s and their receptors, neuropilins (NRPs) and plexins (PLXNs) were initially studied because of their involvement in repulsive axon guidance. To date, SEMA3 signaling is recognized to be of crucial importance for re-vascularization, the immune response and remyelination. The purpose of this review is to summarize and discuss how SEMA3s modulate these processes that are all crucial components of the tissue response to injury. Most of the functions for SEMA3s are achieved through their binding partners NRPs, which are also co-receptors for a variety of other molecules implicated in the above processes. The most notable ligands are members of the vascular endothelial growth factor (VEGF) family and the transforming growth factor family. Therefore, a second aim is to highlight the overlapping or competing signaling pathways that are mediated through NRPs in the same processes. In conclusion, we show that the role of SEMA3s goes beyond inhibiting axonal regeneration, since they are also critical modulators of re-vascularization, the immune response and re-myelination. Class III semaphorins (SEMA3s), apart from their classical axon repulsive properties, exert regulatory functions in a variety of non-neuronal cells associated with CNS trauma.Neuropilins (NRPs) are pleiotropic receptors involved in multiple signaling pathways controlling tissue remodeling following CNS trauma.Semaphorin signaling is regulated at several levels, including receptor complex formation, proteolytic cleavage and interaction with extracellular matrix molecules.Interfering with class III semaphorin signaling might be beneficial for axonal re-growth, re-vascularization, re-myelination and manipulation of the immune response following CNS trauma.The semaphorin family consists of secreted and transmembrane glycoproteins that are involved in many cellular functions. Semaphorins (SEMAs) are subdivided into eight classes based on amino acid sequence similarities and structural features. The first two classes of SEMAs are found in invertebrates, class III till VII belong to vertebrates, while class VIII semaphorins are only expressed by viruses  injuries due to their presence in neural scar tissue. The expression of SEMA3s following CNS trauma is suggested to be important in both the acute and sub-acute/chronic phases of scar formation. To date there is data showing Sema3A mRNA up-regulation as early as 1 day after CNS penetrating lesions  and a small CSPG called Decorin. ChABC digests the chondroitin sulphate (CS)— glycosaminoglycan (GAG) chains of CSPGs, while decorin suppresses multiple repulsive proteoglycans in the extracellular matrix. Among the repulsive molecules that could be affected from treatment with the above approaches is SEMA3A which in earlier in vitro, but was unable to enhance axonal regeneration and functional recovery after SCI in vivo  from Sema3A-induced apoptosis after optic nerve axotomy  enhances axonal regeneration in an in vitro model of cerebral ischemia. VEGF rescued cultured CNS neurons from hypoxia and glucose deprivation induced apoptosis  and receptor complexes consisting of NRPs and VEGFRs , there is degeneration of the vascular architecture and the hypoxic regions display dysregulated hyper revascularization. Interestingly, the newly formed blood vessels are not capable to reach the neural retina. The inflammatory and ischemic stress induced expression of Sema3A by RGCs prevented revascularization of the eye since RGC-derived Sema3A repelled blood vessels and inhibited EC growth , which in turn, inactivates integrin receptors. Secondly, Sema3E induces the activation of ADP-ribosylation factor 6 (Arf6) positive vesicles that promote internalization of integrin receptors. In contrast to these functions of SEMA3s as inhibitors of angiogenesis and integrin functions, Sema3C promotes angiogenesis in vitro by activation of integrins  and their receptors are key modulators of the immune response  secrete cytokines that can be toxic for CNS neurons, while the non-classical anti-inflammatory type 2 macrophages (M2) can promote neuroregeneration across neuronal repulsive substrates  signaling, which is NRP1-independent, exerting anti-proliferative and immunosuppressive effects that induce fibrosis and non-canonical signaling pathway which is NRP1-dependent and can antagonize canonical signaling  As mentioned earlier in this review blockage of SEMA3s signaling might favor revascularization and thus the innate ability of the organism for effective clearance and recycling of the excessive inflammatory cells through the circulatory pathway; and (2) NRP1 receptors of inflammatory cells may become available for TGF-β signaling, which suppress the immune response perhaps more sufficiently than SEMA3s. Therefore, the role of SEMA3s in modulating the immune response after traumatic CNS injury needs to be investigated in more detail.Trauma of the CNS induces mechanical forces that cause loss of OLs and demyelination of axons at and surrounding the injury site. The loss of myelin sheaths decreases the conduction velocity of action potentials and could result in axonal degeneration. The reestablishment of myelin sheets around demyelinated axons and around newly sparsely formed regenerating axons, a process called remyelination, is therefore important to regain neuronal function after brain and spinal cord injuries  axonal regeneration; (2) re-vascularization; (3) immune response; and (4) re-myelination (Figure  axonal rIn the sub-chronic and chronic phase of CNS injury SEMA3s may inhibit axonal regeneration by inducing apoptosis of neurons and by preventing axons to penetrate scar tissue. They may also restrict re-vascularization by SEMA3B inhibition of the recruitment of pericytes that normally stabilize new blood vessels and by SEMA3A mediated elevation of the vascular permeability of blood vessels even when the expression of VEGFA declines. This increased vascular permeability may have a direct impact on the chronic immune response where SEMA3s are shown to inhibit the proliferation of recruited T-lymphocytes and antagonize the synthesis of cytokines such as IL-2, IL-4, IL-10 and IFNγ. Furthermore SEMA3A induces apoptosis of microglia that express NRP1 and thereby influence the survival and axonal regeneration capacity of injured CNS neurons. Additionally SEMA3A induces cell death of M2 macrophages and could thereby induce a “pro-inflammatory” M1 macrophage environment in the scar. An intriguing but yet not fully established hypothesis is that SEMA3s might modulate TGF-β non-canonical signaling via NRPs at inflammatory cells near the scar tissue. Finally, re-myelination is also affected by SEMA3s at the chronic stages of injury. In particular SEMA3A inhibits the recruitment and differentiation of OPCs and inhibits the mobility of migrating Schwann cells. In contrast, SEMA3F promotes OPC migration and remyelination. Lastly, SEMA3A signaling via NRP1 might antagonize furin-processed SEMA3F, EGF and PDGF signaling pathways that all posses the ability to stimulate remyelination.Since the discovery of Sema3A in 1993, SEMA3s signaling and their receptors were increasingly recognized to be involved in key processes influencing axonal outgrowth and guidance. However, substantial evidence indicates that SEMA3s also participate in cell-to-cell communication systems that underlie angiogenesis, inflammation and re-myelination. It will be necessary to combine the knowledge obtained from the research fields of vascular biology, cancer biology, immunology and glial biology in order to better understand the role of SEMA3s signaling in CNS trauma. A combinatorial approach involving these fields of research may be necessary in order to move closer towards a therapy for traumatic CNS injury. SEMA3s are an important target in strategies that aim to repair traumatic spinal cord and brain injuries, however the sometimes overlooked pleiotropic nature of their NRP receptors needs to be addressed/understood in a more thorough fashion in future studies.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.


