Phenotypic and genotypic characterization of enteroaggregative Escherichia coli isolates from pediatric population in Pakistan

Enteroaggregative Escherichia coli (EAEC) are a leading cause of diarrhea among children. The objective of this study was to define the frequency of EAEC among diarrheal children from flood‐affected areas as well as sporadic cases, determine multidrug resistance, and evaluation of virulence using an in vivo model of pathogenesis. Stool samples were collected from 225 diarrheal children from 2010 to 2011 from flood‐affected areas as well as from sporadic cases in Pakistan. Identified EAEC isolates were characterized by phylogrouping, antibiotic resistance patterns including the extended‐spectrum beta lactamase spectrum, single nucleotide polymorphism detection in gyrA and parC, and virulence potential using wax worm, G. mellonella. A total of 35 (12.5%) confirmed EAEC isolates were identified among 225 E. coli isolates. EAEC isolates displayed high resistance to tetracycline, ampicillin, and cefaclor. A total of 34.28% were ESBL positive. Single nucleotide polymorphism detection revealed 37.14% and 68.57% isolates were positive for SNPs in gyrA (A₆₆₀‐T₆₆₀) and parC (C₃₃₀‐T₃₃₀), respectively. Phylogrouping revealed that B2 phylogroup was more prevalent among all EAEC isolates tested followed by D, A, B1, and non‐typeable (NT). Infection of G. mellonella with EAEC showed that killing infective dose was 100% higher than E. coli DH5 alpha control. EAEC are prevalent among Pakistani children with diarrhea, they are highly resistant to antibiotics, and predominantly fall into B2 phylogroup. Epidemiologic surveillance of EAEC and other E. coli pathotypes is critical to assess not only the role of these pathogens in diarrheal disease but also to determine the extent of multidrug resistance among the population.     

Safety of a physical therapy protocol for women with preeclampsia: a randomized controlled feasibility trial

Objective: To assess the feasibility and safety of a physiotherapy protocol applied to pregnant women with preeclampsia. Methods: Randomized, controlled, single-blind feasibility study, with 24 hospitalized pregnant women with preeclampsia. The intervention group received one session of the physiotherapy. The control group remained under the routine care of the hospital. The primary outcomes were Doppler velocimetry, cardiotocography, and maternal–fetal hemodynamics. Secondary outcomes were pain and anxiety assessed before and after the interventions. A mixed effects linear regression model was used, and the data were compared with the level of significance at 5%. Results: The baseline characteristics of the participants were homogeneous between groups. Resistance index of the Middle Cerebral Artery (MAC) and Umbilical Artery (UA) and cardiotocography did not change significantly. The systolic blood pressure (SBP) increased 4.90 mmHg in the control group and 0.22 mmHg in the intervention group. The diastolic blood pressure (DBP) increased 1.34 mmHg in the control group and decreased 0.40 mmHg in the intervention group. The middle bood pressure (MBP) increased 4.66 mmHg in the control group while there was a decrease of 0.09 mmHg in the intervention group, without statistical difference. Heart rate (HR) decreased 0.94 bpm in the control group; whereas, in the intervention group, there was an increase of 6.30 bpm. The pain reduced clinically 2 points after the intervention. The anxiety reduced clinically in both the groups (?1.26 in the intervention group and ?2.17 in the control group). Conclusion: The protocol applied in the intervention group is feasible and safe for both mother and fetus. Both groups showed clinical reduction in the levels of anxiety; whereas, pain was clinically reduced in the intervention group.     

Recurrent hemi-hypoesthesia in a young patient: A clinical challenge

Takayasu’s arteritis (TA) is a chronic inflammatory disease of unknown etiology characterized by granulomatous vasculitis of medium and large arteries, principally the aorta and its branches. It is a rare disorder but neurological manifestations are common.The authors present a case of a 39-year-old woman that presented with several episodes of hemi-hypoesthesia that lasted several days and coursed with complete recovery. She had already preliminary workup for most neurological diseases that presented at her age range that had been normal.We emphasize in this clinical case the atypical presentation of TA with neurological symptoms of somewhat vague and prolonged course with a negative preliminary workup. We believe it is important to keep a high index of suspicion in such cases.     

Infliximab partially impairs the anti‐Mycobacterium tuberculosis immune responses of severe psoriasis patients with positive tuberculin skin‐test

Background Infliximab and etarnecept are now widely used for treating severe psoriasis. However, these drugs, especially infliximab, increased the risk of tuberculosis reactivation. Surprisingly, epidemiological data suggest that the tuberculosis rate in patients taking infliximab in São Paulo State, Brazil, is similar to that of some developed, non‐endemic countries. Objective The aim of this study was to better understand the effect of infliximab on Mycobacterium tuberculosis (Mtb) immune responses of psoriasis patients in an endemic setting (Brazil). Methods We evaluated the tuberculosis‐specific immune responses of severe psoriasis patients and healthy individuals, both tuberculin skin test (TST) positive, in the presence/absence of infliximab. Patients had untreated severe psoriasis, no co‐morbidities affecting the immune responses and a TST >10 mm. Healthy TST⁺ (>10 mm) individuals were evaluated in parallel. PBMC cultures from both groups were stimulated with different Mycobacterium tuberculosis (Mtb) antigens (ESAT‐6, 85B and Mtb lysate) and phytohemagglutinin, with or without infliximab (5 μg/mL). Parameters evaluated were TNF‐α, IFN‐γ and IL‐10 secretion by ELISA, overnight IFN‐γ ELISpot and lymphocyte proliferative response (LPR). Results Infliximab almost abolished TNF‐α detection in PBMC supernatants of both groups. It also significantly reduced the LPR to phytohemagglutinin and the Mtb antigens as well as the IFN‐γ levels secreted into day 5 supernatants in both groups. There was no concomitant exaggerated IL‐10 secretion that could account for the decreases in these responses. ELISpot showed that, contrasting with the central‐memory responses above, infliximab did not affect effector‐memory INF‐γ‐releasing T‐cell numbers. Conclusions Infliximab affected some, but not all aspects of the in vitro antituberculosis immune responses tested. The preserved effector‐memory responses, putatively related to exposure to environmental mycobacteria, may help to explain the lower than expected susceptibility to tuberculosis reactivation in our setting.     

High variation in hypoxanthine determination after analytical treatment of vitreous humor samples

In the field of legal medicine the correct determination of the time of death is of great importance because an error in calculating the post-mortem interval (PMI) could be crucial in a criminal investigation. The quantification of hypoxanthine (Hx) concentration in the vitreous humor (VH) as a means to estimate PMI is useful when dealing with a recent time interval of death and has several advantages over other biological matrices. However, due to its viscous nature, VH has to be liquefied prior to its analysis, and the different procedures to liquefy it may cause alterations in Hx concentration, and hence in estimation of PMI. The aim of this study was to determine differences in Hx concentration after applying several pre-analytical treatments to different aliquots of the same VH sample. Enzymatic digestion, sonication, centrifugation and heat were the liquefying methods applied to reduce the viscosity of 105 VH samples. Analysis was performed using solid phase extraction and liquid chromatography in tandem with mass spectrometry. Statistically significant differences in the measured Hx concentrations were found between enzymatic digestion and the other three methods analyzed (sonication, centrifugation, and heat). Additionally, high data dispersion was found under heat treatment but was not observed for centrifugation or sonication, where similar and more moderate results were found. We conclude that high variations in Hx determination are observed when enzymatic digestion or heat treatment is used and we therefore recommend sonication or centrifugation to quantify Hx concentration in the VH.