In areas approaching malaria elimination, human mobility patterns are important in determining the proportion of malaria cases that are imported or the result of low-level, endemic transmission. A convenience sample of participants enrolled in a longitudinal cohort study in the catchment area of Macha Hospital in Choma District, Southern Province, Zambia, was selected to carry a GPS data logger for one month from October 2013 to August 2014. Density maps and activity space plots were created to evaluate seasonal movement patterns. Time spent outside the household compound during anopheline biting times, and time spent in malaria high- and low-risk areas, were calculated. There was evidence of seasonal movement patterns, with increased long-distance movement during the dry season. A median of 10.6% (interquartile range (IQR): 5.8–23.8) of time was spent away from the household, which decreased during anopheline biting times to 5.6% (IQR: 1.7–14.9). The per cent of time spent in malaria high-risk areas for participants residing in high-risk areas ranged from 83.2% to 100%, but ranged from only 0.0% to 36.7% for participants residing in low-risk areas. Interventions targeted at the household may be more effective because of restricted movement during the rainy season, with limited movement between high- and low-risk areas. The sIn areas approaching malaria elimination, human mobility patterns are important in determining the relative proportions of malaria parasites that are imported or the result of low-level, endemic transmission –13. PopuMany methods to measure human mobility have been explored to describe the impact of human movement on malaria transmission . Long-diMobile phone data are a convenient and accurate method for measuring human movement ,15. HoweOn both long- and short-distance spatial scales, human movement can be measured at a fine spatio-temporal resolution using commercially available GPS data loggers to describe individual movement patterns –25. TheiCommercially available GPS data loggers were used to determine movement patterns among a population of rural, agrarian participants selected from a longitudinal cohort study of malaria epidemiology in Southern Province, Zambia. These analyses aid in explaining the micro-epidemiology of malaria transmission and the risk of imported malaria as elimination is achieved and sustained . Knowled2.2.1.2 in area. The single rainy season lasts from November through to April, followed by a cool dry season from April until August and a hot dry season from August through to November. Malaria transmission peaks during the rainy season with the highest incidence of clinical malaria typically occurring in April [Anopheles arabiensis [The study was conducted in the rural catchment area of Macha Hospital in Choma District, Southern Province, Zambia, 70 km from the nearest town of Choma and approximately 1200 kmin April . The priabiensis ,33. The abiensis . Artemisabiensis ,36 and iabiensis . LLINs wabiensis  and moreSatellite images were used to develop a sampling frame for the random sampling of households to recruit and enrol individuals into longitudinal and cross-sectional surveys of malaria parasitaemia starting in 2008 . The ide2.2.Criteria for selection of the GPS devices were developed to accommodate the study population, ensure participants would not be responsible for charging the devices and to protect privacy. These criteria included size, weight, water resistance, battery life, memory size, programming capabilities, motion detection and validity. IgotU® GT-600 (Mobile Action Technology) GPS loggers were selected as they were shown to be accurate (point accuracy of 4.4 m and line accuracy of 10.3 m) and acceptable in a study conducted in Iquitos, Peru . These dAll participants enrolled in the existing longitudinal cohort who were 13 years and older were invited to participate during bi-monthly study visits. A non-random convenience sample was selected during study visits from October 2013 through to August 2014. The study staff aimed to enrol 12 participants per month and have at least 10 complete the full month. Up to three participants per household were permitted to participate with no more than two individuals participating concurrently in a single month. Enrolled participants were requested to carry the GPS data logger at all times they were active for a one-month period. This allowed for a full year of data collection to assess seasonal movement patterns.Serial numbers of the GPS data loggers were matched to participant unique identification numbers. The power button was locked and the GPS data loggers were password-protected, so that only study staff could access the data. Geographical position was logged every 2.5 min. The loggers were programmed to be motion activated and hibernate when not in motion to conserve battery life. To prevent data loss owing to limited battery life, participants carried one logger for two weeks at which time the device was exchanged for a fully charged logger during a household visit by the study team.Data collected from the loggers contained date, time, longitude and latitude. Study staff maintained a monthly record of the date and time the devices were distributed and collected.The observed rainfall collected at the study site using a HOBO weather station  was graphed to document seasonal rainfall patterns.2.3.After each two-week collection period, data were downloaded from each device using the @trip software . The unique participant and household identification numbers were added manually. The data were checked for inconsistent logging and device errors, such as battery failure or unrealistic locations. Raw data for each two-week period were uploaded to a secure REDCap (Research Electronic Data Capture) server . The mon2.4.rcGIS  for pre-processing and analysis. Pre-processing was done by removing erroneous data points based on unrealistic changes in shape, speed or direction in the movement tract using a software extension developed for GPS-based trajectory analysis in ArcScene by Qi & Du [rcScene software extension [Movement data coordinates were projected into UTM Zone 35S, WGS 1984 and imported into A Qi & Du . The clextension  with a bThe movement density maps created to display the movement trajectory density for each participant were overlaid on a satellite image of the study area with the enumerated households to represent the cumulative amount of time each participant spent in different areas. Density maps for participants were aggregated up to the month of collection to visually evaluate seasonal trends in movement patterns. For each month, movement densities were normalized based on the overall range to have all months on the same scale for comparisons. Short- and long-distance movement patterns were evaluated by overlaying the respective density maps on the satellite imagery. Long-distance movements were defined as those that left the study area and short-distance movements those that did not leave the study area. A three-dimensional density map, with the third dimension representing the amount of time spent at a location, was created and overlaid on a previously published malaria risk map of the study area  to visua2.5.To create movement trajectories for each participant, time was converted from date, hour, minute and second format to a numeric format. The total time participants carried a GPS data logger for each two-week time period was calculated independently to permit inclusion of individuals who only carried the logger for the first two-week period or experienced battery failure during a two-week period. The time elapsed between two consecutively logged geographical locations was then calculated. To account for differences in the total amount of time recorded by the GPS data logger for each participant, the proportion of time spent in each logged geographical location was calculated. These movement trajectories, containing logged geographical locations with calculated proportions of time spent in each location, were spatially joined to the locations of participants’ households. The proportion of time and density of time spent in logged geographical locations were plotted against distance from the household to determine the distribution of movement patterns and activity space in relation to the household compound.The proportion of time participants spent within their household compound was also calculated. A typical household compound in the study area has one or more domestic structures with several smaller structures, such as cooking houses or animal kraals. A household compound was defined as a grouping of these structures that function as a family unit. During each study visit, geographical coordinates of the household were collected from the front entrance of the main domestic structure using a tablet computer. To account for the household compound layout, and error owing to the limits of spatial resolution of the GPS logger and the tablet used to collect the GPS coordinates, the household was defined as a 100 m circular buffer around the measured household coordinates. Participant movement trajectories were spatially joined to the household buffer. This allowed for the movement to be defined as being at or away from the household compound. The proportion of time (median and interquartile range (IQR)) spent away from the household was plotted by month to determine if there was a seasonal trend. A Kruskal–Wallis test was used to determine if there were statistically significant differences between the median proportions of time spent away from the household by month.A. arabiensis, is known to have exophilic feeding behaviour, the amount of time spent away from the household compound during peak biting times was estimated [A. arabiensis were estimated to be between 19.00 and 6.00 hours in the study area [As the primary vector, stimated ,33. We wThe proportion of time spent in areas of high malaria risk was calculated using a previously constructed malaria risk map . The ris3.p = 0.04), but there were no differences by sex (p = 0.71) (p = 0.01) but not different by sex (p = 0.85)  . The con = 0.85) .Table 1The long-distance movement density maps, which display movement patterns outside the study area, showed visual evidence of seasonal patterns in population movement . There w2 study area, did not show visual evidence of seasonal mobility patterns , with a median of 10.6% (IQR: 5.8–23.8) of time spent away from the household compound. This decreased by nearly half during peak anopheline biting times to a median of 5.6% (IQR: 1.7–14.9) of time spent away from the household compound (figure 5b).Movement density in areas of high and low malaria risk was mapped . These mfigure 5The amount of time spent in areas of high malaria risk was dependent on whether the household compound was in an area of high malaria risk . The perp = 0.4) areas (p = 0.001)  and low malaria risk  (4) areas . During = 0.001) .4.Residents of rural, southern Zambia primarily spent time close to their household compound, with frequent short movements around their household and infrequent longer trips to distant locations that included shorter movements around these locations. Long-distance movement patterns showed clear seasonality. During the rainy season, participants did not travel far from their household compound, presumably to stay closer to their farms but perhaps also because roads became impassable. As the rainy season ended, participants began to travel further from their household compound and stayed there for longer durations.The convenience sample of 69 participants for this study was small; however, a large amount of data was generated on movement patterns over a 1 year period that included both rainy and dry seasons. The demographic characteristics of this sample was older but resembled eligible participants in the parent longitudinal cohort. As the longitudinal cohort is a representative sample, the movement patterns observed in the GPS logger study are probably representative of adults in the underlying population. Additionally, these patterns are probably generalizable to similar rural, agrarian populations in sub-Saharan Africa.The long-distance movement patterns at the end of the