Growing demand of resources increases pressure on ecosystem services (ES) and biodiversity. Monetary valuation of ES is frequently seen as a decision-support tool by providing explicit values for unconsidered, non-market goods and services. Here we present global value transfer functions by using a meta-analytic framework for the synthesis of 194 case studies capturing 839 monetary values of ES. For 12 ES the variance of monetary values could be explained with a subset of 93 study- and site-specific variables by utilizing boosted regression trees. This provides the first global quantification of uncertainties and transferability of monetary valuations. Models explain from 18% (water provision) to 44% (food provision) of variance and provide statistically reliable extrapolations for 70% (water provision) to 91% (food provision) of the terrestrial earth surface. Although the application of different valuation methods is a source of uncertainty, we found evidence that assuming homogeneity of ecosystems is a major error in value transfer function models. Food provision is positively correlated with better life domains and variables indicating positive conditions for human well-being. Water provision and recreation service show that weak ownerships affect valuation of other common goods negatively (e.g. non-privately owned forests). Furthermore, we found support for the shifting baseline hypothesis in valuing climate regulation. Ecological conditions and societal vulnerability determine valuation of extreme event prevention. Valuation of habitat services is negatively correlated with indicators characterizing less favorable areas. Our analysis represents a stepping stone to establish a standardized integration of and reporting on uncertainties for reliable and valid benefit transfer as an important component for decision support. Goods and services obtained from nature–ecosystem services (ES)–are essential for human well-being , 2. ManyDespite this critique ES are valued in prominent assessments of natural capital , 18, 19,A first major critique refers to benefit transfer in its basic form. Benefit transfer averages monetary values (point estimates) from study sites and transfer them to a similar unsampled area by accounting for land use/land cover types only , 28, 29.A second critique originates from the complexity and heterogeneity of human-environmental systems. Due to the variation in site characteristics, e.g. socio-economic or biophysical feature , 31, andWith this publication we i) provide a conceptual base supporting the establishment of a standardized integration and reporting on uncertainties of benefit transfer. Building on this we ii) assess the transferability of monetary values of ES and identify major sources of uncertainty by using meta-analytical value transfer functions.We generated a spatially explicit database of 194 globally distributed cases studies covering 839 monetary values of ES from peer reviewed data collections , 36. We We combined two, peer reviewed databases with monetary valued ES , 36, seer >0.75 or r< -0.75) and Spearman’s rho (ρ>0.75 or ρ< -0.75), for two categorical covariates chi-square test and for testing categorical and continuous one-way ANOVA. From 291 covariates 93 remaining were combined with the maps of standardized monetary values by overlay operation in ArcGIS. Combined maps were used as input for the value transfer functions . A BRT computes the relative influences (importance) of covariates for a BRT model, i.e. they identify major determinants of the variance in monetary values of ES. BRT also provide elasticity curves  that account for non-constant marginal value changes over distinct socio-ecological conditions and thus quantify the change of monetary values in response to an alteration in one covariate (i.e. ceteris paribus) ; see S2 We tested different parameter of the BRT algorithm such as learning rate, tree complexity, minimal number of observations in terminal nodes and number of trees see , and choStatistical significant value transfer functions could be computed for 12 ES types based on 839 monetary values . Most important covariates were quantified and monetary values in unsampled areas extrapolated . If therIn an additional step confidence intervals were estimated to examine generalization failure of value transfer functions from training data . We reruFor the discussion of the results we conceptualized three major sources of uncertainty of value transfer functions: (i) Sample error, such as measurement error in input studies for value transfer functions or publication selection bias; (ii) errors originating from statistical estimation of BRT models (model performance and suitability of chosen approach for benefit transfer); and (iii) transfer error from generalization that encompasses distortions due to value transfer without fully accounting for site and study characteristics. Only covariates with >1%  were analyzed for the six groups of covariates. Additionally, in a fourth point we provide information of the spatial application of the value transfer functions.All studies considered in the synthesized database  come froThe final 12 cross-validated value transfer functions explain from 18% (water provision) to 44% (food provision) of the variance of monetary values . FurtherResults from the quantification of the importance of covariates for the value transfer functions indicate which site and study characteristics need to be considered in order to minimize transfer error from generalization. In The application of the value transfer function for spatial extrapolation of values in unsampled areas results in a map shown in In the following section we discussed the results in detail for 6 out of 12 ES. Food and water provision, climate and extreme events regulation as well as recreation and habitat service were selected, because they represent the highest variance of monetary values from each ES group , see Available input data: With 158 monetary values the largest data set is available for food provision. The majority of case studies examine fish provision (51%) in lower latitudes (93%) and in coastal ecosystems or inland wetlands (69%). Lower latitudes  are most likely food-insecure regions with a high vulnerability to climate change . CoastalModel uncertainty: The value transfer function shows the lowest uncertainty and explains 44% of the variance in the data. Estimations of the confidence intervals show that for 91% of the terrestrial earth surface monetary values can be computed with low and medium uncertainty (1 to 80 Int.