Prevalence, drivers and surveillance of antibiotic resistance and antibiotic use in rural China: Interdisciplinary study.

This study aimed to characterise antibiotic prescribing and dispensing patterns in rural health facilities in China and determine the community prevalence of antibiotic resistance. We investigated patterns and drivers of antibiotic use for common respiratory and urinary tract infections (RTI/UTI) in community settings, examined relationships between presenting symptoms, clinical diagnosis and microbiological results in rural outpatient clinics, and assessed potential for using patient records to monitor antibiotic use. This interdisciplinary mixed methods study included: (i) Observations and exit interviews in eight village clinics and township health centres and 15 retail pharmacies; (ii) Urine, throat swab and sputum samples from patients to identify potential pathogens and test susceptibility; (iii) 103 semi-structured interviews with doctors, patients, pharmacy workers and antibiotic-purchasing customers; (iv) Assessment of completeness and accuracy of electronic patient records through comparison with observational data. 87.9% of 1123 recruited clinic patients were prescribed antibiotics (of which 35.5% contained antibiotic combinations and >40% were for intravenous administration), most of whom had RTIs. Antibiotic prescribing for RTIs was not associated with presence of bacterial pathogens but was correlated with longer duration of infection (OR = 3.33) and presence of sore throat (OR = 1.64). Fever strongly predicted prescription of intravenous antibiotics (OR = 2.87). Resistance rates in bacterial pathogens isolated were low compared with national data. 25.8% of patients reported antibiotics use prior to their clinic visit, but only 56.2% of clinic patients and 53% of pharmacy customers could confirm their prescription or purchase included antibiotics. Diagnostic uncertainty, financial incentives, understanding of antibiotics as anti-inflammatory and limited doctor-patient communication were identified as key drivers of antibiotic use. Completion and accuracy of electronic patient records were highly variable. Prevalence of antibiotic resistance in this rural population is relatively low despite high levels of antibiotic prescribing and self-medication. More systematic use of e-records and in-service training could improve antibiotic surveillance and stewardship in rural facilities. Combining qualitative and observational anthropological methods and concepts with microbiological and epidemiological investigation of antibiotic resistance at both research design and analytic synthesis stages substantially increases the validity of research findings and their utility in informing future intervention development.

Mindcraft, a Mobile Mental Health Monitoring Platform for Children and Young People: Development and Acceptability Pilot Study.

BACKGROUND: Children and young people's mental health is a growing public health concern, which is further exacerbated by the COVID-19 pandemic. Mobile health apps, particularly those using passive smartphone sensor data, present an opportunity to address this issue and support mental well-being. OBJECTIVE: This study aimed to develop and evaluate a mobile mental health platform for children and young people, Mindcraft, which integrates passive sensor data monitoring with active self-reported updates through an engaging user interface to monitor their well-being. METHODS: A user-centered design approach was used to develop Mindcraft, incorporating feedback from potential users. User acceptance testing was conducted with a group of 8 young people aged 15-17 years, followed by a pilot test with 39 secondary school students aged 14-18 years, which was conducted for a 2-week period. RESULTS: Mindcraft showed encouraging user engagement and retention. Users reported that they found the app to be a friendly tool helping them to increase their emotional awareness and gain a better understanding of themselves. Over 90% of users (36/39, 92.5%) answered all active data questions on the days they used the app. Passive data collection facilitated the gathering of a broader range of well-being metrics over time, with minimal user intervention. CONCLUSIONS: The Mindcraft app has shown promising results in monitoring mental health symptoms and promoting user engagement among children and young people during its development and initial testing. The app's user-centered design, the focus on privacy and transparency, and a combination of active and passive data collection strategies have all contributed to its efficacy and receptiveness among the target demographic. By continuing to refine and expand the app, the Mindcraft platform has the potential to contribute meaningfully to the field of mental health care for young people.

Pythium and Phytopythium species associated with weeds collected in vegetable production fields in Brazil.