rainy season and during the dry season seemed to be consistent with long-distance foraging patterns used when searching for heterogeneously distributed food ,40. ThisMalaria prevalence has declined dramatically in parts of southern Zambia, but the region remains receptive to malaria transmission and clinical cases typically occur each year throughout the rainy season, increasing at the end of the rainy season in April. This seasonal increase in clinical malaria cases coincides with increased population mobility. While malaria prevalence is low in the study area, some surrounding areas have higher malaria prevalence. Movement to these areas for extended periods and travel back home may result in imported infections. However, this may only be important at the end of the rainy season in the month of April, as this marks the beginning of the dry season when vector populations are insufficient to maintain transmission. Thus, these long-distance, seasonal movement patterns may result in imported infections at the end of the rainy season but are unlikely to facilitate transmission during the dry season.Participants spent approximately 5% of time away from their household compound during peak biting times. However, the spatial resolution of the GPS data loggers and satellite imagery limited the ability to determine if participants were inside a domestic structure. The malaria risk map used to determine areas of high and low risk was developed when malaria prevalence was approximately 9%, and parasite prevalence subsequently dropped to less than 1% at the time of this study. However, as the map predicted malaria risk based on ecological features, qualitatively these findings remain valid for determining areas of high and low malaria risk. For short-distance movement patterns, the proportion of time spent in areas of high malaria risk was strongly dependent on whether the participant's household compound was in an area of high malaria risk. Individuals who resided in areas of higher and lower malaria risk did not spend much time in areas of the opposite risk. However, even a small amount of time spent in a high malaria risk area could result in infection and the introduction of parasites into low malaria risk areas, propagating local transmission. Therefore, malaria elimination interventions implemented at the household level, such as insecticide-treated nets, indoor residual spraying and reactive case detection, may benefit from the less frequent, long-distance movement during the rainy season.5.The human mobility patterns observed in this study can be described as circulatory rural–rural movement , althoug


Recent study showed periostin play a pivotal role in abnormal liver triglyceride (TG) accumulation and in the development of obesity-related liver fat accumulation. However, little is known regarding whether periostin plays a key role in the heightened prevalence of NAFLD and other metabolic phenotypes among large-scale populations. A cross-sectional sample of 8850 subjects aged 40 yr or older from China were evaluated in this study. Serum periostin was measured by ELISA methods. The diagnosis of NAFLD by liver ultrasonic examination. Among overweight and obese subjects, NAFLD subjects had higher serum periostin levels than those without NAFLD . Periostin was associated with a higher risk for NAFLD  among overweight and obese subjects after confounder adjustment. Furthermore, periostin levels among overweight and obese subjects were correlated with aspartate aminotransferase , alanine aminotransferase , waist circumference , homeostasis model assessment index-insulin resistance  and fasting plasma insulin , TG . Elevated circulating periostin level was associated with an increased risk of having NAFLD and insulin resistance among overweight and obese individuals. Obesity is tightly associated with an increased risk of NAFLDet al. revealed that periostin is evidently upregulated in obese rodents and humans livers tissueet al. demonstrated that periostin is highly expressed in methionine-choline-deficient (MCD) diet-induced NASH micePostn−/− mice dramatically lower than wild type mice after administered the MCD diet1213Periostin is a secreted cell adhesion protein of fasciclin family67Postn−/− mice showed specifically impaired pancreatic regeneration in the islet β-cellFurthermore, Thus, the purpose of this study is to examine the association between serum periostin levels and NAFLD as well as other metabolic phenotypes in Chinese people.The biochemical and clinical parameters stratified by NAFLD were shown for lean, and overweight and obese are presented in Circulating periostin was significantly and positively correlated with WC, fasting insulin, AST and HOMA-IR among all subjects . Especially in overweight and obese, circulating periostin levels were positively correlated with WC, fasting serum insulin, triglycerides, AST, ALT, GGT and HOMA-IR  .In this study, we demonstrated that higher periostin levels were significantly associated with increased risk of having NAFLD among overweight and obese subjects. Moreover, increased circulating periostin levels were also significantly correlated with increased insulin resistance, particularly among overweight and obese subjects.et al. has showed that periostin as a potent regulator of hepatic lipid accumulation via activation of the JNK/c-Jun pathway, which prevented expression of PPARα, in obesity mouse primary hepatocytesAbnormal triglyceride accumulation in liver is recognized as the hallmark of NAFLD. Hepatic lipid accumulation in NAFLD impairs insulin signaling that contributes to abnormal hepatic metabolism3Previous study also suggests that ChREBP, a transcription factor which plays an important role in the induction of glucose-regulated genes in liver19et al.We observed a significant positive correlation between circulating periostin levels and fasting plasma insulin, and insulin resistance assessed by HOMA-IR in overweight and obese individuals, but not in normal weight participants. Moreover, Lu As we best known, this is the first study specifically aimed at exploring the relationship between circulating periostin concentrations and NAFLD in a large sample population. The confounding effects have been minimized because most potential covariates were carefully adjusted. However, several limitations should be addressed. The cause-effect inference can not be drawn because of the cross-sectional nature of the current study. In addition, liver biopsies, the gold standard for diagnosed fatty liver, were not available in this study. The NAFLD diagnosis was based on ultrasound imaging, which means that NAFLD patients in our study were in at least moderate stage of the disease. Therefore, we failed to assess the correlation between circulating periostin and mild-stage NAFLD in this study. However, given the several advantages of ultrasound imaging, including portability, low cost, and simplicity of use, made it further applicable and acceptable for investigating the incidence, prevalence, and risk factors of NAFLD, this technique is the most widely used noninvasive method to detect hepatic lipid accumulation in epidemiological investigations and clinical practice.In summary, our study demonstrated that increased circulating periostin was markedly associated with an increased presence of NAFLD in Chinese overweight and obese subjects. Further experimental and longitudinal investigations are expected to determine the role of periostin in the development of NAFLD.In 2011, China lunched a national survey of Risk Evaluation of cAncers in Chinese diabeTic Individuals: a lONgitudinal (REACTION) study, which was conducted among 259,657 adults, aged 40 years and older in 25 communities across mainland China, from 2011 to 2012Age, gender, medical hospital and life habits were collected by trained physicians. The smoking habit was defined as never or current (smoking regularly in the past 6 months). The history of drinking was also collected.2 defined as normal weight, BMI ≥24.0 kg/m2 defined as overweight or obesityOvernight fasting and 2 h OGTT blood samples were collected for analysis. The details of anthropometric measurements including height, weight, waist circumference, hip circumference were carried by trained medical worker. Blood pressure was obtained with an automated electronic device . Obesity was defined according to the standard for Chinese individuals: subjects with BMI < 24.0 kg/mAll subjects were assessed after overnight fasting for at least 10 h, Overnight fasting and 2 h OGTT blood samples were collected in tubes containing EDTA and were centrifuged at 4 °C and stored at −80 °C until analysis. The fasting glucose, glucose 2 h after oral glucose tolerance test, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and γ-glutamyltranspeptidase (GGT), creatinine, total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL) cholesterol and high-density lipoprotein (HDL) cholesterol were measured on an automatic analyzer . Hemoglobin A1c was determined by HPLC method . Circulating C-reactive protein and Interleukin-6 (IL-6) was measured by ELISA kit . Fasting insulin was determined by RIA . Insulin resistance was measured by the homeostasis model of assessment for insulin resistance (HOMA-IR)The circulating adiponectin, CRP and periostin were duplicated measured by ELISA kit  according to manufacturer’s recommendation.Abdominal ultrasound examination was performed after overnight fasting, by two expert physician, who blinded to the clinical and biochemical parameter of subjects, with a 3.5-MHz convex probe and a high-resolution B-mode scanner . Diagnosis of fatty liver based on increased hepatic echogenicity compared to renal cortex252 tests, respectively. Spearman correlation test was used to determine the association between circulating periostin concentrations and the study variables. To investigate the associations between circulating periostin concentrations and NAFLD, the multivariate adjusted logistic regression analyses were performed to assess the OR for the risk of NAFLD. Statistical analyses were performed using the statistical software package SPSS, version 13.0 for Windows . A two-sided P value < 0.05 was considered to be significant.Results were expressed as means ± SD for normally distributed variables and as median (interquartile range) for skewed distribution variables. The skewed distribution variables were log transformed to approximate normality before analysis. Comparisons between the continuous variables and frequencies were performed using the Mann-Whitney U test, 2-sample t test and χHow to cite this article: Yang, Z. et al. Circulating periostin in relation to insulin resistance and nonalcoholic fatty liver disease among overweight and obese subjects. Sci. Rep.6, 37886; doi: 10.1038/srep37886 (2016).Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