-$-2007 per ha), due to a high number of data points .Importance of covariates: Most important covariates are climate indicators (22%), followed by geographic and nature endowment (15%) and valuation methods (15%). Further influential are social covariates such as better life domains of human well-being (12%) and religion (3%) as well as the economic covariate agricultural subsidies (3%).2 per grid cell). Variance explained by distance to sea (9%) is a logical consequence from the distribution of studies and the focus on fish as food resource. In more landlocked areas the importance of fish in the food supply reduces and so does the monetary valuation of fish. Moreover, the type of biome explains 6% of the variance, but does only partly coincide with the land use classification used on the case studies itself. These show that coastal wetlands, coastal systems and cultivated areas (aquaculture) are most valued, and confirm previous findings from [Ecology: Covariates indicating climate and those on geographic and nature endowment show low values in areas with prevailing unfavourable growing conditions  and low human-induced alteration of ecosystems  <11%) or high amounts of alternative food products , followed by benefit transfer (17%) and group valuation (5%). With group valuation significant lower values are derived. Most studies (75%) based on group valuation were conducted for fish in India, a country where fish consumption represents only 2% of protein intake , which mSociety: The valuation of food increases with a lower unemployment rates (<10% of labour force) and a high number of people in working age (15–59 years), good education system  as well as with improving sustainable well-being measured by years of life satisfaction achieved per unit of resource used (Happy Planet Index). These covariates directly relate to the better life domains of the Organisation for Economic Co-operation and Development (OECD), which are essential metrics for human well-being . This shEconomy: With increasing subsidies in agricultural sector the value for food provision decreases. Subsidies distort markets by promoting the production of agricultural commodities beyond market demand, thus, they encourage farmers and fishermen to rely on them instead of consumer wants .Application of value transfer function: Spatial extrapolation of values shows for food the highest uncertainties and lowest values are in India and parts of Africa . On contAvailable input data: For water provision one of the smallest datasets was available (26 case studies). Most of the case studies were conducted in climate sensitive lower latitudes (63%). Climate change is affecting the hydrologic cycle and directly impacts the water resource base, usage, and management, in particular in lower latitudes .Model uncertainty: The value transfer function for water provision shows the highest uncertainties and explains 18% of the variance . ConfideImportance of covariates: Most influential are ecosystem-based covariates that indicate biodiversity and water availability (67%), followed by type of biome (3%). Further important are covariates of social and economic indicators (22%).2 per grid cell), in particular, agricultural frontier areas of cropland foster high values for water provisioning . Third, deforestation explains 3% of the variance and is positively correlated with water values. Water availability, moreover, depends on the spatial (3%) and temporal allocation of water resources (2%). The more unequal rivers and lakes are distributed in an area the higher is the value for water provision. Seasonal variability or long-term climatic changes cause extended periods of droughts or water abundance. The type of biome explains 3% of the variance. However, the applied land use classification system differs from the actual biome type of the case studies. Taking case study based classification into account coastal wetlands and freshwater (rivers/lakes) show highest values, confirming [Ecology and society: Areas of high biodiversity threat and conservation value are positively correlated with monetary values and explain 21% of the variance. Biodiversity and water supply are strongly interrelated , 53. Drinfirming , 28, 29.Economy: The portion of privately owned forests is positively correlated with water provision value (5%). This confirms that weak ownerships might affect valuation of other common goods negatively . SimilarApplication of value transfer function: Spatial extrapolation of values shows the most certain and highest values in areas of high population growth and increasing pressures on water security, for instance in China, India, Java, eastern USA, south Mexico and Western Central America, south coast of Western Africa, and Mediterranean Europe . TransfeAvailable input data: From a total of 36 case studies , carbon sequestration was most frequently examined (68%), followed by other greenhouse gases (<2%) or remain unspecified (24%). The majority of studies were carried out in tropical and temperate forests (25%) as well as inland and coastal wetlands (45%). These biomes are seen as regions highly suitable for carbon sequestration , 59. MorModel uncertainty: With 38% of variance explained in monetary values the transfer model for climate regulation represents the third best prediction performance . For mosImportance of covariates: The most important covariates are ecosystem-based variables of nature threats (49%). Also relevant but explaining less of the variance are covariates indicating input measures of climate sensitive economic sectors (20%) and other economic variables (6%), followed by scale (7%), methods (6%) and social variables (4%),3) exist. Furthermore, the economic covariate inequality of income (6%) shows that the more unequal income is distributed the higher the value for climate regulation.Ecology and economy: Covariates of nature threats show that high values are associated with areas of high risk that unique biodiversity will soon