Lenka Veverková

The anatomical relationship between the saphenous nerve and the great saphenous vein

Objective Damage to the saphenous nerve (SN) has been a known complication during varicose vein surgeries. We tested whether a better knowledge of the anatomy of the SN and the great saphenous vein (GSV) can prevent such damage. Methods We conducted a morphological and histological examination on 86 limbs from 43 cadavers in order to analyse the anatomical interrelation between the SN and the GSV in the lower leg and we also measured the distance between the nerve and the vein in a sample of 42 sections from three parts of the lower leg. Results The anatomical relationship between the SN and the GSV is varied and the two structures run close to each other so a better knowledge of their anatomy in itself proved insufficient in preventing damage to the SN. Conclusion However, in the case of endovenous laser therapy and radiofrequency ablation tumescent anaesthesia decreases the risk of damage to the SN.

Overexpression of c-Myb is associated with suppression of distant metastases in colorectal carcinoma

The MYB gene codes for the c-Myb transcription factor maintaining proliferation of colon epithelial progenitors, thus controlling colon development and homeostasis. This gene is overexpressed in early phases of colorectal cancer (CRC) tumorigenesis. The aim of this study was to examine the expression of c-Myb in CRC tissue samples both at the messenger RNA (mRNA) and protein levels and to evaluate their associations with clinicopathological characteristics in a group of 108 CRC patients. Statistically significant negative association was found between the frequency of the c-Myb-positive tumor cells assessed by immunohistochemistry and the presence of distant metastases (p < 0.01) but not tumor differentiation, tumor stage, lymph node involvement, vascular invasion, tumor localization, age, and gender of the patients. Although the c-Myb protein level in the tumor tissue correlated with its mRNA level, no significant association between MYB mRNA and any clinicopathological characteristics was observed. We conclude that albeit overexpression of c-Myb is considered as an important factor contributing to early phases of CRC tumorigenesis, it may later have negative effect on tumor cell dissemination as observed recently in breast cancer as well. Further studies are required to explain the role of c-Myb during formation of CRC distant metastases.

Neovascularization after ischemic conditioning of the stomach and the influence of follow-up neoadjuvant chemotherapy thereon

Introduction Esophagectomy and reconstruction remain the optimal treatment for patients with resectable esophageal cancer. Neovascularization after ischemic conditioning of the stomach before esophagectomy is a laparoscopic procedure which may potentially reduce gastric conduit ischemia. Aim To investigate the influence of ischemic conditioning on neovascularization along the greater curvature of the stomach and to explore the effect of neoadjuvant chemotherapy on neovascularization after ischemic conditioning. Material and methods Staging laparoscopy was performed before the main resection procedure; during this procedure ischemic conditioning was performed. Samples taken from the human stomach were divided into 3 groups: group A – patients after ischemic conditioning with a delay of 30–45 days after left gastric artery (LGA) ligation (n = 4); group B – patients who were undergoing neoadjuvant chemotherapy with a delay of 90–140 days after left gastric artery ligation (n = 4); and control group C – patients without ischemic conditioning (n = 7). Results After ischemic conditioning with a delay of 30–45 days, the count of neovessels along the greater curvature of the stomach increased from 5.4 ±0.7 in the control group to 17.5 ±0.9 in a low-power field of view (LPF) in group A and increased still further on average to 19.8 ±10.4 in group B. Conclusions Left gastric artery ligation only is a sufficient procedure for ischemic conditioning of the stomach. Neovascularization along the greater curvature is a continuous process that depends on delay time. Neoadjuvant therapy has no influence on the effect of neovascularization.