This study aimed to identify Pythium and Phytopythium species from weeds collected in vegetable fields and test their pathogenicity. Weeds with symptoms of damping-off, root rot or wilt were sampled in the Brazilian states of Ceará, Goiás and Pernambuco, as well as in the Distrito Federal, for isolation and identification of the causal agents. Once isolated, colonies with typical Pythium and Phytopythium characteristics grew in selective V8 medium. Procedures for species identification included morphology and amplification of the ITS and Cox II regions, which were compared with other accessions available at GenBank. The phylogenetic relationships among the isolates and pathogenicity to their original hosts were evaluated. Six Pythium species were identified: P. aphanidermatum, P. oopapillum, P. orthogonon, P. ultimum var. ultimum, P. myriotylum and P. sylvaticum, and two species of Phytopythium, Phy. chamaehyphon and Phy. oedochilum. In the pathogenicity tests, the 10 weed hosts showed symptoms of damping-off or root rot after inoculation, with exception of Portulaca oleraceae in which none of the isolates was pathogenic. Therefore, common weeds in vegetable fields areas can host different Pythium and Phytopythium species and play an important role in the epidemiology of vegetable diseases, in particular on pathogen survival and population increase.

SARS-CoV-2 serosurvey among adults involved in healthcare and health research in Guinea-Bissau, West Africa.

OBJECTIVES: Many African countries have reported fewer COVID-19 cases than countries elsewhere. By the end of 2020, Guinea-Bissau, West Africa, had <2500 PCR-confirmed cases corresponding to 0.1% of the ∼1.8 million national population. We assessed the prevalence of SARS-CoV-2 antibodies in urban Guinea-Bissau to help guide the pandemic response in Guinea-Bissau. STUDY DESIGN: Cross-sectional assessment of SARS-CoV-2 antibody in a cohort of staff at the Bandim Health Project. METHODS: We measured IgG antibodies using point-of-care rapid tests among 140 staff and associates at a biometric research field station in Bissau, the capital of Guinea-Bissau, during November 2020. RESULTS: Of 140 participants, 25 (18%) were IgG-positive. Among IgG-positives, 12 (48%) reported an episode of illness since the onset of the pandemic. Twenty-five (18%) participants had been PCR-tested between May and September; 7 (28%) had been PCR-positive. Four of these seven tested IgG-negative in the present study. Five participants reported that somebody had died in their house, corresponding crudely to an annual death rate of 4.5/1000 people; no death was attributed to COVID-19. Outdoor workers had a lower prevalence of IgG-positivity. CONCLUSIONS: In spite of the low official number of COVID-19 cases, our serosurvey found a high prevalence of IgG-positivity. Most IgG-positives had not been ill. The official number of PCR-confirmed COVID-19 cases has thus grossly underestimated the prevalence of COVID-19 during the pandemic. The observed overall mortality rate in households of Bandim Health Project employees was not higher than the official Guinean mortality rate of 9.6/1000 people.

Species diversity, novel interactions and absence of well-supported host-guided phylogenetic groupings of Neotropical Alternaria isolates causing foliar lesions in Solanaceae.

AIM: To report the characterization of 120 Alternaria isolates inducing early blight-like foliar lesions in nine species of five Solanaceae genera collected across all macrogeographical Brazilian regions. MATERIAL AND RESULTS: Phylogenetic relationships were assessed via analyses of the Alternaria alternata allergenic protein-coding, glyceraldehyde-3-phosphate dehydrogenase and the calmodulin gene sequences. Most of the tomato isolates were placed into the Alternaria linariae cluster, whereas most of the potato isolates were grouped with Alternaria grandis. Novel host-pathogen interactions were also reported. Seventeen isolates were selected for morphometrical characterization, and a subsample of 13 isolates was employed in pathogenicity assays on tomato, potato, eggplant, scarlet eggplant, Capsicum annuum, Datura stramonium, Physalis angulata and Nicotiana tabacum. Eleven isolates were able to induce foliar lesions in tomatoes but none in C. annuum. Potato was susceptible to a subgroup of isolates but displayed a subset of isolate-specific interactions. Morphological traits were in overall agreement with molecular and host range data. CONCLUSION: Alternaria linariae and A. grandis were confirmed as the major causal agents of tomato and potato early blight, respectively. However other Alternaria species are also involved with early blight in solanaceous hosts in Brazil. SIGNIFICANCE AND IMPACT OF THE STUDY: The diversity and host-specific patterns of the Alternaria isolates from Solanaceae may have practical implications in establishing effective early blight genetic resistance and cultural management strategies especially for tomato and potato crops.

