Stenotrophomonas maltophilia is a multidrug resistant opportunistic pathogen, which is normally present in hospital settings and has very high mortality rates.A prospective study was conducted over a period of two years. The specimens were processed by Gram staining and aerobic culture. The bacteria were isolated using standard protocols. The extent of antibiotic resistance of commonly used antimicrobials and biofilm production were studied in the isolates.Stenotrophomonas maltophilia were isolated. The maximum sensitivity (93.8%) of these isolates was noticed for cotrimoxazole. 63.7% of strains were strong biofilm producers. The group given pathogen specific antibiotic showed better prognosis .A total of 80 strains of Stenotrophomonas maltophilia is important to avoid therapeutic failures.Early diagnosis and proper management of cases infected with  Stenotrophomonas maltophilia is a ubiquitous, aerobic, non-fermentative, biofilm producing Gram negative bacillus which bears a close resemblance to the Pseudomonas species and is often linked with the respiratory infections in humans , endocarditis and meningitis , corticosteroids or immunosuppressant therapy and patients previously treated with antibiotics  in healthcare environment . All the patients’ samples whose cultures grew S. maltophilia during the study period were included in the study.The study is a hospital based prospective observational study which was carried out in the Department of Microbiology at a tertiary care hospital in Karnataka over a period of 24 months .Culture for all the samples was done on sheep blood agar and MacConkey agar followed by incubation at 37°C for 18–24 hours in 5–10% COPseudomonas aeruginosa ATCC 27853 was used as quality control strain. Considering the inherent resistance of S. maltophilia to beta lactam antibiotics, quinolones, carbapenems and aminoglycosides, final reporting of sensitivity to clinicians was done for cotrimoxazole and levofloxacin.Antimicrobial susceptibility testing was performed according to Clinical Laboratory Standards Institute (CLSI) guidelines  by KirbyS. maltophilia isolates was interpreted following the CLSI guidelines 2012 . The strips were stored in the refrigerator at 2–8°C when not in use. For MIC testing, the E strips were first brought at room temperature. Bacterial suspension was made from 2–4 colonies of the test isolate and were inoculated into peptone water for 4 hours at 35 ± 2°C. The bacterial suspension was adjusted to 0.5 McFarland Standard by diluting with peptone water and inoculated onto Mueller Hinton Agar (MHA) plates by lawn culture. MIC was determined for ceftazidime, ciprofloxacin and cotrimoxazole using E strip method. A single E strip was placed at the centre of the lawn cultured plate of MHA with the help of a sterile forcep. The MHA plates were incubated for 24–48 hours at 37°C. The MIC for the nes 2012 .Trypticase soy broth (TSB) medium with 1% glucose was prepared and the cultures were inoculated and incubated at 37°C for 24 hours. With a fresh TSB with 1% glucose medium, a 1:100 dilution was made. 0.2 ml of this dilution was added in the wells of 96 well polystyrene flat bottom tissue culture plate and incubated at 37°C for 24 hours. The broth was removed by tapping and the wells were washed with 0.2 ml phosphate buffer saline (pH 7.2) four times. To fix the biofilm, 2% sodium acetate was added to the wells followed by staining with 0.1% w/v crystal violet. Then, excess stain was removed followed by washing with deionized water. After drying the plates, optical density was measured at 570 nm. Isolates were divided into weak, moderate and high biofilm producers based on the optical density values as < 0.120, 0.120–0.240 and > 0.240 respectively.Data analysis was done using SPSS version 16. For demographic data, descriptive analysis was used and results represented as frequency (percentages). Continuous variables were expressed as mean ± standard deviation (SD). Independent Sample t-test was utilised to analyse the duration of hospital stay of overall and ICU patients. Biofilm production and antibiogram data was represented using frequency (percentages). All reported values were two sided (α = 0.05) with a confidence interval of 95%. P value <0.05 was considered significant.S. maltophilia were identified from various samples, with maximum number from sputum  followed by endotracheal aspirate , blood , wound swab , two each (2.5%) from BAL and pus and one (1.25%) from the tissue. Male to female ratio of study cases was 52:28. Most of the patients  were in the age group of 41–60 years. Predominant presenting symptoms were observed as respiratory tract infections (81.3%). The detailed clinico-demographic profile of the study population is shown in In the study period, 80 isolates of S. maltophilia was isolated as a single pathogen in 48 patients (60% of the cases). The other pathogens that grew with this organism in descending order were Acinetobacter baumannii (15%), Klebsiella pneumoniae (6.3%), Pseudomonas aeruginosa (6.3%), Enterococcus spp. (3.8%), Staphylococcus aureus (2.5%) followed by Escherichia coli, Moraxella spp., Aspergillus spp., Enterobacter cloacae, Serratia spp. all contributing to 1.3%.Results of susceptibility testing done by Kirby-Bauer disk diffusion method revealed that cotrimoxazole (93.8%) was the most sensitive antibiotic followed by ciprofloxacin (88.8%) and piperacillin-tazobactam (32.5%) .The results of E-strip test method are shown in The results of biofilm production revealed 63.7% were strong biofilm producers and 36.3% moderate biofilm producers. Among 80 patients, 39 (48.75%) were on empirical antibiotic therapy and the most common empirical antibiotic used was meropenem followed by β-lactam - β-lactamase combinations, ceftriaxone, amikacin etc. Pathogen specific antibiotic was instituted in 43 (53.75%) patients whereas 37 (46.25%) patients received non pathogen specific antibiotic. In the present study with a sample size of 80, males contributed 65% with the male: female ratio 52:28. The predominance of males is probably due to the behavioural and socioeconomic factors in India, where males in larger proportion to females are involved with outdoor activities and females often don’t present in the early course of the illness.S. maltophilia. Our study is in close association with a study done by Gopalakrishnan R that also showed the mean age group of infected patients as 62.4 years  of our patients were between the age group of 41–60 years and about 27 (33.7%) patients were in age group of 61–80 years. This can be attributed to a weekend immune system response observed in older age group which rendered them more susceptible to infections by .4 years .S. maltophilia.Present study shows that respiratory tract is the most common system involved (81.3%) followed by blood stream infection (16.3%) and skin/soft tissue infection (10%). The predominance of respiratory tract involvement is also cited by Brooke et al. in their review paper . PulmonaS. maltophilia but there are conflicting reports of such evidence , followed by ciprofloxacin (88.8%) and piperacillin-tazobactam (32.5%). Another study done by Rutter et al. . Empirical treatment was opted for 46.25% and meropenem, to which S. maltophilia infection. The present study noticed the mortality rate of 11.25% in the study population. In a study done by Jang TN et al.  were observed to be moderate or strong biofilm producers. Earlier Flores T et al. have reported that 53% of their isolated strains were moderate/strong biofilm producers . AbilityN et al. , mortaliN et al.  reportedS. maltophilia as an important nosocomial pathogen having capability of inherent resistance to multiple antimicrobials and stresses the importance of its early diagnosis along with timely management with pathogen specific antibiotic for better prognosis of patients.To conclude, the study highlights the need for awareness about


V1.7 and NaV1.9, play in pain signalling in man. Gain of function mutations in NaV1.7 cause painful conditions while loss of function mutations cause complete insensitivity to pain. Only gain of function mutations have been reported for NaV1.9. However, while most NaV1.9 mutations lead to painful conditions, a few are reported to cause insensitivity to pain. The critical roles these channels play in pain along with their low expression in the CNS and heart muscle suggest they are valid targets for novel analgesic drugs.Chronic pain is a global problem affecting up to 20% of the world’s population and has a significant economic, social and personal cost to society. Sensory neurons of the dorsal root ganglia (DRG) detect noxious stimuli and transmit this sensory information to regions of the central nervous system (CNS) where activity is perceived as pain. DRG neurons express multiple voltage-gated sodium channels that underlie their excitability. Research over the last 20 years has provided valuable insights into the critical roles that two channels, Na Pain is an important warning system to guard against tissue damage and disease. Pathological pain, however, has no warning value and has huge economic, social and personal costs to society. Chronic pain is a global problem affecting up to 20% of the world’s population , 50. SenInflammation and nerve injury sensitise DRG neurons and result in decreased pain thresholds and/or intense pain. This can be in part due to increased voltage-gated sodium channel (VGSC) activity resulting in increased excitability of DRG neurons , 104. VGThis review aims to clarify the roles of two VGSC channels expressed selectively, though not exclusively, in primary sensory neurons in pain pathways, and in the light of evidence from genetic mouse models and mutations in man. We discuss the usefulness of these channels as potential drug targets, and suggest that while our present understanding of function has grown more complex, targeting these channels either alone or in combination may still provide a strategy for analgesic development, potentially even for chronic use.There are two reasons why VGSCs are attractive targets for analgesic drugs. Firstly, VGSCs are required for the firing of action potentials in DRG neurons; therefore, blocking their activity will reduce pain signalling in painful conditions even if they were not the primary or only contributor to increased firing. For example, sensitisation of primary transducing channels, like the transient receptor potential (TRP) channels, is often involved in many forms of pathological pain , 90, 105v1.7, Nav1.8 and Nav1.9) meet the above criteria. Not surprisingly, many pharmaceutical companies are developing and testing subunit specific Nav1.7, NaV1.8 and NaV1.9 blockers as analgesics [Secondly, a few of the VGSC α-subunits expressed in DRG are either exclusive to or enriched in DRG neurons that signal pain, with little expression in other DRG neurons, the CNS, skeletal and heart muscles. Blockers for these subunits would therefore be expected to produce analgesia without detrimental side effects. DRG neurons express many of the cloned α-subunits ; howeveralgesics , 128.V1.9 channel activates over a negative range of membrane potentials close to the resting potential and generates a persistent current. Evidence suggests that it is powerfully regulated by G protein pathways, in a unique way. Therefore, when