Strength of selection pressure is an important parameter contributing to the complexity of antibiotic resistance evolution.

Revealing the genetic changes responsible for antibiotic resistance can be critical for developing novel antibiotic therapies. However, systematic studies correlating genotype to phenotype in the context of antibiotic resistance have been missing. In order to fill in this gap, we evolved 88 isogenic Escherichia coli populations against 22 antibiotics for 3 weeks. For every drug, two populations were evolved under strong selection and two populations were evolved under mild selection. By quantifying evolved populations' resistances against all 22 drugs, we constructed two separate cross-resistance networks for strongly and mildly selected populations. Subsequently, we sequenced representative colonies isolated from evolved populations for revealing the genetic basis for novel phenotypes. Bacterial populations that evolved resistance against antibiotics under strong selection acquired high levels of cross-resistance against several antibiotics, whereas other bacterial populations evolved under milder selection acquired relatively weaker cross-resistance. In addition, we found that strongly selected strains against aminoglycosides became more susceptible to five other drug classes compared with their wild-type ancestor as a result of a point mutation on TrkH, an ion transporter protein. Our findings suggest that selection strength is an important parameter contributing to the complexity of antibiotic resistance problem and use of high doses of antibiotics to clear infections has the potential to promote increase of cross-resistance in clinics.

The effects of COVID-19 fear and anxiety on symptom severity, sleep quality, and mood in patients with fibromyalgia: a pilot study.

BACKGROUND: During the COVID-19 pandemic, individuals faced psychological stress caused by fear and anxiety due to the high transmission and mortality rate of the disease, the social isolation, economic problems, and difficulties in reaching health services. Fibromyalgia (FM) is a chronic centralized pain sensitivity disorder. Psychological, physical and/or autoimmune stressors were found to increase FM symptoms. This pilot study aimed to evaluate the COVID-19 fear and anxiety level, and to examine their effect on disease severity, sleep quality, and mood in FM patients compared to control group. METHODS: This pilot study conducted as a cross-sectional study, and included 62 participants. Participants were divided into two groups: FM patient group (n = 31) and control group (n = 31). Symptom severity, sleep quality, and mood were determined using the Revised Fibromyalgia Impact Questionnaire (FIQR), Pitsburg Sleep Quality Index (PSQI), and Hospital Anxiety Depression Scale (HADS), respectively. In order to evaluate the level of COVID-19 fear and anxiety, the Fear of COVID-19 Scale (FCV-19S) and Coronavirus Anxiety Scale (CAS) were used compared to control group. RESULTS: FIQR, PSQI, HAD-A, HAD-D, FCV-19S and CAS scores were significantly higher in the FM group (p = 0.01). A positive significant correlation was found between FCV-19S and CAS results and FIQR, PSQI, and HAD-anx results in FM patients (p < 0.05). CONCLUSION: This pilot study showed that, the individuals with FM can be more affected by psychological stress, and this situation negatively affects the symptom severity, sleep quality, and mood in FM patients, so these patients should be closely monitored in terms of psychological stressors and their effects during pandemics. More studies with more participants are necessary to describe the challenges lived by fibromyalgia population.

Evaluation of the effects of the COVID-19 pandemic on children with cerebral palsy, caregivers' quality of life, and caregivers' fear of COVID-19 with telemedicine.

BACKGROUND: The sudden and unexpected pandemic changed the daily routine of the children with cerebral palsy (CP) and their caregivers. AIMS: This study aimed to investigate the impact of the novel coronavirus (COVID-19) pandemic on the utilization of health and rehabilitation services and the general health and physical status of children with CP. In addition, the second aim of the study was to examine the effects of the COVID-19 pandemic on caregivers' quality of life (QOL) and their fear of COVID-19. METHODS: The utilization of children health and rehabilitation services during the pandemic, the general health and physical status of the children during the pandemic, and the children and caregivers' history of COVID-19 infections were questioned. Furthermore, the caregivers' level of fear of COVID-19 and their QOL were examined. RESULTS: One hundred twenty caregivers were contacted by phone, and 94 (78.33%) caregivers agreed to participate in the study. Sixty-three of 94 children (67.1%) did not attend their routine control check-up during the pandemic. Twelve children (12.8%) discontinued their physical therapy sessions during the pandemic. Caregivers physical and mental QOL significantly decreased during the pandemic (p < 0.05). The median of caregivers' Fear of COVID-19 scale (FCV-19S) was 17.5 (7-35). CONCLUSION: We think that more attention should be given to telerehabilitation and telemedicine services of the clinicians who deal with the children with CP, and their caregivers in order to prevent the negative effects of future pandemic periods.