Relationship between dental and periodontal health status and the salivary microbiome: bacterial diversity, co-occurrence networks and predictive models.

The present study used 16S rRNA gene amplicon sequencing to assess the impact on salivary microbiome of different grades of dental and periodontal disease and the combination of both (hereinafter referred to as oral disease), in terms of bacterial diversity, co-occurrence network patterns and predictive models. Our scale of overall oral health was used to produce a convenience sample of 81 patients from 270 who were initially recruited. Saliva samples were collected from each participant. Sequencing was performed in Illumina MiSeq with 2 × 300 bp reads, while the raw reads were processed according to the Mothur pipeline. The statistical analysis of the 16S rDNA sequencing data at the species level was conducted using the phyloseq, DESeq2, Microbiome, SpiecEasi, igraph, MixOmics packages. The simultaneous presence of dental and periodontal pathology has a potentiating effect on the richness and diversity of the salivary microbiota. The structure of the bacterial community in oral health differs from that present in dental, periodontal or oral disease, especially in high grades. Supragingival dental parameters influence the microbiota's abundance more than subgingival periodontal parameters, with the former making a greater contribution to the impact that oral health has on the salivary microbiome. The possible keystone OTUs are different in the oral health and disease, and even these vary between dental and periodontal disease: half of them belongs to the core microbiome and are independent of the abundance parameters. The salivary microbiome, involving a considerable number of OTUs, shows an excellent discriminatory potential for distinguishing different grades of dental, periodontal or oral disease; considering the number of predictive OTUs, the best model is that which predicts the combined dental and periodontal status.

A three-decade survey of Brazilian Fusarium oxysporum f. sp. lycopersici races assessed by pathogenicity tests on differential tomato accessions and by molecular markers.

AIM: Physiological race determination of 143 Fusarium oxysporum f. sp. lycopersici (FOL) isolates collected along 30 years in major tomato-producing regions of Brazil. MATERIALS AND RESULTS: Physiological races were determined via root-dipping inoculation of differential tomato accessions and by the PCR-based marker system of Hirano and Arie (2006). According to pathogenicity/virulence assays, five race 1, 23 race 2 and 115 race 3 isolates were identified. FOL race 1 and 2 isolates prevailed up to early 2000s. Afterwards, the large majority of the isolates was classified as the invasive race 3. Novel reports of race 3 were done in five states, thus expanding its geographical distribution. Using this PCR-based marker system, a precise discrimination was observed for all race 3 isolates. However, all race 1 and 2 isolates displayed only the cosmopolitan race 1-specific amplicon pattern. CONCLUSION: The development and/or validation of novel race-specific marker systems are necessary to allow a precise discrimination of the potentially endemic Brazilian FOL race 2. SIGNIFICANCE AND IMPACT OF THE STUDY: The present characterization of isolates indicates that distinct evolutionary mechanisms are acting to select new FOL races and/or genetic variants across agroecosystems around the globe.

Impact of microaeration bioreactor on dissolved sulfide and methane removal from real UASB effluent for sewage treatment.

