Amniotic fluid soluble CD93 is elevated in the presence of intra-amniotic inflammation in preterm prelabor rupture of the fetal membranes.

OBJECTIVE: To determine the soluble form of CD93 (sCD93) concentration in amniotic fluid from pregnancies complicated by preterm prelabor rupture of membranes (PPROM) based on the presence of microbial invasion of the amniotic cavity (MIAC) and/or intra-amniotic inflammation. METHODS: A total of 144 women with a singleton pregnancy complicated by PPROM were included in this study. Amniotic fluid samples were obtained by transabdominal amniocentesis. MIAC was determined by the combination of cultivation and non-cultivation techniques. Intra-amniotic inflammation was characterized as a concentration of interleukin-6 3,000 pg/mL in amniotic fluid. Women were categorized in the following groups: i) intra-amniotic infection (both MIAC and intra-amniotic inflammation), ii) sterile intra-amniotic inflammation (intra-amniotic inflammation per se), iii) colonization of the amniotic cavity (MIAC per se), and iv) negative amniotic fluid (without both MIAC and intra-amniotic inflammation). Levels of sCD93 in amniotic fluid were assessed by ELISA. RESULTS: A difference in the levels of sCD93 in amniotic fluid was found among the groups of women with intra-amniotic infection, sterile intra-amniotic inflammation, colonization of the amniotic cavity, and negative amniotic fluid (intra-amniotic infection: median 22.3 ng/mL, sterile intra-amniotic inflammation: median 21.0 ng/mL, colonization of the amniotic cavity: 8.7 ng/mL, negative: median 8.7 ng/mL; P < 0.0001). CONCLUSIONS: Intra-amniotic inflammation in PPROM, irrespectively of the presence or absence of MIAC, is associated with the elevation of the level of sCD93 in amniotic fluid.

Improved laboratory diagnostics of Streptococcus pneumoniae in respiratory tract samples through qPCR.

The aim of this study was to test the detection performance of the cpsA, lytA and ply genes through qPCR in the identification of Streptococcus pneumoniae in respiratory tract samples. Specificity was tested on a panel of 128 streptococci and other bacteria DNA samples. The qPCR assay was tested on a total of 51 respiratory tract samples from patients with community-acquired pneumonia (CAP). The specificity of the cpsA, lytA and ply genes was 100%, 100%, and 86%, respectively. The quantitative assessment, based on lytA, determined a cutoff value of ~2x104, 4x102 and 4x102 DNA copies per 1 mL of valid sputum, tracheal aspirate and bronchial aspirate samples, respectively. The results from the present study suggest that qPCR detection of all three genes would be optimal in the accurate detection of Streptococcus pneumoniae.

Characterization of KPC-Encoding Plasmids from Enterobacteriaceae Isolated in a Czech Hospital.

Ten Enterobacteriaceae isolates collected in a Czech hospital carried blaKPC-positive plasmids of different sizes (∼30, ∼45, and ∼80 kb). Sequencing revealed three types of plasmids (A to C) with the Tn4401a transposon. Type A plasmids comprised an IncR backbone and a KPC-2-encoding multidrug resistance (MDR) region. Type B plasmids were derivatives of type A plasmids carrying an IncN3-like segment, while type C plasmids were IncP6 plasmids sharing the same KPC-2-encoding MDR region with type A and B plasmids.

The sensitivity and specificity of Abbott Panbio™ COVID 19 Ag Rapid test in the context of four SARS-CoV-2 variants.

Rapid antigen tests for the detection of SARS-CoV-2 are commonly used for the diagnosis of Covid-19. Previously published data showed a wide range of sensitivity and specificity of RATs, but these studies were performed on relatively small numbers of samples and using only limited numbers of virus variants. The aim of the study was to evaluate the main parameters of a commonly used RAT for 4 different virus variants in comparison with PCR. Material and methods: A set of 2874 samples obtained from Covid-19 patients were examined both by PCR and RAT. Two commercial PCR kits (Generi Biotech, Diana Biotechnologies) and one RAT - Abbott Panbio™ COVID 19 Ag Rapid - were compared for their sensitivity and specificity in samples positive for one of the four different SARS-CoV-2 variants - B.1.258 (n = 496), Alpha (n = 645), Delta/Delta+ (n = 687), and Omicron (n = 1046). Results: The sensitivity of Panbio™ COVID19 Ag Rapid test varied from 80.0 % in Omicron to 88.92 % in Alpha variants. The specificities of the RAT for all variants reached above 93 %. Statistically significant differences were found between the results from RAT assay in select virus variants. In addition, significantly higher sensitivity (p < 0.05) was detected in samples with higher viral loads than in those with lower. Conclusion: Despite the different sensitivity and specificity of Panbio™ COVID19 Ag Rapid test (Abbott ®) for different SARS-CoV-2 variants, this test sensitivity was proven to be always above the 80 % suggested by WHO, which makes it suitable for common use, regardless of the virus variability.

