Although antidepressants are commonly used in patients with schizophrenia, meta-analytic guidance on the efficacy and safety of antidepressant augmentation evaluated as a single clinical strategy in patients with insufficient response to antipsychotic monotherapy is missing.Systematic literature search of PubMed/MEDLINE/PsycINFO/Cochrane Library without language restrictions from database inception until 07/20/2015 for randomized, double-blind, efficacy-focused trials comparing adjunctive antidepressants vs placebo to antipsychotics in schizophrenia. Random effects meta-analysis in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) standard to evaluate the efficacy and safety of antidepressant augmentation of antipsychotics in schizophrenia.In 48 studies , antidepressant augmentation outperformed placebo regarding total symptom reduction =-0.36, 95% confidence interval (CI)=-0.57, -0.15, p=0.001), driven by negative , but not positive (p=0.284) or general (p=0.118) symptom reduction. Significant improvements extended to core negative symptoms avolition/apathy  and anhedonia/asociality . In predefined subgroup-analyses, superiority regarding negative symptoms was confirmed in studies augmenting first-generation antipsychotics (FGAs) , but not second-generation antipsychotics . Uniquely, superiority in total symptom reduction by noradrenergic-and-specific-serotonergic-antidepressants  was not driven by negative (p=0.467), but by positive symptom reduction . Antidepressants did not improve depressive symptoms more than placebo . Bupropion was superior to placebo regarding smoking cessation =6). Except for more dry mouth  and dizziness , antidepressants were not associated with more adverse effects or all-cause/specific-cause discontinuation than placebo.For schizophrenia patients on stable antipsychotic treatment adjunctive antidepressants are effective for total and particularly negative symptom reduction, and bupropion helps smoking cessation. However, effects are small-to-medium, differ across individual antidepressants, and negative symptom improvement seems restricted to the augmentation of FGAs.


The ovary is the main regulator of female fertility. Granulosa cell dysfunction may be involved in various reproductive endocrine disorders. Here we investigated the effect of insulin resistance on the metabolism and function of ovarian granulosa cells, and dissected the functional status of the mitogen-activated protein kinase signaling pathway in these cells. Our data showed that dexamethasone-induced insulin resistance in mouse granulosa cells reduced insulin sensitivity, accompanied with an increase in phosphorylation of p44/42 mitogen-activated protein kinase. Furthermore, up-regulation of cytochrome P450 subfamily 17 and testosterone and down-regulation of progesterone were observed in insulin-resistant mouse granulosa cells. Inhibition of p44/42 mitogen-activated protein kinase after induction of insulin resistance in mouse granulosa cells decreased phosphorylation of p44/42 mitogen-activated protein kinase, downregulated cytochrome P450 subfamily 17 and lowered progesterone production. This insulin resistance cell model can successfully demonstrate certain mechanisms such as hyperandrogenism, which may inspire a new strategy for treating reproductive endocrine disorders by regulating cell signaling pathways. The granulosa cell (GC), the oocytes’ companion cell, ignites a continuous cross-talk between the somatic and germ cell compartments . GC dysfDexamethasone (Dex) is a classical glucocorticoid, which triggers unbalanced cellular glucose metabolism, interferes with insulin sensitivity and induces IR by enhancing the function of pancreatic islet β cells, resulting in the inhibition of glucose uptake –8. ThereConventional mitogen-activated protein kinases (MAPKs) include p44/42 MAPK, c-Jun N-terminal kinase (JNK) and p38 MAPK . The p44In this study, we focused on the GC-based IR model and investigated the role of the p44/42 MAPK signaling pathway in this model. Our model may illustrate possible mechanisms of IR in GCs, and provide further insight into the possible therapeutic development of reproductive endocrine disorders.3-week-old female wild-type Institute of Cancer Research (ICR) mice  were injected with pregnant mare serum gonadotropin (PMSG)  (10 U/10 g weight) 48 h before sacrifice. Mice were housed in separate cages under 21°C with a 12 h light/12 h dark cycle. Free access to a standard diet and water supply was provided. Mice were killed by cervical dislocation. Next, killed mice were soaked with 75% ethanol  for 15 min followed by removal of bilateral ovaries. Ovaries were soaked with 10 ml Dulbecco's Modified Eagle's medium (DMEM)-Ham's F12 nutrient medium (F12) with 2% penicillin -streptomycin (Pen-Strep)   in a petri dish after removal of the surrounding adipose tissue and cleaning of bloodstains. Follicles were separated and punctured by micro-forceps to release GCs under a microscope, and the remaining ovarian theca tissue was discarded. GC suspension was transferred to a centrifuge and was centrifuged at 1000 rpm for 5 min. Next, supernatant was discarded and GCs were suspended in DMEM-F12 with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific) and 1% Pen-Strep (culture media), and then transferred to a 25 ml cell culture flask and placed in an incubator at 37°C, 5% carbon dioxide [As IR correlates to glucose intolerance and metabolic disorders , measuriCulture media were collected and centrifuged at 1000 × g for 15 min. Supernatants were collected for measuring glucose concentrations with the Glucose Oxidase Method assay kit . Briefly, 5 μl sample was incubated with 295 μl regent consisting of two components from the kit (R1:R2 = 4:1) for 20 minutes in 37°C. The OD values at 570 nm were measured to determine glucose concentrations of the samples.GC proliferative activity was measured by the methylthiazolyldiphenyl-tetrazolium bromide (MTT) cell proliferation and cytotoxicity assay kit .Cells were incubated in the radio-immunoprecipitation assay (RIPA) lysis buffer (Beyotime Biotechnology) for 30 min on ice for complete lysis. For every 10 ml RIPA buffer, one phosphatase inhibitor mini tablet (Thermo Fisher) was added. The lysate was centrifuged at 13,000 rpm for 15 min at 4°C for harvesting the total protein which was denatured at 95°C for 10 min. Protein concentration was measured by a BCA protein assay kit (Thermo Fisher). Equal amounts (20 μg) of total protein were separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by transferring to a polyvinylidenediflouride membrane  which was blocked with 5% albumin bovine V . The membrane was incubated with a specific antibody overnight at 4°C . AntibodEstradiol (E2), testosterone (T) and progesterone (P4) were measured by enzyme-linked immunosorbent assay (ELISA) .Data are shown as the mean + standard error of mean (SEM) and were analyzed with Graphpad prism 5.01  for statistical significance using one-way analysis of variance (ANOVA) among groups. A p value of 0.05 or less is considered statistically significant.The most important characteristic of IR is the decrease in insulin sensitivity . We firsTo further demonstrate the effect of Dex on IR, We measured Dex-mediated regulation of STAT3 and phosphor-STAT3 as elevated STAT3 phosphorylation is suggested to correlate to IR –33. TotaPrevious studies demonstrated the relations between IR and the downregulation of the phosphatidylinositol 3 kinase (PI3K)/Akt signaling pathway , 35. HenAs a fundamental downstream pathway of the insulin receptor, the p44/42 MAPK signaling pathway correlates with IR . We therFurthermore, we investigated the other two signaling molecules of the conventional MAPK pathways, JNK and p38 MAPK. Dex elevated phosphor-JNK (p = 0.011) and total p38 MAPK (p = 0.020), which demonstrates that the key molecules on these two routes of the conventional MAPK signaling pathway might be increased in our GC-based IR model. In addition, we observed further increase of phosphor-JNK after the addition of PD98059. We hypothesize that inhibition of the p44/42 MAPK signaling pathway leads to a compensatory increase in the JNK signaling pathway .Dex shares the common side effect such as metabolic disorders with other glucocorticoids . FurtherAccording to the two-cell-two-gonadotropin theory, aromatase can transform T into E2 in GCs , which mIn this study, the steroidogenic activities of GCs were evaluated. The concentrations of T, E2 and P4 secreted from GCs treated for 4 h with Dex in the presence or absence of PD98059 were measured, as these hormones are important in the female endocrine system , 38, 39.In this research, we undertook to uncover the pathological changes in dysfunctional GCs by creating an IR cell model and shed light on the importance of the inhibition of the p44/42 MAPK signaling pathway in the GC IR model, which may be achieved through regulating CYP17 and androgen expression. The IR cell model can successfully demonstrate certain mechanisms such as hyperandrogenism, which may inspire a new strategy for treating reproductive endocrine disorders through regulating the cell signaling pathways.Theoretically, ovarian theca cells are required to produce T, and GC is expected to convert T into E2 . TherefoPrevious studies reported that in human HepG2 cells and rat cardiomyocytes, the p44/42 MAPK signaling pathway is up-regulated after IR , which cA study has shown that IR activates conventional MAPKs, including p44/42 MAPK, JNK and p38 MAPK . Our resBesides the MAPK signaling pathway, the PI3K/Akt signaling pathway is another important signaling pathway down-regulated by the insulin receptor . It is rOur data confirmed that in the GC IR model, CYP17 is elevated as a result of the up-regulation of the p44/42 MAPK signaling pathway, which may facilitate T elevation. Inhibition of the p44/42 MAPK signaling pathway can down-regulate CYP17. Nevertheless, whether the concentration of androgen can be down-regulated by inhibiting the MAPK signaling pathway needs to be further clarified. IR can reduce the P4 concentration owing to the loss of GC viability. On the other hand, the concentration of P4 was further decreased after suppressing the p44/42 MAPK signaling pathway . Taken tS1 FigGCs were incubated in the absence (Con) or presence of Dex for 48 h (GD) or Dex for 48 h with PD98059 added 4 h before the end of the incubation (GDP). Relative density ratios were calculated by setting the control group value as one. Data are expressed as the mean + SEM. All data presented are representative of at least three separate experiments. *p < 0.05.(TIF)Click here for additional data file.S2 FigGCs were incubated in the absence (Con) or presence of Dex for 48 h (GD) or Dex for 48 h with PD98059 added 4 h before the end of the incubation (GDP). Relative density ratios were calculated by setting the control group value as one. Data are expressed as the mean + SEM. All data presented are representative of at least three separate experiments. *p < 0.05.(TIF)Click here for additional data file.S3 FigFor normal GCs, P4 is converted from cholesterol followed by partial conversion into T through catalyzation of CYP17. External T is transferred into GC and converted into E2 along with internal T, which is later secreted through catalyzation of aromatase. For the GC IR model, up regulation of the p44/42 MAPK signaling pathway leads to the elevation of the CYP17 level while reducing T intake, resulting in an increased concentration of T. Inhibition of the p44/42 MAPK signaling pathway by PD98059 results in the down-regulation of CYP17.(TIF)Click here for additional data file.