it is activated, it contributes to setting the resting membrane potentials of neurons expressing it [v1.7 generates a transient Na+ current, but has a relatively slower rate of inactivation near the resting potential (slow closed-state inactivation) allowing the channel to generate persistent currents and making it a so-called threshold channel [v1.8 has a relatively depolarised activation voltage (~ − 20 mV) compared with Nav1.7 and Nav1.9 [v1.8 activation comes after and perhaps subsequent to the activation of Nav1.9 and Nav1.7 channels. Nonetheless, Nav1.8’s depolarised inactivation and more rapid recovery from inactivation allow it to contribute to repetitive firing, for example [V1.9 and Nav1.7 subunits since their biophysical properties allow them both to influence pain thresholds through setting the membrane potential and action potential threshold in DRG neurons.The three α-subunits differ in their biophysical properties that determine their role in neuronal excitability . The NaVssing it . Nav1.7  channel , 25. Navd Nav1.9 ; thus, N example . This rev1.7 was cloned from PC12 cells in 1997 [V1.8 and NaV1.3 channels were already under the spotlight and their role in pain was actively being investigated. NaV1.8 was cloned in 1996 and its strong expression in medium and small sensory neurons (the sizes of most nociceptors) made it the best and most obvious target for analgesic drug development [V1.8 knockout mouse was reported 3 years later and although it showed a pain deficit, its phenotype was compromised by a compensatory upregulation of Nav1.7 [V1.8 by antisense oligonucleotides in adult rats inhibited neuropathic pain [v1.8, the expression pattern of Nav1.3 does not suggest it would be a useful drug target. Nav1.3 is expressed throughout the nervous system and its expression is highest during embryonic development and decreases postnatally [v1.3 is the only channel that is re-expressed in DRG following nerve injury and diabetes [v1.3 do not show any deficits in pain phenotype [Na in 1997 . At thatf Nav1.7 . Howeverhic pain , 129. Thtnatally . Howeverdiabetes . This mahenotype .v1.7 became the focus of the pain field in 2004 with the publication of two papers [SCN9A (the gene coding for Nav1.7) as the cause for a rare inherited pain condition known as primary erythromelalgia (PEM). PEM symptoms start at early age with episodes of pain in the extremities  that are triggered by exposure to heat or walking [v1.7 in most nociceptors [v1.8 promotor [v1.7 in nociceptors [v1.7 in mouse proved to be lethal [v1.7 global KOs to survive to adulthood [v1.7 ablation in adult mice causes pain deficits without detrimental effects [Nao papers , 125. Thiceptors . The conpromotor . The comiceptors  excited iceptors , 128. A e lethal . Global dulthood . Inducin effects .v1.7 null mice. While pain can be triggered by mechanical pressure on the feet , conditional null mutants showed a complete loss of pain to mechanical pressure. While PEM patients showed signs of inflammatory pain , conditional null showed a complete loss of inflammatory pain. This helped support the hypothesis that blocking Nav1.7 in humans would significantly reduce pain signalling. However, the mouse study raised significant questions. Firstly, is the role of Nav1.7 in pain signalling in humans as critical as it is in mice, or in other words, would the loss or block of Nav1.7 in humans lead to the all or none loss of pain seen in mice? Furthermore, if this is the case, then would the loss or block of Nav1.7 in humans result in lethality (as it did in mice)? These questions were critical for the validity of Nav1.7 as a drug target. These questions were answered in 2006 when it was reported that a loss of function mutation in SCN9A causes complete insensitivity to pain (CIP) [v1.7 plays as a critical role in pain signalling in humans as it does in mice. Importantly, loss of Nav1.7 did not lead to lethality nor any significant disability (CIP patients are anosmic due to the expression of Nav1.7 in the olfactory epithelia [SCN9A in the same year (2006). Paroxysmal extreme pain disorder , was found to be caused by a gain of function mutation in the Nav1.7 channel [v1.7 is a critical player in pain signalling, catapulting it to the top of the list of analgesic drug targets. Since then, human geneticists have identified scores of mutations causing PEM, PEPD, CIP and small fibre neuropathy (SFN).Remarkably, the symptoms of PEM patients complemented the phenotype of the conditional Nain (CIP) . In CIP pithelia ). A seco channel . The fouSCN9A gene. The condition was first mapped to SCN9A in 2004 by Yang et al. [v1.7, involving a shift of the activation voltage to hyperpolarised potentials [Primary erythromelalgia is an autosomal dominant condition caused by a mutation in the g et al. . The prog et al. . Since tg et al. , 21. Alltentials , and whetentials . FurtherV1.7 [Treatment for PEM patients includes avoidance of the conditions that trigger pain . Patients typically resort to foot lifting, cooling feet by fans or immersing them in water or iced water to reduce or relief pain. Although immersion in cold water is effective for mild cases, it can result in ulceration and maceration of foot skin leading to infection , 116. ReV1.7 , 133.SCN9A that alter the biophysical properties of the Nav1.7 channel [v1.7 channels showed that they have normal activation voltages (unlike PEM mutations). In contrast, PEPD mutations cause a depolarising shift in inactivation voltages with incomplete channel inactivation, leading to a persistent current and increased excitability [SCN9A mutations that are found in PEPD patients. PEPD mutations tend to cluster in domains III and IV of the channel protein, Fig. Paroxysmal extreme pain disorder , is caused by gain of function mutations in  channel . There a channel ). PEPD itability , 119. Tav1.7 [v1.8 [v1.9 [Small fibre neuropathy (SFN) is caused by damage to thinly myelinated and unmyelinated nerve fibres. SFN is often characterised by late onset, bilateral burning pain to the hands and feet. SFN is also associated with disturbances to autonomic functions like sweating, dryness in the eyes or mouth and disturbance to bowel and bladder functions . Autonomv1.7 , Nav1.8 .7 [v1.8  and Nav1.8 [v1.9  channelsv1.7 channels in SFN patients, listed in Table v1.7 channel mutations linked to SFN are not localised to a particular region within the channel but many are clustered in the first intracellular loop between domains I and II, Fig. SCN9A mutations [SCN9A mutations [Characterisation of Nautations ; howeverutations .Table 3Mv1.7 in the human brain is poorly characterised, there is emerging evidence that Nav1.7 plays some role in modulating excitability in the brain. It is known that CIP patients suffer from the loss of the sense of smell due to expression of Nav1.7 in the olfactory epithelia [SCN9A in patients with various types of epilepsies, Table v1.7 function has no detrimental effect on the brain, altered or increased Nav1.7 function does. Therefore, further research is needed to provide insights onto the type of cells that express Nav1.7 (types of neurons? any in glia?) in the brain. Furthermore, knock-in models will help to explore how the mutations cause epilepsy rather than act as modifiers to changes in other genes . Finally, it is intriguing that a few mutations (e.g. Q10R) cause PEM in some patients, and epilepsy in others. This may suggest that variations in the functional expression of other genes or epigenetic changes influence the biological consequences of mutations in Nav1.7.Although the expression of Napithelia  and a papithelia . HoweverSCN9A loss of function mutations cause an autosomal recessive CIP [Complete insensitivity to pain is characterised by loss of all pain sensations throughout patient’s life. sive CIP , 34. Sevsive CIP , 97; howsive CIP .Table 5Sv1.9 (gene name SCN11A) is a tetrodotoxin-resistant (TTX-r), so-called persistent Na+ current, with clear evidence for functional expression in nociceptive primary sensory neurons in the dorsal root ganglia (DRG) and trigeminal  [+ axons in the sciatic nerve , an, anv1.9  the gut , 102. Th SCN11A)  had initve e.g. , 47, 95), 95v1.9 ve  Na+ current [+ current in sensory neuron cell bodies that has ultra-slow activation and inactivation kinetics. It gives rise to a persistent, non-inactivating current operating over the negative portion of its activation membrane potential range, allowing it to act as a ‘threshold channel’, and to contribute to setting the membrane potential. Its unusual kinetic properties and negative activation range produce ‘plateau potentials’ that amplify applied or transduced sub-threshold depolarisations and massively prolong them in duration . With tSCN11A, Table About 20 mutations have been reported for v1.9 currents and the negative activation voltage dependence make the channel functionally unique. It is proposed to act as a threshold channel in peripheral nociceptors, so gain of function mutations associated with facilitated activation would be expected to give rise to painful neuropathy, because the threshold for action potential generation is reduced. Indeed, SCN11A mutations result in two painful conditions, familial episodic pain [SCN11A-related small fibre neuropathy experience pain, tingling and numbness in their arms and legs. Patients may experience diarrhoea which is consistent with expression of Nav1.9 in the gut [The persistent nature of Nadic pain , 93, 130dic pain , 57, 63.SCN11A gain of function mutations cause a complete insensitivity to pain [v1.9 expressing neurons. This prolonged depolarisation causes rapidly gating Na+ channels (e.g. Nav1.7 and Nav1.8) to enter the inactivated state [v1.7 and Nav1.8 in nerve endings leads to an overall decrease in excitability. It was noted that the mutations that lead to CIP are those that produced the largest shift in the activation threshold of the channel, whereas those that lead to familial episodic pain and painful small fibre neuropathy cause smaller shifts, Table SCN11A CIP mutations are all localised to transmembrane segment 6, Fig. v1.9 is expressed in the IB4+ subset of neurons and not in all DRG neurons (at least in rodents). Therefore, a depolarising block in this subset of neurons alone is not expected to cause a complete loss of pain. Second, NaV1.8 which is expressed in most nociceptors (i.e. in same neurons as Nav1.9) is a channel known to operate at more depolarised membrane potentials and can maintain excitability, even in the face of a depolarised membrane potential [Surprisingly, a few  to pain , 79, 98. to pain , althouged state . Since totential , 134.Figv1.7 and Nav1.9 play critical roles in pain signalling. The link between genotype and phenotype for mutations in both channels is poorly understood. Symptoms manifest in the extremities (mainly in the feet/legs) in most human conditions. In Nav1.7 channel mutations, there is a link between mutations that cause enhancement of activation to PEM and mutations that cause incomplete inactivation to PEPD. The physiological or microanatomical basis for these associations in terms of nerve ending function is only partly understood. In NaV1.9 channel mutations, a clear understanding of why most gain of function mutations cause painful conditions while those affecting segment 6 cause insensitivity to pain is also