Genotypic and phenotypic characterization of a Salmonella Typhimurium strain resistant to pulsed electric fields.

Pulsed Electric Fields (PEF) technology is regarded as one of the most interesting alternatives to current food preservation methods, due to its capability to inactivate vegetative microorganisms while leaving the product's organoleptic and nutritional properties mostly unchanged. However, many aspects regarding the mechanisms of bacterial inactivation by PEF are still not fully understood. The aim of this study was to obtain further insight into the mechanisms responsible for the increased resistance to PEF of a Salmonella Typhimurium SL1344 variant (SL1344-RS, Sagarzazu et al., 2013), and to quantify the impact that the acquisition of PEF resistance has on other aspects of S. enterica physiology, such as growth fitness, biofilm formation ability, virulence and antibiotic resistance. WGS, RNAseq and qRT-PCR assays indicated that the increased PEF resistance of the SL1344-RS variant is due to a higher RpoS activity caused by a mutation in the hnr gene. This increased RpoS activity also results in higher resistance to multiple stresses (acidic, osmotic, oxidative, ethanol and UV-C, but not to heat and HHP), decreased growth rate in M9-Gluconate (but not in TSB-YE or LB-DPY), increased ability to adhere to Caco-2 cells (but no significant change in invasiveness) and enhanced antibiotic resistance (to six out of eight agents). This study significantly contributes to the understanding of the mechanisms of the development of stress resistance in Salmonellae and underscores the crucial role played by RpoS in this process. Further studies are needed to determine whether this PEF-resistant variant would represent a higher, equal or lower associated hazard than the parental strain.

A Holistic Approach to Managing Secondary Dysphagia Following Prolonged Intubation and Tracheostomy: A Case Report.

The ability to swallow and maintain the airway is a critical rehabilitation milestone for patients with swallowing disorders following prolonged tracheal intubation. Tracheostomy and dysphagia often coexist in critically ill patients and given the medical complexity analyzing the evidence to optimize swallowing assessment and management is challenging. It takes a holistic approach to dealing with issues in a critical care patient as we also need to deal with issues other than medical. We present a case of a 68-year-old gentleman who was admitted to the critical care unit following a double barrel ileostomy and had multiple complications and organ dysfunction requiring prolonged supportive management, tracheostomy, and mechanical ventilation. After recovering from primary illness and complications, he had a swallowing disorder (secondary dysphagia), which was managed successfully over the next month. The case highlights the need for screening, a multidisciplinary team, empathy, and effort as a part of a holistic management approach.

Ultrasound-Guided Right Internal Jugular Vein Cannulation by Operators of Different Experience: A Randomized, Pilot Study.

BACKGROUND AND AIM: Currently, ultrasound-guided (US-guided) internal jugular vein (IJV) cannulation is the recommended technique. However, it has a learning curve and might be unsafe in inexperienced hands. The present study aimed to compare the performance and complications with two levels of experience in performing US-guided right IJV cannulation. METHODS: With informed consent, 108 procedures were performed after random allocation into two groups based on operator experience. An operator with experience in performing 30 or more ultrasound-guided IJV cannulation was considered an expert. The rate of successful cannulation, the time needed, number of attempts, and complication rate were measured. Quantitative continuous variables were compared using the unpaired student's t-test, and the chi-square test or Fisher's-exact test was used for the comparison of qualitative variables; P-value < 0.05 was considered significant. RESULTS: The successful cannulation rates were 100% versus 94.44% in the expert and non-expert groups, respectively; (P=0.0803). The mean time for successful cannulation and the percentage of patients who required ≥ two attempts were significantly lower in the expert group (33.28 seconds and 12.96% versus 95.42 seconds and 61.12%). Although the incidence of carotid artery puncture and hematoma (7.4% and 5.56%) was higher in the non-expert group, it was not statistically different; (P=1.00). CONCLUSION: US-guided right IJV cannulation has a learning curve, and procedures as many as 30 US-guided IJV cannulation need to be observed and performed under the guidance to achieve it.