Cytotoxic immunity relies on specialized effector T cells, the cytotoxic T cells, which are endowed with specialized cytolytic machinery that permits them to induce death of their targets. Upon recognition of a target cell, cytotoxic T cells form a lytic immune synapse and by docking the microtubule-organizing center at the synaptic membrane get prepared to deliver a lethal hit of enzymes contained in lytic granules. New insights suggest that the directionality of lytic granule trafficking along the microtubules represents a fine means to tune the functional outcome of the encounter between a T cell and its target. Thus, mechanisms regulating the directionality of granule transport may have a major impact in settings characterized by evasion from the cytotoxic response, such as chronic infection and cancer. Here, we review our current knowledge on the signaling pathways implicated in the polarized trafficking at the immune synapse of cytotoxic T cells, complementing it with information on the regulation of this process in natural killer cells. Furthermore, we highlight some of the parameters which we consider critical in studying the polarized trafficking of lytic granules, including the use of freshly isolated cytotoxic T cells, and discuss some of the major open questions. Cytotoxic T lymphocytes (CTLs) armed with lytic machinery exert a non-stop patrol of our body to identify and eliminate target cells with potentially “dangerous” phenotype. CTLs bear T cell receptors (TCRs) through which they bind antigenic peptides presented on the major histocompatibility complex (MHC) molecules of target cells. In this way, they successfully discriminate between healthy cells and those presenting non-self peptides, typically of neoplastic or microbial origin. As consequence of non-self recognition, CTLs attack and lyse malignant and infected cells. In this context, an impaired functioning of CTLs may lead to immune evasion of tumors and the insurgence of chronic infections. Hence, defining the mechanisms underlying CTL-mediated killing could provide essential insights into our understanding of immune pathology.The immune synapse of CTLs represents a highly organized system of intercellular communication. Its assembly is initiated upon CTL recognition of a cognate target, toward which the CTL rapidly polarizes Figure A. WithinThe polarization and exocytosis of lytic granules at the immune synapse are driven by local signaling, which triggers two major polarization events, namely (i) the polarization and docking of the MTOC at the synapse and (ii) the polarization of lytic granules toward the docked MTOC, followed by granule exocytosis or “degranulation,” which results in the focal release of granule contents. Notably, at variance with the mechanisms regulating MTOC translocation, the signaling events that regulate lytic granule polarization remain as yet to be fully characterized. Understanding how granule convergence at the synapse is regulated could shed substantial insight into the mechanisms driving CTL dysfunction.Here, we present an overview of the mechanisms that orchestrate the polarized transport of lytic granules in CTLs and incorporate some relevant information on the signaling of natural killer (NK) cells for a comparative analysis of polarizing signals in these two types of cytotoxic lymphocytes. We also discuss recent experimental evidence describing dysfunctional non-polarized immune synapses. Finally, we comment on the parameters of experimental setups that we consider important for the study of polarized trafficking in CTLs and highlight the major open questions concerning its orchestration.Many of the signaling pathways contributing to the assembly of the CTL synapse have been dissected Table . Loss-ofSignaling initiated upon target recognition by the CTL is dictated by the TCR, which recognizes antigenic peptides presented by MHC class I molecules on the target and by costimulatory signals triggered through the engagement of CTL surface receptors by various ligands expressed on the target. Signaling studies on planar lipid bilayers coated with defined molecular ligands allowed to establish that essentially only two types of molecules may be triggered on a CTL to induce synapse formation . These a+ T cell synapse formation is that the TCR and some of the costimulation/inhibitory molecules are spatially segregated during synapse assembly. This stands true also for the mature immune synapse of CTLs. Upon the initiation of CTL synapse assembly, the TCRs and proximal signaling molecules of the TCR pathway concentrate into the central area of the synapse [known as the central supramolecular activation cluster (cSMAC)], while LFA-1 and other adhesion molecules segregate into a ring which surrounds the cSMAC [peripheral supramolecular activation cluster (pSMAC)] (An important notion that has emerged from the first molecular studies of CD4(pSMAC)] –12. The (pSMAC)] . The thr(pSMAC)] , the pSM(pSMAC)] , while t(pSMAC)] .Interestingly, the formation of a mature immune synapse with signaling molecules arranged in the SMAC pattern does not constitute a pre-requisite for successful killing by CTL. It has been shown that as few as two to three peptide/MHC complexes on the surface of the target can trigger cytotoxicity , 16. In In this review, we will summarize the current knowledge on the TCR and costimulatory signaling pathways in CTLs and discuss their contribution to the regulation of polarized synaptic secretion Figure B. For thProximal TCR signaling at the cSMAC is initiated by the two Src family kinases Lck and Fyn . Lck act2+ signaling, and cytoskeletal reorganization , which recruits numerous signaling effectors to form a multiprotein complex termed the LAT signalosome . The LATnization .2+ influx (2) leading to the formation of the lipid second messengers diacylglycerol (DAG) and inositol 1,4,5-trisphosphate (IP3). The local DAG gradient promotes the synaptic recruitment of protein kinase C (PKC) isoforms, namely novel PKCs including PKCθ, PKCε, and PKCη, which participate in the proximal signaling inducing MTOC polarization  membrane and activates Ca2+ release from the ER into the cytoplasm , the most potent Ca2+-releasing second messenger known, is also produced following TCR stimulation and crucially contributes to the signaling pathway operating during CTL degranulation  , CD2 al, and CD12, which promote the recruitment of LFA-1 to the immune synapse and help it to acquire a stable high-affinity conformation [reviewed in Ref.  from PIP2, which has been identified as a key event promoting the spatial segregation of F-actin to the dSMAC ring . Thus, WASP could be recruited to the synapse by two mechanisms: through LAT-SLP76 signaling  and throThe SLAM family members have been extensively studied in CTL functioning due to their involvement in the X-linked lymphoproliferative (XLP) syndrome , 94. XLPAlthough SLAM receptors are important for CTL-mediated cytotoxicity , 104, thInterestingly, studies on human and murine NK cells have demonstrated that LAT is constitutively associated with 2B4 in glycolipid-enriched membrane microdomains and has a critical role in SLAM-mediated cytotoxicity –110. Conin vitro after their isolation from human or mouse donors. CTL expansion in this case is usually required due to a relatively low percentage of effector cytolytic cells among the total pool of CD8+ T cells [in healthy human donors 10–15% . It has become evident that culturing CTLs in IL-2-supplemented medium may alter signaling requirements for CTL killing when compared to freshly isolated effector CTLs. In particular, this knowledge emerged from studies on CTLs from patients and mice with genetic mutations that affect CTL functioning. For instance, function-disrupting mutations in Munc18-4, which regulates lytic granule fusion with the plasma membrane, abolish cytotoxicity in freshly isolated CTLs due to a loss of their ability to degranulate  O+ cells . Effecto11 and PLCβ, instead of Lck and PLCγ1, at least as shown for CD4+ T cells  that are able to cross-link the MHC class II molecules on target cells and the TCRs on CTLs  . In this T cells , 125. Neely used , 35, 52.The second system of polyclonal stimulation commonly employed to work with human CTLs involves the use of Fc receptor-bearing target cells  loaded with CD3-specific monoclonal antibody. Antibody-coated targets are recognized by the CTLs through direct binding of the TCR to the antibody, thus promoting cytotoxic killing in the so-called “redirected” assay . Of noteAlthough both systems appear to work well with freshly isolated CTLs, the use of MHC class I-peptide system is obviously more physiologically relevant as it may allow to assess the functioning of CTLs with natural ligands of different affinity and specificity. The solution to overcome the limitation of rare individual clones in the total CTL population came with the development of systems to clone and express recombinant TCR receptors, derived from naturally occurring CTL clones, in polyclonal freshly isolated CTLs . For insDirectionality of lytic granule transport along microtubules represent fine means to regulate immune synapse assembly in CTLs and NK cells, thus representing a basic factor in immune regulation. However, the mechanisms defining granule ability to travel bidirectionally along microtubules and, most importantly, the signaling pathways regulating this process are largely elusive. To address this question of outstanding scientific importance, we believe that future research on CTLs should focus on some specific issues. First, it would be of high relevance to identify surface receptors that trigger the signaling pathways driving granule translocation toward the MTOC. So far, it emerges that TCR triggering alone could not be sufficient to induce granule polarization, at least in freshly isolated human CTLs , suggestin vitro expanded CTL blasts (Second, lysosomes have emerged as important signaling hubs implicated in the regulation of multiple cellular processes including metabolic signaling and autophagy . Along tL blasts , 137. WeFinally, it is becoming widely accepted that stimulation with cytokines is capable of reorganizing the molecular and signaling network of granule trafficking and secretion in cytotoxic lymphocytes . HoweverAK and VZ wrote the initial draft for the manuscript which was then revised and contributed by CB.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. The reviewer AA declared a past collaboration with one of the authors CB to the handling editor.