Plasmodium falciparum is a global health concern. Myanmar stands at the frontier of artemisinin-resistant P. falciparum. Myanmar also has the highest reported malaria burden in Southeast Asia; it is integral in the World Health Organization’s plan to eliminate malaria in Southeast Asia, yet few epidemiological data exist for the general population in Myanmar.The spread of artemisinin-resistant Plasmodium falciparum positive samples were tested for genetic mutations in the K13 region that may confer artemisinin resistance. Estimated type-specific malaria PCR prevalence and seroprevalence were calculated, with regression analysis to identify risk factors for seropositivity to P. falciparum. Data were weighted to account for unequal selection probabilities.This cross-sectional, probability household survey was conducted in Phyu township, Bago Region , during the wet season of 2013. Interviewers collected clinical and behavioural data, recorded tympanic temperature and obtained dried blood spots for malaria PCR and serology. Plasmodium falciparum was the most common species (n = 19. 1.1%) and five (26%) P. falciparum samples harboured K13 mutations. Plasmodium knowlesi was detected in 1.0% (n = 16) and Plasmodium vivax was detected in 0.4% (n = 7). Seroprevalence was 9.4% for P. falciparum and 3.1% for P. vivax. Seroconversion to P. falciparum was 0.003/year in the whole population, but 16-fold higher in men over 23 years old (LR test p = 0.016).1638 participants were sampled (500 households). Weighted PCR prevalence was low  and most cases were afebrile (93%). P. falciparum artemisinin resistance, the presence of P. knowlesi and discrete demographic risk groups present opportunities and challenges for malaria control. Responses targeted to working-age men, capable of detecting sub-clinical infections, and considering all species will facilitate malaria elimination in this setting.This is the first population-based seroprevalence study from central Myanmar. Low overall prevalence was discovered. However, these data suggest endemic transmission continues, probably associated with behavioural risk factors amongst working-age men. Genetic mutations associated with  Plasmodium falciparum resistant to artemisinin, the world’s front-line anti-malarial , and less likely for those with higher SES , and of Indian origin [AOR 0.3 (0.1–0.8)]. There was a strong association between seropositivity and forest exposure in the unadjusted model, which was lost after adjusting for gender, age, ethnicity and SES  2.1 1.–3.4, p =P. falciparum, there was a clear peak in profile likelihood plot at age 23 years, indicating a change in SCR  whilst a single SCR fit better for women (p = 0.548) ], with no evidence for a change in SCR.Seropositivity to P. falciparum causes 75% of clinical malaria in Myanmar and P. vivax 25% [P. falciparum mono-infection comprised 45% of infections and P. vivax 12%  with current infection were afebrile and none reported symptoms. The three individuals with raised temperatures tested negative on RDT, possibly signifying a low-density parasitaemia  which may be associated with mild clinical symptoms . HoweverP. falciparum and P. vivax in the general population in Bago region. There was a notable increase in exposure to infection in men aged over 23, who experience a SCR 16-fold higher than the general population. Because the antibodies detected by these methods persist for an unknown period of time, there are two explanations indistinguishable by serology alone: malaria epidemiology may have changed 23 years ago such that men now aged over 23 were historically exposed to more malaria, or there may be ongoing behavioural risk factors specific to those aged over 23. The best working hypothesis is that the change in SCR is behavioural; it is confined to a well-defined epidemiological group (men of working age) and this population is overrepresented in contemporary clinic-based studies suggesting they remain at increased risk of malaria [The serological data support the PCR data and suggest that there has been ongoing low-level transmission of Plasmodium knowlesi is particularly associated with forest malaria, due to the outdoor biting habits of P. knowlesi vectors and the natural primate reservoirs living in forests [Forest work is an established risk factor for malaria transmission . Plasmod forests . The queThe Myanmar Ministry of Health offers malaria prevention, diagnosis and treatment to the general population without distinction. This approach is reflected in the high level of bed-net ownership and might contribute to the low levels of transmission observed. However, these data suggest that a strategy tailored to those at highest risk and with asymptomatic infection might be required to eliminate malaria since bed-nets will not protect those working in forest plantations at dawn and dusk . TargeteThis is the first study in Myanmar to combine detailed population-level epidemiological data with highly sensitive molecular techniques. The strength and novelty of this study also lie in the size of this probability sample from an important but unstudied population, with generalizability to Bago Region and central Myanmar. However, higher diagnostic sensitivity has been achieved with the technique of high volume ultra-sensitive qPCR  though tP. knowlesi. A higher proportion of K13 mutations among P. falciparum cases were observed than has previously documented in this region, and working age men appear at significantly higher risk of malaria exposure when compared to the general population, which needs further research. However, the public health implications are clear; the WHO goal of malaria elimination in this region will require identification and treatment of asymptomatic carriers among high-risk populations in order to control artemisinin resistance and successfully eliminate malaria in this setting.Malaria transmission in this area was low. However, there were low numbers of asymptomatic carriers who probably play an important role in maintaining local transmission, and many were infected with


Plasmodium spp. infections are common in this region and may represent an important reservoir of transmission that must be targeted for malaria elimination.Myanmar has the heaviest burden ofmalaria in the Greater Mekong Sub-region. Asymptomatic A mass blood survey was conducted among 485 individuals from six villages in Kayah State, an area of endemic but low transmission malaria in eastern Myanmar. Malaria infection was screened by rapid diagnostic test (RDT), light microscopy and real-time polymerase chain reaction (PCR), and its association with demographic factors was explored.Plasmodium spp. infection was 2.3% (11/485) by real-time PCR. Plasmodium vivax accounted for 72.7% (8/11) and Plasmodium falciparum for 27.3% (3/11) of infections. Men were at greater risk of infection by Plasmodium spp. than women. Individuals who worked as farmers or wood and bamboo cutters had an increased risk of infection.The prevalence of asymptomatic A combination of RDT, light microscopy and PCR diagnostics were used to identify asymptomatic malaria infection, providing additional information on asymptomatic cases in addition to the routine statistics on symptomatic cases, so as to determine the true burden of disease in the area. Such information and risk factors can improve malaria risk stratification and guide decision-makers towards better design and delivery of targeted interventions in small villages, representative of Kayah State. Plasmodium falciparum, continued political, financial and scientific commitment is required if the ambitious goal of malaria elimination is to be realized [Myanmar has the heaviest burden of malaria among the Greater Mekong Sub-region (GMS) countries. Although the long-term trend showed decreasing malaria morbidity and mortality in Myanmar from 2001 to 2013 [realized .Plasmodium parasites in asymptomatic carriers using strategies such as mass drug administration may play a critical role in the elimination of malaria [Asymptomatic malaria infections are usually undetectable and rarely treated. As a major source of gametocytes for local mosquito vectors, such infections contribute to the persistence of malaria transmission –10. Sign malaria .Despite recent improvement of malaria surveillance in Myanmar, the current data on malaria only represent symptomatic cases, which may be the ‘tip of the iceberg’ of the entire infected population. In addition, because symptomatic cases have mostly been detected by microscopy and rapid diagnostic test (RDT), which have limited detection sensitivity, patients with low-density parasitaemia have likely been missed by these surveys. This study aimed to determine the malaria situation in an eastern state of Myanmar, where malaria has been highly endemic in the past. Here a malariometric survey was conducted using microscopy, RDT and real-time polymerase chain reaction (PCR) assays to screen malaria infections in asymptomatic carriers and to explore associated risk factors of residual malaria parasitaemia among local residents.This study was conducted in Kayah State in eastern Myanmar, which borders Mae Hong Son Province in Thailand. Kayah has a tropical climate with average temperatures ranging from 16 °C in December to 30 °C in April. The rainy season is from May to late October with annual rainfall of approximately ~3000 mm. Malaria transmission is low, unstable and peaks from May to August, coincident with the rainfalls . There aPlasmodium spp. infections. The number of participants enrolled was based on the expected prevalence of malaria infection of 10% in Kayah State, according to the previous study in Myanmar [A community-based, cross-sectional study was conducted to determine the prevalence of asymptomatic  Myanmar . ParticiPlasmodium species in participant samples. This RDT uses histidine-rich protein 2 (HRP 2) to detect P. falciparum and lactose dehydrogenase (LDH) for Plasmodium vivax, Plasmodium ovale and Plasmodium malariae. This RDT has been shown to detect 99 and 94.3% of test samples with P. falciparum and P. vivax, respectively, at a density of 200 parasites/μl of blood and 100% of test samples with at a density of 2000 parasites/μl of blood for both species, with 1.5 and 0.7% false positive rates, respectively [A CareStart™ Malaria HRP2/pLDH (Pf/PAN) Combo RDT was used to detect Plasmodium asexual forms or gametocytes were found after counting 500 WBCs. Thin blood films were examined for Plasmodium species by counting the number of parasites per 5000 red blood cells (RBCs). For quality control purposes, a second experienced microscopist randomly selected 5% of the slides for re-examination.Thick and thin blood smears were made on the same slide, air dried and transported to the laboratory of the 100-bed hospital in Loikaw Township. The slides were stained with 3% Giemsa for 10 min and screened for the presence of plasmodial infections. The slides were read by a local microscopist who was blinded to the individual RDT results. Parasitaemia and gametocytaemia were determined from thick blood films by counting the number of parasites per 200 white blood cells (WBCs). A slide was classified as negative if no Plasmodium [P. vivax, P. ovale, and P. malariae as previously described [P. falciparum, the varATS qPCR assay was used [Plasmodium knowlesi was detected with . All qPCR assays were performed with iTaq™ Universal Probe Supermix 2× on a CFX96 real-time PCR detection system.Whole blood samples, stored at −20 °C in microtainers were transferred on dry ice to the laboratory at the Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand. DNA extraction from 200 to 300 μl of each blood sample was carried out using the Favorgen 96-Well Genomic DNA Extraction Kit, following the manufacturer’s recommendations. Malaria parasite infection was first identified by the QMAL assay, a genus-specific probe-based quantitative PCR (qPCR) targeting a conserved region of the 18S rRNA genes of asmodium , by usinescribed . For P. was used . Plasmod2) test. An error probability  of <0.05 was considered statistically significant.Data were checked for completeness and consistency, and entered into an SPSS 17.0  database. Descriptive statistics and differences in distributions were evaluated using the Chi square  and the Defense Services Medical Research Centre (DSMRC), Myanmar (IRB/2016/31). Written informed consent was obtained from all adults willing to participate in the study, and a parent provided consent for participants under 18 years of age. Malaria-positive participants were treated as per the national treatment guideline.A total of 485 samples were collected in this study, 54.4% (264) of which were from male participants. Adolescent and adult participants were predominant and only 8.6% (42/485) were children younger than 10 years old. The median age of participants was 32 years (6 months–65 years). More than 80% (420) of participants had a primary education or higher. The most frequent occupation among respondents  was forest-related work . More than 70% of respondents (377) had at least three family members in their household. Nearly all respondents reported using bed nets while sleeping at night and 18.1% (87) of bed nets used were insecticide treated. The remaining bed nets were ordinary bet nets  by RDT and 1.44% (7/485) by microscopy. Of the seven microscopy-positive samples, five were positive for P. falciparum, one for P. vivax and one for mixed  infections.The prevalence of malaria infection was detected by RDT, microscopy and qPCR. In this study, infections were considered asymptomatic when a participant’s