Krzysztof Kaczka

Acinetobacter Baumannii Nosocomial Infections

UNLABELLED Nosocomial infections caused by strains Acinetobacter baumannii strands are a growing clinical problem. The occurrence of multidrug-resistant strands is observed and that limits the ways of therapy considerably. The aim of the study was to determine the rate of infection and susceptibility spectrum of the species Acinetobacter baumannii isolated from patients treated at Maria Skłodowska-Curie Memorial Hospital in Zgierz with particular emphasis on surgical wards. MATERIALS AND METHODS The material consisted of Acinetobacter baumannii isolates were obtained from samples of materials from patients treated at Maria Skłodowska-Curie Memorial Hospital in Zgierz from January to December 2011. Isolated bacterial strains were cultured at microbiological substrates. Isolates were identified to species using the VITEK 2 GN card (bioMérieux) and Vitek 2 automated system (bioMérieux). Susceptibility towards antibiotics of particular strains was determined by the means of AST NO 93 card. In the case of resistance towards carbapenem, the MIC was marked by E-test with Mueller Hinton substrate. The occurrence of MBL was verified by the means of disc system with Mueller Hinton substrate. RESULTS We have shown that total number of Acinetobacter baumannii infections at hospital was 140 (10,31% of total results of cultures). Percentage of Acinetobacter baumannii infections at wards: Intensive Care Unit 48%, Surgical Departments 20%, Internal Diseases Department 16%, Neurology 13%, other wards - 3%. The susceptibility percentage of Acinetobacter Baumannii against antibiotics: colistin 90%, imipenem 64%, meropenem 43%, ampicillin-sulbactam 28%, amikacin 27%, gentamicin 24%, cefepime 9%, ceftazidime 7%, ciprofloxacin 7% CONCLUSIONS Acinetobacter baumannii infections are a significant proportion of nosocomial infections. Most relate to surgical wards and ICUs. Acinetobacter baumannii is resistant against most antibiotics. The highest percentage of sensitivity demonstrated for colistin and carbapenems.

Detection of lymph node metastases of papillary thyroid cancer—comparison of the results of histopathology, immunohistochemistry and reverse transcription-polymerase chain reaction—a preliminary report

BackgroundThe range of lymphadenectomy in differentiated thyroid cancer remains still a matter of controversy because of the lack of reliable diagnostic methods for nodal metastases, other than histopathology.AimTo compare the results of detection of lymph node metastases of papillary thyroid cancer by conventional histopathology and immunohistochemistry with the results of reverse transcription–polymerase chain reaction for thyroglobulin mRNA.Patients and methodsEach of 166 cervical lymph nodes obtained from 21 patients was divided into two halves: one was used for conventional histopathology and immunohistochemistry, the other part was investigated by molecular examination.ResultsWe obtained different results from examination of the lymph nodes in six (28.6%) patients. In four patients (19.1%) reverse transcription–polymerase chain reaction (RT-PCR) was more sensitive in detection of positive lymph nodes; in two patients (9.5%) it revealed fewer metastasised lymph nodes than did histopathology. The rest of the patients did not have any differences: 12 (57.1%) of them had negative lymph nodes and three (14.3%) had positive lymph nodes in all examinations.Conclusions(1) Thyroglobulin (Tg) RT-PCR is an appropriate method of detection for thyroid cancer cells. (2) In combination with histopathology, it might help to qualify patients’ nodal status better.