lacking, although insensitivity to pain is hypothesised to be caused by reduced excitability due to a depolarising block [In the past 20 years, mouse models and human genetics have confirmed that Nang block , 122.v1.8 and Nav1.9 in mice does not lead to lethality or any observable detrimental effects. Ablation of Nav1.7 in human [v1.7 is still elusive, despite efforts by several pharmaceutical companies [v1.7 inhibition required for analgesia in vivo. Given that CIP carriers have normal pain phenotype, an Nav1.7 blocker may need to reduce channel activity to a level well below 50% to produce analgesia.Nonetheless, available evidence confirms a critical role for both channels in pain earning them a position in the list of potential drug targets. Ablation of Nain human  and in ain human  does notompanies , 71, 128SCN9A CIP raises the question of whether the CIP phenotype is a direct consequence of the loss of Nav1.7 [v1.7 reduces the excitability of DRG neurons per se. Deletion of Nav1.7 causes an increase in action potential threshold in small-diameter DRG neurons [v1.7 causes a major decrease in DRG neuron responsiveness to the VGSC opener veratridine [v1.7 KO mice [v1.7 KO mouse to contribute to the observed reduced excitability of DRG [V1.7 deletion were not reversed by the opioid receptor antagonist naloxone [V1.7 has a profound effect on the excitability of DRG neurons .The contribution of the endogenous opioid system to the phenotype of the f Nav1.7 , 97. Sev neurons . Deletioatridine . Mechani KO mice . No chany of DRG . Furthernaloxone . Therefov1.9 loss of function mutations may indicate that its loss does not lead to a major phenotype in humans, or at least does not make people go to the doctor, raising doubts as to whether a blocker would lead to a major analgesic effect. Finally, considering that nociceptors express at least two of the peripheral VGSC subunits , an effective analgesic strategy may ultimately result from a combination of blockers against these subunits to have additive and synergistic effects on nociceptors. The effectiveness of various drug combinations to reduce neuronal excitability can only be measured in DRG neurons because they express the target VGSCs at biologically relevant levels. Equally important, for the evaluation of any drug combination, is the potential effects on non-nociceptors as well as nociceptors. We recently described a relevant assay [v1.7 and Nav1.8 blockers produced a reduction in the excitability of DRG neurons close to that measured in Nav1.7 KO [The lack of reported Nant assay  and provav1.7 KO . Changin


Background and aimGram-negative rods (GNR) are the most common pathogens associated with urinary tract infections (UTI). The resistance of these gram-negative rods to various antibiotics is increasing with time. The study aimed to determine the pattern of resistance to antibiotics in GNR causing urinary tract infection in adults.Material and methodsThis is a cross-sectional study conducted during six months (1st December 2019 to 1st June 2020) among adult patients admitted to Russells Hall Hospital Dudley, UK. Urine cultures of 156 patients admitted with urinary tract infection were collected and reviewed. Sources of urine included midstream urine (MSU), catheter specimen urine (CSU), and others from nephrostomy bags and urine bags. Sensitivity and resistance were checked using Clinical and Laboratory Standards Institute (CLSI) guidelines. Results were analyzed using SPSS version 13. ResultsAltogether 156 patients were included in the study. Males were 40.4% of the patients were males, and 59.6% were females. The mean age was 78 with a standard deviation (SD) of 13.15. Most of the samples (67.3%) were urine MSU, 23.1% catheter urine, and 9.6% were others, like from nephrostomy bags or unspecified. Resistance to amoxicillin was found in 61.7%, trimethoprim in 36.2%, nitrofurantoin in 13.2%, ciprofloxacin in 25.6%, fosfomycin in 10.7%, co-amoxiclav in 36.2%, gentamicin in 12.8%, piperacillin-tazobactam in 7.1%, cephalexin in 44.4%, and meropenem in 0% of patients.ConclusionResistance to gram-negative rods causing urinary tract infection is increasing; a particular concern is increased resistance to beta-lactams, trimethoprim, and quinolones. Urinary tract infection (UTI) refers to the invasion and multiplication of microorganisms in any part of the urinary system. Urinary tract infections are the leading cause of presentation and admissions of the adult population to hospitals worldwide. UTIs account for more than 8.3 million office visits and more than one million hospital visits worldwide . A UTI iThis is a cross-sectional study conducted during six months (1st December 2019 to 1st June 2020) among adult patients admitted to Russells Hall Hospital Dudley, UK. The sample size was 156 patients with a positive urine culture. The patient was included in the study if the patient had a positive urine culture with gram-negative rods, aged 18 to 96, and male or female gender. Exclusion criteria included the presence of positive urine culture in the last 30 days; the presence of ureterosigmoidostomy, suprapubic catheterization, previous urinary stricture, urethrovaginal or vesicovaginal fistula; positive urine culture with gram-positive cocci, gram-negative cocci or fungi; antibiotics not effective against gram-negative rods.One hundred and fifty-six patients presenting to the Russells Hall Hospital, Dudley, UK, were selected according to inclusion and exclusion criteria. Data was collected as a part of the clinical audit after approval from hospital authorities and clinical supervisor. Urine samples of patients were placed in culture media for an incubation period of 24 hours at 37°C in order to isolate the uropathogen. Only gram-negative rods were considered for the collection of data. Isolated bacteria were tested against antibiotics in vitro on agar plates. Minimum inhibitory concentration (MIC) of antibiotic required to inhibit the growth of strain were found and compared to Clinical and Laboratory Standards Institute (CLSI) guidelines. Resistance patterns to antibiotics were seen, including penicillins , cephalosporin , aminoglycosides (gentamicin), quinolones (ciprofloxacin), trimethoprim, nitrofurantoin, fosfomycin, and carbapenems (meropenem). Data was collected using performa.SPSS version 13  was used for the analysis of data. For qualitative variables like gender, sensitivity, and resistance to antibiotics, frequency and percentage were calculated. For quantitative variables like age, mean and SD were calculated.A total of 156 urine cultures were analyzed. Out of 156 samples, 63 (40.4%) were from males, and 93 (59.6%) were from females. The mean age of patients admitted with gram-negative rods associated with UTI was 78 , as indicated in Figure One hundred and five samples were urine MSU (67.3%), catheter urine - 36 23.1%), and other samples, including samples from nephrostomy, unspecified or from urine bag, were 15 (9.6%). E. coli was most commonly seen in urine cultures, 82 out of 156 samples (52.6%), Proteus in 17 (10.9%), Klebsiella in 14 (9.0%), Pseudomonas in 7 (4.5%), and other species, including Citrobacter, Morganella, Enterobacter and unspecified species were in 21 (13.5%) samples . Out of 149, 92 GNR species were resistant (61.7%), and 57 sensitive (38.3%). Trimethoprim was tested for 149 samples; 93 strains were sensitive (62.4%), 54 resistant (36.2%), and two had intermediate sensitivity (1.3%). Sensitivity to nitrofurantoin was tested in 111 GNRs for Pseudomonas, Klebsiella, Enterobacter, and Morganella, which are naturally resistant to nitrofurantoin. Ninety-six strains out of 111 were sensitive (86.5%), and 15 resistant (13.5%). Ciprofloxacin showed 74.4% sensitivity (116 out of 156 ) and 25.6% resistance (40 out of 156). Sensitivity to fosfomycin was tested in 149 samples; 133 samples were sensitive (89.3%) and 16 resistant (10.7%). Co-amoxiclav was tested in 149 GNRs; 95 were sensitive (63.8%) and 54 resistant (36.2%). Gentamicin had an overall 84% sensitivity (131 out of 156), 12.8% resistance (20 out of 156), and five strains (3.2%) showed intermediate sensitivity. One hundred and thirty-five samples showed sensitivity to piperacillin-tazobactam (86.5%), 11 were resistant (7.1%), and 10 were intermediate (6.4%). Sensitivity to meropenem was tested in 152 samples, and all the strains (152) were found sensitive (100%). Sensitivity to cephalexin was checked in 90 samples, 50 were found sensitive (55.6%), and 40 were resistant (44.4%).In summary, the resistance to antbiotics in descending order is as follows: amoxcillin 61.7%, cephalexin 44.4%, co-amoxiclav 36.2%, trimethoprim 36.2%, ciprofloxacin 25.6%, nitrofurantoin 13.5%, gentamicin 12.8%, fosfomycin 10.7%, piperacillin-tazobactam 7.1% and meropenem 0%.Our study has shown that the adult population most affected by UTI is the elderly population with a mean age of 78 with SD of 13. As this study was conducted in admitted patients only, the data will tend to miss the younger patients who usually get UTIs and are not admitted, especially in the case of sexually active males and females with UTIs. Our data matches with the figures released by UK essays, which showed the chance of getting infected by UTI in women is 20% between 60-65 years age group and 20-25% over age 80. In males, the prevalence is 3% between 65-70 years and 20% above 80 years . Our stuIn our study, 67.3% specimens were urine MSU, whereas 23.1% were catheter and 9.6% of sources included nephrostomy specimens and other unspecified sources. Although our study was not aimed to compare the incidence of UTI in catheterized and non-catheterized individuals, it has proved that the incidence of UTI is significantly higher in the catheterized individuals than in non-catheterized . Our stuOur study concluded that resistance to amoxicillin is 61.7%. A study from Africa in 2016 showed that resistance to amoxicillin was more than 70% only in Ecoli strains . AmoxiciThe first-line of antibiotics for lower UTI is nitrofurantoin and trimethoprim, according to NICE guidelines. The resistance to trimethoprim was 34% in the UK in 2016 . NICE guOur study showed 12.8% resistance to gentamicin and 7.1% resistance to piperacillin-tazobactam. Gentamicin is the first-line intravenous (IV) antibiotic used in patients admitted with sepsis secondary to UTI; however, impairment in renal functions and level monitoring are the few demerits of the use of gentamicin. Gentamicin resistance again is much higher in the developing world. In India, a study done in 2012 showed that all gram-negative rods, including E. coli, showed more than 50% resistance to gentamicin . This agIn our study, no uropathogen was found resistant to meropenem; however, CDC has reported around 13,100 new carbapenem-resistant Enterobacteriaceae cases in hospitalized patients in the United States alone which were 11,800 cases in 2012 . In the Resistance to antibiotics is increasing. Even in the case of UTI, oral antibiotics are showing much more resistance than before. A particular concern is much higher antibiotic resistance in Asia and Africa secondary to overuse and lack of antibiotic prescription control and availability of over the counter antibiotics. Immediate steps are required to control antibiotics prescription and the use of antibiotics only and when required.