Two bioreactors were investigated as an alternative for the post-treatment of effluent from an upflow anaerobic sludge blanket (UASB) reactor treating domestic sewage, aiming at dissolved sulfide and methane removal. The bioreactors (R-control and R-air) were operated at different hydraulic retention times (HRT; 6 and 3 h) with or without aeration. Large sulfide and methane removal efficiencies were achieved by the microaerated reactor at HRT of 6 h. At this HRT, sulfide removal efficiencies were equal to 61% and 79%, and methane removal efficiencies were 31% and 55% for R-control and R-air, respectively. At an HRT of 3 h, sulfide removal efficiencies were 22% (R-control) and 33% (R-air) and methane removal did not occur. The complete oxidation of sulfide, with sulfate formation, prevailed in both phases and bioreactors. However, elemental sulfur formation was more predominant at an HRT of 6 h than at an HRT of 3 h. Taken together, the results show that post-treatment improved the anaerobic effluent quality in terms of chemical oxygen demand and solids removal. However, ammoniacal nitrogen was not removed due to either the low concentration of air provided or the absence of microorganisms involved in the nitrogen cycle.

Arbuscular mycorrhiza influences carbon-use efficiency and grain yield of wheat grown under pre- and post-anthesis salinity stress.

Soil salinity severely affects and constrains crop production worldwide. Salinity causes osmotic and ionic stress, inhibiting gas exchange and photosynthesis, ultimately impairing plant growth and development. Arbuscular mycorrhiza (AM) have been shown to maintain light and carbon use efficiency under stress, possibly providing a tool to improve salinity tolerance of the host plants. Thus, it was hypothesized that AM will contribute to improved growth and yield under stress conditions. Wheat plants (Triticum aestivum L.) were grown with (AMF+) or without (AMF-) arbuscular mycorrhizal fungi (AMF) inoculation. Plants were subjected to salinity stress (200 mm NaCl) either at pre- or post-anthesis or at both stages. Growth and yield components, leaf chlorophyll content as well as gas exchange parameters and AMF colonization were analysed. AM plants exhibited a higher rate of net photosynthesis and stomatal conductance and lower intrinsic water use efficiency. Furthermore, AM wheat plants subjected to salinity stress at both pre-anthesis and post-anthesis maintained higher grain yield than non-AM salinity-stressed plants. These results suggest that AMF inoculation mitigates the negative effects of salinity stress by influencing carbon use efficiency and maintaining higher grain yield under stress.

Comparison of Traditional and Web-Based Medical Student Teaching by Radiology Residents.

PURPOSE: Web-based learning tools are increasingly available for use and have been described in the pedagogical literature. However, rigorous comparisons between traditional learning methods and newer collaborative online tools have not been performed. Herein, we describe a web-based curriculum hosted on the collaborative Radiopaedia.org website. This curriculum was compared with traditional learning tools in a randomized, controlled fashion. MATERIALS AND METHODS: Medical students rotating through inpatient pediatrics were offered a 1-hour case-based learning session led by radiology residents. Students were randomized to receive online (Radiopaedia.org) versus traditional supplemental educational materials (reading material covering the same content) for review before the in-class session. A 15-point questionnaire was administered at two different points during the rotation: at the beginning of the clinical rotation and at the end of the classroom session. RESULTS: Fifty-eight students were approached for enrollment and a total of 47 (81%) consented to participation and completed the study requirements. Students who completed the web-based module had higher mean knowledge scores (74%) compared with those who were provided the traditional learning material (68%) (P = .06). Specifically, they demonstrated increased knowledge of the ACR Appropriateness Criteria and the "silhouette sign." CONCLUSIONS: A randomized, controlled, nonblinded evaluation of a novel radiology curriculum intervention hosted on Radiopaedia.org demonstrates improved test scores compared with traditional teaching methods.

Anticancer Properties of Essential Oils and Other Natural Products.

Essential oils are secondary metabolites with a key-role in plants protection, consisting primarily of terpenes with a volatile nature and a diverse array of chemical structures. Essential oils exhibit a wide range of bioactivities, especially antimicrobial activity, and have long been utilized for treating various human ailments and diseases. Cancer cell prevention and cytotoxicity are exhibited through a wide range of mechanisms of action, with more recent research focusing on synergistic and antagonistic activity between specific essential oils major and minor components. Essential oils have been shown to possess cancer cell targeting activity and are able to increase the efficacy of commonly used chemotherapy drugs including paclitaxel and docetaxel, having also shown proimmune functions when administered to the cancer patient. The present review represents a state-of-the-art review of the research behind the application of EOs as anticancer agents both in vitro and in vivo. Cancer cell target specificity and the use of EOs in combination with conventional chemotherapeutic strategies are also explored.