be lost (30%) and high risk of erosion (19%). Covariates indicating input measures of climate sensitive economic sectors (e.g. energy production and water business) are positively correlated with monetary values for climate regulation. Most important covariates are: proportion of electricity production from hydroelectric sources (12%), annual mean of solar radiation (6%) and the water storage capacity of dams per country (2%). Values are high, for instance, in areas of lower latitudes where up to 99% of electricity is produced by hydroelectric power plants and where big artificial constructed water reservoirs , followed by direct market prices (16%) and avoided costs (11%). For direct market prices the highest values and for benefit transfer and replacement costs the lowest ones can be observed.Society: Social covariates of relevance are population density per country (2%) or the age of population (2%). Population density is negative correlated with monetary values. Surprisingly, case studies in countries with on average older population report higher values of climate regulation. One might hypothesize older people may have made in their lifetime perceiving changes in climatic conditions and associated consequences, thus, valuing climate regulation service higher than younger persons. This might be an indication for the “shifting-baseline” hypothesis, i.e. shifts of the reference points of human perception for estimating changes .Application of value transfer function: Applying the value transfer function shows that the most certain and highest values are computed for areas under threat of habitat alteration due to climate change or other land degradation processes. Examples are areas like the Sahel Zone, tropical islands and mountains, Mediterranean ecosystems, Eastern USA and parts of Europe . On contAvailable input data: The majority of case studies (82%) were conducted in areas of high vulnerability to extreme events and sites which are increasingly exposed to extreme events due to climate change –63. In 4Model uncertainty: The value transfer function for prevention of extreme events represents medium prediction performance and explains 21% of the variance in monetary values . TransfeImportance of covariates: The impact of extreme events depends on both ecological conditions and societal vulnerability. We found that with 64% the highest explanatory power is represented by covariates showing the inherent conditions of ecosystems, followed by socio-economic characteristics (33%) and covariates dealing with the spatial and temporal scale (3%).Anthropogenic pressure on the ecosystem: Extreme event prevention is valued high in areas with advanced anthropogenic alteration of ecosystems characterized by highly changed biomass (HANPP 13%), high agricultural induced soil erosion rates (10%), degraded freshwater resources and riverine biodiversity (10%), dense infrastructure (5% market access), dense settlements with major markets (4% Anthromes) and high population density (3%). Hence, the risk awareness to weather extremes and natural hazards increases with number of people potentially effected and higher level of land-use and degradation. The more risk aware a society is, the more weight it places on strategies that preserve or build ecosystem resilience, and the higher the value it would allocate to ecosystem configurations that are more robust . This isSpecies richness and unemployment: We also found that areas with high species richness (4%) and high level of conservation priority (3%) show high valuation for extreme event prevention. These areas are characterized by high population, the highest population growth rates globally , 56 and Application of the benefit transfer function: Using the value transfer model for spatial estimations of extreme event prevention show that the most certain and highest valuations are situated in densely populated areas with advanced anthropogenic altered environments . The larAvailable input data: The database for the ES recreation services captures with 96 the most case studies. The majority of studies were conducted in major non-religious and Christian sites (73%) as well as lower latitudes (77%). Studies in developing countries  < 4024 US-$) are underrepresented (26%). Subcategories of recreation services investigated are tourism (41%), hunting (<1%), ecotourism (<1%) and unspecific recreation (47%).Model uncertainty: The value transfer function for recreation services is with 33% variance explained the fourth most certain model . ConfideImportance of covariates: With 32% ecosystem-based covariates are the most influential group. In comparison with other value transfer functions, however, this is the lowest value . Further2 and a soil moisture index of 1 to 1.6. This corresponds to areas in the (sub-) tropics, see Ecology: More homogeneous environments receive lower values for recreation. The value transfer function shows that areas are low valued with intensive agricultural use (6%), a high number of people living on degraded land (2%) and low number of terrestrial protection areas (4%) and biodiversity (2%) as well as low ethnic diversity (1%). Surprisingly there is a negative effect of marine protection areas (4%) and length of coastlines (1%). This is due to the low number of case studies carried out in coastal areas (17%) and marine protection areas (16%). Furthermore, covariates on land use/land cover types explain 9% of the variance and show that wetlands and freshwater areas (river/lakes) are highest valued. The most important climatic covariates are solar radiation (3%) and soil moisture (2%). The latter one is strongly correlated with precipitation. Highest monetary values are shown for areas with 100 to 190 W/mEconomy and society: Economic covariates show that recreation is highly valued in wealthy countries. These countries are characterized by high market access (3%), low GDP growth (3%), low consumer prices (2%), positive trade balance and a high level of export to import ratio (3%). In wealthy countries the access to a greater share of ES and substitutes is provided, thus, the basis for recreation given. However, in areas with a high unemployment rate (6%) recreation values are high, too. We hypothesize these are sites where tourism is already