We present a nanocomposite strain sensor (NCSS) to develop a novel structural health monitoring (SHM) sensor that can be easily installed in a composite structure. An NCSS made of a multi-walled carbon nanotubes (MWCNT)/epoxy composite was installed on a target structure with facile processing. We attempted to evaluate the NCSS sensing characteristics and benchmark compared to those of a conventional foil strain gauge. The response of the NCSS was fairly good and the result was nearly identical to the strain gauge. A neuron, which is a biomimetic long continuous NCSS, was also developed, and its vibration response was investigated for structural damage detection of a composite cantilever. The vibration response for damage detection was measured by tracking the first natural frequency, which demonstrated good result that matched the finite element (FE) analysis. Composites are becoming crucial structural materials in high technology industries. Composites offer excellent strength/stiffness-to-weight, resistance to corrosion and chemical attack, good electrical behavior, and typically provide sustainable and low maintenance. Due to such advantages of composites, the Boeing 787 designed carbon fiber reinforced composite airframe and primary structure saves 20% more fuel than conventionally similar airplanes . DespiteMore sophisticated structural health monitoring (SHM) and nondestructive inspection (NDI) techniques are required to detect defects of composites than metals. Traditional NDI of composites include visual inspection, automated tap tests, ultrasonic inspection, radiography, and thermography. To improve the efficiency of the visual inspection conducted with the naked eye, some techniques have been developed such as impact sensitive coatings, liquid penetrants, and magnetic particles. The vibration analysis in terms of natural frequency change is reported as an effective method compared to ultrasound transmission techniques for filament wound carbon fiber reinforced plastic (CFRP) tubes . Some teA promising sensor should be light, small, and easily installed in a structure with high sensitivity to cover a large target area for an SHM. A sensor made of carbon nanotubes (CNTs) might meet such demands due to their unique properties ,5. CNTs However, the CNT strain sensor is still in early development for SHM. The CNT strain sensing performance by means of voltage responses need to be systematically assessed for structural deterioration monitoring under various loading conditions. Therefore, we report the multi-walled carbon nanotube (MWCNT) composite strain sensing properties under static and dynamic loading conditions and experimentally studied the composite to develop a neuron as a continuous strain sensor. The long continuous strain sensor was named a neuron . Large sThe artificial neuron was fabricated with MWCNT and epoxy. Epoxy  was used as a matrix for the nanocomposite because it shows excellent mechanical strength and is easily fabricated as a composite. Commercially-obtained MWCNTs  were used for the nanocomposite filler. Though single-walled carbon nanotubes (SWCNTs) have excellent electromechanical properties, because of the high cost and difficult incorporation into polymers at high loading, this study used an MWCNT composite to develop a continuous strain sensor. The piezoresistive properties of MWCNT have been greatly improved due to the successful development of the synthesis process. The base MWCNT solution was prepared using a similar process to the one described in our previous study, and the piezoresistive characteristics of both PMMA and epoxy composites are nearly similar .The mixed MWCNT solution was sprayed with an airbrush on a patterned surface of a structure as shown in The piezoresistive characteristics of the nanocomposites were investigated to determine the variations of electrical properties under structural deformations. The film type specimens were fabricated to investigate strain sensing properties under static and dynamic loading conditions and to calibrate neurons. The short length specimens were fabricated in the same way as the nano-spray process, with the change in CNT wt. % fraction set as 0.7, 2, 3, and 10. Specimens were installed on a cantilever and connected to wires with silver conductive epoxy on the surface to build the electrodes of the NCSS. The resistance changes of the NCSS electrodes were measured with a multi-meter  and the cantilever beam deflection was measured using a laser displacement sensor  as shown in NR) with Equation (1):The measured displacements were converted to strains by cantilever beam theory, and the resistance changes were normalized (R0 is the resistance without any displacement and sR is the measured sensor resistance when the beam is strained. gS) of the fabricated NCSS and corresponds to the gauge factor of the sensor determined by Equation (2):aε is the axial strain due to the beam deflection. To investigate the linearity and gauge factor of the MWCNT piezoresistivity, the relationship between the content ratio of MWCNT and the gauge factor of the strain sensors was investigated as shown in Here, The piezoresistivity shows fairly linear bidirectional strain response over various contents of NCSS. The piezoresistive mechanism of CNT composites strongly relate to electrical contact resistance change of the fillers in the matrix and tunneling effects of the fillers ,23. The To benchmark to a conventional strain gauge, a foil strain gauge  was installed with the NCSS on the same cantilever, as illustrated in To evaluate the sensing stability of the sensors, we simultaneously measured the no loading sensor outputs and step responses of NCSS and the strain gauge under static loads to determine the response performance of the sensors, as shown in The NCSS voltage output was as stable as that of the foil strain gauge. Under a no loading condition, the maximum absolute deviation of the NCSS voltage output (0.022 mV) was 22% greater than that of the commercial strain gauge (0.018 mV). It was confirmed that very stable voltage outputs were acquired from NCSS through improvements in both the electrical properties of the nanocomposites and signal processing. Generally, it is difficult to obtain such a stable voltage output from the CNT composite sensors due to resistance drift of the composite resistance originating from the inherent electrical changing property of CNTs according to surroundings . HoweverTo assess the linearity of the NCSS, the output voltage of the sensor was measured according to the deformation of the cantilever. In this test, the end point linearity was 3.8% full scale (FS) non-linearity, which was fairly linear for a strain sensor, and the response performances of the two sensors were nearly identical. The dynamic characteristics were also tested, as shown in The NCSS was confirmed for vibration monitoring of the structures. The phase discrepancy between NCSS and the foil strain gauge on The NCSS was validated to measure the reliable static and dynamic strain information under loading and deformed structural conditions. The NCSS made of the MWCNT/epoxy shows linear strain responses compared to those of the foil strain gauge. Thus, it is expected that it can be used as a new strain sensor. It is necessary to investigate the thermal and other characteristics of the NCSS. The MWCNT/epoxy based strain sensor can be fabricated as various types and applied to structural SHM.A strain gauge conventionally measures the point strain, but the neuron can measure the averaged strain along the length of the structure where it was installed. Damages such as corrosion and deterioration can be represented as a change in the cross section of a beam segment. Therefore, this study assumes the beam shown in A FE model with ANSYS  was constructed for the glass fiber cantilever beam to study the dynamic response capacity of the neuron and establish a baseline FE model for subsequent damage assessment based on the vibration response of the neuron. Damage detection of the cantilever beam was performed using the vibration data from the neuron. The local damage, such as a crack, corrosion, or other defect, reduced the local stiffness of the structure that affects the global dynamic characteristics of the structure ,22. Ther2) shown in Element 2  was studied to develop a novel SHM sensor that could be easily installed in a composite structure. The NCSS was fabricated with epoxy and MWCNT as a matrix and nano filler, respectively. The NCSS was patterned on a structure using a spray-on technique, and this made it easy to apply it to existing structures for SHM applications. We accomplished very stable voltage outputs from the NCSS through improvements of both electrical properties of the nanocomposite and signal processing. This study evaluated the NCSS sensing characteristics by benchmarking to those of a conventional foil strain gauge. Under a no loading condition, even though a maximum absolute deviation of the NCSS voltage output was greater than the foil strain gauge, the NCSS voltage output was as stable as that of a commercial sensor. For the linearity test of the NCSS, the end point linearity was 3.8% FS non-linearity, which is quite linear for a strain sensor. The NCSS also showed good performance for vibration monitoring of the structures. The dynamic characteristics of the NCSS voltage output response were nearly identical at the 28 Hz excitation test. The NCSS power spectra proved that it can be available for vibration based NDI techniques. The NCSS neuron was developed as a continuous strain sensor. The vibration response of the neuron was investigated for structural damage detection of a composite cantilever. The measured vibration response and damage detection by tracking the first natural frequency showed good results that matched the FE analysis. The spray type, long continuous strain sensor is anticipated to be a versatile sensor in mechanical and civil engineering because of its static and dynamic strain measurement with a longer range and less required data acquisition.This study confirmed that NCSS might be able to measure reliable information under loading and deformed structural conditions. NCSS can provide assurance of the operational health of a composite structure without the need for actuators or complex wave propagation analysis. The NCSS artificial neuron may effectively save the cost of SHM by reducing the number of channels of data acquisition compared to point sensors covering the same area. Thus, it may be applied to the state diagnosing system for composite structures. It is necessary to perform studies on the investigation of thermal and other characteristics of the NCSS. We plan to present more details on the strain sensing characteristics of NCSS with sophisticated experimental results including environmental conditions.