Decubitus as a Cause of Death even in the 21st Century

Cil: Cilem studie bylo analyzovat výskyt dekubitů u pacientů hospitalizovaných a podstupujicich operaci na 1. chirurgicke klinice u sv. Anny v roce 2015. Metodika: Retrospektivni analýza administrativnich dat z nemocnicniho informacniho systemu a z dat specialniho elektronickeho nastroje (I-hojeni.cz). Použit byl statistický software R, statisticka analýza byla provedena pomoci Pearsonova chi-kvadratu na hladině významnosti 0,05. Výsledky: Z celkoveho poctu 3 937 hospitalizovaných podstoupila větsina operacni výkon (n = 3 431; 86,4 %). Nově se vytvořil dekubitus u 31 pacientů. Průměrný věk pacientů byl 82,6 let, jednalo se větsinou o ženy (18 : 13). Body Mass Index (BMI) byl průměrně 25,37; delka hospitalizace 26,67 dni. Průměrný stupeň dekubitu byl 1,64. Pět pacientů mělo vice než jeden dekubitus. Nejcastěji se vyskytovaly dekubity na patě, nasledně na hýždich a sakru. Nezjistili jsme statisticky významný vztah mezi BMI a vznikem dekubitů (p > 0,05). Věk pacientů a delka hospitalizace statisticky významně souvisely s výskytem dekubitů (p = 0,02493). Zavěr: Přes vsechna preventivni opatřeni vznikaji dekubity u chirurgicky lecených pacientů i ve 21. stoleti a jejich lecba je nakladna a dlouha.

Comparison of Two Non-Invasive Methods of Microbial Analysis in Surgery Practice: Incision Swabbing and the Indirect Imprint Technique

BACKGROUND A variety of methods exist to take samples from surgical site infections for cultivation; however, an unambiguous and suitable method has not yet been defined. The aim of our retrospective non-randomized study was to compare two non-invasive techniques of sampling material for microbiologic analysis in surgical practice. We compared bacteria cultured from samples obtained with the use of the swab technique, defined in our study as the gold standard, with the indirect imprint technique. METHODS A cotton-tipped swab (Copan, Brescia, Italy) was used; the imprints were taken using Whatman no. 4 filter paper (Macherey-Nagal, Duren, Germany) cut into 5×5 cm pieces placed on blood agar in a Petri dish. To culture the microorganisms in the microbiology laboratory, we used blood agar, UriSelect 4 medium (Bio-Rad, Marnes-la-Coquette, France), and a medium with sodium chloride (blood agar with salt). After careful debridement, a sample was taken from the incision surface by swab and subsequently the same area of the surface was imprinted onto filter paper. The samples were analyzed in the microbiology laboratory under standard safety precautions. The cultivation results of the two techniques were processed statistically using contingency tables and the McNemar test. Those samples that were simultaneously cultivation-positive by imprint and -negative by swabbing were processed in greater detail. RESULTS Over the period between October 2008 and March 2013, 177 samples from 70 patients were analyzed. Sampling was carried out from 42 males and 28 females. One hundred forty-six samples were from incisions after operations (21 samples from six patients after operation on the thoracic cavity, 73 samples from 35 patients after operation on the abdominal cavity combined with the gastrointestinal tract, 52 samples from 19 patients with other surgical site infections not included above) and 31 samples from 11 patients with no post-operative infection. One patient had a sample taken both from a post-operative and a non-post-operative site. Coincidently, the most frequent cultivation finding with both techniques was a sterile one (imprint, 62; swab, 50). The microorganism cultivated most frequently after swabbing was Pseudomonas aeruginosa (22 cases), compared with Escherichia coli when the filter paper (imprint) was used (31 cases). The imprint technique was evaluated as more sensitive compared with swabbing (p=0.0001). The κ statistic used to evaluate the concordance between the two techniques was 0.302. Of the 177 samples there were 53 samples simultaneously sterile using the swab and positive in the imprint. In three samples colony- forming units (CFU) were not counted; 22 samples were within the limit of 0-25×10(1) CFU/cm(2), 20 samples within the limit of 25×10(1)-25×10(2) CFU/cm(2), five within the limit of 25×10(2)-25×10(3) CFU/cm(2), and three of more than 25×10(4) CFU/cm(2). CONCLUSIONS The hypothesis of swabbing as a more precise technique was not confirmed. In our study the imprint technique was more sensitive than swabbing; the strength of agreement was fair. We obtained information not only on the type of the microorganism cultured, but also on the number of viable colonies, expressed in CFU/cm(2).

