The analysis of factors increasing the odds for type 2 diabetes mellitus remission following re-do bariatric surgery after laparoscopic sleeve gastrectomy- cohort study.

INTRODUCTION: Metabolic/bariatric surgery is the only proven treatment for type 2 diabetes mellitus (T2D) with curative intent. However, in a number of patients, the surgery is not effective or they may experience a relapse. Those patients can be offered re-do bariatric surgery (RBS). PURPOSE: The study aimed to determine factors increasing the odds for T2D remission one year after RBS following primary laparoscopic sleeve gastrectomy. METHODS: A multicenter retrospective cohort study was conducted between January 2010 and January 2020, which included 12 bariatric centers in Poland. The study population was divided into groups: Group 1- patients with T2D remission after RBS (n = 28) and Group 2- patients without T2D remission after RBS (n = 49). T2D remission was defined as HBA1c < 6.0% without glucose-lowering pharmacotherapy and glycemia within normal range at time of follow-up that was completed 12 months after RBS. RESULTS: Fifty seven females and 20 males were included in the study. Patients who achieved BMI < 33 kg/m2 after RBS and those with %EBMIL > 60.7% had an increased chance of T2D remission (OR = 3.39, 95%CI = 1.28-8.95, p = 0.014 and OR = 12.48, 95%CI 2.67-58.42, p = 0.001, respectively). Time interval between primary LSG and RBS was significantly shorter in Group 1 than in Group 2 [1 (1-4) vs. 3 (2-4) years, p = 0.023]. CONCLUSIONS: Shorter time interval between LSG and RBS may ease remission of T2D in case of lack of remission after primary procedure. Significant excess weight loss seems to be the most crucial factor for T2D remission.

Multi-Instrumental Analysis Toward Exploring the Diabetic Foot Infection Microbiota.

The polymicrobial nature of diabetic foot infection (DFI) makes accurate identification of the DFI microbiota, including rapid detection of drug resistance, challenging. Therefore, the main objective of this study was to apply matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI TOF MS) technique accompanied by multiply culture conditions to determine the microbial patterns of DFIs, as well as to assess the occurrence of drug resistance among Gram-negative bacterial isolates considered a significant cause of the multidrug resistance spread. Furthermore, the results were compared with those obtained using molecular techniques (16S rDNA sequencing, multiplex PCR targeting drug resistance genes) and conventional antibiotic resistance detection methods (Etest strips). The applied MALDI-based method revealed that, by far, most of the infections were polymicrobial (97%) and involved many Gram-positive and -negative bacterial species-19 genera and 16 families in total, mostly Enterobacteriaceae (24.3%), Staphylococcaceae (20.7%), and Enterococcaceae (19.8%). MALDI drug-resistance assay was characterized by higher rate of extended-spectrum beta-lactamases (ESBLs) and carbapenemases producers compared to the reference methods (respectively 31% and 10% compared to 21% and 2%) and revealed that both the incidence of drug resistance and the species composition of DFI were dependent on the antibiotic therapy used. MALDI approach included antibiotic resistance assay and multiply culture conditions provides microbial identification at the level of DNA sequencing, allow isolation of both common (eg. Enterococcus faecalis) and rare (such as Myroides odoratimimus) bacterial species, and is effective in detecting antibiotic-resistance, especially those of particular interest-ESBLs and carbapenemases.

Screening for volatile biomarkers of colorectal cancer by analyzing breath and fecal samples using thermal desorption combined with GC-MS (TD-GC-MS).

Breath and fecal VOCs, among others, represent a new and encouraging clinical practice for the differential diagnosis of CRC. The purpose of our research was to identify VOCs present in exhaled air and feces of 20 HVs and 15 CRC patients. For collection of gas phase released from feces, emission microchambers were applied. Sorption tubes were used to enrich analytes for both breath and fecal samples. TD technique combined with GC-MS was used at the separation and identification step. The combination of statistical methods was used to evaluate the ability of VOCs to classify control group and CRC patients. Heptanoic acid, acetone, 2,6,10-trimethyldodecane, n-hexane, skatole, and dimethyl trisulfide are observed in elevated amounts in the patients group. The performance of diagnostic models on the tested data set was above 90%. This study is the first attempt to document the using of TD-GC-MS to analyze both breath and fecal samples to search for volatile biomarkers of CRC. A full evaluation of the results described herein requires further studies involving a larger number of samples. Moreover, it is particularly important to understand the metabolic pathways of substances postulated as tumor biomarkers.