The electrochemically prepared PLL/GO sensory platform toward the oxidation of UA and DA exhibited several advantages, including high effective surface area, more active sites and enhanced electrochemical activity. Compared to the PLL-modified GCE (PLL/GCE), GO-modified GCE and bare GCE, the PLL/GO/GCE exhibited an increase in the anodic potential difference and a remarkable enhancement in the current responses for both UA and DA. For the simultaneous detection of DA and UA, the detection limits of 0.021 and 0.074 μM were obtained, while 0.031 and 0.018 μM were obtained as the detection limits for the selective detection of UA and DA, using DPV in the linear concentration ranges of 0.5 to 20.0 and 0.5 to 35 μM, respectively. In addition, the PLL/GO/GCE demonstrated good reproducibility, long-term stability, excellent selectivity and negligible interference of ascorbic acid (AA). The proposed modified electrode was successfully implemented in the simultaneous detection of DA and UA in human blood serum, urine and dopamine hydrochloride injection with satisfactory results.A novel, simple and selective electrochemical method was investigated for the simultaneous detection of dopamine (DA) and uric acid (UA) on a poly( Dopamine  is one of the most important neuro-transmitters that plays an important role in human metabolism . The defUric acid  is the primary end product of purine metabolism. Average level of UA in blood is about 120–450 μM and in urine is about 2 mM. Abnormal levels of UA in human body fluids may lead to the occurrence of gout, hyperuricemia or Lesch-Nyhan syndrome . Thus, tl-lysine) (PLL), because of its good biocompatibility and flexible structure framework, is a well-known biologically friendly polymer that has gained increasing interest, especially when combined with graphene or graphene oxide [2O2 [In order to tackle this issue, many kinds of materials are used to modify the electrode to raise the sensitivity and selectivity in the simultaneous detection of DA and UA ,20,21. Ane oxide . The comne oxide , H2O2 [2ide [2O2 ,29, cancide [2O2  and geneide [2O2 . Althougide [2O2 ,33, no wl-lysine onto the GO-modified GCE in the potential range of −0.8~2.0 V through cyclic voltammetry (CV). Compared to the electrochemical performances of UA and DA at various electrodes, including the PLL-modified GCE (PLL/GCE), untreated GO-modified GCE (untreated-GO/GCE), the modified electrode with GO scanned from −0.8 to 2.0 V for 10 cycles at 50 mV·s−1 (GO/GCE), and the bare GCE, the excellent electrochemical properties at the PLL/GO/GCE were observed as a result of the superior electrochemical activity of the PLL/GO film toward UA and DA. The high electrocatalytic activity of PLL/GO/GCE to the redox reaction of UA and DA is attributed to the good combination of PLL and GO and the synergistic effect of both composites. Fabrication of a novel electrochemical sensor based on the PLL/GO composite film can provide a facile and effective approach to the simultaneous detection of UA and DA.Herein, we report an electrochemical analytical method for the fast, simultaneous detection of DA and UA based on a PLL/GO-modified glassy carbon electrode (PLL/GO/GCE). The sensory platform of the PLL/GO/GCE was constructed by electropolymerization of l-lysine in 0.1 M phosphate buffer solution  at the bare GCE and the untreated-GO/GCE, respectively  and the cathodic peak potential at approximately −0.51 V (peak D), indicating the oxidation and reduction of l-lysine, respectively. The oxidation onset potential (+0.6 V) of l-lysine at the untreated-GO/GCE is much lower than that at the GCE, indicating that the GO sheets as the connecting matrix for PLL can remarkably reduce the oxidation potential of l-lysine. The remarkable redox peaks and lower oxidation onset potential of l-lysine indicate the increased electrochemical polymerization activity of l-lysine at the untreated-GO/GCE. Furthermore, as shown in The first cycles of the cyclic voltammograms (CVs) are recorded for the electropolymerization of 10 mM ectively A. As canl-lysine on the GO-modified GCE.The electrochemical behaviors of DA and UA at various electrodes were investigated by CV in citric acid-phosphate buffer solution . The results are shown in −1, shows one pair of redox peaks of DA and one oxidation peak of UA. Moreover, the peak currents of analytes at the GO/GCE are much larger than those at untreated-GO/GCE (Curve B) and PLL/GCE (Curve D), but still smaller than those at the PLL/GO/GCE (Curve E). It indicates that the treated GO shows better electrocatalysis to the redox reactions of DA and UA than the untreated-GO and PLL. It may be due to the over-oxidation of GO at the high potential, leading to the more oxygen-containing groups produced in carbon sheets. The more oxygen-containing groups are beneficial for the adsorption of UA and DA through hydrogen bonds. In our previous work [The GCE modified with untreated-GO (Curve B) shows the improved peak current densities towards UA and DA attributed to the higher specific surface area of GO, but only a pair of weak redox peaks of DA and one oxidation peak of UA were observed. Therefore, the reactions of both UA and DA are still irreversible at the untreated-GO/GCE. Compared with the bare and untreated-GO modified GCEs, all peak currents at the PLL/GCE increased (Curve D). However, the GO/GCE (Curve C), where the GO was scanned from −0.8 V to 2.0 V for 10 cycles at 50 mV·sous work , the simFor the PLL/GCE (Curve D), the peak current density of UA exhibits approximately 2.5 times enhancement compared to that at the untreated-GO/GCE. The enhanced peak current density and the improved electrochemical activities toward DA and UA can be attributed to the good conductivity and the more active adsorption sites of PLL, which are prone to combination with nitrogen or oxygen atoms of analytes through hydrogen bonding interactions . With thFor the PLL/GCE (Curve D), two redox peaks of DA appear at 280 and 257 mV, with a peak potential separation of 23 mV. These results suggest that PLL can benefit to the increase in electron transfer between DA and the modified electrode. The oxidation potentials of DA and UA were located at 0.280 and 0.408 V, respectively. While for the PLL/GO/GCE, the relative potentials of DA and UA shifted to 0.284 and 0.442 V, respectively. The increased overpotential of PLL/GO/GCE makes the difference between these two oxidation peak potentials increase from 0.128 V at PLL/GCE to 0.152 V at GO/GCE and to 0.158 V at PLL/GO/GCE, which facilitate the simultaneous detection of DA and UA. Thus, the result, i.e., the increase of peak current densities and difference of oxidation potentials, which may be attributed to the good combination of PLL with the more active adsorption sites and GO with a higher specific area and the possible synergistic effect of both composites, favors the simultaneous determination of DA and UA.A is the electrochemical active area of the working electrode, F is the Faraday constant, c is the concentration of the substrate, D is the standard diffusion coefficient in solution, n is the number of transfer electrons, dlQ is the double-layer charge, and other symbols have their acquiescent meanings.The chronocoulometry technique was applied to estimate the electrochemical active surface areas of the bare GCE, PLL/GCE, untreated-GO/GCE and PLL/GO/GCE. According to the Anson equation  below,QQ) and time (t) is shown in Q versus t1/2 shows a linear relationship after background subtraction  [2, respectively. The effective electrochemical active area of the PLL/GO/GCE is twelve times, three times, and two times greater than that of the bare GCE, PLL/GCE and untreated-GO/GCE, respectively. The high active area of PLL/GO/GCE can increase the adsorption capacities of the modified electrodes toward UA and DA, leading to the enhancement of the current response at the PLL/GO/GCE. The large background current of the CV may also be attributed to the high electron transfer efficiency of the PLL/GO/GCE. In addition, the high surface area of the PLL/GO/GCE can also provide a strong evidence that the PLL and GO components were successfully immobilized on the GCE surface.The relationship between charge (traction B. The ef (25 °C) , the eff2, c is the concentration of UA (5 × 10−5 M) or DA (2.5 × 10−5 M), the number of transfer electrons involved in the oxidation of UA or DA are 2 respectively. The calculated diffusion coefficients are 1.03 × 10−5 cm2/s and 4.54 × 10−5 cm2/s for UA and DA, respectively.The chronocoulometry technique was also used to estimate the diffusion coefficient of UA and DA on the PLL/GO/GCE Ca,b. Whe−1, the anodic peak current densities of UA and DA all increase linearly with the increase of scan rate (Jpa to the scan rate (v) was found to be Jpa,UA (μA·cm−2) = 4.1841 v(mV·s−1) + 196.92 (R2 = 0.996) for UA and Jpa,DA (μA·cm−2) = 2.089 v(mV·s−1) + 31.89 (R2 = 0.995) for DA. Moreover, the redox peak potentials of DA shifted positively and negatively, respectively, as the scan rate increased, suggesting that the redox reaction of DA is quasi-reversible [We investigated the influence of scan rate on the peak currents of UA and DA. can rate B. This rversible . The anoEpa,DA (V) = 0.6095–0.058 pH (R2 = 0.995) for DA and Epa,UA (V) = 0.834–0.07399 pH (R2 = 0.995) for UA. The slopes were 58 and 73.99 mV pH−1 for DA and UA, respectively, which are close to the theoretical value of 59 mV·pH−1. It indicates that the electrochemical oxidation of UA and DA at PLL/GO/GCE involves an equal number of protons and electrons. Therefore, the possible reaction mechanisms of UA and DA at the proposed electrode are shown in The influence of buffer pH on the anodic peak potential of UA and DA was investigated by separately determining the CVs of the two compounds in CPS at a pH range of 2.2 to 7.0. As can be seen in −1), followed by electrochemical polymerization of PLL for 10 cycles. The response current density of DA was explored by differential pulse voltammetry (DPV) at various PLL/GO/GCEs. As shown in The amount of GO on GCE was optimized by varying its volume in the range of 2–6 μL (1 mg·mL−1). The peak signal successively increased with increasing electrodeposition scan number up to ten cycles, which is due to the improvement of the active sites in PLL/GO. Further increasing the polymerization cycles led to a decrease in the anodic peak current, suggesting that the PLL film was overly thick, consequently hindering the electron transfer rate and the mass transport process of DA. Therefore, the optimized number of electropolymerization scans was selected as ten for the formation of PLL films in the following experiments.The thickness of the PLL film was a crucial control factor for the oxidation of DA. The amount of PLL electrodeposited onto the surface of the untreated-GO/GCE was controlled by the CV scan. Jpa,D (μA·cm−2) = 22.89 CDA (μM) − 19.52 (R2 = 0.998). The oxidation peak current densities of UA remained nearly constant as the concentration of DA increased, suggesting that the addition of DA does not interfere with the detection of UA. Similar results were obtained during UA determination in the presence of 5 μM DA. The peak current density of DPV was proportional to the concentration of UA over the range of 0.5–20 μM and 20–200 μM, resulting in linear equations of Jpa,UA (μA·cm−2) = 11.41 CUA (μM) + 3.14 (R2 = 0.998) and Jpa,UA (μA·cm−2) = 2.639 CUA (μM) + 199.74 (R2 = 0.997), respectively. The calculated detection limits  were 0.018 and 0.031 μM for DA (0.5–35 μM) and UA (0.5–20 μM), respectively.The selective detection of DA and UA at the PLL/GO-modified GCE was investigated by DPV in a CPS solution containing a mixture of DA and UA. When the concentration of DA changed, the concentration of UA remained constant, and vice versa. As shown in Jpa,DA (μA·cm−2) = 19.72 CDA (μM) − 0.61 (R2 = 0.998). Jpa,UA (μA·cm−2) = 11.50 CUA (μM) + 7.93 (R2 = 0.996), while for 20.0–180 μM, the regression equation is Jpa,UA (μA·cm−2) = 2.27 CUA (μM) + 194.72 (R2 = 0.992). The detection limits for DA and UA are 0.021 and 0.074 μM with the linear range or 0.5–35 and 0.50–20 μM, respectively. The sensitivities of PLL/GO/GCE for DA and UA in the mixture were 19.72 and 11.5 μA·cm−2·μM−1, while the sensitivity for individual DA or UA was 22.89 or 11.41 μA·cm−2·μM−1, respectively, indicating