established and an important economic factor. With increasing portion of privately owned forests the value for recreation is higher (1%). This shows that with the granting of property rights the valuation of common goods might be positively affected  and emphApplication of value transfer function: Applying the value transfer function shows that the most certain and highest values are in the USA, most of the Mediterranean area, Caribbean islands, and Colombia . These aAvailable input data: The database for habitat service contains one of the smallest numbers of case studies .Importance of covariates: The group of ecosystem-based covariates contributes with 90% to the value transfer function . These aEcology and society: Habitat services are valued less in marginal areas, i.e. harsh growing conditions due to low soil quality (24%), arid climate zones with high drought potential (16%) and high UV exposure (26%). Marginal areas are mostly sparsely populated (1%) and characterized by low market access (3%). Also those areas are the home of poor people , 73 withApplication of value transfer functions: Spatial extrapolation shows the most certain values in low valued marginal areas like the highlands and drylands of Asia and sub-Saharan Africa, transition zones of the grassland to deserts of Australia as well as southern Latin Americas Andes Mountains, and the highlands and drylands of North America .Although “money” is seen as a well-known and easily understandable indicator, monetary valuation of ES is ambiguous and shows only a fraction of the multiple characteristics that give utility to the beneficiary . ApplyinThe data availability and the quality of input studies determine the sampling error. We only included valuation studies that were reviewed by experts as a minimal assurance of study quality. Although the peer review approach is often criticized it is also appreciated as an efficient method that increased the scientific progress over the past decades . The majAnother concern with respect to the input data refers to the aggregation of the monetary values. In order to create a comparable data base for spatially explicit value transfer modelling, monetary values need to be disentangled. Only values measured under marginal changes in the socio-ecological system were considered . We utilUtilizing BRT for spatially explicit, meta-analytic value transfer functions have inherent causes of error, too. Although this method is acclaimed to be one of the most sophisticated approaches for benefit transfer , 82 the The spatial application of the value transfer functions in unsampled areas generates transfer errors from generalization, which are determined by uniformity, spatial extent and resolution  as well as temporal aspects , 83. We Regionalization error encompasses extrapolation errors due to generalization from a local case study to a wider region . All valThe major achievement of this study is, first, to clearly distinguish between major sources of uncertainties and to quantify patterns in the influence of different study- and site-characteristics that affect value transfer functions and predicted ES values. Second, the study identifies regions for which sufficient knowledge on our natural capital is available, i.e. a statistically defensible benefit transfer model can be applied in combination with uncertainty values. Third, we provide global maps of the “white spots” on our knowledge on accounting natural capital and ES. This provides guidance for future analysis and concerted action on the assessment of natural capital and ES within the work program of IPBES  and the The integration of the presented value transfer functions in a fully automated spatial benefit transfer tool, such as the Ecosystem Valuation Toolkit , would bS1 FigThis map is the completion of the (TIF)Click here for additional data file.S2 FigTreemaps A) to L) represent models with covariates greater than 1%, their relative influence (or importance) and response. Rectangle groups delimited by bold black lines and numbered from 1 to 7 reflect groups of covariates mentioned in the main article (1) scale, 2) economy, 3) policy, 4) society, 5) ecology, 6) valuation methods, 7) rest <1%). The sizes of rectangles show the relative influence of covariates on a BRT model, in percentage. Rectangle colours illustrate the strength of relationship between monetary values and covariates. Greenish colours symbolize positive correlation and reddish negative, expressed in International-$-2007 per hectare. Multiple colours can occur for nonlinear effects of variables. Bluish colours represent categorical variables and show the maximum range between the levels.(TIF)Click here for additional data file.S3 FigThe graphs show the model performance for twelve parameter configurations. In the table below these configurations are specified. In addition to learning rate (lr), number of trees (nt) and minimal number of observations in the terminal nodes (mintn), there is also the selected model visualized. The selected model represents the final BRT model used for value transfer. It reduces the deviance of residuals in the model (squared error loss) the most and thus explains the variance of monetary values best.(TIF)Click here for additional data file.S1 TableThe table gives an overview of the case study references that were included for the boosted regression trees.(PDF)Click here for additional data file.S2 TableThe table shows sources of covariates either from valuation databases , 36 or f(PDF)Click here for additional data file.S1 Text(PDF)Click here for additional data file.S2 Text(PDF)Click here for additional data file.


Parkinson Disease (PD) is a neurodegenerative disease for which no cure is available yet. It is the second largest neurological disease affecting an estimated 571 per 100,000 people in Europe with rising prevalence due to the aging population  sensory cues from the music as well as from the dance partner  reported no adverse events during dance classes  reported one single fall with no injury , ParkinsonFonds (20094), and EU (ERC-291339). EV received research-funding from ZonMw (630000012), Parkinson Vereniging (2013R16), ParkinsonFonds (20094), and the Hersenstichting (F2013141).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.