Characterization of the salivary microbiome before and after antibiotic therapy via separation technique.

In the present research, the MALDI-TOF MS technique was applied as a tool to rapidly identify the salivary microbiome. In this fact, it has been monitored the changes occurred in molecular profiles under different antibiotic therapy. Significant changes in the composition of the salivary microbiota were noticed not only in relation to the non antibiotic (non-AT) and antibiotic treatment (AT) groups, but also to the used media, the antibiotic therapy and co-existed microbiota. Each antibiotic generates specific changes in molecular profiles. The highest number of bacterial species was isolated in the universal culture medium (72%) followed by the selective medium (48% and 38%). In the case of non-AT patients, the prevalence of Streptococcus salivarius (25%), Streptococcus vestibularis (19%), Streptococcus oralis (13%), and Staphylococcus aureus (6%) was identified while in the case of AT, Streptococcus salivarius (11%), Streptococcus parasanguinis (11%), Staphylococcus epidermidis (12%), Enterococcus faecalis (9%), Staphylococcus hominis (8%), and Candida albicans (6%) were identified. Notable to specified that the Candida albicans was noticed only in AT samples, indicating a negative impact on the antibiotic therapy. The accuracy of the MALDI-TOF MS technique was performed by the 16S rRNA gene sequencing analysis-as a reference method. Conclusively, such an approach highlighted in the present study can help in developing the methods enabling a faster diagnosis of disease changes at the cellular level before clinical changes occur. Once the MALDI tool allows for the distinguishing of the microbiota of non-AT and AT, it may enable to monitor the diseases treatment and develop a treatment regimen for individual patients in relation to each antibiotic. KEY POINTS: The salivary microbiota of antibiotic-treated patients was more bacteria variety MALDI-TOF MS is a promising tool for recording of reproducible molecular profiles Our data can allow to monitor the treatment of bacterial diseases for patients.

Viability study of clinical bacterial strains by capillary electrophoresis and flow cytometry approaches.

One of the challenges medicine faces is the constantly growing resistance of pathogens to various classes of antibiotics. In this study, we investigated the use of capillary electrophoresis (CE) to characterize and assess the physiological states of three clinical bacterial strains-methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive S. aureus (MSSA), and Escherichia coli extended-spectrum ß-lactamases (ESßL)-exposed to different antibiotics. All chosen bacteria are the leading causes of healthcare-associated and hospital-acquired invasive infections in adults. In the first part of the research, it was determined the optimal incubation time of the tested strains with antibiotics, represented as an optimal time of 24 h. In the second part, we have compared two approaches: flow cytometry (FC) as a standard method and CE as a proposed alternative approach. The viability of clinical strains treated with different class antibiotics calculated in CE measurements was strongly correlated (>0.83 for MSSA, >0.92 for ESßL and MRSA) with the viability obtained on the basis of FC measurements. As a result, CE has a chance to become a modern diagnostic method used in clinical practice. The CE cutoff was found to be 50%; above this value, the strain shows resistance to the action of the antibiotic.

The Role of Antibiotics in Endoscopic Transmural Drainage of Post-Inflammatory Pancreatic and Peripancreatic Fluid Collections.