Release kinetics and cell viability of ibuprofen nanocrystals produced by melt-emulsification.

The clinical use of poorly water-soluble drugs has become a big challenge in pharmaceutical development due to the compromised bioavailability of the drugs in vivo. Nanocrystals have been proposed as a formulation strategy to improve the dissolution properties of these drugs. The benefits of using nanocrystals in drug delivery, when compared to other nanoparticles, are related to their production facilities, simple structure, and suitability for a variety of administration routes. High pressure homogenization (HPH) is the most promising production process, which can be employed at low or high temperatures. Ibuprofen nanocrystals with a mean size below 175 nm, and polydispersity below 0.18, have been produced by melt-emulsification, followed by HPH. Two nanocrystal formulations, differing on the surfactant composition, have been produced, their in vitro ibuprofen release tested in Franz diffusion cells and adjusted to several kinetic models (zero order, first order, Higuchi, Hixson-Crowell, Korsmeyer-Peppas, Baker-Lonsdale and Weibull model). Cell viability was assessed at 3, 6 and 24 h of incubation on human epithelial colorectal cells (Caco-2) by AlamarBlue® colorimetric assay. For both formulations, Caco-2 cells viability was dependent on the drug concentration and time of exposure.

Comparative genome analysis and characterization of the Salmonella Typhimurium strain CCRJ_26 isolated from swine carcasses using whole-genome sequencing approach.

Salmonella pathogenicity relies on virulence factors many of which are clustered within the Salmonella pathogenicity islands. Salmonella also harbours mobile genetic elements such as virulence plasmids, prophage-like elements and antimicrobial resistance genes which can contribute to increase its pathogenicity. Here, we have genetically characterized a selected S. Typhimurium strain (CCRJ_26) from our previous study with Multiple Drugs Resistant profile and high-frequency PFGE clonal profile which apparently persists in the pork production centre of Rio de Janeiro State, Brazil. By whole-genome sequencing, we described the strain's genome virulent content and characterized the repertoire of bacterial plasmids, antibiotic resistance genes and prophage-like elements. Here, we have shown evidence that strain CCRJ_26 genome possible represent a virulence-associated phenotype which may be potentially virulent in human infection. SIGNIFICANCE AND IMPACT OF THE STUDY: Whole-genome sequencing technologies are still costly and remain underexplored for applied microbiology in Brazil. Hence, this genomic description of S. Typhimurium strain CCRJ_26 will provide help in future molecular epidemiological studies. The analysis described here reveals a quick and useful pipeline for bacterial virulence characterization using whole-genome sequencing approach.

Phytosterol-enriched milk lowers LDL-cholesterol levels in Brazilian children and adolescents: Double-blind, cross-over trial.

BACKGROUND AND AIMS: Despite evidence of the lipid-lowering effect of plant sterols among adults with hypercholesterolemia, data regarding phytosterol use in children are limited. In this paper, we examined the effects of daily consumption of a phytosterol-enriched milk compound on the lipid profiles of Brazilian children and adolescents with dyslipidemia. METHODS AND RESULTS: This was a randomized, double blind, crossover clinical trial. Twenty eight dyslipidemics outpatients (aged 6-9 years) from an University Hospital were randomly allocated to control or intervention group. The intervention group received milk enriched with 1.2 g/day of plant sterol and the control group received the equivalent amount of skim milk during the period of 8 weeks. Changes from baseline in the mean lipid profile, including total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), and triglyceride (TG) concentrations. Serum lipid profiles, glucose levels, dietary and anthropometric data were determined at weeks 0, 4, 8, 16, and 20. Details regarding the safety and tolerance of phytosterol were obtained, using an open-ended questionnaire. Intention-to-treat analysis were performed, using the proc mixed procedure in SAS. After 8 weeks, the mean concentrations of TC and LDL-C were significantly reduced in the intervention group as compared to the control group with reductions of 5.9% (p = 0.09) and 10.2% (p = 0.002), respectively. In addition, TG concentrations were reduced by 19.7% (p = 0.09). No serious side effects were reported during the study. CONCLUSION: Our results confirm that plant sterols are an effective and safe treatment of infantile dyslipidemia. TRIAL REGISTRATION: RBR-3h7f9k.