[Bacterial infection as a cause of infertility in humans]. / Bakteriální infekce jako prícina neplodnosti u lidí

Microorganisms which are present in the human urogenital tract may be involved in the development of inflammatory changes negatively affecting the genitals in both men and women. Pathological conditions due to inflammatory alterations may result in complete loss of fertility. Infections of the urogenital tract are responsible for 15% of all cases of infertility in couples. Negative impact on the human reproduction is mainly caused by direct damage to the genital tract mucosa by metabolic products of microorganisms or by induction of pro-inflammatory responses of the body. Another mechanism is indirect impact of microorganisms on the genital function. Moreover, the effect of bacteria on spermatogenesis and semen quality is important in men. Infections mainly caused by Chlamydia trachomatis or Neisseria gonorrhoeae represent the greatest risk in terms of permanent consequences for human reproduction. As for other sexually transmitted disorders, such as infections caused by Gardnerella vaginalis, urogenital mycoplasmas or ureaplasmas, the link between infection and infertility has been intensively researched.

Amniotic fluid CD36 in pregnancies complicated by spontaneous preterm delivery: a retrospective cohort study.

OBJECTIVE: The aim of this study was to evaluate CD36 concentrations in amniotic fluid in pregnancies complicated by spontaneous delivery with intact fetal membranes (preterm labor, PTL) and preterm prelabor rupture of membranes (PPROM) with respect to the presence of the intra-amniotic infection. METHODS: A total of 80 women with PPROM and 71 with PTL were included in the study. Amniotic fluid samples were obtained by transabdominal amniocentesis. Amniotic fluid CD36 concentrations were assessed by enzyme-linked immunosorbent assay. Microbial colonization of the amniotic cavity (MIAC) was determined by the cultivation and non-cultivation approach. Intra-amniotic inflammation (IAI) was defined as an amniotic fluid bedside interleukin-6 concentration ≥3000 pg/mL. Intra-amniotic infection was characterized by the presence of both MIAC and IAI. RESULTS: Women with PPROM with intra-amniotic infection had higher amniotic fluid CD36 concentrations than women without infection (with infection: median 346 pg/mL, IQR 262-384 vs. without infection: median 242 pg/mL, IQR 199-304; p = .006) A positive correlation between amniotic fluid CD36 concentrations and interleukin-6 concentrations was found (rho = 0.48; p < .0001). In PTL pregnancies, no statistically significant difference was found in the amniotic fluid level of CD36 between intra-amniotic infection, sterile IAI, and negative amniotic fluid. CONCLUSIONS: The presence of intra-amniotic infection is characterized by higher amniotic fluid CD36 concentrations in pregnancies complicated by PPROM. An amniotic fluid CD36 cutoff value of 252.5 pg/mL was found to be optimal for the prediction of intra-amniotic infection. In PTL pregnancies, no statistically significant change in CD36 concentration was found with respect to the presence of intra-amniotic infection.

A Rare Case of Osteomyelitis of an Ankle Caused by Mycobacterium chelonae.

Mycobacterium chelonae, a rapidly growing nontuberculous mycobacterium, is usually described as a causative agent of soft tissue infections (postsurgical, posttraumatic, posttransplantation, postinjection, catheter infection, etc.), but only rarely as a cause of osteomyelitis. The authors describe a case report of a 72-year-old man with osteomyelitis of the talus. Initially, the infection was assessed as a soft tissue infection, without any osteolytic changes on the X-ray. After cultivation with subsequent targeted molecular typing of the rpoB gene, M. chelonae was identified from the affected tissue. The bone involvement was subsequently detected on MRI and confirmed histologically with findings of the granulomatous tissue and acid-fast bacilli. The patient was initially treated intravenously with a combination of tigecycline, amikacin, and moxifloxacin for 4 weeks, after which the oral combination of doxycycline and moxifloxacin continued. Identification of the infecting pathogen using molecular typing thus helped to establish the correct diagnosis and represents a rarely described case of osteomyelitis caused by M. chelonae.