Background: Although endoscopic treatment of symptomatic post-inflammatory pancreatic and peripancreatic fluid collections (PPPFCs) is an established treatment method, some aspects of endotherapy and periprocedural management remain controversial. The role of antibiotics is one of the most controversial issues in interventional endoscopic management of local complications of pancreatitis. Methods: This study was a randomized, non-inferiority, placebo-controlled, and double-blinded clinical trial to investigate the role of antibiotic prophylaxis in endoscopic transmural drainage in patients with symptomatic non-infected PPPFCs and assess the influence of antibiotic treatment on the results of endotherapy in patients with symptomatic infected PPPFCs.This trial included 62 patients treated endoscopically for PPPFCs in 2020 at our medical center. Patients were divided into two groups; group 1 comprised patients who had received empirical intravenous antibiotic therapy during endotherapy and group 2 comprised patients who did not receive antibiotic therapy during endoscopic drainage of PPPFCs. The end points were clinical success and long-term success of endoscopic treatment. Results: Thirty-one patients were included in group 1 (walled-off pancreatic necrosis [WOPN, 51.6%; pseudocyst, 48.4%) and 31 patients in group 2 (WOPN, 58.1%; pseudocyst, 41.9%) (p=0.6098/nonsignificant statistical [NS]). Infection with PPPFCs was observed in 15/31 (48.39%) patients in group 1 and in 15/31 (48.39%) patients in group 2 (p=1.0/NS). The average time of active (with flushing through nasocystic drainage) drainage in group 1 was 13.0 (6 - 21) days and was 14.0 (7 - 25) days in group 2 (p=0.405/NS). The average total number endoscopic procedures on one patient was 3.3 (2 - 5) in group 1 and 3.4 (2 - 7) in group 2 (p=0.899/NS). Clinical success of PPPFCs was observed in 29/31 (93.5%) patients from group 1 and in 30/31 (96.8%) patients from group 2 (p=0.5540/NS). Complications of endotherapy were noted in 8/31 (25.8%) patients in group 1 and in 10/31 (32.3%) patients in group 2 (p=0.576/NS). Long-term success in group 1 and 2 was reported in 26/31 (83.9%) and 24/31 (77.4%) patients, respectively (p=0.520/NS). Conclusions: The effective endoscopic drainage of sterile PPPFCs requires no preventive or prophylactic use of antibiotics. In infected PPPFCs, antibiotic therapy is not required for effective endoscopic transmural drainage.

Revisional operations among patients after surgical treatment of obesity: a multicenter Polish Revision Obesity Surgery Study (PROSS).

Introduction: Revisional surgery is more technically challenging and associated with increased morbidity and mortality. Nevertheless, the frequency of revisional bariatric surgery (RBS) is increasing. Therefore, investigating this group of patients appears to be currently valid. Aim: The objective of this multicenter study was to collect, systematize and present the available data on RBS after surgical treatment of morbid obesity among Polish patients. Material and methods: This multicenter study included a retrospective analysis of a prospectively maintained database. Outcomes included an analysis of the indications for RBS, the type of surgery most frequently chosen as RBS and the course of the perioperative period of treatment among patients undergoing RBS. Results: The group consisted of 799 patients (624 (78.1%) women, 175 (21.9%) men). The mean age was 38.96 ±9.72 years. Recurrence of obesity was the most common indication for RBS. The most frequently performed RBS procedures were one anastomosis gastric bypass (OAGB) - 294 (36.8%) patients, Roux-en-Y gastric bypass (RYGB) - 289 (36.17%) patients and sleeve gastrectomy (SG) - 172 (21.52%) patients. After primary surgery 63.58% of patients achieved sufficient weight loss, but after RBS only 38.87%. Complications were noted in 222 (27.78%) cases after RBS with GERD being the most common - 117 (14.64%) patients. Conclusions: RBS most often concerns patients after SG. The main indication for RBS is weight regain. OAGB and RYGB were the two most frequently chosen types of RBS. Secondary operations lead to further weight reduction. However, RBS are associated with a significant risk of complications.

Capillary Zone Electrophoresis in Tandem with Flow Cytometry in Viability Study of Various ATCC Bacterial Strains under Antibiotic Treatment.