Also, 2-EH presented apoptosis induction with characteristic hallmarks, such as loss of mitochondrial membrane potential, caspase activation, DNA fragmentation and cell cycle arrest at the S phase. In addition, the presence of autophagosome was detected through L3CB protein staining. We conclude that 2-EH presents differences in cell death endpoints that interestingly differ from the DEHP parent compound. Further studies are needed to establish the molecular pathways responsible for the observed effects.Phthalates have been used in a wide variety of consumer goods. Their versatility as plasticizers has translated into worldwide use in a vast array of consumer products. These compounds can leach into matrices, such as food and liquids that can be routed for human exposure. One of the most used phthalates is Diethylhexyl phthalate (DEHP). Diethylhexyl phthalate and its metabolite 2-ethyl-1-hexanol (2-EH) have demonstrated biological effects which merit further evaluation. In this work, we expand on our previous work with DEHP and screen the 2-EH metabolite for different cell death endpoints such as growth inhibition, apoptosis, autophagy, caspase activation, DNA fragmentation, and cell cycle arrest using fluorophores and the NC3000 instrument. Significant results ( In turn, plastic manufacturing technologies have evolved to modify the characteristics of their products. Phthalates are widely used as plasticizers to increase the flexibility and durability of plastic products. Since the 1920s with the introduction of Diethylhexyl phthalate (DEHP) phtalate ester, compounds have been used extensively in different types of products such as building materials, flooring, piping, food packaging, and medical devices, among others . ExposurDiethylhexyl phthalate is a common phthalate historically used for imparting flexibility to plastic products including medical devices, such as catheters, blood, and IV bags, among others . The higOur previous work  establis2.2.1.2-EH and DEHP were purchased from Sigma Aldrich  and diluted with dimethyl sulfoxide (DMSO) 1% for the working solutions  which were kept in sterile conditions and refrigerated until utilized. Positive controls: DEHP, camptothecin, and chloroquine for autophagy assays were purchased from Sigma Aldrich. DMSO 0.1% and culture media were used as a vehicle negative control.2.2.2.The human lymphoblast cell line TK6 was obtained from the American Type Culture Collection . Cell cultures were maintained in RPMI 1640 culture media with 10% fetal bovine serum (ATCC). Cells were maintained at 37°C with 5% CO2.3.R) lower than 98 was not considered. This growth inhibition dose was used in all subsequent experiments.TK6 Cells were seeded  on fluorometry compatible 96 well plates  then exposed to the experimental compounds at ten different doses (in triplicates) for 24 hours. After the incubation period, the Prestoblue cell viability reagent  was added to all wells. This reagent is reduced by metabolically active cells to its fluorescent form and the fluorescence produced by the reduced form is analyzed using a Fluorostar Optima fluorescence reader  using the 485/520 nm filters. Results were analyzed using the MARS data analysis software (BMG). Data with a coefficient of determination . After the exposure period, cells were stained with annexin V conjugate, and propidium iodide . These stained cells are then analyzed using the Nucleo Counter NC3000 system annexin V assay . Fluorescence of cells is detected and categorized on fluorescence emitted. A one-way ANOVA with post hoc Tukey test was also performed for all cell death assays (GraphPad Prism v.6).The annexin V assay is a common assay used in the detection of apoptosis. This assay measures the migration phosphatidylserine (PS) to the exterior surface of cells, a common event in apoptotic cells . Approxi2.5.6 cells were exposed to the test compounds and the controls at their GI50 doses. Cells suspensions were stained with 200 μg/ml of 5,5’,6,6’-tetrachloro-1,1’,3,3’-tetraethyl benzimidazolo carbocyanine iodide (JC-1). Fluorescence was analyzed using a Fluorostar Optima fluorescence reader  using the 485/520 nm filters. Results were analyzed using the fluorometer’s MARS data analysis software (BMG).Mitochondrial membrane permeabilization (MMP) is a key event in the apoptotic process . A total2.6.Caspase activation has been used as a key endpoint to identify cell death via apoptosis. In brief, apoptosis activation via caspases which can be grouped into two major categories; initiators and effectors. We have selected as endpoints Caspases 3 and 7 as the effectors, and Caspases 8 and 9 as the initiators. Once this signaling is activated, the cell proceeds with activating its enzymatic degradation . Here, c2.7.DNA Fragmentation is another common endpoint used to measure apoptotic responses on cells . This ev2.8.Changes in the cell cycle can provide useful insights into drug effects. Some compounds may cause cells to become arrested in a particular stage . To stud2.9.4 cells were treated using the Premo kit following manufacturer’s specifications and plated in 96 well plates then is analyzed using a Fluorostar Optima fluorescence reader  using the 485/520 nm filter.Autophagy is a key cellular process for maintaining homeostasis, and involves the degradation of subcellular components  17]. Du. Du17]. 2.10.Cathepsins are a group of proteases present in lysosomes and present during autophagy . These e3.3.1.50) is commonly used as a measure of the inhibition of 50% of exposed cells to a substance. The growth inhibition dose of 4.4 μM for 2-EH and 76 μM for DEHP on TK6 cells were determined after a 24-hour exposure (Growth inhibition 50% (GIexposure . The meaexposure .3.2.p < 0.05) induction of apoptosis when compared to the negative vehicle control  differences were detected on all test compounds when comparing to the negative control (m).Mitochondrial health was assessed after exposure to the phthalates and the controls. Statistically significant ( control . Mitocho3.4.3.5.3.5.1.p < 0.05) caspase 3 activation (Activation of effector caspase 3 was detected in the camptothecin positive control and 2-EH samples. Camptothecin samples presented an average of (74.6%) and the 2-EH samples an average of 82.0%. both samples presented statistically significant (tivation . DEHP sa3.5.2.p < 0.05) manner. Negative samples averaged 9.6% of caspase 8 active cells and the camptothecin positive control average 37.6% activation. Phthalate samples presented 25.0% and 84.0% for DEHP and 2-EH respectively  phase . The cam3.5.5.p< 0.05) when comparing them to the negative control. Chloroquine control averaged 1120 FSU while the DEHP and 2-EH averaged 1287 and 1984 FSU respectively  results (Detection of cathepsins presented unexpected results, the negative control and 2-EH did not present any statistically significant ( results . The neg4.50 dose of 4.4 μM shows higher 2-EH toxicity when comparing to DEHP with a 76 μM dose at 24 hours. Both compounds cause cell death however, some of the cell death hallmarks differ between DEHP and 2-EH.The ubiquitous nature of phthalates in the modern world has caused an increase in the study of their biological interactions and potential health effects. Our previous work  presentem are removed by autophagy . It. It23]. utophagy . Still, utophagy  26]. Ca. Cam areutophagy . This miutophagy . The celutophagy  30] [31 [31m are5.Based on our results, 2-EH shows activation of several different cell death events when comparing it to DEHP. The phthalate 2-EH presented higher toxicity with significant activation of caspases, alterations in mitochondrial potential, cell cycle arrest in S phase, the presence of autophagic lysosomes but no cathepsin B activation. Due to the interplay between the autophagic and apoptotic cell death events, further studies are needed to determine the mechanistic details and signaling events. The activation the majority of our tested cell death parameters by 2-EH and its higher toxicity warrants further mechanistic studies; its implications in public health and environmental impacts for these emerging contaminants.


Associations of PM2.5 with risk factors were assessed by mixed-effect generalized estimation equation models adjusted by sex, age, exercise, body mass index, and smoking. In the final model there was further adjustment for country.Cardiovascular risk factor data were obtained from the SUrvey of Risk Factors (SURF) for coronary heart disease (CHD) patients from 10 countries in Europe, Asia, and the Middle-East. Annual average PM2.5 via the postal address of participating hospitals was shown to be feasible, however with several limitations noted.Linkage between cardiovascular risk factor data and PM3 in PM2.5 was significantly associated with decreased BP and increased glucose. After controlling for country, an increase of 10 μg/m3 in PM2.5 was associated with decreased BP and increased LDL . The association with glucose attenuated .Eight thousand three hundred and ninety two patients (30% women) were included. Globally, an increase of 10 μg/m2.5 and cardiovascular risk factors but it is still challenging to interpret these observed associations due to unavailability of potential confounders. After country adjustment, PM2.5 was associated with small increases in LDL and small decreases in BP.It is feasible to link PM- There are limited studies on the association between air pollution and cardiovascular risk factors for patients with established coronary heart disease in low- and middle-income countries;- Data linkage is an efficient and cost-effective method to maximize the use of existing data to investigate more health related research questions;- It is feasible to determine global associations of air pollution and cardiovascular risk factors by data linkage but it is still challenging in terms of interpretation. Several456scarce 8911121314.ations 89. In conttrations . Existinountries . This, t2.5 and cardiovascular risk factors on a global scale is challenging. Data linkage is an efficient and cost-effective method to maximize the use of existing data for more health related research questions [2.5 and cardiovascular risk factors  among patients with established coronary heart disease (CHD) in Europe, Asia, and the Middle East.Conducting targeted studies on the association between PMuestions 17. CurreWe used cardiovascular risk factors from the SUrvey of Risk Factors (SURF). Details have been reported previously 19. Brief2.5 concentrations from the World Health Organization (WHO) database . The database provides estimates of annual average concentration of PM2.5 at a spatial resolution of 0.1° × 0.1°, which is approximately 11 × 11 km at the equator globally. Due to data availability, we used annual average of the year 2014. The estimates are based on the recently developed Data Integration Model for Air Quality [2.5 using satellite retrievals of aerosol optical depth, chemical transport models, population estimates, topography and ground measurements from 6003 stations worldwide. A Bayesian hierarchical model is used to integrate these information sources [We extracted annual average PMhttps://www.eea.europa.eu/data-and-maps/data/airbase-the-european-air-quality-database-7). For the 17 districts in the city of Beijing we also obtained online PM2.5 data from the Beijing Municipal Environmental Protection Bureau for the year 2013.We additionally collected data in 2013 for European centers from countries that report measurements data to the European Environment Agency using the Airbase database -using Google Earth. We first linked PMvailable . For a s3 in PM2.5 were assessed by adjusted mixed-effect generalized estimation equation models. Patient’s characteristics and lifestyles varied country by country and thus, we included all available potential confounding factors related to both cardiovascular risk factors and PM2.5, including sex, age, and individual risk factors  [The associations of cardiovascular risk factors with an increase of 10 μg/mx <BMI>) . All patImputed data were analyzed in the primary analysis. There were about less than 4% missing data for all variables (Appendix Table A). Ten datasets were imputed for missing data with multivariate imputation by chained equations (MICE package in R) . Briefly2.5 levels (China and Saudi Arabia) . We further analyzed associations of PM2.5 retrieved from the Airbase for European countries and the database from the Beijing Municipal Environmental Protection Bureau for China with the same statistical strategy.Because of uncertainty of the shape of the concentration response function at high concentrations, we performed sensitivity analyses excluding the two countries with the highest PMStatistical analyses were performed by using ‘MICE’ and ‘GEEPACK’ packages in R 24. All tWe first describe the collected data and associations between air pollution and cardiovascular risk factors and then summarize the potential limitations of using existing audit data.2.5 level from WHO database was 38.1 μg/m3, ranging from 10.1 μg/m3 in Ireland to 92.7 μg/m3 in Saudi Arabia. Appendix Figure B illustrates the large variation of individual outcome variables, especially within countries.A total of 8392 SURF patients were included. The mean age of all patients was 64.9 years; 29.6% were women; 16% reported current smoker  and the association tended to be stronger  after country adjustment on a global scale. On European level, a similar association between PM2.5 and DBP was observed which became non-significant after country adjustment.Similar results were found for DBP: an increase of 10 μg/m2.5 and lipid levels. Associations of PM2.5 with lipid levels were not statistically significant on a global scale. After controlling for country non-significant associations