Mycobacterioses Induced by Mycobacterium abscessus: Case Studies Indicating the Importance of Molecular Analysis for the Identification of Antibiotic Resistance.

Mycobacterioses are less frequently occurring but serious diseases. In recent years, at a global level, the incidence of mycobacterioses induced by the rapidly growing species Mycobacterium abscessus (M. a.), which is considered to be the most resistant to antibiotics and most difficult to treat, has been on the rise. Correct identification to the level of the subspecies (M. a. abscessus, M. a. massiliense, and M. a. bolletii) and determination of its sensitivity to macrolides, which are the basis of combination therapy, are of principal importance for the management of the disease. We describe five cases of mycobacterioses caused by M. a., where the sequencing of select genes was performed to identify the individual subspecies and antibiotic resistance. The analysis of the rpoB gene showed two isolates each of M. a. abscessus and M. a. massiliense and one isolate of M. a. bolletii. The complete (full length) erm(41) gene responsible for the development of inducible resistance to macrolides was demonstrated in both M. a. abscessus and M. a. bolletii isolates. A partially deleted and non-functional erm(41) gene was demonstrated in M. a. massiliense isolates. The subsequent sequencing of the full length erm(41) gene products showed, however, the mutation (T28→C) in both isolates of M. a. abscessus, causing a loss of the function and preserved sensitivity to macrolides. The antibiotic sensitivity testing confirmed that both the isolates of M. a. abscessus and M. a. massiliense were sensitive to clarithromycin even after prolonged 14-day incubation. The inducible resistance to clarithromycin was maintained only in M. a. bolletii. Thus, the sequence analysis of the erm(41) gene can reliably identify the preservation of sensitivity to macrolides and serve as an important tool in the establishment of therapeutic regimens in cases of infections with M. abscessus.

Dual Infection of an Open Fracture Caused by Mycobacterium setense and Clostridium celerecrescens.

Infections caused by Mycobacterium setense or Clostridium celerecrescens are extremely rare. In this report, for the first time a dual infection with these two pathogens is described. An 18-year-old female suffered multiple injuries, including an open comminuted fracture of the right humeral diaphysis after falling from a fifth-floor balcony in January 2019. Five months after the accident, a fistula appeared in the scar, reaching the bone tissue. M. setense and C. celerecrescens were cultured from sinus swabs and subsequently from perioperative samples. The patient was initially treated with a combination of intravenous antibiotics (ATBs): imipenem, amikacin, and ciprofloxacin. One month after the fracture fixation with a titanium nail, C. celerecrescens was again detected; therefore, metronidazole was added to the therapy. A triple combination of oral (PO) ATBs (trimethoprim-sulfamethoxazole, moxifloxacin, and metronidazole) followed, 8 weeks after the initial intravenous therapy. C. celerecrescens was cultured again two times, most recently in November 2019, when surgical debridement was supplemented by the topical administration of cancellous bone impregnated with vancomycin. Signs of bone healing were found at follow-ups and ATB treatment was finished in March 2020 after a total of 9 months of therapy. To this day, there have been no signs of reinfection. This case thus illustrates the need for a combination of systemic and individualized local therapy in the treatment of complicated cases of dual infections with rare pathogens.

Prevalence and Load of Cervical Ureaplasma Species With Respect to Intra-amniotic Complications in Women With Preterm Prelabor Rupture of Membranes Before 34 weeks.