The aim of this study was to develop an innovative method of examining bacterial survival using capillary zone electrophoresis (CZE) and flow cytometry (FC) as a reference method. For this purpose, standard strains of bacteria from the ATCC collection were used: Enterococcus faecalis ATCC 14506, Staphylococcus aureus ATCC 11632, Klebsiella pneumoniae ATCC 10031, Pseudomonas aeruginosa ATCC 27853, and Escherichia coli ATCC 25922, as well as seven antibiotics with different antimicrobial mechanisms of action. The ratio of live and dead cells in the tested sample in CZE measurements were calculated using our algorithm that takes into account the detection time. Results showed a high agreement between CZE and FC in the assessment of the percentage of live cells exposed to the stress factor in both antibiotic susceptibility and time-dependent assays. The applied measuring system to assess the effectiveness of antibiotic therapy in in vitro conditions is a method with great potential, and the data obtained with the use of CZE mostly correspond to the expected drug sensitivity according to EUCAST and CLSI guidelines.

Culturomics Approach to Identify Diabetic Foot Infection Bacteria.

The main goal of the study was to evaluate the usefulness of the culturomics approach in the reflection of diabetic foot infections (DFIs) microbial compositions in Poland. Superficial swab samples of 16 diabetic foot infection patients (Provincial Polyclinical Hospital in Torun, Poland) were subjected to culturing using 10 different types of media followed by the identification via the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and Biotyper platform. Identified 204 bacterial isolates representing 18 different species-mostly Enterococcus faecalis (63%) and Staphylococcus aureus (44%). Most of the infections (81%) demonstrated a polymicrobial character. Great differences in the species coverage, the number of isolated Gram-positive and Gram-negative bacteria, and the efficiency of the microbial composition reflection between the investigated media were revealed. The use of commonly recommended blood agar allowed to reveal only 53% of the entire microbial composition of the diabetic foot infection samples, which considerably improved when the chromagar orientation and vancomycin-resistant enterococi agar were applied. In general, efficiency increased in the following order: selective < universal < enriched < differential media. Performed analysis also revealed the impact of the culture media composition on the molecular profiles of some bacterial species, such as Corynebacterium striatum, Proteus mirabilis or Morganella morganii that contributed to the differences in the identification quality. Our results indicated that the culturomics approach can significantly improve the accuracy of the reflection of the diabetic foot infections microbial compositions as long as an appropriate media set is selected. The chromagar orientation and vancomycin-resistant enterococi agar media which were used for the first time to study diabetic foot infection microbial profiles demonstrate the highest utility in the culturomics approach and should be included in further studies directed to find a faster and more reliable diabetic foot infection diagnostic tool.

Identification of Bacteria Associated with Post-Operative Wounds of Patients with the Use of Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Approach.

The bacterial infection of post-operative wounds is a common health problem. Therefore, it is important to investigate fast and accurate methods of identifying bacteria in clinical samples. The aim of the study was to analyse the use of the MALDI-TOF MS technique to identify microorganism wounds that are difficult to heal. The most common bacteria are Escherichia coli, Staphylococcus spp., and Enterococcus spp. We also demonstrate the effect of culture conditions, such as the used growth medium (solid: Brain Heart Infusion Agar, Mueller Hilton Agar, Glucose Bromocresol Purple Agar, and Vancomycin Resistance Enterococci Agar Base and liquid: Tryptic Soy Broth and BACTEC Lytic/10 Anaerobic/F), the incubation time (4, 6, and 24h), and the method of the preparation of bacterial protein extracts (the standard method based on the Bruker guideline, the Sepsityper method) to identify factors and the quality of the obtained mass spectra. By comparing the protein profiles of bacteria from patients not treated with antibiotics to those treated with antibiotics based on the presence/absence of specific signals and using the UniProt platform, it was possible to predict the probable mechanism of the action of the antibiotic used and the mechanism of drug resistance.

Problems with identifying and distinguishing salivary streptococci: a multi-instrumental approach.

Aim: The purpose of this study was to create an alternative protocol for the DNA-based identification of salivary microbiota focused on the distinguishing of Streptococcus species. Materials & methods: Salivary bacteria were identified using 16S rDNA sequencing and proteins and lipids profiling using MALDI-TOF/MS as well as FTIR analysis. Results: Most of the isolates belonged to streptococci - mostly the salivarious group indistinguishable by the molecular technique. In turn, MALDI analysis allowed for their fast and reliable classification. Although FTIR spectroscopy demonstrated the correct species classification, the spectra interpretation was time consuming and complicated. Conclusion: MALDI-TOF/MS demonstrated the biggest effectiveness in the identification and discrimination between the salivary streptococci, which could be easily incorporated in the workflow of routine microbiological laboratories.

