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.

Transrectal endoscopic drainage with vacuum-assisted therapy in patients with anastomotic leaks following rectal cancer resection.

BACKGROUND: Surgery is the gold standard for the treatment of malignant tumors of the rectum. Intestinal anastomotic leakage remains a serious complication of colorectal surgery. The efficacy and safety of transrectal endoscopic drainage by vacuum therapy in patients with intestinal anastomotic leakage after surgical treatment of middle and distal rectal tumors were assessed. METHODS: Prospective analysis of treatment outcomes among patients undergoing surgery for middle and distal rectal tumors at the Department of General, Gastroenterological, and Oncological Surgery of the Ludwik Rydygier Collegium Medicum in Bydgoszcz and Nicolaus Copernicus University in Torun from 2016 to 2019 was conducted. RESULTS: Seventy-nine patients with middle and distal rectal tumors underwent laparoscopic resection. Intestinal anastomotic leak was identified in 18 (22.79%) patients [all men, mean age 61.39 (43-86) years] during the postoperative period. Primary protective ileostomy was performed in 8/18 (44.44%) patients. All 18 patients were treated with endoluminal vacuum therapy via transrectal endoscopic drainage. The mean time from surgery to the diagnosis of leakage and initiation of endoscopic treatment was 16 (3-728) days. The mean number of endoscopic procedures per patient was 6 (1-11). The mean duration of endoscopic treatment was 22 (4-43) days. Complications of endotherapy occurred in 2/18 (11.11%) patients treated endoscopically for bleeding from the abscess cavity. Success of endoluminal vacuum therapy was achieved in 17/18 (94.44%) patients. Moreover, 5/18 (27.78%) patients required ileostomy during the endoscopic treatment. The mean follow-up period was 368 (118-724) days. Long-term success of transrectal endoscopic drainage using vacuum-assisted therapy was achieved in 15/18 (83.33%) patients. CONCLUSIONS: Endoscopic rectal drainage using vacuum-assisted therapy is an effective and safe minimally invasive treatment in patients with intestinal anastomotic leaks following resection procedures within the middle and distal rectum.

Outcomes and limitations of endoscopic ultrasound-guided hepaticogastrostomy in malignant biliary obstruction.

BACKGROUND: Transpapillary biliary drainage in ERCP is an established method for symptomatic treatment of patients with unresectable malignant biliary obstruction. Percutaneous transhepatic biliary drainage frequently remains the treatment of choice when the transpapillary approach proves ineffective. Recently, EUS-guided extra-anatomical anastomoses of bile ducts to the gastrointestinal tract have been reported as an alternative to percutaneous biliary drainage. To assess the usefulness of extra-anatomical intrahepatic biliary duct anastomoses to the gastrointestinal tract as endotherapy for unresectable malignant biliary obstruction and to determine factors affecting the efficacy of treatment. METHODS: A prospective analysis of the treatment results of all patients with unresectable biliary obstruction treated with EUS-guided hepaticogastrostomy at our institution in the years 2016-2019. RESULTS: Transmural intrahepatic biliary drainage (EUS-guided hepaticogastrostomy) was performed due to the ineffectiveness of ERCP in 53 patients (38 males, 15 females; mean age 74.66 [56-89] years) with unresectable biliary obstruction. Technical success of EUS-guided hepaticogastrostomy was achieved in 52/53 (98.11%) patients. Complications of endoscopic treatment were observed in 10/53 (18.87%) patients. Clinical success of EUS-guided hepaticogastrostomy was achieved in 46/53 (86.79%) patients. Bismuth type II-IV cholangiocarcinoma, hepatic metastases, ascites, suppurative cholangitis, and high blood bilirubin levels exceeding 30 mg/dL were independent factors for increased complications and inefficacy of EUS-guided hepaticogastrostomy. CONCLUSIONS: In the event of transpapillary biliary drainage proving ineffective, extra-anatomical anastomoses of intrahepatic bile ducts to the gastrointestinal tract provide an effective method for the treatment of patients with malignant biliary obstruction.

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.

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.

Pharmacokinetic study of amoxicillin in human plasma by solid-phase microextraction followed by high-performance liquid chromatography-triple quadrupole mass spectrometry.

Sensitive and selective analytical procedures based on high-performance liquid chromatography with mass spectrometric detection were developed for the determination of amoxicillin in human plasma samples. Samples were prepared by applying in-house manufactured molecularly imprinted solid-phase microextraction probes. The detection of target compounds was performed in multiple reaction monitoring mode. The multiple reaction monitoring detection was operated in the positive electrospray ionization mode using the transitions of m/z 366 ([M + H](+) ) → 349 for amoxicillin and m/z 390 ([M + H](+) ) → 372 for gemifloxacin. The method was validated with precision within 15% relative standard deviation and accuracy within 15% relative error. The method was successfully applied to study of the pharmacokinetics of amoxicillin in human plasma after oral administration of amoxicillin.