Ablation of adult neurogenesis in mice has revealed that young adult-born granule cells (abGCs) are required for some of the behavioral responses to antidepressants (ADs), yet the mechanism by which abGCs contribute to AD action remains unknown. During their maturation process, these immature neurons exhibit unique properties that could underlie their ability to influence behavioral output. In particular, abGCs in the DG exhibit a period of heightened plasticity 4–6 weeks after birth that is mediated by GluN2B-expressing NMDA receptors. The functional contribution of this critical window to AD responsiveness is unclear. Here, we determined the behavioral and neurogenic responses to the AD fluoxetine (FLX) in mice lacking GluN2B-containing NMDA receptors in abGCs. We found that these mice exhibited an attenuated response to FLX in a neurogenesis-dependent behavioral assay of FLX action, while neurogenesis-independent behaviors were unaffected by GluN2B deletion. In addition, deletion of GluN2B attenuated FLX-induced increases in dendritic complexity of abGCs suggesting that the blunted behavioral efficacy of FLX may be caused by impaired differentiation of young abGCs. Adult hippocampal neurogenesis is a process that results in the generation of new neurons in the dentate gyrus (DG) throughout life Gross, . After tDuring the maturation process, abGCs exhibit distinct properties that could underlie their ability to influence behavioral output  of Grin2b. In the presence of Cre, exon 9 gets excised. Western blotting performed using an antibody that targets the C-terminal domain downstream of exon 9 has shown decreased GluN2B expression following Cre recombination, demonstrating that excision of exon 9 results in the complete absence of GluN2B, not merely a truncated protein product  was injected  once per day for 2 days just prior to the start of AD treatment. For dendritic morphology analysis, AD treatment began ~5 weeks after TMX administration and BrdU  was injected 4 times over 8 h the day before AD treatment began. For all mice, FLX (18 mg/kg/day in water) or vehicle  was delivered by oral gavage and in the drinking water (160 mg/mL) at a schedule of 5 consecutive days gavage followed by 2 consecutive days of water, then repeated for the duration of treatment.All behavioral experiments were conducted in male mice 18–24 weeks of age.In the novelty suppressed feeding (NSF) test, chronic AD treatment decreases latency to feed in the center of a novel arena after overnight food deprivation  is a behavioral despair test during which the mobility of mice suspended by their tail is measured. A depressed-like state is characterized by greater immobility while ADs are able to decrease immobility  consisting of both open and closed arms and were given 5 min to explore. The test was video recorded and activity was measured using TopScan software. Their preference for open (anxiogenic) or enclosed (anxiolytic) spaces was used to assess anxiety Hogg, .http://fiji.sc/Fiji) where dendritic length was measured using the freehand trace tool and Sholl analysis was conducted using the Sholl analysis plug-in with parameters previously described , brains postfixed, cryoprotected, and sections (35 μm) of the entire DG were labeled for BrdU, glial fibrillary acidic protein (GFAP), neuronal nuclei (NeuN), doublecortin (DCX), Ki-67, or green fluorescent protein (GFP) , as previously described , while an encoding region immediately preceding the first transmembrane domain of GluN2B is flanked by loxP sites . TMX injection in adult mice leads to deletion of GluN2B and expression of YFP in neural precursor cells (NPCs) and their subsequent progeny  or in an assay of anxiety that was insensitive to FLX in this strain of mice (EPM). While some studies in rodents do find that chronic AD treatment results in an anxiolytic phenotype in the EPM -dependent mechanism. Chronic SSRIs have been shown to increase BDNF in the DG . RH is supported by the National Institute of Mental Health (R37 MH068542 [MERIT]), the National Institute on Aging (R01 AG043688), the National Institute of Neurological Disorders and Stroke (R01NS081203), NYSTEM (C029157), and the Hope for Depression Research Foundation (RGA 11-024). MK was supported by a NIH grant (K01MH099371), a NARSAD Young Investigator Award from the Brain & Behavior Research Foundation, and NYSTEM (C029157).RH receives compensation as a consultant for Roche and Lundbeck. The other 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.


Mortality of ICU patients is a global parameter reported as an end-point in numerous studies. However, causes and characteristics of patients' death are studied only in particular pathologies such as sepsis, cardiac arrest or ARDS , 2)..2]).The aim of our study was to analyse causes and circumstances of death in a general ICU population.We performed a prospective observational study. Every ICU included all death occurring during a month randomised in the year of the study. Demographic data were collected as well as circumstances of death (organ failure and organ support at this time). An organ failure was defined by a SOFA sub-score ≥ 3. Population of the study was dichotomised in expected death  and unexpected death . Data are expressed as median and IQR. Comparisons were made by a Mann-Whitney or chi-squared tests as appropriate. A p value < 0.05 was considered as statistically significant.Ninety-six ICUs included 698 dead patients during the study time. Main characteristics of the population and their comparison between expected (n=473) and unexpected deaths (n=225) are reported in the table.At the time of death, 586 (84%) patients presented at least one organ failure: cardiovascular (58%), respiratory (31%), renal (33%), neurologic (30%), liver (8%) and coagulation (8%). At the same time, an organ support was used in 440 (63%) patients: catecholamines (63%), mechanical ventilation (85%), renal replacement therapy (28%) and liver dialysis (1%). Comparison of these parameters between groups is reported in the figure.Patients who died in ICU presented, most of the time, at least one organ failure. Expected death patients exhibited more neurologic and respiratory failures whereas cardiovascular failure was more prominent in unexpected death. In the latter group of patients, the proportion of organ support was higher corresponding to a greater intensity of care.


The PI3K-AKT pathway has been shown to complement activation of the MAPK pathway in melanocyte transformation in preclinical models. We have utilized clinical specimens and cell lines to examine the regulation, functional role, and clinical significance of the PI3K-AKT pathway in melanoma.All analyses of clinical specimens were performed under protocols approved by the MD Anderson Institutional Review Board. Identities of cell lines were confirmed by STR-fingerprinting, and cell lines were genotyped by sequenom analysis for hotspot mutations. Reverse phase protein array (RPPA) analysis was performed by the MD Anderson RPPA Core Facility.Proteomic analysis of frozen melanoma clinical specimens and cell lines demonstrated that phosphorylation (activation) of AKT (P-AKT) correlated inversely with PTEN protein expression. Confirmatory analysis of PTEN expression by an immunohistochemical (IHC) assay on archival tumor samples identified a linear correlation between PTEN measurement by IHC and RPPA. Significantly increased expression of P-AKT was observed in melanomas with complete loss of PTEN protein expression compared to tumors with any other pattern of PTEN expression. Integrated analysis of BRAF/NRAS mutation status, PTEN expression, and clinical outcomes was performed using lymphadenectomy specimens from patients who underwent surgery for stage IIIB/C melanoma. Complete loss of PTEN expression correlated with shorter overall survival and time to brain metastasis on univariate and multivariate analyses. Testing of human melanoma cell lines demonstrated that cells with baseline or compensatory activation of the PI3K-AKT pathway were generally resistant to apoptosis induction following BRAF or MEK inhibition. Extended molecular profiling found that a subset of melanoma cell lines with de novo and acquired resistance to MEK inhibitors are characterized by increased expression of PGC1alpha and a metabolic phenotype of high oxidative phosphorylation (OxPhos). The resistant high OxPhos cell lines all demonstrated synergistic growth inhibition and apoptosis induction with combined targeting of MEK and mTORC1/2. mTORC1/2 inhibition decreased PGC1alpha expression in vitro and in vivo, and caused cytosolic localization of MITF. Treatment with a MEK inhibitor and an mTORC1/2 inhibitor demonstrated marked synergy in a BRAF-mutant, MEK inhibitor resistant human melanoma cell line in vivo.Activation of the PI3K-AKT pathway correlates with clinical outcomes and resistance to MAPK pathway inhibitors in melanoma. These findings support the rationale to determine the clinical benefit of inhibiting the PI3K-AKT pathway in this disease.