Sentinel lymph node in thyroid tumors – own experience

Aim of the study To determine the feasibility of sentinel lymph node biopsy (SLNB) for the evaluation of the cervical lymph node status in patients with thyroid tumors. Material and methods Twenty-three patients with suspected thyroid cancer were enrolled in the study. 0.5–1.0 ml of 1% Patent Blue dye was injected intratumorally. After SLNB, thyroidectomy and proper lymphadenectomy were performed. Results Sentinel lymph node was detected in 20 (86.9%) patients. Thirty-one SLNs were found – 21 (67.7%) were located in the central neck compartment, 4 (12.9%) in the lateral neck compartment, 6 (19.4%) in the upper mediastinum. The number of SLNs ranged from 1 to 3 (mean 1.6). Sentinel lymph node was positive in 5 (25%) patients, negative in 15 (75%) in the final histopathology. Sentinel lymph nodes were located only in the central neck compartment in 13 patients, and in both the central and lateral neck compartments in 2 patients. In one patient, SLNs were located only in the central neck compartment and upper mediastinum. Three patients had SLNs only in the upper mediastinum, while one had them only in the lateral neck compartment. In one patient a node regarded as SLN was negative, while there were metastases in removed non-sentinel lymph nodes (NSLNs). In two patients, histopathology of SLNs showed that they were actually parathyroid glands. Conclusions Our results confirm that thyroid cancer SLNB is rather easy to carry out. Its performance along with intraoperative examination can help to avoid unnecessary lymphadenectomy. However, it should be kept in mind that parathyroid glands can be stained and removed by mistake during SLNB.

First one-step nucleic acid amplification testing in papillary thyroid cancer lymph nodes - a comparison with histopathology and real-time PCR.

INTRODUCTION The significance of lymph node metastases and the optimal extent of lymphadenectomy remain matters of controversy in papillary thyroid cancer. This study was designed to assess the feasibility and reliability of OSNA and real-time PCR for CK19 and TG mRNA in papillary thyroid cancer lymph nodes evaluation compared to standard histopathology. MATERIAL AND METHODS Each of 92 randomised lymph nodes from 32 papillary thyroid cancer patients were divided into representative parts and assessed using the three studied methods. RESULTS Eighteen (19.6%) lymph nodes from ten (31.3%) patients were positive according to histopathology. When the cut-off value distinguishing metastatic from non-metastatic lymph nodes in the OSNA assay was set at 250 copies per microlitre, the results were positive in 16 (17.4%) lymph nodes from 11 (34.4%) patients. Twenty three (25%) lymph nodes were tested positive in real-time PCR for TG mRNA. Real-time PCR for CK19 mRNA was positive in 18 (19.6%) lymph nodes from 13 (40.6%) patients. No statistically significant differences were noted between the diagnostic accuracy of either molecular method compared to the histopathological examination (p = 0.81). Overall, 20 positive molecular biology results were noted in patients with negative histopathology results. Conversely, in 18 lymph nodes, despite a metastasis finding in histopathology, at least one molecular test yielded a negative result. CONCLUSIONS It was revealed that OSNA is a reliable technique for the evaluation of lymph node metastases in papillary thyroid cancer. This method was shown to have equivalent accuracy to histopathology and real-time PCR.

Early Predictors of Post – Thyroidectomy Hypoparathyroidism

Thyroid surgery is the most commonly performed procedure in the field of endocrine surgery. Studies are still ongoing on the development of a single algorithm for diagnosis and care of patients at risk of postoperative hypoparathyroidism. The aim of the study was to determine the biochemical marker that would allow the most accurate diagnosis of patient groups at risk of developing hypoparathyroidism and to identify risk factors for this disorder. MATERIAL AND METHODS The prospective study included 142 consecutive patients undergoing total thyroidectomy for benign goiter from January 1st 2014 to December 31st 2015. Serum intact parathyroid hormone (iPTH), total calcium (Ca), phosphate (P), and magnesium (Mg) levels have been measured preoperatively and at 1, 6, 24, and 48 h postoperatively. RESULTS Clinical symptoms of hypoparathyroidism developed in 25 (17.6%) of 142 patients. The best diagnostic accuracy for hypoparathyroidism based on ROC curves was obtained for iPTH at 6h (AUC 0.942; 95% CI: 0.866-1.000, p<0.001) and its percentage change from baseline ΔiPTH at 6h (AUC 0.930; 95% CI: 0.858-1.000, p<0.001). In an multivariate analysis, the preoperative Ca level higher by 0.1 mmol/l, and iPTH level higher by 0.1 pmol/l were associated with a lower risk of hypoparathyroidism, by 68% (p=0.012) and 61% (p=0.007), respectively. A 1% decline in iPTH from baseline increased the risk of hypoparathyroidism by 15% (p<0.001). CONCLUSIONS The most reliable markers indicating a high risk of postoperative hypoparathyroidism are the decline in ΔiPTH at 6h by > 65% or iPTH level at 6h <1.57 pmol /l. A postoperative decline in iPTH levels is an independent risk factor for the development of hypoparathyroidism. Preoperative higher concentrations of Ca and iPTH are protective factors for the development of this disorder.