Laser-induced fluorescence spectroscopy in tissue local necrosis detection

The recent effort leads to reliable imaging techniques which can help to a surgeon during operations. The fluorescence spectroscopy was selected as very useful online in vivo imaging method to organics and biological materials analysis. The presented work scopes to a laser induced fluorescence spectroscopy technique to detect tissue local necrosis in small intestine surgery. In first experiments, we tested tissue auto-fluorescence technique but a signal-to-noise ratio didn’t express significant results. Then we applied a contrast dye - IndoCyanine Green (ICG) which absorbs and emits wavelengths in the near IR. We arranged the pilot experimental setup based on highly coherent extended cavity diode laser (ECDL) used for stimulating of some critical areas of the small intestine tissue with injected ICG dye. We demonstrated the distribution of the ICG exciter with the first file of shots of small intestine tissue of a rabbit that was captured by high sensitivity fluorescent cam.

Indirect imprint sampling technique using a filter paper pad for microbial analysis

Compounds present in oil sludge such as polycyclic aromatic hydrocarbons (PAHs) are known to be cytotoxic, mutagenic and potentially carcinogenic. Microorganisms including bacteria and fungi have been reported to degrade oil sludge components to innocuous compounds such as carbon dioxide, water and salts. In the present study, we isolated different bacteria with PAH-degrading capabilities from compost prepared from oil sludge and animal manures. These bacteria were isolated on a mineral base medium and mineral salt agar plates. A total of 31 morphologically distinct isolates were carefully selected from 5 different compost treatments for identifi cation using polymerase chain reaction (PCR) of the 16S rRNA gene with specifi c primers (universal forward 16S-P1 PCR and reverse 16S-P2 PCR). The amplicons were sequenced and sequences were compared with the known nucleotides from the GenBank. The phylogenetic analyses of the isolates showed that they belong to 3 different clades; Firmicutes, Proteobacteria and Actinobacteria. These bacteria identifi ed were closely related to the genera Bacillus, Arthrobacter, Staphylococcus, Brevibacterium, Variovorax, Paenibacillus, Ralstonia and Geobacillus. The results showed that Bacillus species were predominant in all composts. Based on the results of the degradation of the PAHs in the composts and results of previous studies on bacterial degradation of hydrocarbons in oil, the characteristics of these bacterial isolates suggests that they may be responsible for the breakdown of PAHs of different molecular weights in the composts. Thus, they may be potentially useful for bioremediation of oil sludge during compost bioremediation. Unauthenticated Download Date | 10/19/16 10:12 PM 68 O. Ubani, H.I. Atagana, M.S. Thantsha and thermophilic activities. Microbial activities in compost generate high temperature, which increases solubility of contaminants and induces microbial co-metabolic activity (Sheetal 2012). In compost, there is abundance of nutrients and organic matter that have effect on microbial degradation of oil sludge constituents (Sheetal 2012). As the compost matures, the pollutants are degraded, digested, metabolised and transformed into humus as well as inert products such as carbon dioxide, water and salts. In this study, the initial concentrations of the PAHs (2 to 6 rings) detected were between 1.44 mg/kg to 205.81 mg/kg before the co-composting process of oil sludge. The results obtained showed reduction in selected PAHs (66% to 80%) in all co-composting piles over a period of ten months, as shown in Table 3. This result is in agreement with the report from comparison of bioaugmentation and composting for remediation of oil sludge (Ouyang et al. 2005). The results obtained from the compost piles showed that composting can be used to degrade PAHs present in oil sludge, suggesting the existence of active bacterial community in the compost (Katsivela et al. 2003). This study was to determine whether microbial growth and activities were enhanced as well as bacteria community involved in the degradation of oil sludge (PAHs) during the composting period. Culture dependent and non-culture dependent methods have been used to determine the composition of the metabolically active microorganisms that played an important role in the degradation of pollutants (Saman 2010). Culture dependent methods (CDM) often involve proper isolation and in vitro cultivation of microorganisms based on their required growth factors (Saman 2010), while non-culture dependent method is used for identifi cation of uncultivable microorganism. These methods complement each other to enable the identifi cation of more than 90% microorganisms. Perhaps, the most widely used non-culture