We hypothesize that changes of shape of arterial blood pressure (ABP) high-frequency waveform signal can be reflective of body's response to stress, in particular to hemorrhage.To apply Machine Learning (ML) and sequential pattern extraction to ABP waveforms to reliably detect slow bleeding.62 healthy pigs are anesthetized, intubated, ventilated, and instrumented prior to and during a controlled bleeding at a rate of 20 ml/min. ABP is recorded at a 250 Hz rate. To estimate the likelihood of bleeding, our algorithm:(1) Extracts 10 s disjoint intervals of ABP waveform and standardizes the time series to zero mean and unit standard deviation;(2) Discretizes the standardized data into a sequence of symbols each reflective of a particular value of the standardized ABP;(3) Identifies which sequential patterns of symbols are present in the current interval data from among patterns previously extracted from training data using SPADE algorithm ;(4) Feed the identified sequential patterns into previously trained ML classifier (Random Forest) to predict current bleeding status of the subject. The procedure is empirically evaluated using leave-one-pig-out crossvalidation.Figure In addition, our method enables easy interpretation of learned patterns. An example in Figure ML and sequential pattern extraction enables effective monitoring of ABP waveforms for indications of hemorrhage. The presented approach can be used to enhance current practice of hemodynamic monitoring while helping clinicians interpret patterns of patients' physiological response to stress.Work partially supported by NSF (award 1320347) and by NIH (grant NR013912).


Magnetic resonance (MR) conditional pacemakers are increasingly implanted into patients who may need cardiovascular MR (CMR) subsequent to device implantation. While the safety profile appears good, limited data exists regarding the indication and outcome of these examinations for clinical management.CMR and pacing data from consecutive patients with MR conditional pacemakers performed at our centre was retrospectively reviewed. Images were acquired at 1.5T (Siemens Magnetom Avanto). The indication for CMR and any resulting change in management was recorded. The quality of CMR was rated by an observer blinded to clinical details and data on pacemaker and lead parameters collected pre- and post- CMR.45 CMR scans on 42 patients performed between 2011 and 2014 were assessed. All scans were completed successfully with no significant change in lead thresholds or pacing parameters. Steady state free precession (SSFP)cines were non-diagnostic in 6/45 (13%) scans, but diagnostic in all patients when using gradient echo sequences (GRE). Late gadolinium enhancement imaging was performed in 39 patients and appeared less susceptible to artefact than cine images.The CMR data resulted in a new diagnosis in 19 (42%) of examinations, clinical management was changed in a further 14 (31%).CMR in patients with MR conditional pacemakers provided diagnostic or management-changing information in the majority (73%) of our cohort. The use of gradient echo cine sequences, which are less susceptible to metal artefacts, can reduce rates of non-diagnostic imaging.The study was funded by the NIHR Cardiovascular BRU. CER is supported by the British Heart Foundation.