body temperature was <37.5 °C at the time of blood collection and a participant reported no fever within the previous 2 days. The prevalence of Plasmodium falciparum accounted for 27.3% (3/11) and P. vivax for 72.7% (8/11). No mixed infection was detected by PCR. qPCR confirmed parasite DNA in 100% of RDT-positive samples and in 85.7% (6/7) of microscopy-positive samples. qPCR detected additional asymptomatic parasitaemia in six RDT-negative samples and five microscopy-negative samples. Plasmodium ovale, P. malariae and P. knowlesi were not found in this study.Molecular screening by qPCR detected parasite DNA in 2.3% (11/485) of blood samples. P. falciparum by PCR. The rest appeared to be healthy with no signs of malaria-like symptoms, including fever, chills and headache.Only one out of 11 participants with infections presented with fever >37.5 °C; this was confirmed as Plasmodium infections was found in males than females. The odds of being asymptomatic were about ten times higher among men than among women. Most qPCR-positive asymptomatic Plasmodium infections were found in adults. Among the positive samples, the youngest participant age was 17 years old. A higher prevalence of asymptomatic Plasmodium species infection was associated with forest-related occupations for malaria infection. Individuals who worked as farmers or wood and bamboo cutters had increased risk of having asymptomatic malaria infection, with an odds ratio approximately ten times greater than participants whose occupation were not forest-related. There were no statistically significant difference in infections by education, family size, or use and types of bed nets  –27. In tP. falciparum and P. vivax infections were identified. The prevalence was similar to the values (1.9%) found in a previous study conducted in Mae Hong Son, Thailand, which was also determined by PCR [Plasmodium vivax was the most prevalent species among asymptomatic cases in this study, similar to the distribution of malaria species reported in other works [P. falciparum [Because of generally lower parasitaemia in asymptomatic infections, a more sensitive test is critical for identifying asymptomatic carriers. Using PCR, asymptomatic d by PCR . The resd by PCR . Plasmoder works , 12. Howlciparum . The difAge is considered one of the most important factors that correlate with protective immunity in malaria-endemic areas . Young cPlasmodium infections than females [Many previous studies have found that males are more likely to have  females , 31–33.  females , the risThis work is among a very few studies , 16 repoPlasmodium spp. infections and their contribution to the dynamics of malaria transmission and to the incidence of symptomatic infections.A combination of RDT, light microscopy and PCR diagnostics was used to identify asymptomatic malaria infections. Risk factors were identified to improve malaria risk stratification and to guide decision-makers towards better design and delivery of targeted interventions in small villages, representative of Kayah State. In addition to the routine statistics on symptomatic cases, information on asymptomatic cases is useful for determining the true burden of disease in the area. Further studies, including entomological aspects, genotyping of asexual and sexual parasites, and immune system evaluation, are needed to better understand the epidemiology of asymptomatic


Plasmodium falciparum infection is characterized by extensive genetic diversity. Describing this diversity provides important information about the local malaria situation. This study was conducted to evaluate the extent of genetic diversity of P. falciparum in Pawe district, North West Ethiopia, using the highly polymorphic merozoite surface protein 2 gene.In malaria endemic regions, P. falciparum attending Pawe Health Centre were collected from September to December 2013. Genomic DNA was extracted using the Chelex method and analyzed by length polymorphism following gel electrophoresis of DNA products from nested PCR of msp2 (block 3), targeting allelic families of FC27 and 3D7/IC.Atotal of 92 isolates from patients with uncomplicated msp2 alleles, 11 corresponding to the 3D7/ IC and 11 to the FC27 allelic family. The frequency of isolates of the msp2 3D7/IC allelic familywas higher (51%) compared to FC27 (49%). The majority of the isolates (76%) contained multiple infections andthe overall mean multiplicity of infection was 2.8 (CI 95% 2.55–3.03). The heterozygosity index was 0.66 for msp2. There was no statically significant difference in the multiplicity of infection by age or parasite density.There were twenty-two different P.falciparum polymorphismsare extensive in Northwest Ethiopia and most of the infections are composed of multiple clones.The results of this study show that  Plasmodium falciparum and 36% were Plasmodium vivax [Plasmodium falciparum  is the most virulent and associated with the most malaria-related deaths [Despite declining trends, malaria remains an important global public health problem. According to the World Malaria Report 2015, there were about 214 million cases of malaria and an estimated 438,000 deaths. Of these, mostly (90%) occurred in sub-Saharan Africa [P.falciparum are associated with an immune response and have been identified aspotential targets for ablood-stage malaria vaccine [msp2 gene has two major allelic families, FC27 and 3D7/IC based on variation in length and sequence in the central region [msp2 block 3 is the most polymorphic and could serve alone in describing the diversity of parasite populations. P. falciparum parasite populations are often characterized by extensive genetic diversity in areas with high transmission intensity [P. falciparum and multiplicity of infection (MOI) may be useful to describe the level of malaria transmission [Themerozoitesurfaceprotein (MSP-1) and (MSP-2) are of  vaccine . MSP2 is vaccine . The mspl region . The mspntensity . Therefosmission ,12 and tP. falciparum has been extensively studied in various parts of the world [et al. demonstrated moderate to high genetic polymorphism of P. falciparum populations in southwest Ethiopia which revealed moderate transmission in the area [P. falciparum in the country. Since that time, malaria prevention and intervention programs have been scaled up [P. falciparum isolates from Pawe northwest, Ethiopia.The genetic diversity of he world –15, but the area . Since tcaled up . Hence, P. falciparum is the dominant malaria species.The study sample was conducted in the rural town of Pawe, Benishangul-Gumuz Regional state in the northwest of Ethiopia . The stuP. falciparum infected blood spot samples were collected during a therapeutic efficacy study ofarthemeter-lumefantrine (Coartem®) from September to December, 2013. The samples used in this study were from patients aged three to 60 years, presenting to the local Health Center. Inclusion criteria: all patients consented, were febrile, axillary temperatures ≥37.5°C and positive for asexual P. falciparum andwereresidents withinthecatchmentarea. The blood was spotted onto 903®filter paper (Schleicher & Schuell BioScience), air-dried and individually placed into plastic bags with desiccant, before being transported to the Malaria Research Laboratory  at Addis Ababa and kept at -20°C prior to molecular analysis.A total of 92 ® (Bio-Rad Laboratories CA) method [Genomic DNA was extracted from dried blood spots using the Chelex-100) method , with a P. falciparummsp2 gene block 3 was used as a genetic marker for the genotyping of parasite populations. Genotyping the msp2 gene was undertaken in a two-step reaction, primary amplification of the entire polymorphic gene segments of block 3, followed by a nested reaction targeting the allelic type-specific regions within this block [2, and 0.02 units per ul of Taq polymerase  and PCR buffer . In the first round reaction, 4μl of genomic DNA was added as a template. In the nested reaction, 2μl of the first PCR product was added. Each amplification profile consisted of initial denaturation at 94°C for 3 min, followed immediately by 30 cycles at 94°C for 1 min; 50°C for 35 seconds, and 68°C for 2.5 minutes. The final cycle was prolonged extension at 72°C for 3 min. PCR reaction mixtures were incubated in a thermal cycler . Allelic specific positive control 3D7 and DNA-free negative controls were included in each set of reactions. The nested PCR productsseparated on 2% ethidium bromide-stained agarose gels were visualized under ultraviolet (UV) trans-illumination and photographed. The fragment size was estimated in relation to a 50 bp DNA ladder (Boehringer Mannheim Marker VI). Presence of more than one genotype was considered to be a polyclonal infection, while the presence of a single allele was considered as a monoclonal infection. Alleles in each family were considered the same if fragment sizes were within a 20 bp interval [The polymorphic region from the is block . All reainterval .The study was ethically approved by the Scientific and Ethical Review Office (SERO) of the Ethiopian Public Health Institute (EPHI). Written informed consent was obtained from the parents or guardian prior to recruitment.msp2 allelic frequency was calculated as the proportion of the allelic family out of the alleles detected in the isolates. The multiplicity of infection (MOI) was estimated by the average number of PCR fragments per infected individual. Proportion was compared for significance using the Chi-square test. Spearman's rank correlation coefficient was calculated to assess the possible associations between MOI and geometric mean parasite density and age. P value of ≤ 0.05 was considered indicative of statistical significance.Data were analyzed using SPSS version 16 . The P. falciparum collected samples ranged between 237 parasites to 380,270 parasites/ μl of blood with a geometric mean of 20,491  parasites per μl as shown in Of the total 92 samples, 54 (58.7%)were from males and38 (41.3%)were from females. The patients’ ages ranged from three years to 60 years (mean age: 15.78 ± 1.062 years). The asexual parasitaemia for msp2 alleles and the overall mean multiplicity of infection was 2.8 (CI 95%2.55–3.03). No statistical difference was observed between multiplicity of infection according to age or parasite density (P = 0.411) as shown in Figs msp2 locus. Ninety isolates that successfully identified for the presence of MSP2 allelic families (FC27 and 3D7/IC) were classified according to the size of their amplified fragment  and 11 alleles belonged to 3D7/IC type 250–650 bp) , suggesting a large genotype diversity within the alalysia ,25. In cionrates ,27. The P.falciparum parasite population in this localized area. This finding is similarto reports fromthe RepublicofCongo [Furthermore, the present study demonstrated that three quarters of the isolates (76%) harbored multiple genotypes. This suggests a relatively high diversity of the cofCongo  and BurkcofCongo . This fiP. falciparum, due to unavailability of entomological data in the study area. The genetic diversityand MOI reported in the present study confirms a high level of local malaria transmission.Apparent multiplicity of infection (MOI) is often used as proxy for transmission and higher MOI reported in some areas may be related to thetransmission intensity . The MOIThe results of this study show thatage has no associationon multiplicity of infection, similar to reports from others studies . OnthecoP. falciparum isolates in the study area, however, the msp2 gene has immunologic effects under selective pressure [The present study results shows high allelic diversity in pressure . Moreovepressure , 38. FurP.falciparum isolates from northwest Ethiopia, found isolates to have ahighgeneticpolymorphismand that most infections were from multipleclones. This result underlines the fact that assessment of parasite diversitycan play an important role in the evaluation of malaria control interventions. Therefore, furtherinvestigationindifferent transmission setting is required toassess the changes in the genetic parameters and to evaluatemalaria control interventions in the country.This study of S1 Fig(DOCX)Click here for additional data file.S2 Fig(DOCX)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.