dependent method is the molecular method. This usually involves direct DNA extraction from samples, PCR amplifi cation and nucleotide sequence for identifi cation of microbial diversity based on the analysis of the 16SrRNA gene sequences (Mokno-Tlili et al. 2009, Inceoglu et al. 2010). It is a reliable method that can be used to detect diversity, quantity and sometimes viability of microbial contents of a sample (Inceoglu et al. 2010, Saman, 2010). However, it seems that combination of both methods is always essential in isolation and characterisation of bacterial isolates. The aim of this study was to isolate, identify and characterise bacterial consortium present in a compost system with potentials to degrade PAHs present in oil sludge. Materials and methods Soil Garden soil was air-dried and analysed to determine the soil type, organic carbon content, total nitrogen content, total phosphorus content (C; N and P), soil pH and water holding capacity (WHC), using concentrated acid digestion method (CADM) with Induction Coupled Plasma (ICP-OES Optima 4300 DV, Perkin Elmer, Waltham, MA, USA). Oil sludge Oil sludge from a refi nery was soxhlet extracted with dichloromethane as the solvent and characterized by gas chromatography/mass spectrometry (GC/MS) method (US EPA 827Asymptomatic urinary tract infections resulting from poor diagnosis in pregnant women poses a serious threat of complications during pregnancy. This study reports the prevalence of Urinary Tract Infections (UTIs) and the bacteriological causative agents among pregnant women attending antenatal clinics at a Tertiary Care Hospital in Ile Ife, South western Nigeria. 335 mid-stream clean catch urine samples were collected from pregnant women and cultured for the presence of bacterial pathogens. All the isolates were subjected to antibiotic sensitivity testing on Mueller Hinton agar by modified Kirby-bauer disk diffusion technique using CLSI, guidelines. Data were analyzed using SPSS for Windows version 16. A total of 190 showed significant bacterial growth while 145 showed no significant bacterial growth. Bacterial agents isolated included Escherichia coli showing the highest occurrence of [60(31.5%) followed by Klebsiella spp and Staphylococcus aureus with prevalences of [49(25.7%) and [32(16.8%) respectively. Other organisms implicated are Proteus mirabilis [20(10.5%), Coagulase Negative Staphylococci [24(12.3%) and Pseudomonas aeruginosa with the least prevalence of [5(2.6%). Resistant to antibiotics such as amoxycilin 64%, gentamicin 52.6%, erythromycin 62.5%, ceftazidime 69%, cefotaxime 74%, ceftriaxone 79.6% while a high sensitivity to tetracycline (88.5%), nitrofurantoin (96.3%), imipenem (100%) respectively. This study indicated a high prevalence of UTIs (56.7%) in pregnant women though most of them showed no clinical manifestation. Therefore, urine microbial screening should be included in the routine antenatal checkups for pregnant women to detect the asymptomatic infections to reduce its risk to pregnancies.Introduction There exists a variety of methods how to take samples from surgical wounds for cultivation; nevertheless, an unambiguous and most suitable procedure has so far been not defined with satisfactory precision. The aim of this presentation is to introduce an indirect imprint technique using filter paper pad for microbial analysis of wounds (shows principles of collecting, storaging, transporting, evaluation of samples and interpreting obtained information with all its advantages and disadvantages) and compare it with routinely used swab Method Imprints were carried out using the filter paper Whatman No. 4, cut to a size of 5 x 5 cm and previously sterilized in an autoclaved at 120 oC for 20 minutes and aseptically attached to 24 hour-old blood agar in a Petri dish, for the swabs, we used sterile cotton tips made by the Italian company COPAN; After careful debridement, a sample was taken from the wound surface by the swab and subsequently the same area of the investigation surface was imprinted onto a filter paper. Sheet of filter paper was taken from Petri dish with blood agar by sterile tweezers and put on to surface of wound. After 10 seconds paper became wet and was moved back to the Petri dish and then transported to a microbial laboratory as soon as possible. In the microbial laboratory a filter paper was removed from Petri dish and put on to new blood agar for 10 seconds and then to UriSelect 4 medium and after then to NaCl (salt) blood agar. The cultivation was done in the atmosphere with 10 % CO2 and temperature 37 oC. The final control of Petri dishes happened 48 hours later. The cultivation results of the two techniques were statistically processed using contingency tables and McNemar test. Those samples that were simultaneously cultivation positive in the imprint and negative in the swab were processed in greater detail. Results Result of an indirect imprint was semi-quantitative, one part of the result was type of microbe cultivated from wound and other was density of them accounted by colony forming unit (CFU), an estimate of viable bacterial numbers in wound, the results were given as CFU/cm2. 