that the interference between DA and UA in the mixture could be ignorable. The PLL/GO/GCE can be utilized to simultaneously detect the DA and UA with good selectivity and high sensitivity.DA and UA were simultaneously detected by changing their concentrations using the PLL/GO/GCE at pH = 5.0. When the concentrations of DA and UA changed synchronously, the oxidation peak current densities increase accordingly A. The reThe range of linearity, detection limit and sensitivity of this proposed method were compared with some other reports for DA and UA determination; the results were listed in The stability and reproducibility of the PLL/GO/GCE were also studied. The stability of the constructed electrode was investigated by storing at 4 °C for 10 days. The DPV curves were measured and compared with the original curves. The results showed that the oxidation peak currents of DA and UA decreased less than 6.5%, indicating that the proposed electrode is very stable A. The re+ (100 mM), K+ (50 mM), Ca2+ (5 mM), Fe3+ (5 mM), Cl− (100 mM), Zn2+ (5 mM), Mg2+ (5 mM), NO3− (50 mM), SO42− (50 mM), HCO3− (50 mM), CH3COO− (50 mM), glutamic acid (400 μM), alanine (400 μM), aspartic acid (400 μM) and tryptophan (200 μM) did not interfere with the determination  (The possible interferences of some inorganic ions on the determination of 10 μM DA and 100 μM UA were studied by DPV. The results showed that Nathan 5%) C, indicaAA always coexists with DA and UA in human body fluids, and the concentration of AA is much higher than that of UA and DA. The oxidation peak potential of AA is similar to that of DA and UA at the bare electrode. As shown in All of these advantages of the proposed sensor based on PLL/GO/GCE result from the combination of graphene and PLL, which takes full advantages of each of their merits and integrates their unique properties.DA and UA often coexist in real biological samples, such as serum and urine. Therefore, human blood serum and urine were chosen as the actual samples to investigate the possibility of the application of the proposed method by measuring the concentration of UA. All of the urine and blood serum samples were diluted by 100-fold and 5-fold, respectively, using CPS (pH 5.0). The standard addition technique was utilized to measure the recovery by the proposed DPV method. The summarized results for the analysis of human blood serum and urine samples are given in To ascertain the performance of the proposed approach for the determination of DA in dopamine hydrochloride injection, the PLL/GO/GCE was applied to the detection of DA in a pharmaceutical product. The sample was diluted by 5-fold with CPS (pH 5.0), and then a different dilution of the sample was added into the CPS (pH 5.0) for detection. The results are shown in 2HPO4 and 0.1 M citric acid. The PBS was prepared with the stock solutions of 0.2 M NaH2PO4 and 0.2 M Na2HPO4. De-ionized water with a resistivity of 18 MΩ cm was used throughout the experiments.Uric acid, dopamine and ascorbic acid were obtained from Aladdin Industrial Corporation . Graphite powder was purchased from Sinopharm Chemical Reagent Co., Ltd., Shanghai, China. All other chemicals were of analytical reagent grade. The CPS with different pH values was mixed with stock solutions of 0.2 M Nad = 3 mm), a platinum wire, and a saturated calomel electrode (SCE) are used as the working electrode, the counter electrode and the reference electrode, respectively. All potential values were reported with respect to the reference electrode. All measurements were performed in an electrolytic cell with 10 mL of solution. The solution was bubbled with high-purity nitrogen for 15 min to remove the oxygen, and a nitrogen atmosphere was maintained over the surface of the solution throughout the measurements. The pH measurements were made on a pHS-2C digital pH meter . The EDS maps were recorded using an FEI Quanta 250F. All of the experiments were performed at ambient temperature (25 ± 0.5 °C).All electrochemical experiments were performed with a CHI660D electrochemical system . The electrochemical measurements were performed in a single-compartment cell with a conventional three-electrode system, where a bare or modified GCE  was dropped onto the surface of the cleaned GCE and dried at room temperature for two hours. The obtained electrode is denoted untreated-GO/GCE. Secondly, the PLL film was electropolymerized onto the surface of untreated-GO/GCE by cyclic potential sweeping from −0.8 V to 2.0 V in 0.1 M PBS (pH 8.0) containing 0.01 M l-lysine at a scan rate of 50 mV·s−1. The amount of PLL deposited was controlled by the cycling number. The resulting electrode is denoted PLL/GO/GCE. After polymerization, the modified electrode was rinsed with de-ionized water to remove any physically adsorbed substances and dried in air for the following experiments. For comparison, a PLL/GCE was prepared with a bare GCE by the same method. In addition, an untreated-GO/GCE was scanned for 10 cycles in 0.1 M PBS by cyclic potential sweeping from −0.8 V to 2.0 V at 50 mV·s−1. The electrode was named as GO/GCE, where GO was overoxidized during CV scanning.GO was synthesized from natural graphite powder using a modified Hummers method as described elsewhere . Then, t−1. DPV measurements were recorded the following conditions: initial potential, −0.2 V; final potential, 0.6 V, pulse width, 0.05 s; pulse amplitude, 50 mV.CV and DPV measurements were performed with three electrodes in CPS. CV measurements were conducted under the following conditions: initial potential, −0.2 V; high potential, 0.6 V; low potential, −0.2 V; and scan rate, 100 mV·sBlood samples of healthy volunteers were obtained from the Nanjing University of Science and Technology Hospital. Fresh human blood samples were centrifuged to remove precipitating materials for 20 min at 5000 rpm. The serum collected was diluted 5-fold with CPS (pH 5.0) and 10 mL of the diluent was placed in the electrolytic cell to determine UA by the proposed DPV method.The human urine samples were collected and diluted 100-fold with CPS (pH 5.0) before measurement. No other pretreatment was performed. The dopamine hydrochloride injection  was purchased from the Nanjing University of Science and Technology Hospital. The sample was diluted 5-fold with CPS (pH 5.0).l-lysine on the surface of GO sheets, which was successfully used as a sensory platform for the fast simultaneous detection of DA and UA. The calculated diffusion coefficients are 1.03 × 10−5 cm2/s and 4.54 × 10−5 cm2/s for UA and DA, respectively. The PLL/GO-modified GCE exhibited high electrocatalytic activity toward the electro-oxidation of DA and UA by significantly amplifying the peak currents. The larger peak separation between DA and UA achieved by CV or DPV at the PLL/GO/GCE allowed for their simultaneous detection. The proposed sensor showed acceptable sensitivity, good reproducibility, long-term stability, good selectivity and negligible ascorbic acid interference. Moreover, the modified electrode was also successfully utilized to the analysis of DA and UA in real samples with satisfactory results.We fabricated a PLL/GO-modified electrode by the electrochemical polymerization of


Human T-lymphotropic virus 1 (HTLV-1) infection has been associated with ATL and inflammatory diseases but remains a neglected health problem. HTLV-1 associated diseases were originally described as sporadic entities, but family aggregations have been reported. Viral, genetic, immunological and behavioral factors were used to explain family clusters, but until now a clear explanation remains uncertain. In the present study we report, for the first time, a family cluster of diseased persons presenting the infection across three generations associated with FAS -670A/G polymorphism. HumanT-lymphotropic virus 1 (HTLV-1) is a human retrovirus that causes lifelong infection . Recent Although the infection has been associated with inflammatory and lymphoproliferative diseases  it remaiHuman T-lymphotropic virus 1 (HTLV-1) associated diseases were originally described as sporadic entities, but family clusters have been long reported worldwide . MembersMBL2 and FAS genes, respectively, were strongly associated with HTLV-1 infection. ∗BMBL2 allele was associated with higher proviral load, a viral biomarker of disease outcome. Similarly, FAS -670A/G single nucleotide polymorphism allele ∗G was more frequent among HTLV-1 asymptomatic infected persons and allele ∗A was more prevalent among those with clinical neurological symptoms diagnosed as HTLV-1 associated myelopathy/Tropical spastic paraparesis (HAM/TSP). The present study aimed to investigate if a family aggregation of HTLV-1 infection could be associated with FAS and MBL2 gene polymorphisms.The search for host immunogenetic markers, possibly associated with HTLV-1 infection and progression to disease was pursued by our research group in the Amazon region of Brazil and other groups elsewhere . PolymorThe study was approved by the Research Ethics Committee of the University Hospital João de Barros Barreto from the Universidade Federal do Pará (process No. 2061/2005). Participants were fully informed about the research objectives, and those who agreed to participate signed an informed consent form.Figure 1 and Table 1 describe a three generation family, from the Northern of Brazil (Amazon region), which included six members . HTLV-1 infection was investigated in the plasma and cells of blood samples. An enzyme immune assay was used for the detection of antibodies to HTLV-1/2 in the plasma. Positive samples were submitted to DNA extraction from PBMC using the purification kit GFX for genomic DNA  and assayed using nested PCR and qPCR to confirm the infection, as previously described  was an asymptomatic married HTLV-1 carrier, who breastfed her two children and both were also asymptomatic HTLV-1 carriers.MBL2 gene polymorphism analysis showed the presence of AA wild genotype in three subjects. The allele MBL2∗B (rs1800450), carried by heterozygous genotype (AB), was observed in two sisters (second generation) and in one male of the third generation .∗G; three of them were asymptomatic homozygous GG and two were symptomatic heterozygous AG, one with a clinical diagnosis of uveitis and the other with HAM/TSP. One of the females with HAM/TSP was a carrier of AA genotype. The allele ∗G was more frequent among the asymptomatic persons, but the allele ∗A, was present among the three symptomatic sisters.The investigation of FAS -670A/G polymorphism showed that five members of the family (83%) carried the allele ∗B in the exon 1 of the MBL2 gene has been reported with the occurrence of immunodeficiency and chronic infectious diseases  carry the genotype AA, suggesting that there are other contributing factors for the progression of disease, apart from MBL2 gene SNP.MBL is an important acute-phase serum protein involved in the innate immune response that can trigger complement activation , and an diseases . In our ∗G was more frequent among the asymptomatic persons. Allele ∗A, was present in three symptomatic sisters, supporting our previous report that showed higher frequency of allele ∗A among HAM/TSP patients as compared to HTLV-1 asymptomatic persons (FAS gene (FAS -670A/G polymorphism was investigated and the allele  persons . The phyFAS gene . Our preFAS gene . Farre eFAS gene . MoreoveIt is important to point out that although familial aggregation of HTLV-1 infection and disease is not a recently described characteristic of the natural history of the virus, there are few attempts to characterize associated biomarkers, which favor the clustering of cases and define the outcome of the infection. FAS -670A/G polymorphism seems to be a reliable indicator to pursue involving larger numbers of other families with HTLV-1 aggregation. This would certainly help to assist toward prevention of the virus spread.Conceived and designed the experiments: AV, MS, and RI; wrote the paper: AV, IC-V, and RI; assisted with editing the paper: MQ, AS, and IC-V; performed the experiments: BS, CC, and MQ; analyzed the data: AV, BS, RI, and MS; contributed reagents/materials/analysis tools: AV, MS, and RI.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. At the time of acceptance, the handling Editor declared a shared affiliation, though no other collaboration, with the authors.