Emerging evidence suggests that arterial stiffness, an important marker of cardiovascular health, is associated with alcohol consumption. However, the role of longer‐term consumption patterns in the progression of arterial stiffness over time remains unclear. A longitudinal cohort design was used to evaluate the association between alcohol consumption over 25 years and subsequent changes in arterial stiffness.II cohort study of British civil servants, in which participants completed repeat pulse wave velocity assessments of arterial stiffness across a 4‐ to 5‐year interval. Repeated alcohol intake measurements were used to categorize participants into alcohol consumer types, accounting for longitudinal variability in consumption. Sex‐stratified linear mixed‐effects modeling was used to investigate whether drinker types differed in their relationship to pulse wave velocity and its progression over time. Males with consistent long‐term heavy intake >112 g of ethanol/week had significantly higher baseline pulse wave velocity  than those who drank consistently moderately (1–112 g of ethanol/week). Male former drinkers showed significantly greater increases in arterial stiffness longitudinally compared to consistently moderate drinkers . All associations were nonsignificant for females after adjustment for body mass index, heart rate, mean arterial pressure, diabetes mellitus, high‐density lipoprotein, and triglycerides.Data  were drawn from the Whitehall This work demonstrates that consistently heavy alcohol consumption is associated with higher cardiovascular risk, especially among males, and also provides new insights into the potential impact of changes in drinking levels over time. It discusses the additional insights possible when capturing longitudinal consumption patterns in lieu of reliance on recent intake alone.URL: http://www.clinicaltrials.gov. Unique identifier: NCT02663791. Cardiovascular diseases remain the leading cause of mortality, accounting for 30% of global deaths.Arterial stiffness, which occurs where the vessel wall lacks elasticity, is 1 indicator of cardiovascular health that may be directly affected by alcohol consumption. This stiffness alters arterial responsiveness to pressure variations and is indicative of both functionally and structurally adverse changes.Research into alcohol consumption and PWVStudies examining associations between alcohol and arterial stiffness have predominantly relied on cross‐sectional data, yet such designs mask longitudinal variability in consumption levels and hinder comparisons of former drinkers to nondrinkers.Two earlier cohort studies have shown that daily consumption of ≥23 g of ethanol is associated with increased incidence of arterial stiffness over a 9‐year period.Using repeat alcohol assessments covering more than 2 decades, we aim, in the current study, to more accurately capture complexity in how drinking behaviors are associated with arterial stiffness than has been previously achieved. The primary aims of this study are to examine whether long‐term patterns of alcohol consumption are independently associated with a baseline assessment of PWV and with longitudinal change in PWV. Secondary aims include an examination of whether short‐term intake levels show comparable associations with PWV.The data have been sourced from the Whitehall II cohort study. This study incorporates longitudinal assessments of 10 308 UK civil servants, originally recruited between 1985 and 1988. These recruits comprised 6895 males (67%) and 3413 females, with an overall age range of 34 to 56 years. Arterial stiffness is deemed a surrogate endpoint for cardiovascular disease, as outlined in the introduction in the main article. Consequently, participants with a previous history of cardiovascular disease were excluded from the analytic sample, leading to the exclusion of 897 participants. Clinical assessments were repeated at 4‐ to 5‐year intervals. PWV measurement was introduced to the clinical protocol at phase 9, with assessments taking place in 2007–2009. Three thousand eight hundred sixty‐nine participants had a baseline PWV assessment during this time. Follow‐up testing took place at phase 11, during 2012 and 2013. Of the participants who underwent a PWV assessment at phase 9, 3130 also had a successful PWV assessment at phase 11. The participant selection process is illustrated in Figure http://www.ucl.ac.uk/whitehallII/data-sharing.Approval for the Whitehall II study was received from the University College London Medical School Committee on the ethics of human research, and participants gave written informed consent. Whitehall II data are available to bona‐fide researchers for research purposes. The Whitehall II data‐sharing policy is available at The outcome of interest is carotid‐femoral PWV, measured in meters per second (m/s). Research has shown that carotid‐femoral PWV offers a more‐sensitive arterial stiffness measurement than PWV evaluation at other sites,PWV was assessed using applanation tonometry . Path length was ascertained using both the carotid‐sternal notch distance and femoral‐sternal notch distance. Participants underwent up to 3 PWV measurements at each phase to ensure assessment accuracy, and the mean of these was taken as their PWV measurement for that phase.Self‐reported alcohol consumption was assessed at phases 1, 3, 5, 7, and 9. Participants reported the number of glasses of wine, pints of beer/cider, and measures of spirit/liqueur consumed in the week preceding each assessment. These values were then converted into ethanol volumes. In terms of conversion ratios, 8 g of ethanol was assumed for each measure of spirit and 16 g for each pint of beer/cider.Weekly consumption levels for all beverage types combined were then used to categorize intake according to the current UK guidelines.Utilizing these self‐reported weekly ethanol intake data, participants' pattern of long‐term alcohol consumption across phases 1, 3, 5, 7, and 9 was then derived. Definitions and counts for this long‐term intake categorization are presented in Table To account for potential confounds, known demographic and clinical risk factors for arterial stiffness were selected for inclusion in the modeling. The covariates comprised standard demographic characteristics, including age,To account for the existing cross‐sectional evidence that sex may moderate the alcohol‐PWV relationship, the analyses in the current study were stratified by participant sex.To address missing data, multiple imputation by chained equations was undertaken using the R “mice” package , with 100 imputations performed for both the male‐only and female‐only data subsets. The outcome variable was included in the imputation model, but not itself imputed; auxiliary variables were also utilized to improve precision and adherence to the missing at random assumption. Sensitivity analyses were undertaken to examine whether complete case analyses produced comparable results.P<0.05 was used for inferring significance.All tests of statistical significance were 2‐tailed, and a threshold of Of the total 3869 participants in the analysis sample, 73.7% were male. As shown in Table The majority of the male (94.3%) and female (88.5%) participants were of a white ethnicity. The most common socioeconomic position among females was medium (48.7%) and among males was high (57%). Few among either sex were current smokers, whereas the majority  failed to meet the WHO's recommended weekly exercise levels. Among both cohorts, 1 in 10 had type II diabetes mellitus.Stable nondrinkers, regardless of sex, were the most likely to have never smoked , whereas stable heavy drinkers had the highest proportion of current smokers . Former drinkers  were most likely to get insufficient exercise. For both sexes, diabetes mellitus was most common among nondrinkers.P<0.001). The female mean was consistently lower, progressing from 8.2 m/s (SD=1.9) at baseline to 8.7 m/s (SD=2.3) by the follow‐up, also a significant increase (t(779)=10.9; P<0.001).The mean PWV at baseline for males was 8.5 m/s (SD=2.0), which significantly increased to 9.1 m/s (SD=2.4) by the follow‐up (t(2349)=19.5; P=0.012; female, b=0.66 m/s; 95% CI=0.01–1.32; P=0.048). Female participants with an unstable pattern of moderate consumption  also had higher PWV than those who consistently drank moderately.Table P=0.045). With the female participants, however, the associations for the stable heavy  and unstable moderate consumers  were no longer statistically significant, although their effect estimates remained larger than that observed with the male stable heavy drinkers.Following adjustment for additional demographic, lifestyle, and clinical factors, the PWV of male stable heavy drinkers remained significantly higher at baseline . Interestingly, these former drinkers had shown a large drop in association with baseline PWV following adjustment for lifestyle and clinical characteristics, yet these covariate adjustments had no impact on the magnitude of the longitudinal association observed in this group or any of the long‐term drinker types. With the female participants, no significant longitudinal effects were found, the largest effect estimate in the maximally adjusted model being for the stable nondrinkers .These models also enabled the evaluation of differences between the long‐term drinker types in terms of the subsequent change in PWV. Among males, regardless of covariates included in the model, former drinkers showed significantly greater increases in PWV over time relative to stable moderate drinkers . No longitudinal effects were observed for the female sample. Results for the recent alcohol intake types are provided in Table A secondary aim of this study was to determine whether recent intake levels showed similar patterns of association to arterial stiffness. In the male and female samples, participants without any recent intake and those with recent heavy intake showed no significant difference in baseline PWV compared to recent moderate drinkers, regardless of the extent of covariate adjustment. In terms of longitudinal change, male participants with no recent reported intake had significantly higher PWV than those with recent moderate consumption . All drinker types, regardless of sex, experienced increases in their PWV from baseline across a subsequent 4‐ to 5‐year interval, but only male former drinkers showed significantly accelerated progression .Using over 25 years of prospective alcohol consumption data, this study has provided new insights into the relationship between drinking and arterial stiffness. Male participants who habitually consumed heavy volumes of alcohol had significantly higher PWV than consistently moderate consumers , but this was no longer significant after additional adjustment for clinical covariates . This difference from the male sample may be attributable to a disparity in statistical power because of the much smaller number of female stable heavy drinkers. However, it may also be attributable to sex differences in how the alcohol‐PWV relationship is confounded by other clinical characteristics. Increased high‐density lipoprotein has previously been shown to be associated with lower PWV in males, but not females, whereas triglycerides were a significant predictor of PWV in both sexes.The finding that male stable heavy drinkers had higher baseline PWV than stable moderate consumers is consistent with existing researchP=0.015), but not clinical covariates . Research into variable alcohol intake patterns has shown that triglyceride levels may attenuate the relationship between such variable intake levels and cardiovascular disease risk.There was also evidence in the female sample that unstable moderate drinkers had higher baseline PWV compared to the stable moderate group. This effect was again significant after adjustment for demographic and lifestyle characteristics . This association was also found by Matsumoto et al,When drinker type was defined according to recent intake only, the sole significant effect was for increased PWV progression among male nondrinkers  is funded by the UK Medical Research Council/Alcohol Research UK (MR/M006638/1) and European Research Council (ERC‐StG‐2012‐309337_AlcoholLifecourse). The UK Medical Research Council , British Heart Foundation (RG/13/2/30098), and the National Institutes of Health  have supported the Whitehall II Study data collection.The alcohol life‐course project (None.