Analysis of clinical significance of equivocal thyroid cytology with a special consideration for FLUS category — five years of new classification of FNA results

INTRODUCTION The diagnostic category of follicular lesion of undetermined significance (FLUS) was intended to allow selection of cases with low risk of malignancy from all smears with indeterminate, suspicious cytology (ISC), which can potentially take advantage from repeat fine-needle aspiration (rFNA). Aim of the study was a comparison of the risk of malignancy related to FLUS nodules and other nodules with ISC: suspected follicular neoplasm (SFN) and suspected malignancy (SM), as well as analysis of the usefulness of assessing ultrasonographic malignancy risk features (UMRF) in nodules with ISC. MATERIAL AND METHODS We analysed UMRF, rFNA, and results of histopathological examination (H) in 441 FLUS, 135 SFN, and 72 SM nodules. RESULTS The frequency of exposing cancer in H in FLUS nodules was 5.9%, and when cytological follow up was also included it was 2.9%. rFNAs made the diagnosis more precise in 72.7% of FLUS, and in 5.2% it was diagnosis/suspicion of cancer. The incidence of cancer in SFN nodules was 8.2%, in SM nodules with suspicion of papillary cancer - 61.1%, and in nodules with suspicion of other or unspecified malignancy - 53.8% (p < 0.0001 FLUS vs. both groups). The presence of calcifications is the only independent UMRF for nodules with ISC (OR 4.7). Features of importance are also microcalcifications (OR 3.8), especially in the SM group, and taller-than-wide-shape (OR 2.2). FLUS and SFN nodules are characterised by particularly low value of assessing suspicious margins; analysis of hypoechogenicity is of low value in SFN nodules, like suspected vascularisation in SFN and SM nodules. CONCLUSIONS The risk of cancer in FLUS and SFN nodules is lower than in SM nodules. rFNAs of FLUS nodules make the diagnosis more precise in more than 70% of cases and are effective in revealing cancers. UMRFs present variable diagnostic value depending on the subcategory of ISC.

One-step nucleic acid amplification testing in medullary thyroid cancer lymph nodes: a case series.

Introduction Locoregional relapse in medullary thyroid cancer (MTC) may be caused by nodal micrometastases. Medullary thyroid cancer lymph nodes have not yet been evaluated by one-step nucleic acid amplification (OSNA). Therefore, the aim of this study was to detect MTC cells by OSNA in cervical lymph nodes and compare the obtained outcomes with conventional histopathology. Material and methods Twenty-one randomized, unenlarged lymph nodes from 5 patients with MTC were examined by histopathology and OSNA. Lymph nodes were divided into four representative blocks by a sterile, single use, special cutting device in the same way as in the clinical protocol study performed by Tsujimoto et al. Two blocks were used for histopathology and immunohistochemistry, 2 for OSNA. Results Positive results of histopathology and OSNA were revealed in 4 patients. The outcomes of OSNA and histopathology were corresponding in 3 patients. Positive histopathology results of 2 lymph nodes from 2 patients were confirmed by OSNA. In 1 patient there were only negative results of both examinations. One-step nucleic acid amplification failed to detect metastasis in 1 lymph node in 2 patients although it did not change the TNM status in these patients. There were no false positive results in the OSNA test. Conclusions One-step nucleic acid amplification may be an alternative method to histopathology in detecting nodal involvement in MTC. Further studies should evaluate the sensitivity and specificity of OSNA and the impact on staging in MTC.

Cholelithiasis – always infected?