Objectives: To determine the prevalence and load of Ureaplasma spp. DNA in the cervical fluid of women with singleton pregnancies complicated by preterm prelabor rupture of membranes (PPROM) with respect to intra-amniotic infection, sterile intra-amniotic inflammation, and colonization of the amniotic fluid. Methods: A total of 217 women with PPROM between gestational ages 24 + 0 and 33 + 6 weeks were included in this study. Paired amniotic and cervical fluid samples were collected at the time of admission via transabdominal amniocentesis and using a Dacron polyester swab, respectively. Microbial invasion of the amniotic cavity was diagnosed using a combination of culture and molecular biology methods. Intra-amniotic inflammation was determined based on the concentration of interleukin-6 in the amniotic fluid. Based on the presence or absence of these conditions, the women were stratified into the following subgroups: intra-amniotic infection (with both), sterile intra-amniotic inflammation (with inflammation only), colonization (with microorganisms only), and negative amniotic fluid (without either). The Ureaplasma spp. DNA load in the cervical fluid was assessed using PCR. Results: Ureaplasma spp. DNA in the cervical fluid was found in 61% (133/217) of the women. Women with negative amniotic had similar prevalence of Ureaplasma spp. DNA in cervical fluid (55%) to those with sterile intra-amniotic inflammation (54%) but lower than those with intra-amniotic infection (73%) and colonization (86%; p < 0.0001). Women with negative amniotic fluid had a lower load of Ureaplasma spp. DNA in their cervical fluid (median: 4.7 × 103 copies of DNA/ml) than those with intra-amniotic infection (median: 2.8 × 105 copies DNA/ml), sterile intra-amniotic inflammation (median: 5.3 × 104 copies DNA/ml), and colonization (median: 1.2 × 105 copies DNA/mL; p < 0.0001). Conclusion: In conclusion, in PPROM at <34 weeks, the presence of intra-amniotic infection, sterile intra-amniotic inflammation, or colonization of the amniotic fluid was associated with a higher prevalence and/or load of Ureaplasma spp. DNA in the cervical fluid than the absence of intra-amniotic complications.

Francisella tularensis caused cervical lymphadenopathy in little children after a tick bite: Two case reports and a short literature review.

Although Francisella (F.) tularensis is a well-described and understood zoonotic pathogen, its importance in Central Europe is relatively minor and, as such, tularaemia may be missed in the differential diagnosis. The annual incidence of tularaemia in the Czech Republic is relatively stable with up to 100 reported cases per year, except in the epidemic years 1998 and 1999 with 225 and 222 reported cases, respectively. It is, however, higher in comparison with the neighbouring countries. The common route of transmission in Central Europe is handling infected animals. Tularaemia is not commonly recognized as a tick-borne disease. Here we report two rare cases of a tick bite-associated ulceroglandular form of tularaemia in 2.5-year-old and 6.5-year-old children presenting with cervical lymphadenopathy. The unusual and interesting features of those cases are the young age and relatively uncommon route of transmission suggesting possible changes in the epidemiology of tularaemia in the Czech Republic. Therefore, the infection with F. tularensis should be considered in the differential diagnosis after a tick bite even in infants.

Intra-amniotic inflammation and birth weight in pregnancies with preterm labor with intact membranes: A retrospective cohort study.

Objective: To assess the association between newborn birth weight and the presence of intra-amniotic infection, presence of sterile intra-amniotic inflammation, and absence of intra-amniotic inflammation in pregnancies with preterm labor with intact membranes. Methods: A total of 69 pregnancies with preterm labor with intact membranes between gestational ages 22 + 0 and 34 + 6 weeks who delivered within seven days of admission were included in this retrospective cohort study. Transabdominal amniocentesis to determine the presence of microorganisms and/or their nucleic acids in amniotic fluid (through culturing and molecular biology methods) and intra-amniotic inflammation (according to amniotic fluid interleukin-6 concentrations) were performed as part of standard clinical management. The participants were further divided into three subgroups: intra-amniotic infection (presence of microorganisms and/or nucleic acids along with intra-amniotic inflammation), sterile intra-amniotic inflammation (intra-amniotic inflammation alone), and without intra-amniotic inflammation. Birth weights of newborns were expressed as percentiles derived from the INTERGROWTH-21st standards for (i) estimated fetal weight and (ii) newborn birth weight. Results: No difference in birth weights, expressed as percentiles derived from the standard for estimated fetal weight, was found among the women with intra-amniotic infection, with sterile intra-amniotic inflammation, and without intra-amniotic inflammation (with infection, median 29; with sterile inflammation, median 54; without inflammation, median 53; p = 0.06). Differences among the subgroups were identified in the birth weight rates, expressed as percentiles derived from the standard for estimated fetal weight, which were less than the 10th percentile (with infection: 20%, with inflammation: 13%, without inflammation: 0%; p = 0.04) and 25th percentile (with infection: 47%, with inflammation: 31%, without inflammation: 9%; p = 0.01). No differences among the subgroups were observed when percentiles of birth weight were derived from the birth weight standard. Conclusions: The presence of intra-amniotic inflammatory complications in pregnancies with preterm labor with intact membranes prior to the gestational age of 35 weeks was associated with a higher rate of newborns with birth weight less than the 10th and 25th percentile, when percentiles of birth weight were derived from the standard for estimated fetal weight.