To compare the effectiveness of constraint induced movement therapy versus motor relearning programme to improve motor function of hemiplegic upper extremity after stroke.A sample of 42 patients was recruited from the Physiotherapy Department of IPM&R and Neurology OPD of Civil Hospital Karachi through non probability purposive sampling technique. Twenty one patients were placed to each experimental and control groups. Experimental group was treated with Constraint Induced Movement Therapy (CIMT) and control group was treated with motor relearning programme (MRP) for three consecutive weeks. Pre and post treatment measurements were determined by upper arm section of Motor Assessment Scale (MAS) and Self Care item of Functional Independence Measure (FIM) Scale.Intra group analysis showed statistically significant results  in all items of MAS in both groups. However, advanced hand activities item of MAS in MRP group showed insignificant result . Self-care items of FIM Scale also showed significant result  in both groups except dressing upper body item  in CIMT group and grooming and dressing upper body items  in MRP group showed insignificant p-values.CIMT group showed more significant improvement in motor function and self-care performance of hemiplegic upper extremity as compared to MRP group in patients with sub-acute stroke assessed by the MAS and FIM scales. Thus CIMT is proved to be more statistically significant and clinically effective intervention in comparison to motor relearning programme among the patients aged between 35-60 years. Further studies are needed to evaluate CIMT effects in acute and chronic post stroke population. Stroke is the third leading cause of death and the most common cause of disability in the developed and developing countries. In Pakistan it is the leading cause of mortality and morbidity after cancer and ischemic heart disease.3Rehabilitation of upper extremity is a challenge, various therapeutic strategies like neurodevelopmental technique (NDT), splinting, biofeedback and electrical stimulation are designed, however the best method to improve upper limb function is still not clear.9This study is a randomized controlled trial carried out to evaluate the effectiveness of CIMT over motor relearning programme. Constraint induced movement therapy (CIMT) is proved to be effective in western countriesIn this randomized controlled trail 42 males and females stroke patients having first attack of stroke and hemiplegic upper extremity had functional level of ≥ 20 degrees of wrist extension and ≥10 degrees of extension of all digits were included. Subjects were between 35-60 years of age and diagnosed with ischemic or hemorrhagic strokes confirmed by CT/MRI scan. Patients were excluded if they had cognitive deficit, recurrent or cerebellar strokes, severe aphasia, balance and equilibrium problems, severe shoulder pain affecting therapy or other neurological deficits.The research was started after approval from institutional review board (IRB) of Dow University of Health Sciences (DUHS). Sample was recruited from Physiotherapy department of Institute of Physical Medicine & Rehabilitation (IPM&R) and Neurology OPD of Civil Hospital Karachi through non-probability purposive sampling technique. Informed consent was taken from the patients. Then patients were randomly allocated to CIMT group and MRP group by using computer-generated random numbers kept in sealed envelopes. Each group had 21 patients. After baseline assessment, patients in CIMT group were asked to perform tasks only with the hemiplegic upper extremity with the unaffected hand restraint in a mitt. The patients in MRP group were asked to perform tasks with both affected and unaffected upper extremities. Tasks were attempted in different positions like supine lying, sitting and standing.th visit. All the exercises were demonstrated to the patients by the therapist and were performed under the supervision of the therapist. Details of exercises are given in Both interventions consisted of 6 sessions per week, two hours duration of each session, for three consecutive weeks. Pre and post treatment evaluation of motor function of hemiplegic upper extremity was determined by using upper arm section of Motor Assessment Scale. Upper limb section of MAS includes, Upper arm function item, hand movements item and advanced hand activities item. An activity of daily livings (ADLs) was measured by using self care item of Functional Independence Measure Scale (FIM) which includes 5 items  for quantitative data. A Non parametric Wilcoxon Signed Ranks test was conducted to find the significance of interventions used within the groups . To test the significance of changes in scores on each of the outcome measures  between the groups , a Non parametric Mann Whitney test was used both at the baseline and at the end of 3A sample of 42 patients participated in the study and no patient dropped out. There were total 28 male participants (66.67%) and 14 (33.33%) female participants in both groups, the distribution of males and females in each groups was almost equal. The mean age of the participants was 49.67 years (SD = 7.01) in CIMT group and 49.47 years (SD = 8.19) in MRP group. Total patients who suffered from ischemic stroke were 32 (76.2%) and number of patients having hemorrhagic stroke were 10 (23.8%) respectively. At the end of third week, mean scores significantly increased in all items of both outcome measures (MAS and FIM) in both CIMT and MRP groups. However, mean value in CIMT group was greater than in MRP group, showing better improvement in CIMT group  & III. Ird week. Post treatment P-value showed statistically significant results  in all items of MAS and FIM Scale accept dressing upper body item.For inter-group analysis a non parametric Mann Whitney test was used at baseline and at the end of 3This study showed the effectiveness of constraint induced movement therapy (CIMT) over motor relearning programme (MRP) to improve motor function of hemiplegic upper extremity in sub-acute stroke patients. The affected patients in the present study were between two weeks to three months post stroke. CIMT initiated within 2 weeks of stroke onset is more effective in upper extremity rehabilitation.12This study results are in agreement with Myint et al. (2008)9In the present study an alteration was made from the original protocolCIMT is considered to attain its beneficial effects by 2 associated but independent mechanisms: overcoming learned nonuse and use-dependent neural plasticity.18The second mechanism by which CIMT has been established to give good functional results is use-dependent neural plasticity.20CIMT group showed more significant improvement in motor function and self-care performance of hemiplegic upper extremity as compared to MRP group in patients with sub-acute stroke assessed by the MAS and FIM scales. Thus CIMT is proved to be more statistically significant and clinically effective intervention in comparison to motor relearning programme among the patients aged between 35-60 years. Further studies are needed to evaluate CIMT effects in acute and chronic post stroke population.


Maturity Onset Diabetes of the Young 3 (MODY3) is the most common monogenic form of diabetes, characterized by early age of onset (before the age of 25), autosomal dominant transmission and severe defect in insulin secretion ,2. MODY bmf gene expression knockout on beta cell mass and islet organisation in a transgenic mouse model of MODY3.To study the role of Bmf (Bcl-2 modifying factor) in the pathogenesis of MODY3, immunohistochemical staining of male and female pancreatic islets for insulin- and glucagon-positive expression utilising confocal microscopy followed by Image J analysis were used to investigate the effect of Bmf gene expression knockout was observed to significantly increase beta cell mass in DN-HNF1a-expressing transgenic male pancreas (p=0.04) but had no observable effect on transgenic female pancreas. Surprisingly, islet disorganisation was seen in both male and female transgenic mice and was not rescued by bmf knockout.Data generated from this study indicate a possible role for Bmf in beta cell mass reduction and thereby the pathogenesis of MODY3 but demonstrates no effect on islet organisation. These data can be built upon by further research to examine in greater detail how Bmf contributes to the characteristic loss of beta cell mass in MODY3 and in mediating DNHNF1a-induced apoptosis.