Heterogeneity of SOS response expression in clinical isolates of Escherichia coli influences adaptation to antimicrobial stress.

In recent years, new evidence has shown that the SOS response plays an important role in the response to antimicrobials, with involvement in the generation of clinical resistance. Here we evaluate the impact of heterogeneous expression of the SOS response in clinical isolates of Escherichia coli on response to the fluoroquinolone, ciprofloxacin. In silico analysis of whole genome sequencing data showed remarkable sequence conservation of the SOS response regulators, RecA and LexA. Despite the genetic homogeneity, our results revealed a marked differential heterogeneity in SOS response activation, both at population and single-cell level, among clinical isolates of E. coli in the presence of subinhibitory concentrations of ciprofloxacin. Four main stages of SOS response activation were identified and correlated with cell filamentation. Interestingly, there was a correlation between clinical isolates with higher expression of the SOS response and further progression to resistance. This heterogeneity in response to DNA damage repair (mediated by the SOS response) and induced by antimicrobial agents could be a new factor with implications for bacterial evolution and survival contributing to the generation of antimicrobial resistance.

Azole and Amphotericin B MIC Values against Aspergillus fumigatus: High Agreement between Spectrophotometric and Visual Readings Using the EUCAST EDef 9.3.2 Procedure.

The EUCAST EDef 9.3.2 procedure recommends visual readings of azole and amphotericin B MICs against Aspergillus spp. Visual determination of MICs may be challenging. In this work, we aim to obtain and compare visual and spectrophotometric MIC readings of azoles and amphotericin B against Aspergillus fumigatussensu lato isolates. A total of 847 A. fumigatussensu lato isolates (A. fumigatus sensu stricto [n = 828] and cryptic species [n = 19]) were tested against amphotericin B, itraconazole, voriconazole, posaconazole, and isavuconazole using the EUCAST EDef 9.3.2 procedure. Isolates were classified as susceptible or resistant/non-wild type according to the 2020 updated breakpoints. The area of technical uncertainty for the azoles was defined in the updated breakpoints. Visual and spectrophotometric (fungal growth reduction of >95% compared to the control, read at 540 nm) MICs were compared. Essential (±1 2-fold dilution) and categorical agreements were calculated. Overall, high essential (97.1%) and categorical (99.6%) agreements were found. We obtained 100% categorical agreements for amphotericin B, itraconazole, and posaconazole, and consequently, no errors were found. Categorical agreements were 98.7 and 99.3% for voriconazole and isavuconazole, respectively. Most of the misclassifications for voriconazole and isavuconazole were found to be associated with MIC results falling either in the area of technical uncertainty or within one 2-fold dilution above the breakpoint. The resistance rate was slightly lower when the MICs were obtained by spectrophotometric readings. However, all relevant cyp51A mutants were correctly classified as resistant. Spectrophotometric determination of azole and amphotericin B MICs against A. fumigatussensu lato isolates may be a convenient alternative to visual endpoint readings.

Therapeutic potential of selanyl amide derivatives in the in vitro anticholinesterase activity and in in vivo antiamnesic action.

The present study evaluated the in vitro acetylcholinesterase (AChE) inhibitor activity of two new selanyl amide derivatives in cerebral structures of mice. Our results demonstrated that N-(2-(3-(phenylselanyl)propoxy)phenyl)furan-2-carboxamide (1) and N-(2-(3-(phenylselanyl)propoxy)phenyl)thiophene-2-carboxamide (2) inhibited the in vitro AChE activity in mice. Another objective was to assess the effect of the best AChE inhibitor in an amnesic model induced by scopolamine (SCO) in male Swiss mice. The involvement of AChE activity and lipid peroxidation in the cerebral structures was investigated. Our results showed that compound 1 (10 mg/kg, intragastrically) attenuated the latency to find the escape box and the number of holes visited in the Barnes maze task, without altering the locomotor and exploratory activities in an open-field test. Compound 1 protected against increasing in lipid peroxidation levels and AChE activity caused by SCO in the cerebral cortex and hippocampus of mice. In conclusion, the present study evidenced the in vitro anticholinesterase effect of two new selanyl amide derivatives in the cerebral structures of mice. Moreover, compound 1, a selanyl amide derivative containing a furan ring, demonstrated antiamnesic action due to its antioxidant and anticholinesterase activities in cerebral structures.

