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BackgroundEnvironmental water samples are increasingly recognised as an important reservoir of antimicrobial resistance (AMR) genes. Polymerase chain reaction (PCR) and next generation sequencing (NGS) offer a potentially inclusive surveillance platform for a wide range of AMR genes. However, molecular methods are dependent upon the extraction of DNA of high yield and quality. Current options for DNA extraction from complex environmental matrices for downstream molecular applications are either expensive or low yielding. We present here a novel magnetic bead-based DNA extraction method, for the detection of antimicrobial resistance genes (ARGs) from river water in Malawi, named MagnaExtract. MethodsMagnaExtract involves initial filtration of 250ml freshwater, followed by an overnight incubation of the filter in 15ml buffered peptone water (BPW), common procedure in microbiology laboratories. 200{micro}l is then taken for a boil (95{degrees}C) and spin step and mixed with magnetic beads to bind DNA. Following washes with ethanol, the DNA is eluted in nuclease-free water. To determine the effectiveness of this method, 98 freshwater samples were collected from two rivers in Southern Malawi, and DNA was isolated using the MagnaExtract method, two commercial Qiagen (Germany) kits; PowerWater and DNeasy Blood and tissue, alongside a boil and spin of BPW, and a boil and spin from bacterial isolate grown on agar media. All samples were screened with a high-resolution melt (HRM) PCR panel previously validated for the detection of third generation cephalosporin and carbapenem ARGs. We compared the DNA yield obtained using all extraction methods, as well as the identification of each ARG. ResultsDNA yield using MagnaExtract was statistically greater than both boil and spin methods and DNeasy Blood & Tissue (Qiagen, Germany). DNA yield was slightly lower than using PowerWater (Qiagen) but the difference was not statistically significant. MagnaExtract was the only method to identify ARGs in all 98 water samples compared with PowerWater (n=82), DNeasy (n=95) boilate of BPW (n=75) and boilate of bacterial isolate (n=87). The most commonly detected ARG was OXA-48 (n=93). In addition, we found overnight incubation in non-selective enrichment broth (BPW) to promote the growth of bacteria harbouring extended spectrum beta lactamase (ESBL) genes and reduction in the detection of carbapenemase genes. ConclusionThe MagnaExtract approach offers a simple, affordable, high yielding DNA extraction method for the detection of ARGs isolated from river water samples.

BackgroundLow- and middle-income countries (LMICs) have high morbidity and mortality from drug-resistant infections, especially from enteric bacteria such as Escherichia coli. LMICs have varying infrastructure and services in the community to separate people from human and animal waste, creating risks for ESBL-Enterobacterales (ESBL-E) transmission. Limited data exist from Southern Africa on the prevalence of ESBL-E the community.Methods and findingsIn this longitudinal cohort study we took a one-health approach to investigating prevalence and distribution of ESBL-E in urban, peri-urban and rural Malawian households between May 2018 and October 2020. We described human health, antibiotic usage (ABU), health seeking behaviour, structural and behavioural environmental health practices, and animal husbandry at these households. In parallel, human and animal stool and diverse environmental samples were collected and cultured to identify presence of ESBL E. coli and ESBL K. pneumoniae. Univariable and multivariable analysis was performed to determine associations with human ESBL-E colonisation.We recruited 300 households, totalling 841 visits, and a paucity of environmental health infrastructure and materials for safe sanitation was noted across all sites. In total, 11,975 samples were cultured and ESBL-E were isolated from 41.8% (n=1190) of human stool and 29.8% (n=290) of animal stool samples. Animal species with particularly high rates of ESBL-E colonisation included pigs (56.8%, n=21) poultry (32.5%, n=148) and dogs (58.8% n= 30). ESBL-E were isolated from 66.2% (n=339) of river water samples and 46.0% (n=138) of drain samples. Urban areas had greater ESBL-E contamination of food, household surfaces, floors and the external environment, alongside the highest rates of ESBL-E colonisation in humans (47.1%, n=384) and animals (55.1%, n=65). Multivariable models illustrated that human ESBL E. coli colonisation was associated with the wet season (aOR = 1.66, 95%CrI: 1.38-2.00), living in urban areas (aOR = 2.01, 95%CrI: 1.26-3.24), advanced age (aOR = 1.14, 95%CrI: 1.05-1.24) and in households where animals were observed interacting with food (aOR = 1.62, 95%CrI: 1.17-2.28) or kept inside (aOR = 1.58, 95%CrI: 1.00-2.43). Human ESBL K. pneumoniae colonisation was also associated with the wet season (aOR = 2.23, 95%CrI: 1.63-2.76.ConclusionWe identified extremely high levels of ESBL-E colonisation in humans and animals and contamination of the environment in Southern Malawi. Urbanisation and season are key risks for ESBL-E colonisation, perhaps reflecting environmental contamination as toilets overflow in high population density areas in heavy rains in the wet season. Without adequate efforts to improve environmental health, ESBL transmission is likely to persist in this setting.