Identification of In Vitro Metabolites of Amoxicillin in Human Liver Microsomes by LC-ESI/MS.

Amoxicillin (AMOX) metabolism in human liver microsomes was studied in vitro using liquid chromatography-mass spectrometry (LC/MS). Amoxicillin was incubated with human liver microsomes along with NADPH, and the reaction mixture was analyzed by LC/MS to obtain the specific metabolic profile of the studied antibiotic drug. Positive electrospray ionization was employed as the ionization source. An ACE C18-column (4.6 mm × 150 mm, 3 µm) was implemented with acetonitrile and water (+0.1 % formic acid) in isocratic mode as the mobile phase at the flow 0.4 mL min-1. The chemical structures of metabolites were proposed on the basis of the accurate mass measurement of the protonated molecule as well as their main product. Six phase I and one phase II metabolites were detected and structurally described. The metabolism of AMOX occurred via oxidation, hydroxylation and oxidative deamination, as well as through combination of these reactions. Compound M7, with glucuronic acid was also observed as phase II metabolite. Neither sulfate nor glutathione conjugates were detected. This study presents novel information about the chemical structure of the potential AMOX metabolites and provides vital data for further pharmacokinetic and in vivo metabolism studies.

Endoscopic pancreatogastric anastomosis in the treatment of symptoms associated with inflammatory diseases of the pancreas.

BACKGROUND: The outflow of pancreatic juice into the duodenum is often impaired in pancreatic inflammatory diseases. The basis of interventional treatment in these cases is anatomical transpapillary access of the main pancreatic duct during endoscopic retrograde cholangiopancreatography (ERCP), which ensures the physiological outflow of pancreatic juice into the lumen of the digestive tract. However, in some patients, anatomical changes prevent transpapillary drainage of the main pancreatic duct. Surgery is the treatment of choice in such cases. AIM: To evaluate the effectiveness and safety of endoscopic pancreaticogastrostomy under endoscopic ultrasound (EUS) guidance. METHODS: Retrospective analysis of treatment outcomes of all patients with acute or chronic pancreatitis who underwent endoscopic pancreatogastric anastomosis under EUS guidance in 2018-2023 at the Department of General, Gastroenterological and Oncological Surgery, Ludwik Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, Poland. RESULTS: In 9 patients [7 men, 2 women; mean age 53.45 (36-66) years], endoscopic pancreatogastric anastomosis under EUS guidance was performed because of the lack of transpapillary access during ERCP. Narrowing of the main pancreatic duct at the head of the pancreas was observed in 4/9 patients (44.44%). Pancreatic fragmentation (disconnected pancreatic duct syndrome) was diagnosed in 3/9 patients (33.33%). In 2/9 patients (22.22%), narrowing of the pancreatoenteric anastomosis was observed after pancreaticoduodenectomy. Technical success of endoscopic pancreaticogastrostomy was observed in 8/9 patients (88.89%). Endotherapeutic complications were observed in 2/9 patients (22.22%). Clinical success was achieved in 8/9 patients (88.89%). The mean follow-up period was 451 (42-988) d. Long-term success of endoscopic pancreatogastric anastomosis was achieved in 7/9 patients (77.78%). CONCLUSION: Endoscopic pancreaticogastrostomy under EUS guidance is an effective and safe treatment method, especially in the absence of transpapillary access to the main pancreatic duct.

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.

Small bowel segment with Meckel's diverticulum volvulus related to short mesodiverticular band: a case report.

BACKGROUND: Meckel's diverticulum is a remnant of the omphalomesenteric duct and occurs in only about 2% of people. Mesodiverticular band is the congenital remnant of the vitelline artery and is an even less often occurring phenomenon. PRESENTED CASE: We present the case of a 56-year-old Caucasian male who was admitted to the emergency department with a very intense, sudden abdominal pain, without past abdominal surgery history. Contrast enhanced computed tomography showed a possibly ischemic closed loop of the small intestine. Urgent laparotomy was performed, during which bloody content in the peritoneal cavity and torsed loop of the small intestine with Meckel's diverticulum were found. The bowel loop and Meckel's diverticulum were ischemic. At the tip of Meckel's diverticulum there was a broken fibrous band extending to mesentery with pulsating artery. We did segmental resection of small intestine including Meckel's diverticulum and primary end-to-end anastomosis. The patient had an unremarkable postoperative hospital stay and was discharged home after 5 days. CONCLUSION: In our case, we describe a patient with the volvulus of a segment of small bowel and Meckel's diverticulum, which eventually led to small bowel obstruction and ischemia. It was a very rare case that required urgent surgical treatment.