Multigravid women seemed to have the highest levels (p = 0.039). In infants, the level was highest in cord blood and decreased before stabilizing after 18 months (p<0.001). For children, a high level of sHLA-G was associated with malaria infection during the follow-up (p = 0.02) and low birth weight (p = 0.06). The mean level of sHLA-G during infancy was strongly correlated with the mother’s level during pregnancy (<0.001), and not only at delivery. Moreover, mothers with placental malaria infection had a higher probability of giving birth to a child with a high level of sHLA-g (p = 0.006). High sHLA-G levels during pregnancy might be associated with immune tolerance related to placental malaria. Further studies are needed but this study provides a first insight concerning the potential role of sHLA-G as a biomarker of weakness for newborns and infants.Human leukocyte antigen (HLA) G is a tolerogenic molecule involved in the maternal-fetal immune tolerance phenomenon. Its expression during some infectious diseases leading to immune evasion has been established. A first study conducted in Benin has shown that the production of soluble HLA-G (sHLA-G) during the first months of life is strongly correlated with the maternal level at delivery and associated with low birth weight and malaria. However sHLA-G measurements during pregnancy were not available for mothers and furthermore, to date the evolution of sHLA-G in pregnancy is not documented in African populations. To extend these previous findings, between January 2010 and June 2013, 400 pregnant women of a malaria preventive trial and their newborns were followed up in Benin until the age of 2 years. Soluble HLA-G was measured 3 times during pregnancy and repeatedly during the 2 years follow-up to explore how sHLA-G evolved and the factors associated. During pregnancy, plasma levels of sHLA-G remained stable and increased significantly at delivery ( Human leukocyte antigen-G (HLA-G) is a nonclassical HLA class I antigen that differs from the others HLA class I molecules in its limited polymorphism, its restricted tissue distribution and its particular expression . Alternain vitro fertilization ), 9.05% of them had a LBW, and 14 newborns were preterm. During the study, 12.16% of children developed a malaria infection between 6 and 24 months. There is no infection at birth in newborns.The general characteristics of the study population are presented in p<0.001).Blood was sampled in 85% of mothers for the three ANVs, and 98% were sampled at least twice. sHLA-G was described by month of pregnancy after grouping the first 3 months . The sHLp = 0.036) than primigravid women. The sHLA-G level seemed lower in women who took mefloquine for IPTp (p = 0.05) and those with placental malaria (p = 0.04). In contrast, there is no effect of age, peripheral malaria, foetus gender and LBW. In multivariate analysis, gravidity and IPTp remained significant , but the association of sHLA-G with placental malaria became marginally significant (p = 0.07).At the first ANV, univariate analysis showed that multigravid women had a higher sHLA-G level (p = 0.009)  had a higher sHLA-G level than the youngest (= 0.009) .p = 0.086 and p = 0.07, respectively)  .p<0.001)  .p = 0.02) (p = 0.06)  was not associated with the children’s sHLA-G level. In contrast, children infected by malaria at least once during the follow-up had a higher sHLA-G level ( = 0.02) . We iden = 0.06) .p<0.001) between each mother’s cross-sectional measurement and her child’s levels throughout the follow-up (p<0.001). Children born to a mother with very low profile had the lowest level, whereas children born to a mother with very high profile had the highest mean sHLA-G level.As described above, the sHLA-G level in children is correlated with their mother’s level. There were a strong correlation (ollow-up  and an ap<0.001). For an increase in the mother’s profile from “very low” to “low”, this OR was 3.05 (CI95 = [1.75–5.31]). The OR was 17.59 (CI95 = [9.54–32.41]) for an increase from “very low” to “very high”  occurred at different times for women . It could be that multigravid subjects produce more HLA-G and have a better aptitude for acceptance of semiallogenic foetal tissue. In other studies, a gravidity effect was not found , 16, butThe source of sHLA-G in newborns is not precisely known and documented to date. We hypothesize that this HLA-G could be produced by the organs of newborns himself such as thymus or umbilical cells  or immunP. falciparum could upregulate the HLA-G secretion by stimulating cytokines such as IL-10 and IFN-γ, which are well-known HLA-G inducers. HLA-G inhibits the function of T, NK, dendritic cells, neutrophils and B cells through direct interaction with ILT2 and/or ILT4 receptors [P. falciparum mediated by HLA-G may allow the parasite to escape the immune system and be responsible for higher susceptibility to infection. We have shown recently that high level of sHLA-G is associated with a higher probability to develop a malaria attack in the following weeks but also that the presence of malaria infection can increase sHLA-G that could lead to immune evasion [Malaria infection  in mothers has no effect on the sHLA-G level throughout pregnancy. This might be due to the substantial increase of sHLA-G in pregnancy, which could result in not detecting a malaria effect. It is noteworthy that pregnant women received two supervised doses of Intermittent Preventive Treatment against malaria, as recommended by WHO in all endemic areas, and that both the incidence of clinical malaria and parasitemia were low, for all IPT groups . Indepeneceptors , 43–47. eceptors , 9. Thereceptors . It has eceptors . Consequ evasion , 50. Ove evasion , 42 and The association between HLA-G and LBW shows that the evolution of sHLA-G during the first 2 years of life strongly differs between LBW children and others. LBW is correlated with infants’ morbidity and mortality , 21 and et al., even though the sHLA-G level in cord blood was lower than at term [Our results also confirmed the strong correlation between the mother’s sHLA-G levels at delivery and the cord blood level . Compara at term . However at term , 35, it  at term .Mother/child resemblance could have consequences from a public health point of view. If it is confirmed that high levels of sHLA-G are associated with LBW and a higher risk for malaria , the mother’s sHLA-G level during pregnancy could be a useful biomarker of frailty in children before delivery.It is accepted that children born from placental malaria are more susceptible to malaria during infancy, suggesting that this could stem from immune tolerance , 52, 53.


Indoor residual spraying (IRS) is used as part of the integrated vector management strategy for the control of malaria in Lagos, Nigeria. The purpose of this study was to compare the malariometric indices of children under 5 years old living in IRS-implementing and non-IRS-implementing communities of Lagos, Nigeria.The study was a community-based, comparative, cross-sectional study of 480 children under five recruited using a multi-stage sampling method. Data on each child were collected using a household questionnaire administered to the consenting care-giver of each selected child. Each child underwent a comprehensive physical examination. On-the-spot malaria rapid diagnostic testing and haemoglobin estimation to assess parasitaemia and anaemia, respectively, were also carried out. Risk factors for parasitaemia and anaemia were identified using multivariate logistic regression.p < 0.001). There was no significant difference in anaemia, spleen rate and fever in the IRS-implementing group  and the non-IRS-implementing group , respectively. Residing in an IRS-implementing community was associated with lower odds of parasitaemia . Sleeping under a bed net was the only factor associated with anaemia (p < 0.01).A total of 238 children were studied in the IRS-implementing group while 242 children were studied in the non-IRS-implementing group. The IRS -implementing community had a lower level of parasitaemia (1.3 %) compared to the non-IRS-implementing community (5.8 %) (IRS has led to a reduction in the level of parasitaemia in the under-fives in the study areas. Indoor residual spraying (IRS) is one of three main interventions recommended by the World Health Organization for the control of malaria. It is also one of the strategies recommended to achieve roll back malaria (RBM) updated targets. IRS is the application of long-lasting chemical insecticides on the walls and roofs of all houses and domestic animal shelters, in order to kill the adult mosquito vectors that come in contact with these surfaces. The intervention involves repeated spraying over a number of years , 2. NigeIn Nigeria, malaria constitutes a major public health problem as it has remained endemic despite control efforts by government and the civil society. Malaria is a preventable disease, affects all age groups, those particularly at risk are pregnant women and children, especially under-fives, and almost 110 million cases are reported every year. It is responsible for 60 % of outpatient visits to hospitals annually, 11 % of maternal deaths and 30 % of childhood deaths .Varying results have been obtained from the different vector control methods used for the control of malaria in Nigeria . EvidencLagos State, in addition to expanding coverage of ITNs, has embraced IRS as part of its IVM strategy in some local governments. Unlike ITNs that are individual-level interventions, IRS is a community-level intervention. There is limited information on the impact of IRS on malaria morbidity in vulnerable groups such as under-five children especially in high-burden countries such as Nigeria.This study was a community-based, comparative cross-sectional survey which compared the malaria indices of under-fives in IRS-implementing and non-IRS-implementing communities in a district of Lagos, southwest Nigeria.Lagos State consists of 20 local government areas (LGAs), 16 of which are urban and four rural . The LGAPrior to this study, IRS with pyrethroid insecticides had been carried out in Iwerekun thrice over the previous 5 years with an estimated coverage of 80 %.The study population comprised children under the age of 5 years who had been resident in the community for a minimum of 6 months prior to the study and whose caregivers consented to participating in the study. Children who were acutely ill, determined based on information from care-giver or a medical report, were excluded.A minimum sample size of 218 children in each community was calculated using the formula for the comparison of proportions for two independent groups  and baseA multi-stage sampling technique was used for this study: in the first stage, one ward in which IRS was being