The authors conducted a systematic review and meta-analysis to determine whether arthrocentesis or arthroscopy combined with platelet-rich plasma (PRP) or platelet-rich growth factor (PRGF) injection compared with no injection or saline injection (control group) or hyaluronic acid (HA) injection reduced pain and increased maximum mouth opening (MMO) in patients with temporomandibular joint (TMJ) osteoarthritis (OA).The authors used the Cochrane Library, Embase, PubMed, Web of Science, Google Scholar databases and hand searched reference lists through May 4, 2018, to identify randomized controlled trials and controlled trials including patients with TMJ OA receiving injections (PRP or PRGF versus other). The authors assessed the risk of bias according to the Cochrane guidelines.The authors screened 36 abstracts. They included 5 studies (3 randomized controlled trials and 2 controlled trials) with a total of 285 patients with TMJ OA in this review. The authors assessed all 5 studies as being at high risk of bias. The quality of evidence was very low owing to statistical heterogeneity, small sample size, or high risk of bias. Meta-analyses with 2 studies showed a visual analog scale pain improvement from baseline of -2.778 units (0-10 scale, 0 = no pain, 10 = worst pain) favorable to PRP or PRGF compared with findings in control groups (95% confidence interval [CI], -3.504 to -2.052; P.001) and an improvement of -0.968 favorable to PRP or PRGF compared with findings in HA groups (95% CI, -1.854 to -0.082; P = .032). The authors found no significant increase in MMO in those receiving PRP or PRGF compared with that in the control or HA groups.Although the results of the included studies showed that arthrocentesis or arthroscopy with PRP or PRGF, saline, or HA injections all reduced pain and increased mouth opening, the evidence was of very low quality. Further studies are needed to confirm these preliminary results showing that PRP or PRGF with arthrocentesis or arthroscopy significantly improved pain but did not increase MMO compared with findings in the control or HA groups.

Background The authors conducted a systematic review and meta-analysis to determine whether arthrocentesis or arthroscopy combined with platelet-rich plasma (PRP) or platelet-rich growth factor (PRGF) injection compared with no injection or saline injection (control group) or hyaluronic acid (HA) injection reduced pain and increased maximum mouth opening (MMO) in patients with temporomandibular joint (TMJ) osteoarthritis (OA). Types of Studies Reviewed The authors used the Cochrane Library, Embase, PubMed, Web of Science, Google Scholar databases and hand searched reference lists through May 4, 2018, to identify randomized controlled trials and controlled trials including patients with TMJ OA receiving injections (PRP or PRGF versus other). The authors assessed the risk of bias according to the Cochrane guidelines. Results The authors screened 36 abstracts. They included 5 studies (3 randomized controlled trials and 2 controlled trials) with a total of 285 patients with TMJ OA in this review. The authors assessed all 5 studies as being at high risk of bias. The quality of evidence was very low owing to statistical heterogeneity, small sample size, or high risk of bias. Meta-analyses with 2 studies showed a visual analog scale pain improvement from baseline of −2.778 units (0-10 scale, 0 = no pain, 10 = worst pain) favorable to PRP or PRGF compared with findings in control groups (95% confidence interval [CI], −3.504 to −2.052; P Conclusions and Practical Implications Although the results of the included studies showed that arthrocentesis or arthroscopy with PRP or PRGF, saline, or HA injections all reduced pain and increased mouth opening, the evidence was of very low quality. Further studies are needed to confirm these preliminary results showing that PRP or PRGF with arthrocentesis or arthroscopy significantly improved pain but did not increase MMO compared with findings in the control or HA groups.

An annotated guide to reference materials for social studies multimedia programs is provided. (DE)

Two instructors explain the whys, hows, and advantages of emphasizing primary prevention theories and strategies in their baccalaureate course in mental health-psychiatric nursing. (Author)

Discusses the role of the teacher, the school, and the community in child catechesis, and urges the development of catecheses that will help children to become mature and responsible adults. (AN)

What is a palindrome? It is a number, word, or phrase that reads the same backward or forward. The Greek word palindromos means “running back again.” The Greek poet Sotades is generally credited with creating the first word palindrome around 300 BC. The earliest references to a numeric palindrome, 12345654321, are found in Indian Sanskrit mathematical literature written around AD 850.

Leishmaniases are parasitic diseases that spread in many countries with a prevalence of 12 million cases. There are few available treatments and antimonials are still of major importance in the therapeutic strategies used in most endemic regions. However, resistance toward these compounds has recently emerged in areas where the replacement of these drugs is mainly limited by the cost of alternative molecules. In this paper, we reviewed the studies carried out on antimonial resistance in Leishmania. Several common limitations of these works are presented before prevalent approaches to evidence antimonial resistance are related. Afterwards, phenotypic determination of resistance is described, then confronted to clinical outcome. Finally, we detail molecular mechanisms and targets involved in resistance and already identified in vitro within selected mutant strains or in clinical isolates.

This paper is an attempt to offer a preliminary study of a Christian tradition of allegorizing the zodiac which is found in certain literary texts and artistic representations. What prompted the investigation from the artistic point of view was an examination of the twelfth-century baptismal font in the church of St Peter at Hook Norton, Oxfordshire, which is decorated with a mixture of selected signs of the zodiac and scriptural images (plate i). It raises the question of how early was the tradition in which the zodiac was linked with baptism in Christian thought, and what other connections there might be. So the question I should like briefly to illustrate here is the connection between Christian decorations which feature the zodiac, particularly in the medieval period, and an allegorical tradition which goes back to the early Church.