Chronic administration of gonadotropin releasing-hormone improves learning in old gonadectomized rats.

The aim of this study was to determine whether chronic administration of GnRH improves performance of learning tasks and expression of spinophilin in the hippocampus of gonadectomized old rats. Eighteen-month-old male Wistar rats were used and divided into three groups: control (intact rats); gonadectomized; and gonadectomized + GnRH. The latter group was injected intramuscularly with GnRH (100 nM) twice a day for five weeks. The learning tasks we used were the novel object recognition task (NOR), elevated T-maze (ETM) and active avoidance test (AAT). Results showed that in NOR and ETM learning tasks, gonadectomized rats treated with GnRH had a significantly better performance than gonadectomized rats without treatment. GnRH-treated gonadectomized rats displayed performance comparable to that of intact rats. Furthermore, the expression of spinophilin in the hippocampus of gonadectomized rats treated with GnRH increased with respect to untreated gonadectomized rats. In conclusion, the chronic administration of GnRH improves learning in old gonadectomized rats. It is possible that the mechanism could involve a greater number of dendritic contacts associated with a higher expression of spinophilin.

Uncontrollable chronic stress affects eating behavior in rats.

The issue of whether the decrease in food intake induced by inescapable shock is due to the uncontrollability of the stressor or the shock per se has not yet been settled. Besides, whether food intake is differentially affected by an uncontrollable chronic stressor has been explored only by a few studies. Thus, we evaluated the effects of chronic escapable or inescapable electric shocks on eating behavior. Rats were exposed to shock sessions for 20 days in two occasions separated by baseline sessions with no shock in an ABAB design. Results showed a reduction in food and water intake and body weight gain during stress periods, especially with inescapable shocks. The findings support a close link between learned helplessness, chronic stress, and eating behavior.

Comparison between Vitek MS, Bruker Biotyper, Vitek2, and API20E for differentiation of species of the genus Raoultella.

Rapid and reliable identification of microorganisms in the clinical laboratory is essential for an early and accurate diagnosis guiding timely therapy. However, conventional methods are sometimes unreliable and show controversial outcomes. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been reported as a rapid and reliable method for identification of bacteria and fungi isolated from clinical samples. Members of the genus Raoultella are increasingly recognized as clinically relevant. There are difficulties in their identification at the species level since sequencing the 16S rRNA or the rpoB genes does not show conclusive results. The aim of this study has been to compare two MALDI-TOF MS systems (Vitek MS and Bruker Biotyper) with Vitek2 and API20E systems for differentiation of Raoultella species. A collection of 97 clinical isolates of Raoultella species was identified with Vitek MS, in parallel with Vitek2 and API, and finally with Bruker Biotyper. Among the two most widely used MALDI-TOF MS platforms, results obtained with Vitek MS were slightly superior to those obtained with the Bruker Biotyper system, with sensitivities and specificities of 98.9/57.9% and 98.8/37.0%, respectively. The current commercial phenotypic identification systems are not optimized for the identification of Raoultella species. Our results indicate that MALDI-TOF-based identification is more accurate and sensitive than that provided by phenotypic methods.

Contribution of serotonergic and nitrergic pathways, as well as monoamine oxidase-a and Na+, K+-ATPase enzymes in antidepressant-like action of ((4-tert-butylcyclohexylidene) methyl) (4-methoxystyryl) sulfide (BMMS).