implemented (Iwerekun 1) and another in which IRS was not being implemented (Orimedu 2) were selected by balloting; in the second stage, the houses in each of the selected wards were enumerated to determine the sampling frame for that ward and this was divided by the sample size to get a sample interval for each of the communities. Systematic random sampling was then used to select compounds consisting of one or more households. Finally, one household was selected in each compound by balloting. In each selected household, only one child under 5 years of age was selected. Where a household had more than one eligible child one was selected by balloting. If there was no eligible child in the selected household the next eligible house was selected.The instrument used was a pre-tested, structured, interviewer-administered household questionnaire, which was adapted from the Nigerian Malaria Indicator Cluster Survey .The questionnaires were administered to the mother of the child or the primary care-giver. One questionnaire was administered to one household in each compound. Care-givers were interviewed on demographic information, malaria preventive practices such as the use of ITNs, and history of febrile illness. The data were collected by trained interviewers and a laboratory technician.Plasmodium falciparum detection using malaria rapid diagnostic test (RDT) kits [First response malaria Ag. P. falciparum (HRP2) card test]. Haemoglobin concentration was assessed using a self-calibrating portable blood haemoglobin spectrophotometre . A new sterile lancet was used to make a single finger prick on the index finger of the non-dominant hand of each child after cleaning with alcohol swab.Each child had a physical examination, and weight and height measurements were taken. Axillary temperature was carried out using a self-calibrating digital thermometer to measure temperature closest to 0.1 °C. Any child found to be febrile was referred to the nearest health facility. Each child was also examined for splenomegaly by the researcher using Hackett’s method . The chiParasitaemia was defined as a positive RDT test result in the presence or absence of fever. Anaemia was defined as haemoglobin measurement of less than 11 g/dl [spleen rate for each community was calculated by dividing the number of children aged 24–59 months with enlarged spleens by the total number of children that were recruited into the study in the same age group. Fever was defined as an axillary temperature greater than 37.5 °C. The socio-economic status of each child was determined using a composite index based on the average score obtained on the occupation and highest educational attainment of both parents [Malnutrition was defined as any child with weight for height (wasted), height for age (stunted) or weight for age (underweight) measurements <2 Z scores below the reference value [ 11 g/dl . Followi parents –16. The  parents . The socce value .t test used to calculate the differences between means. Chi square tests were used to compare categorical variables as appropriate. Multiple logistic regression analysis was carried out to identify the factors associated with parasitaemia and anaemia. Level of significance was set at 5 % (p ≤ 0.05).The data were analysed using SPSS version 20 software. Means and standard deviations were calculated for numerical variables and Student’s p = 0.10) . There was no statistical difference in the spleen rate among those aged 24–59 months in both groups (p = 0.91). At the time of the study 12 (5 %) under-fives were febrile in the IRS community while 21 (8.7 %) were febrile in the non-IRS community, but this difference was not statistically significant (p = 0.12). About a third of under-fives in both communities had slept under a bed net the previous night. Participants in both communities  were reported to have had a fever in the 2 weeks prior to the study. These differences were not statistically significant  was lower than in the non-implementing community (5.8 %) and this was statistically significant  (Table p = 0.01) with children residing in the IRS-implementing community having lower odds of parasitaemia , while children who slept under a bed net the previous night had a lower odds of anaemia . Living in an IRS-implementing area, recent history of fever, socioeconomic status and malnutrition were not associated with anaemia.Table IRS was commenced in Lagos State in a bid for a more robust malaria control response. It is still a relatively new addition to the State’s vector control strategy and spraying has been carried out three times over the past 5 years. This study has shown that there was a lower risk of parasitaemia among children in areas where IRS was being carried out but the same did not apply to the other indices.Although the study was carried out among apparently healthy children, an unexpected finding in was the relatively low prevalence of parasitaemia Table . This waA consistent finding among studies carried out in other African countries comparing malariometric indices between areas of IRS-implementation and non-implementation is a significantly lower prevalence of parasitaemia, anaemia and splenomegaly in the IRS-implementing area compared to the non-IRS-implementing area , 9, 22. p = 0.01)  Table . This suThe study limitation includes the cross-sectional study design which is not appropriate for making causal inferences due to the collection of exposure and outcome data at the same time. The causes of anaemia in this environment are multifactorial, however, this study did not investigate other factors that may affect anaemia, such as haemoglobinopathies, helminthiasis and HIV infection. The HRP-2 based RDTs used for the detection of parasitaemia in this study have a high sensitivity but are also reported to detect recently cleared infections in addition to current infections thus overestimating the prevalence of parasitaemia . The perContinuous efforts are needed to control malaria. As the prevalence of parasitaemia reduces, a robust surveillance system will be required to detect changes in transmission patterns. In addition, reducing transmission will also lead to reduced immunity, leaving the community more vulnerable in the event of sporadic outbreaks from imported cases.Follow-up studies are required to confirm changes to malaria burden as malaria control efforts continue. Furthermore, longitudinal studies, which would allow calculation of malaria incidence and prove causal relationships, are required. Finally, entomological studies would add value to such comparisons. This will help in confirming the success of vector control programmes.This study shows parasitaemia was lower in the IRS-implementing community and this is likely to be due to the impact of IRS. Further studies are required to determine the long-term effect IRS on the burden of malaria in those areas.


Plasmodium falciparum, the commonest cause of severe malaria in children, is an important cause of mortality in developing nations like Nepal. Duodenal perforation in a case of complicated malaria, although a rare entity, can occur in children. Early diagnosis, proper medical treatment, and early surgical repair can be a lifesaving measure in such cases. Here, we report a case of a 5-year-old male child with falciparum malaria complicated by a duodenal perforation that was successively managed with appropriate antimalarial drugs and early surgical repair. P. falciparum can present with unarousable coma, loss of consciousness, metabolic acidosis, hypoglycemia, increased lactate, jaundice, acute kidney injury, cardiac failure, hyperparasitemia, and pulmonary edema [Duodenal perforation in children is a rare surgical emergency often missed or not considered commonly in the differential diagnoses of acute abdomen , 2. Peptry edema . Abdominry edema –7. Rarel3, and platelet count was 90,000 cells/mm3. Peripheral smear (both thick and thin) was positive for malarial parasite  with parasite load of 190/μL. Antigen test for the same was positive for Plasmodium falciparum. Child was started on intravenous artesunate and clindamycin as per WHO guidelines. Plain X-ray abdomen showed gas under diaphragm , intermittent fever associated with loose motions, and vomiting for 3 days. He developed severe abdomen pain associated with distension 2 days prior to admission. There was no history of any drug intake prior to admission except WHO oral rehydration solution. On admission, he had signs of severe dehydration . On examination, child had clinical features suggestive of shock . Abdomen examination revealed distension with features of peritonitis. Child received 3 fluid boluses  and inotropes along with intravenous antibiotics. On investigation, hemoglobin was 10 gm/dL, white blood cells were 25000 cells/mmiaphragm . Child wiaphragm  and surgiaphragm  was nega Helicobacter pylori infection [Peptic ulcers which rarely occur in pediatric age groups can be of primary or secondary etiology. Primary ulcers have a male preponderance , often present at a mean age of 8 years, occur commonly in children having blood group O, and are associated withnfection , drugs (nfection –13. PrimDuodenal perforation in children has been reported comprising various etiologies . LiteratThe majority of children with duodenal perforation present with abdominal pain and signs of peritonitis . Althoug H. pylori infection though an important factor leading to perforation in developing countries was not seen in our case. Therefore, greater awareness of the causes of duodenal perforations is of vast importance; clinicians should be aware of these causes to allow for earlier identification of perforation and to facilitate better patient outcomes. The present case report also emphasizes the need for larger studies with severe malaria and duodenal perforation to find the actual cause and effect relationship.The theory proposed for duodenal perforation in malaria is due to physiological stress which results in secondary peptic ulcer disease and perforation. Malarial parasites mostly infect mature enterocytes of the duodenal villi and the host's inability to eliminate the parasites due to limited systemic and intestinal immunity during illness . Severe Duodenal perforation is successively treated by repair. Laparotomy has been used extensively for its repair for long time. A minimal invasive laparoscopic surgical repair has been proved to be safe and effective in repair of perforation .Duodenal perforation in children with malaria is rare, but a severe and potentially lethal complication can occur in those cases. Awareness of the complication in children with malaria is important for appropriate treatment. Early diagnosis and appropriate medical management with antimalarial drugs along with surgical repair can be a lifesaving measure in such cases.