Withdrawal from bariatric treatment - an analysis under various demographic conditions.

Poland is high on a list of countries affected by epidemics of obesity, a problem that has especially suddenly increased in the post-transformation period. MATERIALS AND METHOD: Documentation of 115 subsequent patients diagnosed with grade 3 obesity, considered eligible for surgical treatment, was analysed since 2015. A subgroup of 68 patients was selected from this group and the included patients who decided to cancel their treatment at various stages. Their history since presentation at the obesity treatment centre, BMI with its history, age, education and place of residence were analysed, followed by a telephone survey. 42 patients were finally contacted. Special attention was placed on the analysis of patients taking into account their place of residence. Patients were divided into inhabitants of rural and urban areas. RESULTS: In the study group of 68 patients, 19 (27.9%) were inhabitants of rural areas and 49 (72.1%) of urban areas. Women accounted for 67.6%, and men for 32.4%. The mean age of patients when they presented for treatment was 43 years. On average, men presented 5 years later compared to women. The mean BMI on qualification for treatment of obesity was 47.6 kg/m2. The mean BMI max - 49.6 kg/m2. There were no differences regarding education in both populations. Despite the lack of differences regarding changes in the BMI since withdrawal from treatment, as many as 63% of patients from rural areas reached the BMI max in this time. CONCLUSIONS: There were demographic differences between the degree of obesity observed among patients receiving bariatric treatment, and inhabitants of urban areas were favoured. Patients from rural areas who withdrew from bariatric treatment and were left without medical care significantly more often achieved their maximum body weight, when compared to those living in urban areas. The active participation of physicians, both specialists and general practitioners, in the life of obese patients is imperative.

Antimicrobial properties of biosynthesized silver nanoparticles studied by flow cytometry and related techniques.

This work reports the effect of silver bionanoparticles (Bio(AgNPs) synthesized by Actinobacteria CGG 11n on selected Gram (+) and Gram (-) bacteria. Flow cytometry, classical antibiogram method and fluorescent microscopy approach was used for evaluation of antimicrobial activity of Bio(AgNPs) and their combination with antibiotics. Furthermore, the performed research specified the capacity of flow cytometry method as an alternative to the standard ones and as a complementary method to electromigration techniques. The study showed antibacterial activity of both BioAgNPs and the combination of antibiotics/BioAgNPs against all the tested bacteria strains in comparison with a diffusion, dilution and bioautographic methods. The synergistic effect of antibiotics/BioAgNPs combination (e.g. kanamycin, ampicillin, neomycin and streptomycin) was found to be more notable against Pseudomonas aeruginosa representing a prototype of multi-drug resistant "superbugs" for which effective therapeutic options are very limited.

Capillary electrophoresis in the diagnosis of surgical site infections.

A surgical site infection (SSI) is an infection that occurs after surgery in the part of the body where the surgery took place. A SSI may range from a spontaneously limited wound discharge within 7-10 days of an operation to a life-threatening postoperative complication, such as a sternal infection after open heart surgery. Most surgical site infections are caused by contamination of an incision with microorganisms from the patient's own body during surgery. From the analytical point of view the complex natures of these samples as well as the low concentrations of analytes require a system with high sensitivity and efficiency. Such situation require a technique such as capillary electrophoresis which is a powerful and versatile separation technique that promises to rival high performance liquid chromatography when applied to the separation of both charged and neutral species. During the study, it has been demonstrated that CZE identifying characteristics of such groups of pathogens such as bacteria G (+) and different species of bacteria G (-) and also develop weekly individual profiles for patients after application of antibiotics. This was done in order to show the impact of antibiotic therapy in change "numbers" of bacteria present in the wound after surgery. The method proved to be the ideal straight specificity in the case of E. coli (100%). Finally analysis of the spectra and the second derivatives of the UV-Vis spectra confirmed the similarity in the profiles and showed that the CZE is a great method for fast screening test in bacterial infection.

A study of interactions between bacteria and antibiotics by capillary electrophoresis.