The present study investigated a possible antidepressant-like effect of ((4-tert-butylcyclohexylidene)methyl) (4-methoxystyryl) sulfide (BMMS) by using the forced swimming test (FST) and the tail suspension test (TST) in Swiss mice. The contribution of serotoninergic, glutamatergic and nitrergic systems in the antidepressant-like activity of BMMS was evaluated. We also examined the involvement of monoamine oxidase (MAO)-A, MAO-B and Na+, K+-ATPase activities in prefrontal cortex of mice. BMMS, (0.1-10 mg/kg, intragastrically (i.g.)) and fluoxetine (32 mg/kg, i.g.) decreased the immobility time in the FST and TST. The anti-immobility effect of BMMS (10 mg/kg, i.g.) in the TST was prevented by the pretreatment of mice with WAY100635 (0.1 mg/kg, subcutaneously (s.c.), a 5-HT1A receptor antagonist), ketanserin (5 mg/kg, intraperitoneal (i.p.), a 5-HT2A/2C receptor antagonist), and partially blocked by ondansetron (1 mg/kg, i.p., a 5-HT3 receptor antagonist). The anti-immobility effect of BMMS (10 mg / kg, i.g.) was not avoided by pretreatment with MK-801 (0.01 mg/kg, s.c. a non-competitive N-methyl D-Aspartate (NMDA) receptor) in the TST. Pretreatment with L-arginine (500 mg/kg, i.p., a nitric oxide precursor) reversed partially the reduction in the immobility time elicited by BMMS (10 mg/kg, i.g.) in TST. BMMS altered Na+,K+-ATPase and MAO-A activities in prefrontal cortex of mice, but was not able to change the MAO-B activity. In conclusion, BMMS exerted an antidepressant-like effect in mice and serotonergic and nitrergic systems are involved in the antidepressant-like action of compound. BMMS modulated MAO-A and Na+, K+- ATPase activities in prefrontal cortex of mice.

Propofol for Treatment-Resistant Depression: A Pilot Study.

Background: We hypothesized that propofol, a unique general anesthetic that engages N-methyl-D-aspartate and gamma-aminobutyric acid receptors, has antidepressant properties. This open-label trial was designed to collect preliminary data regarding the feasibility, tolerability, and efficacy of deep propofol anesthesia for treatment-resistant depression. Methods: Ten participants with moderate-to-severe medication-resistant depression (age 18-45 years and otherwise healthy) each received a series of 10 propofol infusions. Propofol was dosed to strongly suppress electroencephalographic activity for 15 minutes. The primary depression outcome was the 24-item Hamilton Depression Rating Scale. Self-rated depression scores were compared with a group of 20 patients who received electroconvulsive therapy. Results: Propofol treatments were well tolerated by all subjects. No serious adverse events occurred. Montreal Cognitive Assessment scores remained stable. Hamilton scores decreased by a mean of 20 points (range 0-45 points), corresponding to a mean 58% improvement from baseline (range 0-100%). Six of the 10 subjects met the criteria for response (>50% improvement). Self-rated depression improved similarly in the propofol group and electroconvulsive therapy group. Five of the 6 propofol responders remained well for at least 3 months. In posthoc analyses, electroencephalographic measures predicted clinical response to propofol. Conclusions: These findings demonstrate that high-dose propofol treatment is feasible and well tolerated by individuals with treatment-resistant depression who are otherwise healthy. Propofol may trigger rapid, durable antidepressant effects similar to electroconvulsive therapy but with fewer side effects. Controlled studies are warranted to further evaluate propofol's antidepressant efficacy and mechanisms of action. ClinicalTrials.gov: NCT02935647.

Validation of the Scale on Parental Feeding Behaviors (ECOPAL) for Caregivers of Mexican Children.