Comparing the effectiveness of ultrasound guided versus blind genicular nerve block on pain, muscle strength with isokinetic device, physical function and quality of life in chronic knee osteoarthritis: a prospective randomized controlled study.

BACKGROUND: The genicular nerve block (GNB) is demonstrated from several reports to alleviate pain and improve knee functionality in patients with chronic knee osteoarthritis (OA). Ultrasound (US)-guided GNB has been the most used imaging method. This study aimed to compare the effectiveness of US-guided versus blind GNB in the treatment of knee OA. METHODS: This prospective, randomized clinical trial included patients with knee OA based on American College of Rheumatology diagnostic criteria. The patients were evaluated for clinical and dynamometer parameters at the baseline, 4 weeks after treatment, and 12 weeks after treatment. The patients underwent blind injection or US-guided injection. RESULTS: When compared with the baseline, both groups showed significant improvement in pain, physical function, and quality of life parameters. Significant differences were observed between the groups for clinical parameters (30-second chair stand test, 6-minute walk test) in favor of the US-guided group. On the other hand, blind injection was more significantly effective on some parameters of the Nottingham Health Profile. There wasn't any significant improvement in isokinetic muscle strength for either group. CONCLUSIONS: This study demonstrated that both US-guided and blind GNB, in the treatment of knee OA, were effective in reducing symptoms and improving physical function. GNB wasn't an effective treatment for isokinetic muscle function. USguided injections may yield more effective clinical results than blind injections.

High versus Moderate Dosage of Daily and Weekly Administration of Vitamin D Supplements in the Form of Oil Drop in Nursing Home Residents.

OBJECTIVE: To investigate the effectiveness of daily (800 IU), weekly-moderate (5600 IU) and weekly-high (8000 IU) supplementation of Vitamin D in nursing home residents. STUDY DESIGN: A descriptive study. PLACE AND DURATION OF STUDY: Nursing Home, MEVA, Istanbul, Turkey, from July 2016 to July 2017. METHODOLOGY: Nursing home residents were divided into 3 groups for supplementation of Vitamin D: Daily Dose Group (DDG), Weekly Dose Group-moderate (WDG-moderate) and Weekly Dose Group-high (WDG-high). Blood and physical performance tests were done initially to obtain a baseline value and the tests were repeated at 13th and 26th weeks of supplementation. Statistical analysis was conducted only on patients who were able to complete the 6-month-long study. RESULTS: WDG-moderate (5600 IU/week) supplementation is found to be the most effective intervention in our study [25 (OH) D from 23.50 ±12.67 ng/mL to 37.38 ±14.42 ng/mL]. In WDG-moderate, the resulting Vitamin D level was found to reach near-optimum therapeutic levels. Only a limited increase was observed in 25 (OH) D level of DDG and WDG- high at the end of 26 weeks. CONCLUSION: Weekly (5600 IU/week) moderate supplementation of Vitamin D could be more beneficial than weekly (8000/week) high supplementation among nursing home residents. Multi-drug use among nursing home residents may hinder the therapeutic efficiency of Vitamin D administration. Physical performance tests may fail to demonstrate increased performance in mobility after Vitamin D administration in nursing home residents.