This study aims to present results regarding the presence and identification of bacterial strains found in bile and gallstones located in the gallbladder and bile ducts in patients operated on due to cholelithiasis. MATERIALS AND METHODS Bacterial culture was evaluated in 92 patients. There were 54 women (59%) and 38 men (41%) who underwent surgery on account of cholelithiasis and /or gallstones in bile ducts between 2013 and 2014. Bile and gallstone samples were cultured intraoperatively for bacteria; bacterial strains were identified, and their sensitivity to antibiotics was determined. Molecular methods (NGS and Sanger method) were used to separate bacterial strains in one of the gallbladder stones and the results were compared with bacterial strains grown from the bile. RESULTS Bile cultures were positive in 46 patients that is, 50% of the study group. The following bacteria strains were grown: Enterococcus spp. (44%), Escherichia coli (37%) and Klebsiella spp. (35%). Candidiasis accompanied by bacterial infection was detected in 7 patients (15%). Molecular testing of gallstones revealed DNA of Enterococcus spp., Escherichia spp., Streptococcus spp. and Clostridium spp. In the bile culture of the same patient Enterococcus spp. (avium and faecalis) was detected. Conclusion 1. More than one pathogen was grown on samples obtained from 31 patients (70%) with bile infection. 2. The most common pathogens include Enterococcus spp., Escherichia coli and Klebsiella spp. 3. Bacterial infections are often accompanied by a fungal infection (Candida albicans) 4. Bacterial strains grown from a gallstone sample partially corresponded with strains identified in the bile of the same patient.

One-step nucleic acid amplification analysis of sentinel lymph nodes in papillary thyroid cancer patients

Introduction It is essential to look for methods to define the need for central lymphadenectomy for papillary thyroid cancer patients. The aim is to determine the efficacy of one-step nucleic acid amplification (OSNA) and sentinel lymph node (SLN) biopsy in the intraoperative detection of nodal involvement. Material and methods This prospective, experimental study enrolled 49 patients with clinically negative lymph nodes. Intraoperatively, 1% Patent Blue dye was injected intratumorally. Lymph nodes that stained blue were defined as SLNs. They were directly cut into blocks at 2-mm intervals. Nonadjacent blocks were subjected to either the OSNA assay or histological examination. Results Sixty-five SLNs were found in 43 (87.8%) patients. There were 20 (30.8%) histopathologically positive SLNs. According to the OSNA, 22 (33.8%) SLNs were positive. The OSNA results were different from histopathology in 8 (12.3%) SLNs. The OSNA gave a positive result in 5 (7.7%) SLNs, while they were not involved according to the histopathology. However, OSNA upstaged N status from N0 to N1 only in 2 (3.1%) patients. Inverse results (histopathology +, OSNA–) were obtained in 3 (4.6%) SLNs. Positive and negative predictive values (PPV and NPV) for OSNA were 0.77 and 0.93, respectively. The concordance rate between examinations was 85.5%. Conclusions In some patients with clinically negative lymph nodes, OSNA and SLN biopsy may prevent unnecessary central lymphadenectomy. On the other hand, the sentinel lymph node biopsy may reveal the presence of potentially involved sentinel lymph nodes outside the central compartment. These SLNs can also be assessed by means of OSNA.

Jednostopniowa amplifikacja kwasównukleinowych (ang. One-Step Nucleic AcidAmplification – OSNA) – nowa metodawykrywania przerzutów guzów litych dowęzłów chłonnych

Introduction and purpose. None of the applied methods of lymph node assessment is perfect. The need to wait a long time for the results is the disadvantage of postoperative histopathological examination. In the case of a positive outcome of the post-operative histopathology of sentinel lymph node it is necessary to perform a second surgery. Some metastases smaller than 2 mm can be omitted in routine histopathology. In turn, frozen section examination, with relatively high specificity, has lower sensitivity than postoperative histopathology. Molecular assays, using PCR techniques, with high sensitivity, have lower specificity. In addition, except for real-time PCR, all other techniques are not suitable for use as an intraoperative examination due to the long waiting time for the result. OSNA is one of the latest methods of lymph node assessment in solid tumours. The article summarizes world reports on the application of this technique in the evaluation of lymph nodes in patients with solid tumours. Summary of the state of knowledge. The OSNA test is based on mRNA for cytokeratin 19(CK19) amplification. At present, this test is often performed in the search for lymph nodes metastases in patients with breast cancer. Intraoperative evaluation of sentinel lymph nodes in breast cancer, using the OSNA method, allows deciding whether or not to perform lymphadenectomy. In other cancers, the use of the OSNA method has been evaluated in numerous clinical trials. Summary. The OSNA method could become a routine technique for the assessment of lymph nodes. It has a relatively high sensitivity and allows evaluation of lymph nodes in 30 minutes, which enables its use as an intraoperative examination.