Assessment of safe bioactive doses of Foeniculum vulgare Mill. essential oil from Portugal.

This study was designed to evaluate the antifungal activity of Foeniculum vulgare essential oil; concomitantly, the safety of bioactive doses was also unveiled, for the first time, in several mammalian cells. The chemical characterisation was made by GC and GC-MS. Antifungal activity was evaluated against yeasts, dermatophytes and Aspergillus strains and germ tube inhibition assay was evaluated using Candida albicans. Cell viability was assessed by the MTT assay. The main constituents of the oil are E-anetol (47%), α-phellandrene (11%), α-pinene (10.1%) and fenchone (10.8%). The oil was more active against Cryptococcus neoformans and C. albicans (MICs 0.32-0.64 µL/mL) and the filamentation of C. albicans was totally inhibited with 0.08 µL/mL. The oil is safe for keratinocytes, hepatocytes and fibroblasts in concentrations up to 1.25 µL/mL, and to macrophages up to 0.64 µL/mL. These findings highlight safe bioactive concentrations that should be deeper investigated for further application in pharmaceutical industry.

Ibuprofen nanocrystals developed by 22 factorial design experiment: A new approach for poorly water-soluble drugs.

The reduction of the particle size of drugs of pharmaceutical interest down to the nano-sized range has dramatically changed their physicochemical properties. The greatest disadvantage of nanocrystals is their inherent instability, due to the risk of crystal growth. Thus, the selection of an appropriate stabilizer is crucial to obtain long-term physicochemically stable nanocrystals. High pressure homogenization has enormous advantages, including the possibility of scaling up, lack of organic solvents and the production of small particles diameter with low polydispersity index. The sequential use of high shear homogenization followed by high pressure homogenization, can modulate nanoparticles' size for different administration routes. The present study focuses on the optimization of the production process of two formulations composed of different surfactants produced by High Shear Homogenization followed by hot High Pressure Homogenization. To build up the surface response charts, a 22 full factorial design experiment, based on 2 independent variables, was used to develop optimized formulations. The effects of the production process on the mean particle size and polydispersity index were evaluated. The best ibuprofen nanocrystal formulations were obtained using 0.20% Tween 80 and 1.20% PVP K30 (F1) and 0.20% Tween 80 and 1.20% Span 80 (F2). The estimation of the long-term stability of the aqueous suspensions of ibuprofen nanocrystals was studied using the LUMISizer. The calculated instability index suggests that F1 was more stable when stored at 4 °C and 22 °C, whereas F2 was shown to be more stable when freshly prepared.

A general measles vaccination campaign in urban Guinea-Bissau: Comparing child mortality among participants and non-participants.

BACKGROUND: Measles vaccination campaigns targeting children aged 9-59months are conducted every three years in Guinea-Bissau. Studies have demonstrated beneficial non-specific effects of measles vaccine. We compared mortality one year after the December 2012 measles vaccination campaign in Bissau city for children who received campaign measles vaccine with children who did not receive campaign measles vaccine. METHODS: Field workers from Bandim Health Project registered all children living in the Bandim Health Project's study area who received measles vaccination at the campaign posts. Children not seen during the campaign were visited at home and campaign participation status was assessed. We compared mortality rates of participants vs. non-participants in Cox regression models. RESULTS: 5633 children aged 9-59months (85%) received campaign measles vaccination and 1006 (15%) did not. During the subsequent year 16 children died. Adjusted for background factors, the hazard ratio (HR) comparing measles vaccinated versus unvaccinated was 0.28 (95% CI: 0.10-0.77). The benefit was larger for girls (HR: 0.17 (0.05-0.59)) and for children who had received routine measles vaccine before the campaign (HR: 0.15 (0.04-0.63)). CONCLUSIONS: We found indications of strong beneficial non-specific effects of receiving measles vaccine during the 2012 campaign, especially for girls and children with previous routine measles vaccination. Measles vaccination campaigns may be an effective way of improving child survival.