Intra-amniotic infection and sterile intra-amniotic inflammation in women with preterm labor with intact membranes are associated with a higher rate of Ureaplasma species DNA presence in the cervical fluid.

OBJECTIVE: To determine the prevalence of Ureaplasma spp. DNA and its load in the cervical fluid in women with preterm labor with intact membranes (PTL) complicated by intra-amniotic infection (the presence of both microbial invasion of the amniotic cavity and intra-amniotic inflammation) or sterile intra-amniotic inflammation (the presence of intra-amniotic inflammation alone). METHODS: Overall, 115 women with singleton pregnancies complicated by PTL between gestational ages of 22 + 0 and 34 + 6 weeks were included in this study. Paired amniotic and cervical fluid samples were collected at the time of admission via transabdominal amniocentesis using a Dacron polyester swab. Microbial invasion of the amniotic cavity was diagnosed based on a combination of culture and molecular biology methods. Intra-amniotic inflammation was determined based on the concentration of interleukin-6 in the amniotic fluid. Bacterial and Ureaplasma spp. DNA loads were assessed in the cervical fluid using PCR. RESULTS: Intra-amniotic infection and sterile inflammation were identified in 14% (16/115) and 25% (29/115) of the women, respectively. Ureaplasma spp. DNA in the cervical fluid was identified in 51% (59/115) of women. The presence of Ureaplasma spp. DNA in the cervical fluid was higher in women with intra-amniotic infection (75% (12/16)) and sterile intra-amniotic inflammation (76% (22/29)) than in women without intra-amniotic inflammation (36% (25/70); p = .0002). Concurrent presence of Ureaplasma spp. and Mycoplasma hominis DNA was higher in women with intra-amniotic infection (42% (5/12)) than women with sterile intra-amniotic inflammation (7% (2/29)) and women without intra-amniotic inflammation (7% (5/70); p = .001). There were no differences in the load of Ureaplasma spp. DNA in the cervical fluid among women with intra-amniotic infection, sterile intra-amniotic inflammation, and those without intra-amniotic inflammation (median values; infection: 1.2 × 104 copies DNA/mL; sterile: 5.0 × 105 copies DNA/mL; without: 8.4 × 104 copies DNA/mL; p = .18). CONCLUSIONS: In PTL , both forms of intra-amniotic inflammation were associated with a higher prevalence of Ureaplasma spp. DNA in the cervical fluid. The presence of intra-amniotic infection was related to a higher rate of concurrent Ureaplasma spp. and M. hominis DNA in the cervical fluid.

Preterm prelabor rupture of membranes without microbial invasion of the amniotic cavity and intra-amniotic inflammation: a heterogeneous group with differences in adverse outcomes.

OBJECTIVE: The absence of microbial invasion of the amniotic cavity and intra-amniotic inflammation at the time of hospital admission is the most common condition associated with preterm prelabor rupture of membranes (PPROM). Although the intensity of intra-amniotic inflammatory response does not exceed the threshold for the diagnosis of intra-amniotic inflammation in this subgroup of PPROM, whether there could be differences in outcomes concerning the intensity of intra-amniotic inflammatory response remains unclear. Therefore, the main aims of this study on PPROM without microbial invasion of the amniotic cavity and intra-amniotic inflammation were (i) to characterize the association between the intensity of intra-amniotic inflammatory response, measured according to amniotic fluid interleukin (IL)-6 concentrations, and the presence of acute histological chorioamnionitis and acute inflammation in the amnion; (ii) to characterize the association between the intensity of intra-amniotic inflammatory response and fetal inflammatory response, and (iii) to describe the short-term morbidity of infants based on the intensity of intra-amniotic inflammatory response. METHODS: This retrospective study included 131 women with singleton pregnancies with PPROM without microbial invasion of the amniotic cavity and intra-amniotic inflammation between gestational ages of 24 + 0 and 36 + 6 weeks and who had delivered within 72 h of membrane rupture. Microbial invasion of the amniotic cavity was assessed based on a combination of cultivation and non-cultivation methods. Intra-amniotic inflammation was characterized based on the amniotic fluid IL-6 concentration. In addition, a histopathological assessment of the placenta was performed. Fetal inflammatory response syndrome was characterized according to IL-6 concentration in the umbilical cord blood of >11 pg/mL. Based on the quartiles of IL-6 concentrations in the amniotic fluid, these women were divided into four subgroups (from the lowest to the highest IL-6 concentrations). RESULTS: IL-6 concentrations in amniotic fluid were higher in women with acute histological chorioamnionitis (median: 819 pg/mL vs. 520 pg/mL; p = .003) and with acute inflammation of the amnion (median: 1116 pg/mL vs. 533 pg/mL; p = .0002) than in women without these complications. The rates of acute histological chorioamnionitis and acute inflammation of the amnion were the highest in the subgroup with IL-6 concentrations above the 75th percentile in amniotic fluid (chorioamnionitis, p = .02; amnion, p = .0002). No differences in IL-6 concentrations in amniotic fluid were identified between women with and without a fetal inflammatory response syndrome (p = .40). The rate of fetal inflammatory response syndrome did not vary among the amniotic fluid IL-6 quartile subgroups of women. Moreover, no differences were noted in short-term neonatal outcomes among the amniotic fluid IL-6 quartile subgroups. CONCLUSION: A higher intensity of the intra-amniotic inflammatory response, measured by amniotic fluid IL-6 concentrations, is associated with a higher rate of acute inflammatory lesions in the placenta in the subset of PPROM pregnancies without microbial invasion of the amniotic cavity and intra-amniotic inflammation.