Geobacillus stearothermophilus STCC4517 spore suspensions showed survival curves with shoulder phenomena independent of sporulation temperature and pH, whose duration was an exponential function of treatment temperature.

Deviations in linearity in survival curves are common in inactivation kinetics during heat treatment. These might lead one to underestimate how effective thermal treatment is. In previous research we reported a relationship between decimal reduction time values (DT) and shoulder lengths (Sl) of survival curves which was characteristic of each microorganism. However, the impact of other factors such as sporulation temperature and pH of the treatment media on shoulder length is still not known. The objective of this research was to evaluate the effect of sporulation temperature (45, 55 and 65 °C) and pH (4.0, 5.0, 6.0 and 7.0) treatment has on the profile of survival curves and on the relationship between Sl/DT of G. stearothermophilus STCC 4517. The results obtained demonstrated that all the spore suspensions, independently of sporulation temperature and pH, showed survival curves with shoulder phenomena, whose duration was an exponential function of treatment temperature. Although both parameters had a significant effect on heat resistance, the relationship between the shoulder length and DT values was constant at all pHs for spores produced at the optimum sporulation temperature.

The Relationship of Respiratory Rate-Oxygenation (ROX) and Modified ROX Index With High-Flow Nasal Cannula Oxygen Therapy in COVID-19 Patients: An Observational Pilot Study.

Background and aim Respiratory Rate-Oxygenation (ROX) and modified ROX (mROX) indexes have been proposed to detect early high-flow nasal cannula (HFNC) therapy failure. We evaluated the utility and relationship of ROX and mROX indexes in COVID-19 patients started on HFNC oxygen therapy. Methods This pilot study collected data from adult COVID-19 patients requiring HFNC oxygenation from 29 Jan - 29 Jun 2021. The patients were divided into two cohorts based on HFNC therapy success. ROX and mROX were compared using statistical diagnostic testing, including receiver operating characteristics and area under the curve (AUC) using online Epitools (https://epitools.ausvet.com.au/) and MedCalc software (MedCalc Software Ltd, Ostend, Belgium, https://www.medcalc.org/); p<0.05 was considered significant. Results Twenty-seven patients fulfilled the inclusion criteria; 48.15% of therapy failed. The cohort's mean ± standard deviation age was 53.93 ± 10.67 years; 74.1% were male. The accuracy of predicting failure for mean ROX versus mROX at baseline and six-hour values was 59.81 versus 70.68 and 67.42 versus 74.88, respectively (all p>0.05). The AUC for ROX and mROX at baseline and at six hours were statistically indifferent. Only an mROX of 4.05 (mean value) and 3.34 (Youden's J cut-off) had a sensitivity plus specificity at 156% and 163%, respectively. Conclusion Both ROX and mROX at baseline and six hours had fair-to-good accuracies and AUC; the differences were insignificant. Both ROX and mROX had better accuracies at six hours. However, only mROX < 4.05 at six hours fulfilled the sensitivity plus specificity criteria to be a clinically valuable screener.

Individualizing Time-in-Range Goals in Management of Diabetes Mellitus and Role of Insulin: Clinical Insights From a Multinational Panel.

Diabetes mellitus is a global health concern associated with significant morbidity and mortality. Inadequate control of diabetes leads to chronic complications and higher mortality rates, which emphasizes the importance of achieving glycemic targets. Although glycated hemoglobin (HbA1c) is the gold standard for measuring glycemic control, it has several limitations. Therefore, in recent years, along with the emergence of continuous glucose monitoring (CGM) technology, glycemic control modalities have moved beyond HbA1c. They encompass modern glucometrics, such as glycemic variability (GV) and time-in-range (TIR). The key advantage of these newer metrics over HbA1c is that they allow personalized diabetes management with person-centric glycemic control. Basal insulin analogues, especially second-generation basal insulins with properties such as longer duration of action and low risk of hypoglycemia, have demonstrated clinical benefits by reducing GV and improving TIR. Therefore, for more effective and accurate diabetes management, the development of an integrated approach with second-generation basal insulin and glucometrics involving GV and TIR is the need of the hour. With this objective, a multinational group of endocrinologists and diabetologists reviewed the existing recommendations on TIR, provided their clinical insights into the individualization of TIR targets, and elucidated on the role of the second-generation basal insulin analogues in addressing TIR.