Plasmodium infection caused by whole blood or a blood component transfusion from a malaria infected donor to a recipient. Infected blood transfusions directly release malaria parasites in the recipient’s bloodstream triggering the development of high risk complications, and potentially leading to a fatal outcome especially in individuals with no previous exposure to malaria or in immuno-compromised patients. A systematic review was conducted on TTM case reports in non-endemic areas to describe the epidemiological characteristics of blood donors and recipients.Transfusion-transmitted malaria (TTM) is an accidental Plasmodium species, malaria diagnostic method employed, blood donor screening method, incubation period between the infected transfusion and the onset of clinical symptoms in the recipient, time elapsed between the clinical symptoms and the diagnosis of malaria, infection outcome, country of origin of the blood donor and time of the last potential malaria exposure.Relevant articles were retrieved from Pubmed, EMBASE, Scopus, and LILACS. From each selected study the following data were extracted: study area, gender and age of blood donor and recipient, blood component associated with TTM, Plasmodium species were detected in 100 TTM case reports with a different frequency: 45% Plasmodium falciparum, 30% Plasmodium malariae, 16% Plasmodium vivax, 4% Plasmodium ovale, 2% Plasmodium knowlesi, 1% mixed infection P. falciparum/P. malariae. The majority of fatal outcomes (11/45) was caused by P. falciparum whilst the other fatalities occurred in individuals infected by P. malariae (2/30) and P. ovale (1/4). However, non P. falciparum fatalities were not attributed directly to malaria. The incubation time for all Plasmodium species TTM case reports was longer than what expected in natural infections. This difference was statistically significant for P. malariae (p = 0.006). A longer incubation time in the recipient together with a chronic infection at low parasite density of the donor makes P. malariae a subtle but not negligible risk for blood safety aside from P. falciparum.TTM risk needs to be taken into account in order to enhance the safety of the blood supply chain from donors to recipients by means of appropriate diagnostic tools. Plasmodium responsible for a potentially fatal acute febrile illness following invasion and multiplication in human red blood cells (RBCs) during their complex life cycle. Five species of Plasmodium are currently known to cause malaria in humans: the deadliest Plasmodium falciparum and Plasmodium vivax, Plasmodium malariae, Plasmodium ovale, Plasmodium knowlesi. Malaria parasites are naturally transmitted by the infective bites of female Anopheles mosquitoes during their blood meal. Malaria can manifest with severe symptoms leading to a fatal outcome in non-immune individuals, often young children and pregnant women in endemic areas or naïve adults in non-endemic settings, and remains asymptomatic in adults who have acquired a premunition maintained by repeated antigen exposure.Malaria is an infectious disease caused by intracellular protozoan parasites of the genus Plasmodium infection caused by the transfusion of whole blood or a blood component from a malaria infected donor to a recipient, described for the first time by Woolsey in 1911  OR “Transfusion Medicine”[MeSH] OR “Lymphocyte Transfusion”[MeSH] OR “Leukocyte Transfusion”[MeSH] OR “Erythrocyte Transfusion”[MeSH] OR “Blood Component Transfusion”[MeSH]) OR (Transfusion*)) AND  OR (Plasmodi*) OR ). Original research papers were included and additional references retrieved from narrative reviews; restriction to case reports was deemed necessary as the main scope of this systematic review was to investigate in fine details the relevant characteristics of each reported case of TTM. Two independent investigators  screened titles and abstracts, selected articles for full text review, performed the final article selection; a third reviewer (AA) was consulted in case of disagreement in order to reach a consensus. Case reports were excluded if the Plasmodium species. Reference mean values of MTM were drawn from the results shown by Dover and Schultz [The incubation time of each TTM case report was analysed through standard one sample two-tailed t-tests (level of significance α = 0.05) to evaluate the difference between incubation periods of TTM and MTM for each  Schultz . All sta Schultz .The number of selected papers at each step of the screening and criteria for exclusion/inclusion are reported in the flow diagram Fig. ; 100 casP. falciparum, 14 children and 12 adults for P. malariae, 8 children and 6 adults for P. vivax, 1 child and 3 adults for P. ovale, and 2 adults for P. knowlesi. Female versus male ratio was 1:1 for recipients and 1:6 for donors.i.Plasmodium species. The most common Plasmodium species detected in TTM resulted to be P. falciparum (45%) and P. malariae (30%); P. vivax, P. ovale were less frequently observed: 16 and 4% respectively; two TTM were caused by P. knowlesi (2%), and one by a mixed infection P. falciparum/P. malariae. Plasmodium praecox, an avian Plasmodium species, was described in a case report whose infection was acquired in Greece [n Greece .ii.P. falciparum whilst all the other fatalities occurred in individuals infected by P. malariae (2/30) and P. ovale (1/4).Species involved in fatal outcomes. The majority of fatal outcomes (11/45) was indeed caused by iii.Plasmodium species between TTM and MTM. For all species, the mean incubation time in TTM was longer, but the most relevant difference was observed for P. malariae .Incubation period (IP). Table iv.Blood component causing TTM. The vast majority of TTM cases were caused by whole blood and/or RBCs transfusion; however, two TTM cases due to platelets and one TTM case due to plasma only were reported.v.P. malariae occurred in US, ex-Cyprus) and/or PCR . Donor “screening” was in fact in the earlier cases the diagnosis subsequently made on the donor, classically with microscopy. Serology (IFAT) was first reported on donors in 1968 . When reported, serology (most often IFAT) appears to be by and large the most frequent method used for donor screening.Diagnostic method used for screening (if any) and diagnosis. They are also reported in detail in Table The first report of TTM went back to 1911 and the most recent occurred in 2015, both in USA. The age of TTM case reports ranged from premature children to an 85 years old individual. The partitioning of cases in children and adults (≥ 18 years) when age was available resulted in 2 children and 39 adults for Plasmodium infection which may cause morbidity and mortality especially in non-endemic areas where individuals have no premunition to malaria. Given the long-time span, over a century, of the case reports some countries which were endemic several decades ago are now malaria free such as the case of Greece and Italy. Therefore, it was not possible to infer any particular geographical pattern of TTM, whose occurrence may reflect people movements due to historical events as well as the proximity to a malaria endemic areas; an example is provided by the numerous army returnees from Vietnam to USA in the late 1960s who were not identified at the time as potential malaria infected blood donors, and caused an increase of TTM cases in the following years in USA [P. vivax TTM reported by the WHO to have taken place in Spain in 1971 due to a single blood bank in Barcelona [P. falciparum whilst all the other fatalities occurred in individuals infected by P. malariae (2/30) and P. ovale (1/4). However, these other fatalities were not attributable to malaria: two deaths were due to pneumonia and one was due to the complications of a premature newborn. Furthermore, all fatalities caused by P. falciparum were observed in adults and elderly people, which may reflect other co-morbidities or a more severe prognosis of malaria in adults compared to children within non-immune populations [Transfusion-transmitted malaria is an alternative accidental arcelona . Furtherulations .Plasmodium species as reported by Dover and Schultz [P. malariae, which is arguably the species with the longest incubation time and lowest parasite density. No other statistically significant difference was observed possibly due to the limited number of case reports, thus any interpretation must be taken with caution. Moreover, particularly in some cases of P. falciparum, the IP was surprisingly and unusually long, and, although it might explained in theory by an exceedingly small number of parasites inoculated, a reporting error cannot be excluded. Nevertheless, such potential error is expected to have occurred across all TTM cases, thus making the observation still useful to reinforce the need to extend the window of time for a malaria diagnosis in blood transfusion recipients beyond the expected IP. Moreover, according to the reported data none of the TTM cases occurred in individuals with previous history of malaria, thus ruling out the possibility of recrudescence, circulating anti-malarial antibodies , or prophylaxis which might have delayed the onset of symptoms and diagnosis. Interestingly, the incubation time of the only mixed P. falciparum and P. malariae infection was of 15 days, a nearly typical incubation time for the dominant P. falciparum species compared to the milder P. malariae which employs 35 days on average to clinically develop.There are important differences between malaria natural infection and TTM with respect to the incubation time and delayed diagnosis: a longer incubation period was observed for all  Schultz  despite  Schultz . The incP. malariae (N = 30) and P. vivax (N = 16) reinforces the need to consider these other Plasmodium species as a not negligible cause of transfusion-transmitted malaria aside from P. falciparum.Furthermore, the observation that almost half of the TTM cases reported in this systematic review are due to Several layers of complexity underline the risk of TTM in non-endemic areas: on one hand, the limited proportion of potentially infective donors imposes a cost-effective strategy of blood donors screening, on the other hand the accuracy of such screening needs to be optimal for the serious outcomes of TTM in malaria naïve recipients.P. falciparum and several decades in the case of P. malariae [In most non-endemic countries the first step in the blood supply chain is an epidemiological questionnaire to assess the potential donor’s risk to be infective which may result in a deferral for two groups of individuals: (i) those who were born and had lived for several years in malaria-endemic areas and (ii) those who were born and are resident in non-endemic areas but had visited an endemic area. According to the European guidelines individuals are acceptable as blood donors when an immunologic or molecular test for malaria is negative after at least 6 months since their last visit to an endemic area. When these donors have resided for more than 3 months in the endemic area, the deferral time may be longer. However long the deferral does not totally exclude infectious semi-immune individuals: in fact cases of TTM have been linked to donations given more than 5 years after the last potential exposure of the donor to i.Plasmodium species most commonly involved in TTM were, expectedly, P. falciparum and P. malariae, but cases of P. vivax were not infrequent, either. This parasite is not known to remain so long in blood as the two other species, while it shares with P. ovale the phenomenon of hepatic hypnozoites .The ii.P. falciparum and none by P. vivax, a parasite that has long been considered benign, although its potential to cause severe malaria has been repeatedly demonstrated in recent years [Species involved in fatal outcomes. All fatal outcomes attributable to malaria were caused by nt years .iii.The incubation period was longer than the average IP for mosquito-transmitted malaria, which may be a further reason for lack of suspicion and diagnostic delay.iv.Almost all TTM cases were caused by whole blood and/or RBCs transfusion, as expected, but for two cases by platelets and one by plasma only.v.Classical Light microscopy (LM) was used in all cases of TTM for diagnostic purposes. Only in very few cases this was complemented by serology and/or PCR in the more recent period. Serology (IFAT) was the most frequently used method for donor screening.WHO regulations on blood donation needs to be reinforced as many of the TTM case reports observed even in the time span since blood safety guidelines were implemented could have been prevented if those guidelines had been applied with stringency. Thus, different strategies need to be combined in order to ensure the safety of blood transfusions i.e. blood donor screening by appropriate diagnostic tools, which should probably include molecular tests, and possibly parasite inactivation of the blood supply.