remained for TC and HDL; while, an increase of 10 μg/m3 in PM2.5 was associated with an increased LDL level . Weak positive associations of TC and LDL were observed among European participants , which disappeared after adjustment for country. There was no significant association for HDL among European patients with or without adjustment for country.Figure 2.5 exposure data (Appendix Table B) did not alter the main findings.Separate analyses with exclusion of China and Saudi Arabia  is associated with 32.1 mmHg (95% CI 21.6–42.6) and 31.3 mmHg (95% CI 25.4–37.1) increases in SBP and DBP, respectively, after controlling age, sex, BMI, smoking, and drinking habitats [2.5 and BP in each cohort and the pooled results remained non-significant [2.5 could instigate acute autonomic imbalance and then lead BP increases [525We observed an inverse association of PMvated BP 92526. Oted BP 925. For inshabitats . A comprnificant . Studies [9252.5 8 μg/m3 ieases 52529. Howev2.5 may affect lipid levels but the quantity and quality of these studies is still limited and results are not fully consistent [303) of PM2.5 was associated with a higher level of TC  [2.5 had significantly higher levels of TC and LDL than those exposed to filtered air [Some previous evidence suggested that PMnsistent 3031. A l2.5 with glucose in both global and European analyses, although these associations attenuated after country adjustment. These findings are in line with previous studies [2.5 [2.5 to be associated with increased insulin resistance and higher rates of type II diabetes [2.5 with endothelial dysfunction, endoplasmic reticulum stress, insulin signaling abnormalities, and systematic inflammation [533We observed direct associations of PM studies 33. A croies [2.5 . A reviediabetes . Mechanimmation 53334. DifThe current study has piloted feasibility to add air pollution exposure using a coherent methodology to the rich database of clinical observations on cardiovascular risk factors from SURF. Data linkage is a robust, valuable and cost-effective research tool for combining individual level data from different sources for maximizing use of these existing database and increasing amounts of data that are being produced in order to: 1) address clinical research questions that require large sample sizes, detailed data on hard-to-reach population, or specified measurements by using a single dataset; 2) generate evidence with a high level of external validity and applicability; 3) reduce participant burden and avoid duplication of effort 353637. T35362.5 concentrations were assigned to each center, as PM2.5 is a regionally varying pollutant with limited small-scale spatial variation [There are several limitations in this study. SURF records CHD management in daily practice. Unlike other epidemiological studies, physical and laboratory measurements are not standardized. Some potential confounders were not available and thus could not be adjusted for. In addition, SURF collected anonymous data and thus only participating center’s locations at aggregated level were linked to air pollution data instead of individual level, which may not reflect actual exposure at the individual level. However, most routine cardiology visits were conducted in local hospitals with the distance between home and clinic generally being less than 10 km as confirmed by SURF national collaborators for 80%–90% of their patients had their residence near hospitals. The lack of individual addresses resulted in that only PMariation . Finally2.5 exposure data provided by local resource from 2013 and found that the results are broadly similar. Annual average concentrations may vary from year to year due to variations in weather, but the spatial contrasts in air pollution are typically stable over years and as such it may only have limitedly impacted our findings [23Furthermore, the air pollution data from WHO database was not available for 2013, which was our year of interest because this coincides with the year of observation for the SURF study. In sensitivity analysis, we further analyzed association with PMfindings 2324.2.5 mostly varies on a regional scale with limited local variability. In a large monitoring study across Europe, we observed that 81% of the variance was due to between study area variability [While most epidemiological studies of air pollution are based upon more individual exposure assessment, our approach does not invalidate the epidemiological study. First, the selected pollutant PMiability . Second,iability . Fourth,iability . TherefoWe hope that our findings may stimulate linkage studies on cardiovascular risk and disease in primary prevention settings in which relationships may be stronger and the findings less likely to be confounded by medication. Even though clinicians may not be able to change patients’ living environment they should become more aware of the hazards of air pollution and take it into account in their risk assessment and recommendations, such as promoting exercise in less polluted areas.2.5 levels are marginally associated with increases in LDL cholesterol and decreases in BP. The implication is that similar global studies should aim at multiple centers per country with sufficient within country exposure contrast to balance any effects of over adjustment.The current study has demonstrated the feasibility of data. The approach exemplifies the opportunity to assess the impact of the environment on cardiovascular risk factors across large geographic areas. We noted that estimates were highly sensitive to adjustment for country. After country adjustment, PMThe data that support the findings of this study are available from the SURF project, which have been published previously. The references have been included in the current study.The additional files for this article can be found as follows:10.5334/gh.877.s1Appendix Table A.Missing data in SURF.10.5334/gh.877.s2Appendix Table B.Associations between CVD risk factors and PM2.5 retrieved from WHO, airbase for European countries and local government database for China.10.5334/gh.877.s3Appendix Table C.Risk factors, stratified by centres.10.5334/gh.877.s4Appendix Figure A.2.5 distribution.PM10.5334/gh.877.s5Appendix Figure B.Variations of individual outcome variable by country.10.5334/gh.877.s6Appendix Figure C.2.5 and cardiovascular risk factors.Crude association between PM


Multidetector computed tomography (MDCT) angiography is a useful examination to detect the source of the bleeding in patients with hemoptysis. The aim of the study was to prospectively evaluate the role and clinical efficacy of MDCT angiography before bronchial artery embolization (BAE) for the management of hemoptysis, and to investigate the predictors of early recurrence.It is a double-center study which included 57 hemoptysis patients undergoing MDCT angiography prior to BAE from August 2019 to July 2020. A prospective analysis of culprit arteries detected by MDCT angiography allowed an evaluation of the role of this technique. A follow-up was done to assess the efficacy of BAE with preprocedural MDCT angiography and to explore the risk factors of early recurrent hemoptysis.p = 0.022) was associated with an increase in the risk of early recurrence.The accuracy of MDCT angiography in the identification of culprit arteries was as high as 97.5%. The average number of total culprit arteries per patient was 2.75 ± 1.73. Among which, the average numbers of culprit ectopic bronchial arteries (BAs) and non-bronchial systemic arteries (NBSAs) per patient were 0.21 ± 0.41 and 1.04 ± 1.57, respectively. The immediate clinical success rate, total hemoptysis recurrence rate, and early hemoptysis recurrence rate of BAE following MDCT angiography were 94.7, 18.5, 16.7%, respectively. Aspergilloma (HR = 6.63, 95% CI: 1.31–33.60, MDCT angiography should be performed before BAE for the management of hemoptysis. Aspergilloma was an independent predictor for early recurrence. Hemoptysis is a challenging symptom which frequently involves life-threatening conditions, such as malignancies. Therefore, the diagnostic work-up should be as exhaustive as possible. However, the lack of evidence-based guidelines regarding appropriate diagnostic management may lead to inaccurate diagnosis and delayed treatment of the symptom . The comIn patients with hemoptysis, the identification of sources and causes of bleeding with non-invasive methods prior to interventional techniques is essential. To date, there have been many studies reporting that multidetector computed tomography (MDCT) angiography before BAE plays a critical role in the diagnosis of abnormal arteries and the hemoptysis cause –15. ImpoHowever, most reports have been retrospective studies, and the evaluation of efficacy of BAE following MDCT angiography on clinical outcomes is limited. Some literature also has uncertainly recommended MDCT angiography as a regular examination in patients undergoing BAE for hemoptysis –22. TherA prospective, double-center study, including 57 consecutive patients with hemoptysis who were referred for BAE, was conducted at Military Hospital 103 and National Lung Hospital, two national hospitals in Hanoi, Vietnam between August 2019 and July 2020. The classification of hemoptysis grades include massive hemoptysis , moderate hemoptysis (100–300 mL/day), and mild hemoptysis (< 100 mL/day)  and that of categorical variables were presented as numbers and percentages. The baseline characteristics between history and non-history of pulmonary TB groups were compared by using n = 23, 40.4%), active/reactivated TB , and aspergilloma . Notably, of 57 patients, 23 (40.4%) had a history of pulmonary TB, and 21 (36.8%) had massive hemoptysis. The mean ± SD duration from BAE to death or the end day of follow-up was 4.7 ± 3.2 . The baseline characteristics of the participants are reported in Table The study included 57 patients from August 2019 to July 2020. Among whom, there were 40 males (70.2%), and the mean ± SD age at admission was 54.4 ± 16.6 years . The incidences of cardiovascular diseases, diabetes mellitus, and chronic liver diseases were 19.3% (11/57), 8.8% (5/57), 7.0% (4/57), respectively. It is noted that the top three common causes of hemoptysis requiring BAE were bronchiectasis . The average number of culprit orthotopic BAs, ectopic BAs, and NBSAs per patient were 1.54 ± 0.73, 0.21 ± 0.41, 1.04 ± 1.57, respectively. Totally, the average number of culprit arteries per patient was 2.75 ± 1.73 , ostial narrowing (n = 1), and acute or multiple branching (n = 2). As a result, the rate of technical success was 93.0% (53/57). Regarding the clinical outcomes, the rate of immediate clinical success was 94.7% (54/57). Of the 3 patients who experienced significant hemoptysis within 24 h of BAE, one underwent an urgent lobectomy and two continuously received the medical treatment. In patients with immediate clinical success, 10/54 (18.5%) were recurrent bleeding with the follow-up periods ranging from 1 to 12 months. The rate of early recurrence was 16.7% (9/54), and there were 4 deaths because of massive hemoptysis recurrence (n = 3) and severe underlying lung diseases (n = 1).Among enrolled patients received BAE, the technical failure occurred in 4 patients due to artery tortuosity (p < 0.05). The results revealed that aspergilloma  was associated with the early recurrence. Multivariate analysis of three variables, including the etiology of hemoptysis, extent of lung diseases, and grade of hemoptysis, was performed. Our findings also indicated that aspergilloma  was the risk factor of early recurrence for patients who received BAE with preprocedural MDCT angiography.We excluded three patients without immediate clinical success and two patients in the other etiology group, the remaining 52 patients were analysed to explore the risk factors of early recurrence. Table p > 0.05). However, it is noted that the number of total culprit arteries and culprit NBSAs was significantly higher in the group of pulmonary TB history . In contrast, the number of culprit orthotopic BAs and ectopic BAs was similar between two groups .Of all 57 patients, there were 23 40.4%) with and 34 (59.6%) without pulmonary TB history  [Surprisingly, aspergilloma was found as a predictor for early recurrence among hemoptysis patients treated by BAE following MDCT angiography. A recent systematic review by Panda et al. 2017) summarized that technical failure due to missing culprit vessels or embolization inability might be causes of early recurrence 017 summa, 37–41. ctively) , 41. Aboctively) . Therefoctively) . Cavernoctively) .The third question in this research was what the similarities and differences between hemoptysis patients with and without pulmonary TB history were, who received BAE following MDCT angiography. Consistent with the literature , 41, 44,n = 10), and this might be a cause of bias in our results. In addition, the incidence of hemoptysis recurrence might vary by many factors, these findings cannot be extrapolated to all patients.This study has several limitations. Firstly, it has no control group. Therefore, we have compared with previous studies to evaluate the value of MDCT angiography before BAE. But randomised controlled trials will be recommended to gain an insightful picture of this technique. Secondly, only short-term outcomes were covered in the scope of this study. However, it should be noted that the recurrent event could occur from 6 months to 1 year after BAE . Hence, MDCT angiography could be considered as a regular examination prior to BAE for the management of hemoptysis. Aspergilloma was a risk factor for early recurrence. Patients with pulmonary TB history have a significantly higher number of total culprit arteries and culprit NBSAs than patients without pulmonary TB history.