Impact of temperature on Covid 19 in India

The role of temperature in Covid 19 pandemic has been subjected to frequent review. An effort was made to find out such relations from three districts in India. Data were analyzed for 14 weeks. It appears that temperature could impact the spread of the pandemic.

Heat resistance, membrane fluidity and sublethal damage in Staphylococcus aureus cells grown at different temperatures.

In this work the influence of growth temperature (10-42 °C) on Staphylococcus aureus heat resistance was studied, and its relationship with the ability of cells to repair sublethal damages and with membrane fluidity was evaluated. Non-linear, convex from above survival curves were obtained, and therefore a special case of the Baranyi model was used to fit them. For exponential phase cells, heat resistance did not change with growth temperature in the range between 10 and 37 °C, but cells grown at 42 °C were significantly more resistant, showing D58 and shoulder length (sl58) values 2.5 and 4 times greater than the others, respectively. For stationary growth phase cells, an increase in growth temperature above 20 °C resulted in an increase in D58 values, and cells grown at 42 °C also displayed the highest D58 and sl58 values. The increased heat resistance at 58 °C of stationary growth phase cells grown at higher temperatures was coincident with the appearance of a higher proportion of sublethally damaged cells capable of recovery and outgrowth in non-selective medium. Membrane fluidity was measured at treatment temperatures, and it was observed that those cells with more rigid membranes displayed greater heat resistance (Pearson coefficient = 0.969***). Additionally, S. aureus cells whose membrane was fluidized through exposure to benzyl alcohol were notably sensitized against the action of heat, in a concentration-dependent manner. Results obtained in this research indicate that membrane physical state could be an important factor determining the survival capacity of bacterial cells to a heat treatment.

Osmotolerance provided by the alternative sigma factors σB and rpoS to Staphylococcus aureus and Escherichia coli is solute dependent and does not result in an increased growth fitness in NaCl containing media.

The aim of this work was to examine the role of the alternative general stress sigma factors σ(B) and rpoS on the ability of Staphylococcus aureus and Escherichia coli, respectively, to grow in liquid and solid media of different osmolarity. For this purpose, S. aureus strain Newman and its isogenic ΔsigB mutant IK84 and E. coli strain BJ4 and its isogenic ΔrpoS mutant BJ4L1 were grown in media (TSBYE) with different concentrations of NaCl. Growth parameters (lag phase duration, growth rate and maximum number of microorganisms) and limiting growth concentrations (Maximum Non-Inhibitory Concentration - MNIC - and Minimum Inhibitory Concentration - MIC-) were determined. The mechanisms underlying the differences observed between parental and mutant strains were also explored. The absence of the sigma factors σ(B) and rpoS led to a decrease in the MNICs and MICs calculated for S. aureus and E. coli, respectively. Conversely, neither σ(B) nor rpoS provided with increased growth fitness to S. aureus and E. coli cells at NaCl concentrations up to 1.36M and 1M, respectively. The decreased osmotolerance of the σ(B) and rpoS deficient strains, as compared to their parental strains, was compensated by the addition of glycine-betaine (1mM) to the growth medium. It was also observed that the decreased tolerance to NaCl of the mutant strains was coincident with a decreased tolerance to sucrose, KCl, and LiCl but not to glycerol, MgCl2, and CaCl2. Results obtained also demonstrate that the increased osmotolerance of stationary growth phase E. coli cells, as compared to exponential growth phase ones, would be due to the activation of both rpoS-independent and rpoS-dependent mechanisms. This work will help to understand the mechanisms of bacterial resistance to osmotic stress and the role of the alternative sigma factors σ(B) and rpoS in this process.