To assess the bacteria-antibiotic interactions in patients with postoperative wound infections, a simple electrophoretic test was performed. To estimate the effectiveness of the antibiotic therapy and to prepare 3-day profiles of bacteria "quantity" in biological samples, CE was used. As our team demonstrated earlier, the method is easy and fast, sample pretreatment is not necessary, and it is characterized by high selectivity. Finally, the statistically optimal and significant results of the CZE test analysis for detection of Escherichia coli cells was established for migration time lower than 3.5 min. The obtained sensitivity and specificity amounted to 89.5 and 100%, respectively. It is the first application of CZE in the study of medical therapy.

Clinical application of a rapid microbiological test based on capillary zone electrophoresis to assess local skin infection.

BACKGROUND: The basic clinical problem associated with infection treatment is the fact that classic, commonly and routinely used isolation and identification methods are based on long-term processes of a phenotypic analysis of microorganisms. Consequently sometimes, especially in small centres, rapid implementation of antibacterial treatment becomes delayed.The work presents the initial results of rapid microbiological identification based on an original method of capillary zone electrophoresis (CZE). The study involved the analysis of 78 biological samples from post-operative wounds and trophic ulcers. RESULTS: The attempt was made to identify individual bacterial species based on characteristic features of electropherograms achieved. Finally, G(+) cocci type bacteria and different G(-) rods were identified with sensitivity of 88.1% and specificity of 100%. CONCLUSIONS: Based on the clinical trials using an electrophoretic technique in the field of microbiological diagnostics of infected exudate from a post-operative wound it can be concluded that it is a rapid and relatively sensitive method for initial identification of infectious pathogens.

[Value of urease breath test in helicobacter pylori infection diagnostics among children and youth]. / Wartosc mocznikowego testu oddechowego w diagnostyce zakazen helicobacter pylori u dzieci i mlodziezy.

UNLABELLED: One of non-invasive methods of Helicobacter pylori diagnostics is the breath test with urea 13C. THE AIM: of the study was to estimate the sensitivity and specificity of urea breath test (UBT) in Helicobacter pylori (Hp) infection diagnosis among children and youth. PATIENTS AND METHODS: medical records of 100 patients, hospitalised due to gastrointestinal disorders, between 3 and 18 years of age were analysed. In all patients urea breath test, endoscopy of upper gastrointestinal tract with macroscopic assessment of gastric and duodenal mucosa and morphologic assessment of biopsy specimens were performed. RESULTS: positive results of urea breath test were present in twenty-six patients. In five patients (5%) breath test was positive despite the lack of Helicobacter pylori infection in histhopathology samples. This group was 19.23% of all patients who had positive result of breath test. Among 21 (21%) patients result of breath test was correlated with positive result of histhopathology studies. This group was 80.77% of all patients with positive results of breath test. Among the remaining 74 patients (74%) results of breath test were negative. The results of Hp and UBT correlation are as follows: in 64% children: Hp (-), UBT (-), in 5% Hp (-), UBT (+), in 10% children: Hp (+), UBT (-), in 24%, Hp (+), UBT (+) CONCLUSIONS: result of urea breath test should be verified by endoscopy studies with morphologic assessment of specimens to avoid overdiagnosis of Helicobacter pylori infection.

[Cryptogenic hepatic cirrhosis in a 14 years old boy]. / Marsksc watroby o nieustalonej etilogii u 14-letniego chlopca.

Alcoholism is the main etiological factor of hepatic cirrhosis among adults whereas among children the following pathologies lead to hepatic cirrhosis: bile ducts disease (atresia or hypoplasia of bile ducts, progressive familial intrahepatic cholestasis), metabolic disorders (tyrosinemia, hemochromatosis, galactosemia, alpha1 - antitrypsin deficiency, Wilson's disease), chronic viral hepatitis B, delta, C, autoimmune hepatitis. Despite the availability of modern diagnostic methods scanty clinical symptomatology may be a cause of late diagnosis, often in the stage of complications. Presented case of a 14 years old boy is an example of late diagnosis of cryptogenic hepatic cirrhosis.