Giardia duodenalis is one of the most common enteric parasites in domestic animals including dogs. Young animals are more prone to the infection, with clinical manifestations ranging from asymptomatic to acute or chronic diarrhoea. Dogs are primarily infected by canine-specific (C-D) assemblages of G. duodenalis. However, zoonotic assemblages A and B have been increasingly documented in canine isolates, raising the question of whether and to which extent dogs can act as natural reservoirs of human giardiosis.G. duodenalis in dogs in the province of Castellón, Eastern Spain. A total of 348 individual faecal samples from sheltered (n = 218), breeding (n = 24), hunting (n = 68), shepherd (n = 24), and pet (n = 14) dogs were collected between 2014 and 2016. Detection of G. duodenalis cysts in faecal material was carried out by direct fluorescence microscopy as a screening test, whereas a qPCR targeting the small subunit ribosomal RNA gene of the parasite was subsequently used as a confirmatory method.In this cross-sectional epidemiological survey we assessed the molecular diversity of Giardia duodenalis was detected in 36.5% (95% CI: 31.6–41.7%) of dogs. No significant differences in prevalence rates could be demonstrated among dogs according to their sex and geographical origin, but breeding  and sheltered  dogs harboured significantly higher proportions of G. duodenalis. Multi-locus sequence-based genotyping of the glutamate dehydrogenase and β-giardin genes of G. duodenalis allowed the characterization of 35 canine isolates that were unambiguously assigned to assemblages A (14.3%), B (22.9%), C (5.7%), and D (37.1%). A number of inter-assemblage mixed infections including A + B (11.4%), A + D (2.9%), and A + B + D (5.7%) were also identified.G. duodenalis infections detected, but very few of the genotypes identified have been previously documented in Spanish human populations. Although possible, zoonotic transmission between dogs and humans seems an infrequent event in this Spanish region.Data presented here are strongly indicative of high infection pressures in kennelled animals. Zoonotic sub-assemblages AII, BIII, and BIV were responsible for a considerable proportion of the The online version of this article (10.1186/s12917-018-1353-z) contains supplementary material, which is available to authorized users. Pet animals in general and dogs in particular are increasingly regarded as true family members in many homes globally. In the UK in 2016 canine population stood at around 8.5 million, with 24% households owning a dog . HoweverGiardia duodenalis is one of the most commonly detected pathogens associated with diarrhoea in humans and animals, including domestic dogs . Out of the 127 dogs that tested positive by DFAT, 81.1% (103/127) were confirmed by qPCR, whereas 18.9 (24/127) failed to be detected by the latter method. The qPCR-positive samples had cycle threshold (Ct) values ranging from 21.4 to 39.6 . Overall, 68.9% (71/103) of the Giardia-positive isolates by qPCR had Ct values ≥30 . Regarding dog status, detected G. duodenalis prevalences ranged from 20.6%  in hunting dogs to 45.8% (95% CI: 27.9–64.9%) in breeding dogs. Statistically significant differences were found between hunting and breeding dogs (P = 0.017) and between hunting and sheltered dogs (P = 0.003). No obvious differences in the geographical distribution pattern of G. duodenalis was demonstrated (P = 0.152), although the infection was more frequently detected in Alcalatén  and Bajo Maestrazgo , with Alto Maestrazgo  and Plana Alta  harbouring the lowest infection rates.Table G. duodenalis isolates confirmed by qPCR, 34.0% (35/103) were successfully amplified at the gdh and/or bg markers. Multi-locus genotyping data were produced for 28.6% (10/35) of them, whereas 51.4% (18/35) and 20.0% (7/35) of the canine isolates were only amplified at the gdh or the bg loci, respectively. Sequence analyses revealed the presence of assemblages A , B , C , and D (37.1%¸13/35) , A + D , and A + B + D  were also identified  of reference sequence U60984. Finally, the 13 isolates assigned to the assemblage D of G. duodenalis were distributed in nine distinct, including a novel (MF285575), genotypes differing between none and four SNPs in a 402-bp fragment stretching from positions 80–481 of reference sequence U60986. The phylogenetic analysis revealed that our A-D sequences clustered together in well-supported clades with the corresponding assemblage and sub-assemblage reference sequences from NCBI, as they also did with sequences of human and canine origin previously documented in other Spanish studies [Sub-genotyping data of the 21  studies , 21, 33 bg marker are shown in Table bg locus in the present study. A novel C genotype (MF285584) revealing two polymorphic sites at positions 37 (G to A) and 451 (C to T) of reference sequence AY545646 was also found, whereas the three isolates genotyped as assemblage D differed from none to four SNPs in a 486-bp stretch (positions 105–590) of reference sequence AY545647.Sub-genotyping data of the 10 isolates with only single-assemblage infections at the G. duodenalis in Spanish dog populations. Microscopy-based surveys have estimated the prevalence of G. duodenalis at 6–38% in the northeast [G. duodenalis cysts have also been found in 5–18% of faecal droppings and soil samples collected in public parks of the latter region [G. duodenalis [Few epidemiological surveys have been aimed at investigating the occurrence of ortheast , 19, at ortheast , 22, andortheast , 24 of tr region . Overallr region , 35. Comr region , 21, mucodenalis . Of inteodenalis . A potenG. duodenalis infection primarily presented as a sub-clinical, asymptomatic  disease in the surveyed canine communities. Although exact age could not be established, the vast majority of the sampled dogs were young adults or mature animals, suggesting that acquired immunity may play a role in the control and/or severity of the infection [Giardia was reported from dogs older than 1 year which attended at a veterinary hospital in USA during an 11-year period [G. duodenalis cyst counts obtained during DFAT examination, and ii) relatively high Ct values obtained during qPCR. Taken together, these findings indicate that low-to-moderate G. duodenalis burden was the norm in most of the infected dogs detected in the present study. Importantly, shedding of limited cyst numbers would negatively impact the diagnostic sensitivity of the PCR-based methods used for genotyping and sub-genotyping purposes, a fact further aggravated when considering that both gdh and bg markers are single-copy genes. Very similar findings have been previously documented in other canine studies in Spain [Our results indicate that nfection . Indeed,r period . Indirecin Spain , 21.G. duodenalis prevalences were observed among the dog communities investigated here. Sheltered and breeding dogs had a significantly higher prevalence compared to hunting dogs, but not compared to other dog categories. Kennel dogs have been demonstrated to be at higher risk of infection due to continuous exposure to G. duodenalis cysts in kennels with high animal density [G. duodenalis observed in pet dogs, a potentially serious public health concern if infected animals are in close contact with children or immunocompromised individuals. Because none of the G. duodenalis isolates obtained from pet dogs in the present survey could be genotyped at the assemblage level, more research should be conducted to elucidate the actual role of domestic dogs as natural reservoirs of human giardiosis. Finally, hunting dogs exhibited the lowest (20.6%) G. duodenalis infection rate, a figure in the lower range of those (20–30%) reported in similar studies conducted in Spain [Marked differences in  density , 8, 37. in Spain  and ItalG. duodenalis circulating in dogs in the province of Castellón. Assemblages C/D are known to account for ~ 70% of the cases of canine giardiosis documented in European countries [G. duodenalis assemblages. The fact that the vast majority of these mixed intra-assemblage infections were found in kennelled animals was well in agreement with the former observation that these dogs underwent high infection pressures associated with crowded living conditions (see above). In this regard, mixed intra-assemblage infections have been documented in 2–27% of the canine isolates genotyped in Europe [G. duodenalis infections were demonstrated in two independent molecular surveys based on PCR and sequencing analyses targeting sheltered and pet dogs in the north of the country [As anticipated, our genotyping analyses confirmed that canine-specific assemblages C and D, particularly the latter, were the most prevalent genetic variants of ountries , 38, 39,ountries , Germanyountries , Greece ountries , and Spaountries , 19. Howountries , 24, Gerountries , and Norountries  moleculan Europe  and up tn Europe . In Spai country , 21, altgdh and bg loci also revealed exciting molecular data. For instance, AII sequences confirmed the high genetic diversity previously reported in canine isolates of this particular sub-assemblage in other Spanish regions [gdh and bg genes than those of human origin has two important consequences. Firstly, it may supports the existence of genetic exchange, challenging the still widely accepted  notion that Giardia is an organism strictly asexual [Giardia. Additionally, recombination events would be enhanced in epidemiological scenarios characterized by high prevalence rates and elevated infection pressures as those locally described in the present study. Indeed, intra-assemblage recombination has been already demonstrated in AII and B isolates in a highly endemic area in Peru [Sub-genotyping analyses at both the  regions , with vi regions , 45. The asexual . In this asexual , intrage asexual , and nuc asexual  have bee in Peru , 50. Sec in Peru , 11.gdh (MF285566) or the bg (MF285583) genes to those predominantly found circulating in Spanish symptomatic patients [Less surprising was the demonstration of high levels of genetic diversity within the canine isolates assigned to sub-assemblages BIII and BIV, in line with previously reported genotyping data in both canine , 21 and patients , 45. ThiG. duodenalis is common in dogs, typically presenting as a light, asymptomatic infection. Highly endemic foci of disease were detected in breeding kennels and dog shelters where infection pressures were high. Although a significant proportion of the infected dogs harboured potentially zoonotic assemblages and sub-assemblages of G. duodenalis, most of these genotypes seemed to be primarily transmitted within canine cycles and posed, therefore, a limited risk to humans. However, the actual extent of this statement must be corroborated in future molecular epidemiological surveys including human and dog puppy isolates from this Spanish geographical area.Prevalence data presented here are consistent with an epidemiological scenario in which Additional file 1: Table S1.Giardia duodenalis in this study. (DOCX 13 kb)Oligonucleotides used for the molecular identification and characterization of Additional file 2: Figure S2.Giardia duodenalis in DNA isolates from canine faecal samples. (DOCX 31 kb)Histogram of cycle threshold (Ct) values obtained by real-time PCR for the detection of