Parental feeding practices can be crucial to preventing childhood obesity. This study aimed to validate a self-applicable instrument for evaluating the diverse parental feeding behaviors of Mexican caregivers based on the theoretical constructs of coercive control, structure, and autonomy support. The scale's content validity achieved significant values when assessed by expert judges, with moderate intensity in congruence (Kendall's W = 0.462; p = 0.000) and clarity (Kendall's W = 0.369; p = 0.001). The participants were 1185 Mexican adults (32.7 ± 7.6 years of age, 97% women, and 90% mothers) responsible for the main meal of at least one child (4.8 ± 3 years old). The data were subdivided randomly for an exploratory factor analysis (n = 581) and a confirmatory factorial analysis (n = 604). The first analysis grouped the items into 11 factors, with an accumulated variance of 63.9%. In the confirmatory analysis, a 10-factor model showed a better fit (CMIN = 1531.5, p < 0.001, CMIN/df = 2.20, RSEA = 0.045, CFI = 0.92, TLI, 0.91, and NFI = 0.87). The factors in this model were (1) the disposition of non-recommended foods, (2) nutritional education, (3) pressure to eat, (4) praise for healthy eating, (5) monitoring of consumption, (6) structured offer of fruits and vegetables, (7) consumption conditioning, (8) overt restriction, (9) guided choices, and (10) covert restriction. The Cronbach's alpha value was 0.816. Therefore, this scale presents good psychometric properties with which to evaluate the frequency of child caregivers' feeding behaviors in the context of ten different feeding practices in Mexico's urban areas and contributes to the knowledge of current practices in the Mexican population. It also evaluates changes resulting from future interventions that promote eating practices that favor the formation of healthy eating habits.

Quality Improvement in Thyroid Fine-Needle Aspiration Biopsy accuracy at San Juan City Hospital Endocrinology Clinics.

OBJECTIVE: This study aimed to determine if there was a reduction in the amount of non-diagnostic cytopathology results of thyroid fine-needle aspiration (FNA) biopsies performed at San Juan City Hospital (SJCH) endocrinology clinics since the inclusion of a cytopathologist at the clinics. METHODS: This research consisted of a retrospective analysis of thyroid nodule FNA biopsy results performed at SJCH endocrinology clinics. The biopsies analyzed were performed during academic years from July 2017-June 2018 and July 2018-June 2019, a period that reflects one academic year prior and a year after the inclusion of a cytopathologist to the clinics. The patients were classified into "pre group" and "post -group." Descriptive analysis was conducted, taking into consideration variables including sex, age, period, location, size of the nodule, and cytology results. A Chi-square test and Confidence Interval were used to assess the association and estimates between predictors and outcomes. RESULTS: From the 145 thyroid nodules biopsied, a total of 121 nodules (83.4%) resulted in diagnostic cytologic results, while 24 nodules (16.6%) were non-diagnostic. From the "pre group," 57 nodules (78.1%) had a diagnosis, while the other 16 (21.9%) were reported as non-diagnostic. From the "post group", 64 nodules (88.9%) had a diagnosis, while the other 8 (11.1%) resulted in non-diagnostic findings (p-value: = 0.08). Even though results were statistically non-significant, a clear trend towards a decrease in non-diagnostic samples was evident. CONCLUSION: In our study, there was a decrease in the number of non-diagnostic thyroid nodule FNA results after on-site adequacy determination guided by a cytopathologist.

The Challenge of Measuring Sweet Taste in Food Ingredients and Products for Regulatory Compliance: A Scientific Opinion.

The Codex Alimentarius Commission, a central part of the joint Food and Agricultural Organization/World Health Organizations Food Standards Program, adopts internationally recognized standards, guidelines, and code of practices that help ensure safety, quality, and fairness of food trade globally. Although Codex standards are not regulations per se, regulatory authorities around the world may benchmark against these standards or introduce them into regulations within their countries. Recently, the Codex Committee on Nutrition and Foods for Special Dietary Uses (CCNFSDU) initiated a draft revision to the Codex standard for follow-up formula (FUF), a drink/product (with added nutrients) for young children, to include requirements for limiting or measuring the amount of sweet taste contributed by carbohydrates in a product. Stakeholders from multiple food and beverage manufacturers expressed concern about the subjectivity of sweetness and challenges with objective measurement for verifying regulatory compliance. It is a requirement that Codex standards include a reference to a suitable method of analysis for verifying compliance with the standard. In response, AOAC INTERNATIONAL formed the Ad Hoc Expert Panel on Sweetness in November 2020 to review human perception of sweet taste, assess the landscape of internationally recognized analytical and sensory methods for measuring sweet taste in food ingredients and products, deliver recommendations to Codex regarding verification of sweet taste requirements for FUF, and develop a scientific opinion on measuring sweet taste in food and beverage products beyond FUF. Findings showed an abundance of official analytical methods for determining quantities of carbohydrates and other sweet-tasting molecules in food products and beverages, but no analytical methods capable of determining sweet taste. Furthermore, sweet taste can be determined by standard sensory analysis methods. However, it is impossible to define a sensory intensity reference value for sweetness, making them unfit to verify regulatory compliance for the purpose of international food trade. Based on these findings and recommendations, the Codex Committee on Methods of Analysis and Sampling agreed during its 41st session in May 2021 to inform CCNFSDU that there are no known validated methods to measure sweetness of carbohydrate sources; therefore, no way to determine compliance for such a requirement for FUF.