In the Surveillance Summary “Abortion Surveillance — United States, 2016,” data on the number of known previous induced abortions for women having abortions in this reporting year were erroneously included for New York City. These estimates did not meet reporting standards and should have been excluded from this report. When corrected, among women with abortions in the reporting year, the proportion with no previous induced abortions increased, and the proportion with one or more previous induced abortions decreased.40.7% of abortions, and women with no previous abortions accounted for 59.4%.”On page 2, the second sentence of the first paragraph should have read “Women with one or more previous induced abortions accounted for 41 areas that reported the number of previous abortions for women who obtained abortions in 2016 indicate that the majority (59.4%) had no previous abortions, 34.4% had one or two previous abortions, and 6.3% had three or more previous abortions (Table 17). Among the 34 reporting areas***** that provided data for the relevant years of comparison , the percentage of women who had no previous abortions increased 3% (from 57.4% to 59.1%), whereas a 4% decrease occurred among women who had one or two previous abortions, and a 4% decrease occurred among women who had three or more previous abortions from 2007 to 2016. However, the percentage of women who had no previous abortions decreased 1% from 2007 to 2011 (from 57.4% to 56.8%) and then increased 3% from 2012 to 2016 (from 57.6% to 59.1%). By contrast, the percentage of women who had three or more previous abortions increased 4% from 2007 to 2011 (from 6.8% to 7.1%) then decreased 6% from 2012 to 2016 (from 6.9% to 6.5%). The percentage of women who had one or two previous abortions increased 1% from 2007 to 2011 (35.8% to 36.1%) and then decreased 3% from 2012 to 2016 (from 35.5% to 34.5%).” New York City should have been included in the ***** footnote, which lists reporting areas that were not included in these estimates.On page 10, the first paragraph of the first column should have read “Data from the 244,362 (59.4), 100,248 (24.4), 40,972 (10.0), 25,986 (6.3), 411,568 (98.5). The * footnote should have read “Data from 41 reporting areas; excludes 11 areas  that did not report, did not report by the number of previous induced abortions, or did not meet reporting standards.” The total in the ** footnote should have been 417,809.In Table 17, the line for New York City should be deleted. For the total line, the numbers and percentages should have read


Gastrointestinal disorders are frequently reported in patients withParkinson’s disease whose disorders reduce the absorption of nutrients anddrugs, worsening the clinical condition of patients. However, the mechanismsinvolved in modifying gastrointestinal pathophysiology have not yet beenfully explained. To evaluate its effects on gastrointestinal motility and the involvement ofthe vagal and splanchnic pathways. Male Wistar rats  were used and divided into two groups.Group I (6-OHDA) received an intrastriatal injection of 6-hydroxydopamine. Group II (control) received a saline solution under the same conditions. The study of gastric emptying, intestinaltransit, gastric compliance and operations (vagotomy and splanchnotomy) wereperformed 14 days after inducing neurodegeneration. Test meal (phenol red 5%glucose) was used to assess the rate of gastric emptying and intestinaltransit. Parkinson’s disease delayed gastric emptying and intestinal transit at alltime periods studied; however, changes in gastric compliance were notobserved. The delay in gastric emptying was reversed by pretreatment withvagotomy and splanchnotomy+celiac gangliectomy, thus suggesting theinvolvement of such pathways in the observed motor disorders.  Parkinson’s disease compromises gastric emptying, as well as intestinaltransit, but does not alter gastric compliance. The delay in gastricemptying was reversed by truncal vagotomy, splanchnotomy and celiacganglionectomy, suggesting the involvement of such pathways in delayinggastric emptying. Allexperiments were carried out according to the Ethical Principles Guide for theCare and Use of Laboratory Animals, proposed by the Brazilian Society ofLaboratory Animal Science after approval by the local ethics committee (protocolN° 40/2015). All efforts to minimize the number and suffering of the animalswere implemented.® 6 μL syringe and a stereotactic device with the following coordinates (mm): site 1: L: - 2.5, AP: +0.5, V: + 5.0; Location 2: L: - 3, AP: - 0.5, V: + 6.0; and location 3: L: -3.7, AP: - 0.9, V: + 6.5 of bregma, according to the Paxinos and WatsonAtlasThe animals were initially divided into two groups (n=6), they received withketamine  and xylazine  for anesthesia purposes,followed by a stereotaxic surgery. Group I (6-OHDA) received a unilateralintrastriatal injection of 6-OHDA  on the ipsilateral side. GroupII (control) received a saline solution  under the same conditions.The unilateral intrastriatal injection of 6-OHDA or the saline solution wasperformed using a HamiltonThe rotational behavior was evaluated by monitoring the rotations induced byapomorphine (3 mg/kg i.p) which induces the animal to rotate in the oppositedirection of the injury . The number of rotations (360°)around the axis was counted every 10 min for a total period of 60 min,The gastric emptying study was performed 14 days after the neurodegenerationinduction operation. After 12 h of fasting, the animals were fed (via gavage)with 1.5 ml of liquid test meal containing a non-absorbable marker (0.5 mg/ml ofphenol red in 5% glucose solution). After 10, 20 or 30 min, the rats weresacrificed by cervical dislocation and the intestine was exposed by laparotomy,fixed to the pylorus, cardia and ileocecal junction, and then removed. Theintestine was carefully stretched from the stomach to the colon and divided intofour consecutive segments: stomach, proximal small intestine (first 40%), medialsmall intestine (intermediate 30%) and distal small intestine (last 30%). Eachsegment was placed in graduated cylinders containing 100 ml of NaOH 0.1 N. Afterbeing crushed and homogenized for 30 s, the sediment suspension was left for 20min at room temperature. The supernatant (10 ml) was centrifuged for 10 min at2800 rpm and the proteins contained in 5 ml of the homogenate volume wereprecipitated after adding 0.5 ml of trichloroacetic acid (20% w/v). Aftercentrifugation for 20 min at 2800 rpm, 3 ml of the supernatant was added to 4 mlof 0.5 N NaOH. A standard dilution curve was generated to relate theconcentration of phenol red in 0.1 N NaOH with the absorbance (560 nm) obtainedin each segment,After anesthesia with ketamine  and xylazine , theanimals were submitted to laparotomy and a cannula was inserted into theintestine. Its tip was moved to the duodenum 1 cm distal to the pylorus andfixed to the stomach wall; its free end was channeled subcutaneously,externalized and attached to the skin. After two days, the animals were fed(gavage) with the liquid test meal (1 ml) through the duodenal cannula and weresacrificed 20 min later. After exeresis, the intestine was carefully stretchedand removed. Obstructive bandages were placed to obtain five consecutivesegments of the small intestine (~20 cm long). Each segment was homogenized, andthe dye content was determined by spectrophotometry. The data obtained for eachindividual segment was multiplied by the total number of segments and added tocalculate the geometric center of the marker distribution throughout theintestineTo assess the effects of PD on gastric compliance, a barostat system associatedwith a plethysmometer was used. After 14 days of the neurodegeneration inductionoperation, the animals were anesthetized with urethane (1.2 g/kg i.p) followedby a tracheostomy. A balloon catheter (~4 ml) made with the fingertips ofsurgical gloves was inserted orally and positioned in the stomach of the rats.The free end was connected to a glass reservoir , creating a system of communicating vessels filled with a standard ionicsolution  at37° C. Thus, changes in the volume of the reservoir, displayed by theplethysmometer  were considered gastricvolume. After the system was balanced, the stomach was progressively distended,increasing the liquid level in the reservoir 4, 8 and 12 cm above the rat’sxiphoid appendix every 10 min. Gastric volume was recorded every 1 min andcollected at consecutive 10 min intervals,The animals initially fasted for 24 h, with free access to water. Afteranesthesia with ketamine  and xylazine  bothgroups were submitted to median laparotomy. Animals in the truncal vagotomygroup were submitted to the vagus nerve section through an esophageal serotomy~1.5 cm above the cardia. On the other hand, those in the splanchnotomy + celiacgangliectomy group underwent median laparotomy with exposure of the abdominalviscera and the celiac trunk. Then, the celiac ganglion and splanchnic nerveswere sectioned, followed by a 100% alcohol instillation® software version 5.01 . Theresults of gastric emptying and intestinal transit were presented as a histogramthat is representative of the ± SEM. The statistical differences between themeans were analyzed with the Student’s t-test. The gastric compliance resultswere presented in a box chart and wisker plots. The statistical differences inthe means obtained in different groups of animals in the study of the vagal andsplanchnic pathways were analyzed by ANOVA followed by the Bonferroni test. p<0.05 values were considered significant.The graphs and statistical analysis of the results were performed using thePrismaRotations of the animal around the axis on the opposite side of the lesion, inducedby apomorphine, reflect the hypersensitivity and severity of the damage caused bythe intra-striatal lesions produced by the 6-OHDA injectionRetention of the test meal in the stomach of animals in the 6-OHDA group 10 minpostprandial was 24.85% higher than in the control group, whose mean dye retentionwas 53.40±2.80 vs. 42.77±2.65, respectively. Dye retention in the proximal portionof the small intestine of animals in the 6-OHDA group was 18.21% lower than in thecontrol group, whose dye retention averages were 29.19±1.95 and 35.69±1.31,respectively  A.The retention of the test meal in the stomach of animals in the 6-OHDA group 20 minpostprandial was 31.52% (33.50±2.53 vs. 44.06±2.23) higher than the control group.In the proximal portion of the small intestine of animals in the 6-OHDA group, therewas a reduction of 32.68% (26.74±1.65 vs. 18.00±2.55) of dye retention when comparedto animals in the control group B.The retention of the test meal by the stomach of animals in the 6-OHDA group 20 minpostprandial was 33.04% (27.72±1.96 vs. 36.88±2.19) higher than the control group.In the proximal portion of the small intestine of animals in the 6-OHDA group, therewas a 61.86% reduction (22.89±1.03 vs. 8.73±1.76) of dye retention when compared toanimals in the control group  C.The number of fecal boluses released by the animals in the experimental group(6-OHDA) was 6.571±1.88, while the control group (NaCl 0.9%) was 16.03±0.72 .There was a 44.80% delay in the center of mass of the test meal in animals in the6-OHDA group compared to those in the control group. When in animals in the 6-OHDAgroup, the center of mass was observed between segments 2 and 3 (2.95±0.32) of thesmall intestine, while