Assessment of Streptococcus mutans biofilm formation on calcium phosphate ceramics: The role of crystalline composition and microstructure.

Streptococcus mutans is one of the bacteria that initiates the colonization of the pellicle at the tooth surface. It forms a plaque, together with other bacteria, which gradually dissolves the pellicle and leaves the tooth surface unprotected against the acidic oral environment. Calcium phosphate ceramics are excellent synthetic materials for the study of biofilm formation in dentistry because they are comparable to teeth in chemical composition and structure. Calcium phosphates can be processed to achieve a variety of crystalline compounds with biologically relevant ionic substitutions and structures that allow study of the effect of the surface chemistry and the topography independently. In this article, we describe the preparation and characterization of three types of calcium phosphate-based materials as a suitable surface for the formation of the S. mutans biofilm: beta-tricalcium phosphate (ß-TCP); sintered hydroxyapatite (SHA); and calcium-deficient hydroxyapatite (CDHA). The densest biofilms were formed on the surfaces of SHA and CDHA, with no significant differences due to the stoichiometry or microstructure. In contrast, ß-TCP showed a lower susceptibility to S. mutans biofilm formation, suggesting that the crystalline structure is the controlling parameter. Subsequently, SHA was selected to develop a dental biofilm model that allowed study of S. mutans biofilm susceptibility to chlorhexidine and ethanol.

Clinical characteristics of colonization of the amniotic cavity in women with preterm prelabor rupture of membranes, a retrospective study.

To determine the main clinical characteristics of preterm prelabor rupture of membranes (PPROM) complicated by colonization of the amniotic cavity (microbial invasion of the amniotic cavity without intra-amniotic inflammation). A total of 302 women with PPROM were included. Transabdominal amniocentesis was performed and amniotic fluid was assessed. Based of microbial invasion of the amniotic cavity and intra-amniotic inflammation (interleukin-6 ≥ 3000 pg/mL), the women were divided into following groups: intra-amniotic infection, sterile intra-amniotic inflammation, colonization of the amniotic cavity, and negative amniotic fluid. Colonization was found in 11% (32/302) of the women. The most common bacteria identified in the amniotic fluid were Ureaplasma spp. with a lower burden than those with intra-amniotic infection (p = 0.03). The intensity of intra-amniotic inflammatory response measured by interleukin-6 was higher in women with colonization than in those with negative amniotic fluid (medians: 961 pg/mL vs. 616 pg/mL; p = 0.04). Women with colonization had higher rates of acute inflammatory placental lesions than those with negative amniotic fluid. In PPROM, colonization, caused mainly by microorganisms from the lower genital tract, might represent an early stage of microbial invasion of the amniotic cavity with a weak intra-amniotic inflammatory response.

Effect of Restriction of Fluoroquinolone Antibiotics on Clostridioides difficile Infections in the University Hospital Hradec Králové.