Transanal total mesorectal excision (TaTME) in rectal cancer treatment within an expert center.

The objective of this study was to evaluate treatment outcomes in patients who underwent the TaTME procedure for cancer of the middle and low rectum in an expert center. Prospective analysis of the outcomes of all consecutive patients treated using the TaTME technique for cancer of the middle and distal rectum at the our medical center between March 1, 2015, and March 31, 2022. A total of 128 patients (34 women, 94 men; mean age 66.01 [38-85] years) with cancer of the middle and distal rectum qualified for TaTME. TaTME procedures were performed in 127/128 (99.22%) patients. Complications of surgery were observed in 22/127 (17.32%) patients. Negative proximal and distal margins were confirmed in all 127 patients. Complete (R0) resection of the mesorectum was confirmed in 125/127 (98.43%) and nearly complete (R1) resection was confirmed in 2/127 (1.57%) patients. The average follow-up period was 795 days (296-1522) days. Local recurrence was detected during the follow-up period in 2/127 (1.57%) patients. This study showed that the TaTME procedure is an effective and safe method for the minimally invasive treatment of middle and low rectal cancers, particularly within an expert center setting.

Medical or Common Knowledge? Knowledge of Medical Professionals on Obesity Diagnosis Criteria and Treatment.

INTRODUCTION: Proper diagnosis of obesity, its severity, and complications and their effective treatment requires an interdisciplinary healthcare approach. Nevertheless, obesity remains under-identified and undertreated. Academic knowledge concerning obesity pathology, diagnosis, and treatment is advancing. It is not clear whether this translates into clinical practice. The goal of the study was to assess the knowledge of healthcare professionals (HCPs) on obesity and particularly on the criteria for diagnosis as well as for conservative and surgical treatment. METHODS: This cross-sectional study was conducted among active HCPs (N = 184), including physicians, nurses, physiotherapists, and paramedics who had contact with adult patients with obesity. The proprietary research survey, implemented in an online tool, was used to assess knowledge on the diagnosis and treatment of obesity and self-assessment of that knowledge. The analysis was limited to the following: body mass index (BMI) definition, BMI values, visceral obesity definition, bariatric surgery indications, choice of treatment method, role of diet and physical activity, knowledge of obesity pharmacotherapy, length of obesity pharmacotherapy, financing of bariatric procedures, and goals of bariatric treatment. The correct answers were determined according to the Polish guidelines for the diagnosis and treatment of obesity. RESULTS: Half of the respondents (52.2%) were doctors, 20.7% were nurses and midwives, 19.0% were physiotherapists, and 8.2% were other medical professionals. Among questions related to knowledge on obesity, 67.1% of respondents provided correct answers, with respondents answering questions concerning obesity diagnosis correctly more frequently (70.1%) than those concerning methods of treatment (64.6%). The largest number of correct answers was related to the definition of BMI and normal BMI values. The smallest number of correct answers pertained to the diagnostic criteria for visceral obesity and pharmacological treatment of obesity. There was no statistically significant impact of a responder's knowledge levels on the obesity of different HCPs. Workplace and participation in training sessions were found to have the largest impact on the level of knowledge on obesity. HCPs' own assessment of their knowledge on obesity was negatively correlated with their actual level of knowledge. CONCLUSION: The prevalence of overweight and obesity implies that essentially every HCP has daily contact with patients with excessive body weight. Knowledge of BMI values cannot be considered as exclusively medical knowledge: these values were established years ago and are present in widely available sources. Our research showed that 32.9% of HCPs did not have sufficient knowledge about how to diagnose and treat obesity.

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.

The Association of Polish Surgeons (APS) clinical guidelines for the use of laparoscopy in the management of abdominal emergencies. Part I.