in the control group (NaCl 0.9%) it was between segments 4and 5 ; 2.64±0.26 vs. 2.63±0.11 ml (8cmH2O) e 2.96±0.27 vs. 2.77±0.11 ml (12 cmH2O),respectively. Thus, the gastric volume values of animals in the 6-OHDA and controlgroups did not show statistically significant differences, p <0.05  gastric fluidretention, with the results of animals in the vagotomized saline group andvagotomized 6-OHDA group (34.40±1.740 vs. 34.30±2.764) were compared to each other.Pretreatment with vagotomy accelerated gastric emptying (44.78±1.380 vs.34.30±2.764), when the 6-OHDA group was compared with the vagotomized 6-OHDA group.On the other hand, vagotomy did not alter fluid retention in the proximal, medialand distal intestines, respectively A.Surgical pretreatment with splanchnotomy + celiac ganglionectomy (EGc) did not alter(p>0.05) the results of gastric fluid retention in animals in the splanchnotomysaline and splanchnotomy 6-OHDA groups (30.94±1.740 vs. 26.30±2.764). Splanchnotomy+ celiac gangliectomy accelerated gastric emptying (44.78±1.380 vs. 26.30±2.764) ofthe splanchnotomy 6-OHDA group in relation to the non- splanchnotomy 6-OHDA group.On the other hand, splanchnotomy+celiac gangliectomy did not significantly alterfluid retention in the proximal, medial and distal intestines, respectively B.PD was initially described as a central nervous system diseaseThe 6-OHDA-induced neurodegeneration model is one of the most widely used. Thestructural similarity of 6-OHDA with dopamine and noradrenaline, allows 6-OHDA to bequickly captured by dopaminergic and noradrenergic neurons, forming lipid peroxide,as well as, the inhibition of mitochondrial complex I activity, consequently causingthe death of most of these cellsContralateral rotations induced by apomorphine reflect the hypersensitivity and levelof severity of striatal lesions caused by the unilateral injection of 6-OHDAThe rate of gastric emptying is measured by the gastric content delivery rate in theduodenumThese results corroborate with several studies on the effects of PD on gastricemptying in animal models. Studies carried out with rats injured with6-hydroxydopamine reported delayed gastric emptying and constipationPD did not interfere with the animals’ gastric compliance at the different pressuresstudied B. This rThe gastrointestinal tract is largely interconnected with the central nervous systemand receives sympathetic and parasympathetic signs, providing extensive innervationto the plexuses of the enteric nervous systemPD delayed gastric emptying and intestinal transit, whose disorders can affect thebioavailability of drugs and nutrient absorption, worsening the patient’s clinicalcondition. The delay in gastric emptying was reversed after the surgicalintervention of the truncal vagotomy and splanchnotomy and celiac gangliectomy,suggesting the involvement of such pathways in the pathophysiology of the disease.Thus, the 6-OHDA-induced neurodegeneration was presented as a model capable ofmimicking the main symptoms of PD, enabling a greater understanding of the effectsof this disease on gastrointestinal motility, making it possible to diagnosedisorders with greater precision and leading to discoveries of pharmacologicalinterventions for more efficient treatment.PD compromises gastric emptying, as well as intestinal transit, but does not altergastric compliance. These factors together contribute to the worsening of symptomsby interfering in drug therapy absorption. The delay in gastric emptying wasreversed by truncal vagotomy, splanchnotomy and celiac ganglionectomy, suggestingthe involvement of such pathways in delaying gastric emptying.


We report the epidemiology of Lassa fever in Bauchi State, a disease-endemic region, in Nigeria. Since 2015, major increases in Lassa fever attack rate and in the case-fatality rate have occurred in this state. A delay in seeking care by a case-patient for >7 days after symptom onset was the major predictor of death. In recent years, Lassa fever (LF) outbreaks in Nigeria have become more frequent and larger in magnitude; the outbreak in 2018 was described as the largest in history .https://www.cdc.gov/epiinfo/support/downloads.html). We calculated frequencies and proportions and examined the relationship between the outcome variable (death) and the risk factors (including sociodemographics) by using the χ2 test. We included significant variables (p< 0.1) by bivariate analysis and the biologically plausible ones (sex and age) in an unconditional logistic regression model; α = 0.05 was considered the level of significance.We analyzed data by using Epi Info version 7.2 software  case-patients were 15–64 years of age. This age group had the highest age-specific attack rate , and patients <5 years of age had the lowest attack rate . Most (54.0%) patients were male; the attack rate was 1.2 cases/100,000 persons for male sex and 1.1 cases/100,000 persons for female sex. The overall case-fatality rate (CFR) was 54.0% (41/76) and was highest (66.6%) for persons <5 years of age  Figure 12 4.8; p = 0.03), and the attack rate increased from 0.1 cases/100,000 persons during 2015 to 0.4 cases/100,000 persons during 2018 . We found by multivariate analysis that a delay in seeking care for >7 days after onset of illness  or for >24 hours after onset of bleeding  were independent predictors for dying from LF  compared with the Sahel savanna vegetation in the central and northern parts of Africa (>24 hours after onset of bleeding was a strong predictor of death among cases is a concern. LF has some common early symptoms similar to those of other febrile diseases, especially malaria, that are frequently encountered in most LF-endemic settings (Our finding that a delay in seeking care of Additional information on the epidemiology of Lassa fever and factors associated with deaths, Bauchi State, Nigeria, 2015–2018.


Drosophila melanogaster as reference at the whole‐genome level. The results show that SSR numbers (33 076–560 175/genome) and genome sizes (574.57–1342.21 Mb) are significantly positively correlated , but the correlation in individual species varies in these mosquito species. In six types of SSR, mono‐ to trinucleotide SSRs are dominant with cumulative percentages of 95.14%–99.00% and densities of 195.65/Mb–787.51/Mb, whereas tetra‐ to hexanucleotide SSRs are rare with 1.12%–4.22% and 3.76/Mb–40.23/Mb. The (A/T)n, (AC/GT)n and (AGC/GCT)n are the most frequent motifs in mononucleotide, dinucleotide and trinucleotide SSRs, respectively, and the motif frequencies of tetra‐ to hexanucleotide SSRs appear to be species‐specific. The 10–20 bp length of SSRs are dominant with the number of 110 561 ± 93 482 and the frequency of 87.25% ± 5.73% on average, and the number and frequency decline with the increase of length. Most SSRs (83.34% ± 7.72%) are located in intergenic regions, followed by intron regions (11.59% ± 5.59%), exon regions (3.74% ± 1.95%), and untranslated regions (1.32% ± 1.39%). The mono‐, di‐ and trinucleotide SSRs are the main SSRs in both gene regions (98.55% ± 0.85%) and exon regions (99.27% ± 0.52%). An average of 42.52% of total genes contains SSRs, and the preference for SSR occurrence in different gene subcategories are species‐specific. The study provides useful insights into the SSR diversity, characteristics and distribution in 23 mosquito species of genomes.Simple sequence repeats (SSRs) exist in both eukaryotic and prokaryotic genomes and are the most popular genetic markers, but the SSRs of mosquito genomes are still not well understood. In this study, we identified and analyzed the SSRs in 23 mosquito species using  Due to the high mutability of SSRs, which stem from their susceptibility to slippage events in DNA replication, SSRs provide an evolutionary mechanism for faster adaptation in response to environmental stress , of which 1966 SSRs belong to mosquitoes. Some SSR markers have been successfully applied to population studies in Ae. aegypti  enrichment of SSR‐containing genes and of all genes. The SSRs identified in the present study provide potentially important molecular markers for the study of population genetics, genetic mapping and regulatory mechanisms of functional genes in mosquito species. More importantly, this work provides useful insights into structure and distribution characteristics of SSRs as well as their variation patterns among different species.https://www.vectorbase.org/), and the genome sequences and annotation of Drosophila melanogaster (as reference) were from NCBI (http://www.ncbi.nlm.nih.gov/). All these genome sequences were assembled into scaffolds, except for An. nili which was assembled into contigs , was used to identify the SSRs in the 23 mosquito genomes and D. melanogaster investigated in this study. The software has high site detection with accuracy rates, and with export results including the numbers of different types of SSRs and the position and length (motif bp × number of motif) of each 9 was treated as (AG)18.MISA  and abundance  were used to measure the frequency of SSR in genomes  regions of sequence  and the GO annotation numbers obtained, we created a functional classification of SSR‐containing genes and analyzed the number and proportion of each functional category of SSR‐containing genes in comparison to all annotated genes for each species.Based on the identification of the SSRs in different genomic regions, we sorted out SSR‐containing genes from the annotation documents. We then extracted the GO annotation numbers of SSR‐containing genes and of all genes from VectorBase for each species. Using WEGO . This phenomenon was the same as that for 30 marine species reported , Ae. aegypti (1342.21 Mb) and Cx. quinquefasciatus (574.57 Mb), with an average of 1261.62 ± 531.1 Mb (mean ± SD), were much bigger than those of the 20 Anopheles species in Anophelinae (201.88 ± 43.09 Mb), and correspondingly the SSR numbers of these three species (411 573 ± 109 989) were much larger than those of the 20 Anopheles species (111 637 ± 39 542) ; they were significantly correlated for the 20 Anopheles species . For some Anopheles species, there were some exceptions for the correlation between the genome size and SSR number. For example, the genome size of An. sinensis (214.86 Mb) was bigger than that of nine Anopheles species, An. nili (98.58 Mb), An. darlingi (132.94 Mb), An. maculatus (141.20 Mb), An. albimanus (165.33 Mb), An. farauti (175.52 Mb), An. christyi (169.04 Mb), An. mininus A (195.70 Mb), An. culicifacies A (198.03 Mb) and An. atroparvus (217.57 Mb), whereas the SSR number (50 397) of An. sinensis was only larger than that of An. nili (33 076). These differences of genome size reflect the characteristics of the two subfamilies at the genomic level. The SSRs were thought to stem from the slippage events in DNA replication in response of faster adaptation in response to environmental stress  analysis, and they all showed a high level of polymorphism in a An. maculatus population , respectively  to 99.00%  of the total number of SSRs in the 23 mosquito species and in D. melanogaster, whereas the tetra‐ to hexanucleotide SSRs only made up 1.12%  to 4.22%   to 787.51/Mb (An. darlingi), while the density of tetra‐ to hexanucleotide SSRs was only 3.76/Mb (An. nili) to 40.23/Mb (An. darlingi) . For dinucleotide SSRs, the average density (30.82/Mb) was eight times smaller than that of the 20 Anopheles species (237.83/Mb). For trinucleotide SSRs, the average density (33.65/Mb) was three times smaller than that of Anopheles species (97.86/Mb). The average density for all six types of SSRs (378.97/Mb) was about 1.5 times smaller than that of Anopheles species (550.22/Mb). The results showed that the density of different types of SSRs at the genome level was also specific to the Culicinae and Anophelinae subfamilies.The cumulative density of mono‐ to trinucleotide SSRs ranged from 195.65/Mb (D. melanogaster (Table S1). For mononucleotide SSRs, the motif (A/T)n was predominant with an average frequency of 84.83% ± 7.90%, ranging from 70.88% (An. darlingi) to 97.73% (An. nili), while the motif (C/G)n was far rarer with an average of 15.17% ± 7.90% (2.27% in An. nili to 29.12% in An. darlingi) in these genomes. This result is consistent with earlier reports for the rhesus monkey  and Ae. aegypti (31.41%). The result is consistent with an earlier study in An. gambiae, in which the (AC/GT)n was a little more than two times as abundant, much as (AG/CT)n, and (AT/AT)n and (CG/CG)n were rare n predominated, while the (ACG/CGT)n, (ACT/AGT)n, (AGG/CCT)n and (CCG/CGG)n were rare. However, (AAG/CTT)n, (ACG/CGT)n and (AAT/ATT)n are predominant in six bamboo species n and (AAAAC/GTTTT)n were the most frequent pentanucleotide SSRs in six mosquito species , but there was no common motif in the remaining 12 species. For the hexanucleotide SSRs, the motif (AACAGC/GCTGTT)n was predominant in seven mosquito species (average 24.57% ± 5.62%), (AAGTAG/CTACTT)n was dominant in An. mininus A (18.25%), An. maculatus (23.08%) and An. stephensi (17.14%), and there was no common motif in the other species. The high variation of motif occurrence frequency in tetra‐ to hexanucleotide SSRs is consistent with earlier investigations, for example there are four main motifs, (AAAT/ATTT)n (27.14%), (AACTG/CAGTT)n (40.06%), (AGTTC/GAACT)n (39.49%) and (AAACAA/TTGTTT)n (16.35%) in six bovid species n was the most frequent tetranucleotide SSR in 18 mosquito species (average frequency 26.79% ± 10.83%), but not in D. melanogaster  for the 23 mosquito species and Anopheles species, three Culicinae species, and for D. melanogaster  was higher than the AT‐content (47.61%). In the six types of SSRs, the trinucleotide SSRs had the highest GC‐content values , and the mononucleotide SSRs had the lowest GC‐content values  for Anopheles, Culicinae and D. melanogaster groups, respectively. Earlier whole‐genome studies of SSRs for other species have also shown that GC contents are lower than AT contents; for example, six bovid species , exon regions (3.74% ± 1.95%) and UTRs (1.32% ± 1.39%) . This suggests that the distribution of SSRs in different genomic regions is specific for different species. SSRs in different regions show different functions. SSRs in intronic regions can affect gene regulation, messenger RNA splicing, and gene silencing  and in Cx. quinquefasciatus (82.86%), but less than half in Ae. aegypti (29.60%), Ae. albopictus (33.72%) and D. melanogaster (26.79%). Earlier whole‐genome SSR studies have also shown a trinucleotide SSR preference in exon regions in Ap. cerana (66.6% SSRs in exon region), Ap. mellifera (76.7%)  . As a result, an average of 42.52% of total genes contained SSRs in the 21 mosquito genomes. The number of SSR‐containing genes ranged from 2669  in Cx. quinquefasciatus to 8319 (57.14%) in An. coluzzii. In the three main GO categories  in the GO enrichment, the subcategories and percentage genes in most subcategories were highly similar for both SSR‐containing genes and all genes in the 21 mosquito species genomes .For understanding the characteristics of SSR‐containing genes, and the differences of SSR‐containing genes in mosquito genomes, we compared functional annotations between SSR‐containing genes and all genes in 21 species with GO annotation by GO enrichment  which contained SSRs, while the other 17 did not. In the protein tag subcategory of the Molecular Function category, there were 15 mosquito species that contained SSRs, and the remaining six  did not. In the viral reproduction subcategory of the Biological Process category, three species of mosquito  contained SSRs, whereas the remaining 18 did not. The reason that preference for SSR occurrence differed in these subcategories remains to be further studied with a wider range of species. An earlier comparative analysis of GO enrichment between genes containing the motif (ATGTAC/GTACAT)n and all genes in marine species showed that the genes containing the motif are involved in evolution n or (AGC/GCT)n with a length less than or equal to 20 bp would be better molecular markers. Most SSRs are distributed in intergenic regions, and the mono‐, di‐ and trinucleotide SSRs are the main SSRs in both gene regions and exon regions. This study lays an important basis for the better understanding of SSRs and the selection of SSR molecular markers in mosquitoes.The authors declare no conflict of interest.Fig. S1. GO classifications of SSR‐containing genes and all genes in 21 mosquito species.Click here for additional data file.Table S1. The most frequent SSR motifs in the 23 mosquito species and D. melanogaster genomes.Click here for additional data file.


Although much research has focused on socio-demographic determinants of uptake of contraception, few have studied the impact of poor mental health on women's reproductive behaviours. The aim of this study was to examine the impact of poor mental health on women's unmet need for contraception and fertility rate in a low-income country setting.n = 999), 2.5 (n = 971) and 3.5 years (n = 951) post-delivery of index child and number of pregnancies during study period. We tested the association between CMD symptoms, unmet need for contraception and fertility rate.A population-based cohort of 1026 women recruited in their third trimester of pregnancy in the Butajira district in rural Ethiopia was assessed for symptoms of antenatal common mental disorders  using Self-Reporting Questionnaire-20. Women were followed up regularly until 6.5 years postnatal (between 2005 and 2012). We calculated unmet need for contraception at 1 year (s.d. 0.98). There was no prospective association between maternal CMD and number of pregnancies in the follow-up period.Less than one-third of women reported current use of contraception at each time point. Unmet need for birth spacing was higher at 1 year postnatal, with over half of women (53.8%) not using contraception wanting to wait 2 or more years before becoming pregnant. Higher CMD symptoms 1 year post-index pregnancy were associated with unmet need for contraception at 2.5 years postnatal in the unadjusted  and fully adjusted model . During the 6.5 year cohort follow-up period, the mean number of pregnancies per woman was 2.4 (CMD symptoms are associated with increased unmet need for family planning in this cohort of women with high fertility and low contraceptive use in rural Ethiopia. There is a lack of models of care promoting integration of mental and physical health in the family planning setting and further research is necessary to study the burden of preconception mental health conditions and how these can be best addressed. The women were identified by the Butajira Rural Health programme (BRHP) enumerators in the course of their 3-monthly surveillance interviews and, after giving informed consent, were interviewed by the project data collectors , living in the HDSS and in the third trimester of pregnancy during the study recruitment period  was created by adding the following poverty indicators: self-reports of indebtedness, hunger in the last month and perceived lower relative wealth  the husband was not providing enough help, (ii) the relationship was ‘average, bad or very bad’, (iii) they sometimes or often quarrelled or (iv) the woman felt that her husband drank too much alcohol  for the effect of these other variables on unmet need for contraception.Stata version 15 software was used for data analysis  with the corresponding confidence intervals (CIs) for the effects of other variables, including contraceptive use, socio-demographic and obstetric factors on the number of pregnancies the during study period. Missing data were addressed by case-wise deletion.n = 971) at 2.5 years, 92.7% (n = 951) at 3.5 years and 86.8% (n = 891) at 6.5 years (Follow-up rates were 94.6% (.5 years . Women w.5 years .Table 1s.d. ±5.9) at baseline and almost all women were married (99.0%), with 18.2% at baseline and 16.8% at 6.5 years in a polygamous marriage. Only 21.0% of women at the time of the index pregnancy had formal education, whereas over 50% of husbands had attended primary school or above. Over two-thirds of women were Muslim. At baseline mean parity was 3.0 (s.d. ±2.3) whereas at 6.5 years parity was 5.5 (s.d. ±2.1).Mean maternal age for women at time of recruitment into the study was 26.3 years ; 14.2% of all pregnancies at 3.5 years (n = 20).High levels of CMD symptoms (SRQ ≥ 6) were experienced by 15.0% of women at one time point and 6.5% at two or more time points during the follow-up period. At 1 year postnatal, 43 women  were pregnant again. At 2.5 years and 3.5 years, 24.3 and 14.8% of women reported current pregnancy, respectively. Of these, 37.2% at 12 months, 48.7% at 2.5 years and 27.7% at 3.5 years were unwanted pregnancies and 20.8% at 2.5 years and 17.7% at 3.5 years were due to contraceptive failure . Some won = 538, 53.8%) not using contraception and wanting to wait for 2 or more years before the next pregnancy. Unmet need for contraception for limiting was higher at 3.5 years when 206 women (21.6%) reported not wanting to have more children. Total rates of unmet need for contraception were calculated to be 64.0% at 12 months, 53.1% at 2.5 years and 62.7% at 3.5 years . Only a very small proportion reported not using contraception for religious reasons or lack of access (<5%). More than two-thirds of women reported that their husbands approved of contraception.n = 99) than at 3 years  and other subsequent points in the study period. Higher levels of CMD symptoms at 12 months post-index pregnancy were associated with higher levels of unmet need for contraception at 2.5 years in both the crude  and fully adjusted hypothesis-driven models , including potential confounders of maternal and husband education, SES, age, parity at baseline and religion. However, higher levels of CMD symptoms at 3 years were not associated with unmet need for contraception at 3.5 years in either the unadjusted or adjusted model  continued to be significant .Table 4n = 876) was 2.4 (s.d. 0.98), ranging from 0 to 6 pregnancies , particularly in the first year post-delivery  seeks to integrate mental health care into all general health care services WHO, . AlthougTo our knowledge this was the first study to investigate the impact of poor mental health on unmet need for contraception and fertility rate in a rural sample of women from a lower income country. We showed that high level of CMD symptoms at 1 year postpartum is associated prospectively with higher unmet need for contraception independently of SES and educational status. There is a lack of models of care promoting integration of mental and physical health in the family planning setting and further research is necessary to study the burden of preconception mental disorders and how these can be best addressed.