Structural brain changes are associated with response of negative symptoms to prefrontal repetitive transcranial magnetic stimulation in patients with schizophrenia.

Impaired neural plasticity may be a core pathophysiological process underlying the symptomatology of schizophrenia. Plasticity-enhancing interventions, including repetitive transcranial magnetic stimulation (rTMS), may improve difficult-to-treat symptoms; however, efficacy in large clinical trials appears limited. The high variability of rTMS-related treatment response may be related to a comparably large variation in the ability to generate plastic neural changes. The aim of the present study was to determine whether negative symptom improvement in schizophrenia patients receiving rTMS to the left dorsolateral prefrontal cortex (DLPFC) was related to rTMS-related brain volume changes. A total of 73 schizophrenia patients with predominant negative symptoms were randomized to an active (n=34) or sham (n=39) 10-Hz rTMS intervention applied 5 days per week for 3 weeks to the left DLPFC. Local brain volume changes measured by deformation-based morphometry were correlated with changes in negative symptom severity using a repeated-measures analysis of covariance design. Volume gains in the left hippocampal, parahippocampal and precuneal cortices predicted negative symptom improvement in the active rTMS group (all r⩽-0.441, all P⩽0.009), but not the sham rTMS group (all r⩽0.211, all P⩾0.198). Further analyses comparing negative symptom responders (⩾20% improvement) and non-responders supported the primary analysis, again only in the active rTMS group (F(9, 207)=2.72, P=0.005, partial η 2=0.106). Heterogeneity in clinical response of negative symptoms in schizophrenia to prefrontal high-frequency rTMS may be related to variability in capacity for structural plasticity, particularly in the left hippocampal region and the precuneus.

Ziziphora tenuior L. essential oil from Dana Biosphere Reserve (Southern Jordan); Chemical characterization and assessment of biological activities.

ETHNOPHARMACOLOGIC RELEVANCE: Ziziphora tenuior L. (Lamiaceae) is a medicinal plant in Jordan, which is included in various antimicrobial, antiseptic, expectorant and wound healing preparations. It is used for the treatment of cough, stomach ache, dysentery, fever, uterus infection, gut inflammation and painful menstruation. AIM OF THE STUDY: The aim of this study was to assess, for the first time, the chemical composition of the essential oil of Z. tenuior originated from southern Jordan and its antifungal effects against several yeasts. Concomitantly, the mechanisms behind the anti-fungal activity against Candida albicans were also disclosed. Since the Z. tenuior traditional uses are related with inflammatory-associated conditions, the putative anti-inflammatory activity of the oil was also unveiled. Importantly, the potential toxicity of pharmacologically active concentrations was screened in different types of mammalian cells. MATERIALS AND METHODS: Z. tenuior essential oil, isolated by hydrodistillation, was analyzed by gas chromatography, gas chromatography-mass spectrometry and 13C nuclear magnetic resonance spectroscopy. Antifungal activity was evaluated against yeasts, dermatophytes and Aspergillus strains. Germ tube inhibition and biofilm formation assays were evaluated using C. albicans. Assessment of cell viability was made by the MTT assay using different types of mammalian cells, including hepatocytes, keratinocytes and macrophages. The in vitro anti-inflammatory potential of the oil was evaluated by measuring nitric oxide production using lipopolysaccharide-stimulated mouse macrophages. RESULTS: Oxygen-containing monoterpenes are the main oil compounds: pulegone (46.8%), p-menth-3-en-8-ol (12.5%), isomenthone (6.6%) and 8-hydroxymenthone (6.2%). The highest antifungal activity was against Cryptococcus neoformans, with a MIC value of 0.16µL/mL. The oil revealed an important inhibitory effect on germ tube formation with a filamentation inhibition rate higher than 80% at 0.16µL/mL. The amount of the attached biomass was reduced. Importantly, concentrations devoid of toxicity on several mammalian cell types still displayed anti-inflammatory activity (0.16 and 0.32µL/mL). CONCLUSIONS: These findings add significant information to the pharmacological activity of Z. tenuior, thus justifying and reinforcing the use of this plant in traditional medicine. Additionally, the antifungal and anti-inflammatory potential of the oil at non-toxic concentrations, opens new avenues for its further exploitation, for instance in health-care product development.