Introduction: Over the past three decades, almost every type of abdominal surgery has been performed and refined using the laparoscopic technique. Surgeons are applying it for more procedures, which not so long ago were performed only in the classical way. The position of laparoscopic surgery is therefore well established, and in many operations it is currently the recommended and dominant method. Aim: The aim of the preparation of these guidelines was to concisely summarize the current knowledge on laparoscopy in acute abdominal diseases for the purposes of the continuous training of surgeons and to create a reference for opinions. Material and methods: The development of these recommendations is based on a review of the available literature from the PubMed, Medline, EMBASE and Cochrane Library databases from 1985 to 2022, with particular emphasis on systematic reviews and clinical recommendations of recognized scientific societies. Recommendations were formulated in a directive form and evaluated by a group of experts using the Delphi method. Results and conclusions: There are 63 recommendations divided into 12 sections: diagnostic laparoscopy, perforated ulcer, acute pancreatitis, incarcerated hernia, acute cholecystitis, acute appendicitis, acute mesenteric ischemia, abdominal trauma, bowel obstruction, diverticulitis, laparoscopy in pregnancy, and postoperative complications requiring emergency surgery. Each recommendation was supported by scientific evidence and supplemented with expert comments. The guidelines were created on the initiative of the Videosurgery Chapter of the Association of Polish Surgeons and are recommended by the national consultant in the field of general surgery. The first part of the guidelines covers 5 sections and the following challenges for surgical practice: diagnostic laparoscopy, perforated ulcer, acute pancreatitis, incarcerated hernia and acute cholecystitis. Contraindications for laparoscopy and the ERAS program are discussed.

Recommendations for modern perioperative care for elective surgery: consensus of panel of exerts.

INTRODUCTION: Correct surgical technique and perioperative care are two factors that can reduce the number of complications, improve treatment outcomes and shorten the length of hospital stay. The introduction of enhanced recovery protocols has changed the approach to patient care in some centers. However, there are significant differences among centers, and in some the standard of care has remained unchanged. AIM: the goal of the panel was to develop recommendations for modern perioperative care in accordance with current medical knowledge in order to reduce the number of complications associated with surgical treatment. An additional goal was to optimize and standardize perioperative care among Polish centers. MATERIALS AND METHODS: the development of these recommendations was based on a review of the available literature from the PubMed, Medline and Cochrane Library databases from January 1, 1985 to March 31, 2022, with particular emphasis on systematic reviews and clinical recommendations of recognized scientific societies. Recommendations were formulated in a directive form and were assessed using the Delphi method. RESULTS AND CONCLUSIONS: 34 recommendations for perioperative care were presented. They cover aspects of pre-, intra- and post-operative care. Implementation of the presented rules allows to improve the results of surgical treatment.

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.

Development of novel molecularly imprinted solid-phase microextraction fibers and their application for the determination of antibiotic drugs in biological samples by SPME-LC/MS(n).

Novel molecularly imprinted polymer (MIP)-coated fibers for solid-phase microextraction (SPME) fibers were prepared by using linezolid as the template molecule. The characteristics and application of these fibers were investigated. The polypyrrole, polythiophene, and poly(3-methylthiophene) coatings were prepared in the electrochemical polymerization way. The molecularly imprinted SPME coatings display a high selectivity toward linezolid. Molecularly imprinted coatings showed a stable and reproducible response without any influence of interferents commonly existing in biological samples. High-performance liquid chromatography with spectroscopic UV and mass spectrometry (MS) detectors were used for the determination of selected antibiotic drugs (linezolid, daptomycin, amoxicillin). The isolation and preconcentration of selected antibiotic drugs from new types of biological samples (acellular and protein-free simulated body fluid) and human plasma samples were performed. The SPME MIP-coated fibers are suitable for the selective extraction of antibiotic drugs in biological samples.

Desarda versus Lichtenstein technique for primary inguinal hernia treatment: 3-year results of a randomized clinical trial.

BACKGROUND: The Shouldice method and other tissue-based techniques are still acknowledged to be acceptable for primary inguinal hernia repair according to the European Hernia Society guidelines. Desarda's technique, presented in 2001, is an original hernia repair method using an undetached strip of external oblique aponeurosis. This randomized trial compared outcomes after hernia repair with Desarda (D) and mesh-based Lichtenstein (L) techniques. METHODS: A total of 208 male patients were randomly assigned to the D or L group (105 vs. 103, respectively). The primary outcomes measured were recurrence and chronic pain. Additionally, early and late complications, foreign body sensation, and return to everyday activity were examined in hospital and at 7, 30 days, and 6, 12, 24, and 36 months after surgery. RESULTS: During the follow-up, two recurrences were observed in each group (p = 1.000). Chronic pain was experienced by 4.8 and 2.9% of patients from groups D and L, respectively (p = 0.464). Foreign body sensation and return to activity were not different between the groups. There was significantly less seroma production in the D group (p = 0.004). CONCLUSIONS: The results of primary inguinal hernia repair with the Desarda and Lichtenstein techniques are comparable at the 3-year follow-up. The technique may potentially increase the number of tissue-based methods available for treating groin hernias.