The effects of COVID-19 fear and anxiety on symptom severity, sleep quality, and mood in patients with fibromyalgia: a pilot study

Abstract Background: During the COVID-19 pandemic, individuals faced psychological stress caused by fear and anxiety due to the high transmission and mortality rate of the disease, the social isolation, economic problems, and difficulties in reaching health services. Fibromyalgia (FM) is a chronic centralized pain sensitivity disorder. Psychological, physical and/or autoimmune stressors were found to increase FM symptoms. This pilot study aimed to evaluate the COVID-19 fear and anxiety level, and to examine their effect on disease severity, sleep quality, and mood in FM patients compared to control group. Methods: This pilot study conducted as a cross-sectional study, and included 62 participants. Participants were divided into two groups: FM patient group ( n = 31) and control group ( n = 31). Symptom severity, sleep quality, and mood were determined using the Revised Fibromyalgia Impact Questionnaire (FIQR), Pitsburg Sleep Quality Index (PSQI), and Hospital Anxiety Depression Scale (HADS), respectively. In order to evaluate the level of COVID-19 fear and anxiety, the Fear of COVID-19 Scale (FCV-19S) and Coronavirus Anxiety Scale (CAS) were used compared to control group. Results: FIQR, PSQI, HAD-A, HAD-D, FCV-19S and CAS scores were significantly higher in the FM group ( p = 0.01). A positive significant correlation was found between FCV-19S and CAS results and FIQR, PSQI, and HAD-anx results in FM patients ( p < 0.05). Conclusion: This pilot study showed that, the individuals with FM can be more affected by psychological stress, and this situation negatively affects the symptom severity, sleep quality, and mood in FM patients, so these patients should be closely monitored in terms of psychological stressors and their effects during pandemics. More studies with more participants are necessary to describe the challenges lived by fibromyalgia population.

Mechanical slowing-down of cytoplasmic diffusion allows in vivo counting of proteins in individual cells.

Many key regulatory proteins in bacteria are present in too low numbers to be detected with conventional methods, which poses a particular challenge for single-cell analyses because such proteins can contribute greatly to phenotypic heterogeneity. Here we develop a microfluidics-based platform that enables single-molecule counting of low-abundance proteins by mechanically slowing-down their diffusion within the cytoplasm of live Escherichia coli (E. coli) cells. Our technique also allows for automated microscopy at high throughput with minimal perturbation to native physiology, as well as viable enrichment/retrieval. We illustrate the method by analysing the control of the master regulator of the E. coli stress response, RpoS, by its adapter protein, SprE (RssB). Quantification of SprE numbers shows that though SprE is necessary for RpoS degradation, it is expressed at levels as low as 3-4 molecules per average cell cycle, and fluctuations in SprE are approximately Poisson distributed during exponential phase with no sign of bursting.

Fluidic and microfluidic tools for quantitative systems biology.

Understanding genes and their functions is a daunting task due to the level of complexity in biological organisms. For discovering how genotype and phenotype are linked to each other, it is essential to carry out systematic studies with maximum sensitivity and high-throughput. Recent developments in fluid-handling technologies, both at the macro and micro scale, are now allowing us to apply engineering approaches to achieve this goal. With these newly developed tools, it is now possible to identify genetic factors that are responsible for particular phenotypes, perturb and monitor cells at the single-cell level, evaluate cell-to-cell variability, detect very rare phenotypes, and construct faithful in vitro disease models.

Building a morbidostat: an automated continuous-culture device for studying bacterial drug resistance under dynamically sustained drug inhibition.

We present a protocol for building and operating an automated fluidic system for continuous culture that we call the 'morbidostat'. The morbidostat is used to follow the evolution of microbial drug resistance in real time. Instead of exposing bacteria to predetermined drug environments, the morbidostat constantly measures the growth rates of evolving microbial populations and dynamically adjusts drug concentrations inside culture vials in order to maintain a constant drug-induced inhibition. The growth rate measurements are done using an optical detection system that is based on measuring the intensity of back-scattered light from bacterial cells suspended in the liquid culture. The morbidostat can additionally be used as a chemostat or a turbidostat. The whole system can be built from readily available components within 2-3 weeks by biologists with some electronics experience or engineers familiar with basic microbiology.