Metabolic disorders are a group of interrelated conditions which increases the risk of developing heart diseases, stroke, and diabetes. These usually occur as a consequence of deficiency of enzymes involved in biochemical reactions in the body. The dietary habits, lack of physical exercise, stress, and genetic susceptibility leads to an increased risk of developing metabolic disorders. A diet rich in processed food items containing high calories aggravates the production of a purine metabolite, the uric acid (UA). UA functions as an antioxidant, protects against inflammation, aging, and cancer. It exists as urate ions in the circulation and blood level of UA is maintained by a balance between its production in the liver and its excretion by the renal tubules. The regular excretion of UA through the kidneys is necessary to maintain optimum blood levels of UA (3-7 mg/dl). There are various transporters of uric acid present around the renal tubules, which help in reabsorption of UA into the blood. These urate transporters (UT) are proteins coded in the genes. Mutations in these genes may prompt disturbances in UA reabsorption, and could lead to the development of hyperuricaemia, insulin resistance, endothelial dysfunction, diabetes and other metabolic diseases. This paper reviews eight such genes coding for UTs and attempts to unravel the link between the activities of UA, UTs, and the consequences during mutations in the genes coding for the UTs in the development of metabolic disorders. The genes reviewed included SLC2A9, SLC17A1, SLC22A12, SLC16A9, GCKR, LRRC16A, PDZK1, and ABCG2. Metabolic disorders are a cluster of conditions which may include abdominal obesity, high blood pressure, high blood sugar, high serum triglycerides and low serum high-density lipoprotein (HDL). Metabolic diseases are caused due to imbalances in energy utilization and storage, and is associated with risk of developing chronic diseases like the cardiovascular disease and type 2 diabetes mellitus (T2DM) . Among tUA is a heterocyclic compound of carbon, nitrogen, hydrogen, and oxygen, and is an oxidation product of purine metabolism present in the blood circulation. UA acts as an antioxidant and helps in defense against inflammation, aging, and cancer. UA exists as urate anion at physiological pH, and these urate ions are impermeable to membrane and require a transporter. UA levels in the body are maintained by a balance between its production and excretion. The major site of urate formation is in the liver and it is excreted in the proximal tubule of the kidneys, where 90% of it is reabsorbed into the blood, facilitated by various transporters. Kidneys play a major role in regulating UA levels and are responsible for 60–70% of UA excretion and the remaining 30–40% of UA is secreted into the intestine where it is acted upon by bacteria and then eliminated from the body . ElevateHU is strongly associated with insulin resistance, an established risk factor for T2DM. It was estimated that for every mg/dl rise in blood UA, the risk of developing T2DM increases by 20% in adults and 15% in children . HyperurSolute carrier family 2 member 9 (SLC2A9)SLC2A9 is a high-affinity urate transporter present in the apical and basolateral membrane of the proximal convoluted tubule (PCT) of the kidneys. It carries information regarding the synthesis of a protein, glucose transporter 9 (GLUT9). Hence, this gene is also commonly called as GLUT9. GLUT9 is involved in reabsorption of hexoses, but pH variations can make this transporter to carry negatively charged purine derivatives such as urate instead of hexoses . GLUT9 iThe experimental evidence indicates that SLC2A9 is a direct target gene for p53 (apoptotic gene) and plays a key role in reducing reactive oxygen species (ROS) through uric acid transport. Thus, its expression is regulated by p53 gene and is mediated by oxidative stress . The rolMutation in this gene or loss of its activity leads to hyperuricaemia, gout, Alzheimer, and T2DM as shown in Figure Solute carrier family 17 member 1 (SLC17A1)SLC17A1 is a family of nine transporters majorly involved in phosphate transport and organic anion transport. SLC17A1 transporter facilitates the transport of phosphate into the cells via sodium co-transport present in the renal brush border membrane . This ge+/H+ exchanger regulatory factor 1 (NHERF1) by inhibiting phosphate uptake by PCT cells, which in turn inhibits NPT1 gene activity. NHERF1 present in kidney regulates NPT1 activity by stimulating phospholipase-C (PLC) and protein kinase C (PKC) signaling molecules. We know that membrane potential changes lead to sequential events in the signaling molecules to release hormone thus, NHERF1 may cause a change in membrane flux which influences insulin secretion via these PLC and PKA [SLC17A1 also interacts with hepatic nuclear factor 1-alpha (HNF1α) as it is expressed in hepatocytes. The electrogenic nature of this transporter facilitates the clearance of organic anions like urate both from liver and kidneys. It interacts with HNF-1α, which acts as a transcriptional regulator of insulin secretion, bile acid and HDL metabolism, and mutation in SLC17A1 may lead to hypercholesterolemia and hyperglycemia. It was found in experimental studies that this gene activity is regulated by Na and PKA . Thus, NIt was also found that single nucleotide polymorphism (SNP) on SLC17A1 gene or a mutation in this gene not only leads to decreased UA excretion but raises the blood levels of UA leading to hyperuricemia . HyperurSolute carrier family 22 member 12 (SLC22A12)Solute carrier family 22 belongs to an organic anion transporter (OAT) family. There are about 10 members of OAT family transporters located at various regions of renal tubules, and these are involved in the transport of various metabolites, drugs, and toxins . Of thesThe structural analysis of URAT1 revealed an N-glycosylation site as well as phosphorylation sites which helps in its regulation. The phosphorylation of URAT1 by phosphokinase-C (PKC) stimulates uptake of UA by OAT3 and OAT4, which then is reabsorbed with the help of URAT1 across the PCT of kidney. The PKC phosphorylation can also activate phospho inositol-3- kinase (PI3K) pathway which has a role in insulin secretion  and aids in glucose uptake by glucose transporters. The release of insulin from beta cells of pancreas also increases the reabsorption of UA in PCT. Hence mutation in the gene may alter beta-cell function and raised two-hour glucose levels like SLC2A9 gene. So, it can be concluded that URAT1 and GLUT9 together efficiently help in UA reabsorption in PCT . URAT1 pAs URAT1 is also involved in the transport of drugs and pharmacological agents, use of anti-inflammatory, hypertensive drugs could increase the excretion of UA, thereby decreasing its levels in the blood . MutatioSolute carrier family 16 member 9 (SLC16A9)This gene codes for a protein which helps in the transport of monocarboxylic acids like pyruvic acid, lactic acid, etc. Hence, this gene belongs to the family of monocarboxylates which specifically codes for monocarboxylate transporter 9 (MCT9). MCT9 helps in the transport of monocarboxylic acids together with proton-linked transport. The monocarboxylates play a vital role in the cellular and metabolic communications between tissues . MCT9 isA study by Kolz et al. had noted that there is a triangular association between serum UA, MCT9 and metabolites like carnitine, acylcarnitine, and propionyl carnitine . TherefoMutation or loss of this gene activity leads to gout of extra renal origin, obesity, cardiovascular complications, kidney disorders, T2DM, and cancer as shown in Figure Glucokinase regulatory protein (GCKR)This gene codes for a protein glucokinase (GK), which is produced in hepatocytes. The glucokinase is an enzyme which helps in phosphorylation of glucose at C6 position and its activity is dependent on blood glucose levels. It acts as a glucose sensor in response to raised blood glucose levels. GK enzyme moves between nucleus and cytosol, i.e., at high blood glucose levels the GK enzyme moves to cytosol, and forms a complex with microfilaments of hepatocytes and then phosphorylates available glucose molecules in conjugation with the insulin hormone function . The gluDepending on the ATP (adenosine triphosphate) levels of the cell, the glucose 6 phosphate enters a glucose oxidative pathway (hexose monophosphate shunt (HMP) pathway) to produce pentoses, and nicotinamide adenine dinucleotide phosphate (NADPH). Pentoses are utilized in nucleic acid metabolism, and the availability of ribose-5-phosphate increases UA levels. NADPH is used for the biosynthesis of fat . Alternatively, during fasting or energy-deprived state, GK is inactive and thus metabolic pathway proceeds for gluconeogenesis and inactivation of GK during gluconeogenesis prevents a futile cycle (energy of the cell is dissipated as heat) . Its expMutation in this gene leads to hypoglycemia, hypertriglyceridemia, obesity, and cardiovascular risk. Loss of its activity leads to the development of hyperglycemia and maturity-onset diabetes of the young 2 (MODY2). Enhanced activity of HMP pathway increases the availability of pentoses, which in turn leads to gout and kidney diseases as shown in Figure Leucine-rich repeat-containing protein 16 A (LRRC16A)+). The UTS also interacts with scaffolding proteins like PDZK1, NHERF1, and cytoskeletal filaments like actin [This gene encodes a protein called capping protein ARP2/3 and myosin-I linker (CARMIL), which is an inhibitor of actin capping protein (CP). The CP is an essential element of actin cytoskeleton, which binds to the actin filaments and prevents its polymerization (process of converting monomer to polymer). The CP inhibits polymerization of actin filaments by binding to its barbed ends. CARMIL protein is expressed by various tissues, but predominantly in kidneys. In the apical membrane of proximal tubules, urate transportsome (UTS) is formed. UTS is formed by several UA transporter proteins like URAT1, OAT4L, NPT1, and ABCG2 which have a role in the absorption of UA in exchange with sodium ions  and protein kinase C (PKC) signaling molecules. Considering the fact that Insulin hormone is regulated mainly by these signaling molecules, a defect in the gene coding for CARMIL could be associated with gout as shown in Figure Protein domain containing 1 (PDZK1)This gene codes for a scaffolding protein. A scaffolding protein connects membrane proteins and regulatory components, and also regulates its expression on the surface of epithelial cells. PDZK1 protein is expressed on the epithelial cells of kidneys, liver, pancreas, gastrointestinal tract, and adrenal cortex. It acts as a scaffolding protein by interacting with membrane-associated proteins like the MAP-17, and with other cytoplasmic proteins. Hence, PDZK1 has a role in cell proliferation, differentiation and ion transport .