Clostridioides difficile is the most common pathogen responsible for hospital-acquired diarrhea. This complication of antibiotic treatment mainly endangers the health of elder patients. Preventing the development of C. difficile infections (CDI) is still a challenge that needs to be addressed. In our study, the results of 872 C. difficile positive stool samples were used to describe the epidemiological situation affected by a change in the prescription of fluoroquinolone antibiotics. In a total, 93 of strains were typed by polymerase chain reaction (PCR) and capillary gel electrophoresis. Between years 2014 and 2018 the decline in the fluoroquinolones consumption was 69.3 defined daily dose (DDD) per 1000 patient-days (from 103.3 to 34.0), in same period CDI incidence declined by 1.3 cases per 10,000 patient-bed days (from 5.6 to 4.3). Results of epidemiologic and statistical analysis shows that decline in fluoroquinolones consumption has significant influence on CDI incidence and prevalence of hypervirulent strains. In the University Hospital Hradec Králové properly managed antibiotic stewardship policy has reduced CDI incidence by 23.2% and lowered rate of hypervirulent ribotypes 001 and 176.

Isolation of Bordetella trematum from the respiratory tract of a patient with lung cancer: a case report.

We report the case of isolation of Bordetella trematum from the respiratory tract of a patient with lung carcinoma. This gram-negative, opportunistic rod was firstly described in 1996. To date, only several strains of Bordetella trematum have been isolated and reported, mostly from skin and soft tissue infections. The patient was admitted to the ICU of the Pulmonary Department in incipient septic shock with respiratory failure. Intravenous fluid resuscitation and non-invasive ventilation were administered immediately. A broad spectrum antibiotic piperacillin/tazobactam was administered empirically after sampling of material for microbiological examination. The bronchoscopy showed a large cavern of decayed tumour invading into mediastinum. Both sample cultures showed significant quantities of gram-negative non-fermenting bacteria. The isolate was identified using MALDI-TOF MS as Bordetella trematum and the identification was confirmed using 16S ribosomal RNA sequencing. In the last few years, routine bacterial identification using MALDI-TOF MS has enabled correct discrimination of this species. Nevertheless, isolation of Bordetella trematum in clinical samples is still very uncommon, and it is appropriate to confirm the species identification via 16S ribosomal RNA sequencing. To our knowledge, this is the first case of B. trematum isolated from the human respiratory tract since its first description. The clinical significance of Bordetella trematum in the rapid deterioration of the patient's status remains unclear.

Comprehensive proteomic investigation of infectious and inflammatory changes in late preterm prelabour rupture of membranes.

Preterm prelabour rupture of membranes beyond the 34th week of gestation (late PPROM) is frequently associated with the risk of the microbial invasion of the amniotic fluid (MIAC) and histological chorioamnionitis (HCA). Hence, we employed a Tandem Mass Tag-based approach to uncover amniotic fluid proteome response to the presence of MIAC and HCA in late PPROM. Protein dysregulation was associated with only five cases in the group of 15 women with confirmed MIAC and HCA. Altogether, 138 amniotic fluid proteins were changed in these five cases exclusively. These proteins were particularly associated with excessive neutrophil responses to infection, such as neutrophil degranulation and extracellular trap formation. We believe that the quantification of these proteins in amniotic fluid may assist in revealing women with the highest risk of excessive inflammatory response in late PPROM.

Antimicrobial effect of OKCEL® H-D prepared from oxidized cellulose.

The antimicrobial effect of OKCEL® H-D, a topical, absorbable hemostatic textile prepared from oxidized cellulose, was tested. Testing by dilution and diffusion methods was conducted on a spectrum of 27 select microorganisms, including also antibiotic-resistant strains. OKCEL® H-D showed inhibitory effects on nearly all tested bacteria. In testing using the dilution suspension method, the majority of bacteria showed decrease in cell density by 7-8 orders of magnitude after just 6 h of exposure. For clinical isolates of antibiotic-resistant strains, a reduction occurred after 24 h of exposure. In testing the antimicrobial effects of OKCEL® H-D by the dilution method was least effective on spore-forming Bacillus subtilis, for which no antimicrobial effect was detected after 48 h, and on Mycobacterium smegmatis, for which the number of cells decreased by four orders of magnitude only after 24 h. By the diffusion method, inhibition zones were recorded for nearly all test microorganisms except for Staphylococcus aureus, M. smegmatis, and Listeria monocytogenes. No growth beneath the tested OKCEL® H-D material was recorded, however, even for the latter-named bacteria strains, which attests to its good inhibitory effect.