Lingual thyroid gland: it's time for awareness.

SUMMARY: Lingual thyroid (LT) gland is the most common type of ectopic thyroid tissue, but it is an extremely rare presentation. We present a case of a 41-year-old Hispanic female patient complaining of dysphonia and dysphagia. As part of the evaluation, fiber optic flexible indirect laryngoscopy (FIL) was performed which revealed a mass at the base of the tongue. The morphological examination was highly suspicious for ectopic thyroid tissue and the diagnosis was confirmed with neck ultrasound and thyroid scintigraphy. Although the patient presented subclinical hypothyroidism, levothyroxine therapy was initiated with a favorable response which included resolution of symptoms and mass size reduction. Our case portrays how thyroid hormone replacement therapy (THRT) may lead to a reduction in the size of the ectopic tissue and improvement of symptoms, thus avoiding the need for surgical intervention which could result in profound hypothyroidism severely affecting the patients' quality of life. LEARNING POINTS: Benign LT and malignant LT are indistinguishable clinically and radiographically for which histopathology is recommended. THRT, radioactive iodine 131 (RAI) therapy, and surgical excision are potential management options for LT. THRT may lead to size reduction of the ectopic tissue and resolution of symptoms avoiding surgical intervention.

Drug-Induced liver Injury Associated with Severe Cutaneous Hypersensitivity Reactions: A Complex Entity in Need of a Multidisciplinary Approach.

Idiosyncratic drug-induced liver injury (DILI) occasionally occurs in the setting of severe cutaneous adverse reactions (SCARs), including Stevens-Johnson syndrome (SJS), toxic epidermal necrolysis (TEN) and drug reaction with eosinophilia and systemic symptoms (DRESS). This strengthens the proposed immunologic mechanism associated with this adverse reaction. DRESS exhibits the most common association with DILI. SCARs have a wide spectrum of heterogeneous clinical presentations and severity, and genetic predisposition has been identified. In the context of SCARs, DILI present a different clinical picture, ranging from mild injury to acute liver failure. Elucidating the role of DILI in the clinical presentation and outcome of SCARs represents a challenge due to limited information from published studies and the lack of consensus on definitions. The cholestatic and mixed pattern of liver damage typically predominates in the case of DILI associated with SCARs, which is different from DILI without SCARs where hepatocellular is the most common injury pattern. Only a few drugs have been associated with both DILI and SCARs. Is this article, the criteria used for DILI recognition among SCARS have been revised and discussed, along with the drugs most commonly involved in these syndromes as well as the outcome, prognostic factors and the need for a multidisciplinary approach to improve the management of DILI in the context of SCARs.

Hip-related toxicity after prostate radiotherapy: Treatment related or coincidental?