+/H+ exchange regulatory factor 3 (NHERF-3) in the brush border cells of renal tubules. PDZK1 and NHERF 3 together act as scaffolding protein [It was found in experimental studies that PDZK1 increases the expression of the Na protein -36. Alon protein . Another protein . PDZK1 a protein -40.PDZK1 organizes a transportsome and regulates several transporter functions like urate and drug transport. A urate transportsome is formed by linking several UA transporters like ABCG2, SLC22A12, NPT1, which in turn relate to each other by scaffolding proteins like PDZK1, NHERF1 at the apical surface of renal tubules. Mutations in these UA transporters  also leads to hyperuricemia and gout. So, loss of activity of PDZK1 indirectly causes gout .PDZK 1 also stabilizes and prevents the degradation of scavenger receptor class B type 1 (SRB-1) protein on the surface of hepatocytes and endothelial cells. SRB 1 is a cell surface glycoprotein that binds to HDL with high affinity. It also plays an important role in reverse cholesterol transfer function of HDL, and has a role in atheroprotection . PDZK1 aAdenosine triphosphate-binding cassette sub-family G member 2 protein (ABCG2)ABCG2 gene codes for ATP binding cassette (ABC) transporter. ABC transporters utilize energy by hydrolyzing ATP and transports substrates across cell membrane . ABCG2 wABCG2 is an efflux urate transporter seen on the proximal renal tubules. It is a high capacity secretory urate transporter  whose activity is completely dependent on intracellular levels of UA. ABCG2 is also expressed in liver and intestinal cells. Experimental studies on mice have confirmed that dysfunction or loss of activity of ABCG2 increases the urinary excretion of UA due to the action of URAT1 on intestine. This concludes that decreased urate excretion could be via extra-renal pathway. Thus, ABCG2 regulates UA levels via the extra-renal UA excretion, which is the major cause of extra-renal underexcretion, and gout . Later iUric acid is a product of purine metabolism and its reabsorption majorly happens in the renal tubules. There are about eight genes coding for urate transporters involved in its reabsorption process present in various parts of renal tubules. These transporters are arranged in a pocket-like or a scaffolding-type structure around the renal tubules, and the genes coding for them acts in combination to transport the urate and other molecules. Mutation or polymorphism or loss of genes coding for transport causes hyperuricemia, which increases the glycation of proteins, and later could be responsible for the development of metabolic disorders like T2DM. Hence, identification of mutations in the genes encoding urate transporters will play a crucial role in the control and prevention of metabolic disorders.


Fertility is one of the three principal components of population dynamics. High fertility and rapid population growth exert negative influences on economic and social development. This study was aimed to estimate the level and trends of fertility among (15–49 years) old women in kersa demographic surveillance and health research center, kersa district Eastern Ethiopia.The study was conducted at kersa demographic surveillance and health research center in kersa district, Eastern Ethiopia. The study utilized five years follow up data (2008–2012). All women (15 to 49 years old) who have been living at kersa demographic surveillance and health research center in kersa district from 2008 to 2012 were included in the analysis to estimate the level and pattern of fertility. Descriptive statistics was computed to determine level and pattern of fertility.Age specific fertility rate was highest in women of the age group 25–29 years old  in five consecutive years from 2008 to 2012. Total fertility rate didn't show any significant decline during the study period, i.e., 4.3, 4.5, 4.9, 3.5, 4.0 live births per woman throughout her reproductive period (15–49) years old in 2008, 2009, 2010, 2011 and 2012, respectively. On the other hand, general fertility rate declined from 110.3 births per 1000 women in 2008 to 95.9 per 1000 women in the reproductive age in 2012.The total fertility rate was found to be relatively high. Fertility rate is higher in rural residents and illiterate women than in urban residents and literate women. Strong information, education, communication and behavior change communication on family planning should be designed and implemented to prevent unwanted fertility. The total fertility rate (TFR) is the average number of children a woman would bear if fertility rates remain unchanged during her lifetime . Fertilith in 2002 and projected to be third in 2050. The population of Sub-Saharan Africa is growing at 3 % per year .There was no as such any significant decline in the total fertility rate (TFR) in the study period i.e., 4.3, 4.5, 4.9, 3.5, 4.0 live births per woman in her reproductive period (15–49 years old) in 2008, 2009, 2010, 2011 and 2012, respectively. Similarly, the decline in general fertility rate was not as such big, i.e., it declined from 110.3 births per 1000 women in reproductive age in 2008 to 95.9 per 1000 women in the reproductive age in 2012 [Fig. Approximately three fourth of women gave birth before twenty years old. Relatively higher percentage of women gave birth below twenty years old in 2009 (73.7 %) and 74.4 % in 2010. Almost one in three women give birth when their age was twenty years and above. There was no significance difference across age at first birth in the five consecutive years (2008–2012).There was a difference in the total fertility rate pattern between literate and illiterate women. The total fertility rates among illiterate women were approximately two times higher than the total fertility rate of the literate women. The interesting thing here, was that there was consistent decrement of fertility among both literate and illiterate women Fig. .Fig. 3PaThe trends in TFR in the study period didn't show as such big declines which shows that the transition in fertility among the populations of the study area was very slow. The fertility transition was very rapid in urban areas of the study population especially starting from the year 2010 the level of TFR has dropped to replacement level fertility Fig. .Fig. 4TrThe population of the study area grew by 2.9 % per annum during the study period and the crude birth rate changed from 31.4 births per 1000 population in 2008 to 29.8 births per 1000 population in 2012 Figs.  and 6.FiThis study determined the level and pattern of fertility rates in the Kersa demographic surveillance and health research center survey. The findings from this study showed that the population of Kersa Demographic Surveillance and Health Research Center has grown by 2.9 % per annum during the study period  and the crude birth rate has declined from 31.4 births per 1000 population in 2008 to 29.8 births per 1000 population in 2012 Fig.s.  and 6. TThe TFR in this study declined on average by 0.19 % annually. Although a change in TFR over five years was not statistically significant  the direction of change was declining. Absence of a statistically significant change in TFR could be because of shortness of the study period, i.e., only covering five years in which significant changes in factors affecting fertility may not be expected. The decline in TFR observed in this study (from 4.3 in 2008 to 4.0 in 2012) is comparable with findings from Kenya in the years 2002–2008 in which TFR declined from 4.92 to 3.92 .The level of TFR in this study is higher than the TFR of Namibia in 2006 which was 3.6 births per woman , ZimbabwAge specific fertility rate was highest in the 25–29 years old age group in five consecutive years from 2008 to 2012. Age specific fertility rates observed in the two extreme ages. i.e., for adolescents aged (15–19) and for mothers aged > =35 years was lower compared to other age categories. The findings of this study are comparable to the findings of other studies. For example, there was high age specific fertility rate for women 20–24 years old in Zimbabwe, and in other similar studies fertility peaks for ages 25–29 years old both in rural and urban areas . The finAge specific fertility rates, total fertility rates and general fertility rates were higher among rural resident women than urban residents. Similarly, illiterate women had higher fertility rates than literate women. This finding is also apparent in the results in Namibia, that women who live in rural areas have more children than those in urban areas. The effect of place of residence on the total number of children ever born indicates a decrease from 1992 to 2006. In the long run it is anticipated that fertility will decrease among women in the rural areas . In SoutEthiopia is the second populous country in Africa with a growth rate of 2.6 percent per year and total fertility rate of 4.8 children per woman. The Federal Ministry of Health has strived to achieve the MDG number 4 and 5. The goal of the country is to reduce TFR to 4.0 through education of women and universally accessibility of family planning to women in the reproductive ages. Therefore, findings of the current study have clear implications for policy makers, program planners and any concerned bodies in their efforts to design strategies to tackle unwanted fertility.The strength of this study is that it used community based quantitative demographic and health surveillance data for five consecutive years. However, the data are self reported so the accuracy of the response depends on the willingness of the respondent to give trustworthy information. This is a cross sectional study design that can not establish cause and effect relationship.Total fertility rate was declining. Fertility rate is higher in rural residents and illiterate women than urban residents and literate women. Strong information, education, communication and behavior change communication on family planning, and should be designed and implemented to prevent unwanted fertility.