Chemical composition and biological activities of Artemisia judaica essential oil from southern desert of Jordan.

ETHNOPHARMACOLOGIC RELEVANCE: Artemisia judaica L. (Arabic name: Beithran), is a medicinal and aromatic plant growing in the valley bottoms of desert areas, particularly in the southern desert of Jordan nearest to the Jordan-Saudi Arabia borders and in Wadi Araba in the Southern Badia. In Jordan, A. judaica is widely used in traditional medicine being recommended by aboriginal Bedouins in the North Badia region of Jordan as calmative. Furthermore, it is used for the treatment of stomach ache, heart diseases, sexual weakness, diabetes, gastro-intestinal disorders and external wounding. Additionally, other folk medicines of the Arabic region commonly use this aromatic plant for the treatment of inflammatory-related diseases, for instance fungal infections, diabetes, atherosclerosis, cancer and arthritis. AIM OF THE STUDY: Considering the traditional medicinal uses and the lack of scientific studies addressing the cellular and molecular mechanisms behind A. judaica claimed activities, the present study was designed to validate some of the traditional uses ascribed to this species, specifically the antifungal and anti-inflammatory activities of A. judaica essential oil at doses devoid of cytotoxicity to mammalian cells. MATERIALS AND METHODS: Chemical analysis of A. judaica essential oil isolated by hydrodistillation from aerial parts was carried out by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The antifungal activity (minimal inhibitory concentrations and minimal lethal concentrations) was evaluated against yeasts, dermatophyte and Aspergillus strains. In order to deeply explore the mechanisms behind the anti-fungal effect of the essential oil, the germ tube inhibition assay and the biofilms formation assay were evaluated using Candida albicans. The assessment of cell viability was accomplished using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in both hepatocytes and macrophages. Furthermore, the in vitro anti-inflammatory potential of A. judaica oil was evaluated by measuring nitric oxide (NO) production using lipopolysaccharide (LPS)-stimulated mouse macrophages. RESULTS: Oxygen containing monoterpenes are a representative group of constituents (68.7%) with piperitone (30.4%), camphor (16.1%) and ethyl cinnamate (11.0%) as main compounds. The highest antifungal activity of the oil was observed against Cryptococcus neoformans, with a MIC value of 0.16µL/mL. The oil revealed an important inhibitory effect on germ tube formation in C. albicans with 80% inhibition of filamentation at a concentration of 0.16µL/mL. Importantly, the oil also interfered with pre-formed biofilms by reducing the amount of the attached biomass. Furthermore, the essential oil significantly inhibited NO production evoked by LPS on macrophages at concentrations with very low toxicity (0.32µL/mL) or without toxicity (0.16µL/mL) to both macrophages and hepatocytes. CONCLUSIONS: The present study revealed that A. judaica essential oil from Jordan significantly inhibited germ tube formation and disrupted preformed biofilms of C. albicans, emphasizing the therapeutic potential for the treatment of disseminated candidiasis. Additionally, safe concentrations of this essential oil significantly inhibited NO production elicited by LPS in macrophages, highlighting its potential anti-inflammatory activity. Overall, A. judaica bears promising therapeutic potential for further drug development. Importantly, this work also validates some of the traditional uses of A. judaica.