BACKGROUND AND PURPOSE: To evaluate the incidence and predictors of hip toxicity postradiotherapy for localized prostate cancer. METHODS AND MATERIALS: 4067 prostate cancer patients were treated with external beam radiotherapy (EBRT; n=2569; 63%) or brachytherapy with or without supplemental EBRT (n=1508; 27%). 43% (n=1738) were treated with neo-adjuvant and concurrent ADT and 57% (n=2329) with radiotherapy alone. Hip toxicity was defined as moderate or severe pain upon ambulation with or without the need for hip-revision surgery. Median follow-up was 7years (range, 3-21years). RESULTS: One hundred twenty-one (2.7%) patients developed moderate-to-severe hip pain after radiotherapy affecting ambulation. Of these, 73 (60%) required hip replacement secondary to persistent hip pain. Among patients with baseline degenerative joint disease (DJD) changes on scans, 10-year incidence of hip-related toxicity was 11% versus 3% for those without such changes (P<.001). The only variables on multivariate analysis associated with hip-related toxicity post-radiotherapy were baseline DJD on imaging (P<.0001) and prolonged ADT for salvage therapy (P<.0001). CONCLUSIONS: Prostate EBRT or brachytherapy is associated with low incidence of long-term hip-related toxicity. The only variables identified associated with hip toxicity posttherapy was the presence of baseline DJD and prolonged salvage ADT posttreatment for patients developing recurrence.

Analysis of differential selective forces acting on the coat protein (P3) of the plant virus family Luteoviridae.

The coat protein (CP) of the family Luteoviridae is directly associated with the success of infection. It participates in various steps of the virus life cycle, such as virion assembly, stability, systemic infection, and transmission. Despite its importance, extensive studies on the molecular evolution of this protein are lacking. In the present study, we investigate the action of differential selective forces on the CP coding region using maximum likelihood methods. We found that the protein is subjected to heterogeneous selective pressures and some sites may be evolving near neutrality. Based on the proposed 3-D model of the CP S-domain, we showed that nearly neutral sites are predominantly located in the region of the protein that faces the interior of the capsid, in close contact with the viral RNA, while highly conserved sites are mainly part of beta-strands, in the protein's major framework.

Apoyo y barreras sociales en el comportamiento alimentario de mujeres con obesidad / Support and social barriers received and given in the food behavior of obesity women

Resumen Una de las principales causas de la obesidad es la alimentación. La alimentación forma parte del medio social, y algunas de las variables sociales vinculadas con el bienestar y la salud son el apoyo y las barreras sociales. Por lo tanto, el objetivo del presente estudio fue analizar las diferencias en comportamiento alimentario, así como apoyo y barreras sociales recibidas y proporcionadas en mujeres con obesidad vs. normopeso en un contexto natural de alimentación. Participaron cinco mujeres con obesidad y cinco normopeso, quienes fueron videograbadas por cuatro días consecutivos durante el momento de la "comida" en su hogar. El alimento fue registrado y pesado; las conductas de alimentación, de apoyo y las barreras sociales fueron registradas a partir de catálogos de observación sistemática elaborados exprofeso. Los resultados mostraron que las mujeres con obesidad ingirieron más frecuentemente, y en mayor cantidad, bebidas endulzadas, antojitos mexicanos fritos y comida rápida. Además, fueron objeto de más barreras instrumentales que les impedían alimentarse saludablemente. En conclusión, es necesario abordar el medio social en el que la alimentación tiene lugar, ya que el tipo de interacciones sociales pueden jugar un papel importante en la alimentación.

Abstract One of the main causes of obesity is food. Food is part of the social environment and some of the social variables that have been linked to well-being and health are support and social barriers. Therefore, the aim was to analyze the differences on eating behavior, support and social barriers received and given between obesity women vs. normal weight women in the natural feeding context. Five women with obesity and five with normal weight participated, who were videotaped with their consent four consecutive days during the feeding time of the "food" in their home; food was recorded and weighed. We recorded eating behaviors, and behaviors of support and social barriers from a systematic observation catalog prepared exprofeso. The results showed that women with obesity ingested a greater frequency and quantity of sweetened beverages, fried Mexican snacks and fast food. In addition, they received a greater frequency of instrumental barriers to eat healthy. In conclusion, it is necessary to address the social environment in which food takes place, since the type of social interactions may be playing an important role in food.