177 samples from 70 patients were analyzed. Sampling was carried out in 42 men and 28 women. The identical results of both culture methods were in 114 samples and different in 63 samples. The most frequent result of swab was sterile result, in 62 cases; imprint was sterile in 50 cases. Imprint and swab were both negative in 46 cases, and positive in 68 cases. In 15 cases there was a negative imprint and positive swab and in 48 cases there was positive imprint and negative swab. Coincidently, the most frequent cultivation finding with both techniques was a sterile one. The most frequently cultivated microorganism after using swab was Pseudomonas aeruginosa (in 22 cases), compared with Escherichia coli when the filter paper (imprint) was used (in 31 cases). The imprint technique was evaluated as more sensitive compared to swab, at the level of statistical significance P = 0.0001. Of the 177 samples there were 53 samples sterile in the swab and positive in the imprint simultaneously. In three samples CFU was not counted, 22 samples were within the limit of 0 - 25 x 101 CFU/cm2, 20 samples within the limit of 25 x 101 – 25 x 102 CFU/cm2, five within the limit of 25 x 102 – 25 x103 CFU/cm2, and three of more than 25 x 104 CFU/cm2. Conclusion In our study the imprint technique appears as a highly more sensitive than swabbing. We obtain information not only on the type of the microorganism cultured, but also on the number of viable colonies per square unit, expressed in CFU/cm2.Malaria and anaemia during pregnancy is still a major health problem in endemic countries with clinical consequences including death of both mother and child. In Nigeria, statistics shows that as many as 300,000 lives especially those of children and pregnant women are lost annually due to malaria. This study was aimed at assessing the impact of malaria and anaemia among pregnant women living in Calabar South Local Government Area of Cross River State, Nigeria, which is characterized by unstable transmission of malaria. .A total of 664 subjects were enrolled in the study made up of 414 pregnant women attending antenatal clinic in the University of Calabar Teaching Hospital Calabar, Nigeria and 250 age-matched non-pregnant women served as control group. Full blood count was done using PCE-210 automatic cell counter, malaria parasite detection was through examination of peripheral blood smears and malaria parasite count/density was done using WHO standard method (WHO, 1991). Anaemia was significantly(P <0.05) higher among the pregnant women 253(61.1)(Hb<11g/dl) than in the non-pregnant women 96(38.3%)(Hb<12g/dl).The prevalence of malaria parasite infection was 290(70.1%) in pregnant women and 152(60.8%) in the control group. Prevalence of anaemia and malaria parasite was found to be higher in the primigravidae than in the multigravidae. Primigravidae were more susceptible to the parasite especially Plasmodium falciparum with mean parasite density of 1962.50 ± 220.90 (parasite/µl) than the multigravidas with parasite density 446.70 ± 296.90 (parasite/µl). Malaria parasite density increased significantly with gestational age but anaemia was more prevalent in the second trimester than in the other trimesters. There was a negative correlation between haemglobin and malaria parsite density in both pregnant and non-pregnant women (r = -0.1964). The results showed that malaria infection caused by P. falciparum had serious effect on pregnant women living in the study area. Malaria in pregnancy should be recognized as a global priority in health care services.The study advocate the need for pregnant women to undergo routine haemoglobin estimation and early malaria prophylaxis considering the deleterious effects of anaemia on them and their foetus.A identification of coagulase-negative staphylococci (CoNS) is important especially when they are isolated from multiple blood cultures of a patient. Maldi-Tof MS is a new fast and accurate technique which allows examination of protein profiles of bacteria. This study aimed to compare two methods; Maldi-Tof MS (Bruker Daltonics) and BD Phoenix (Becton Dickinson) for identification of CoNS isolates from blood culture. Totally 102 CoNS bloodstream isolates representing 9 species were analyzed in this study. There was 73.5% correlation between two methods. The 10 S.hominis described by Maldi-Tof MS were identified as 3 S.epidermidis, 5 S.saprophyticus, 1 S.capitis, and 1 S.haemolyticus; 12 S.epidermidis identified as 3 S.saprophyticus, 2 S.haemolyticus 1 S.capitis, 1 S.hominis, 1 S.chromotogenes, 1 S.warneri, 1 S.hyicus, 1 S.aureus, and 1 S.capitis; 4 S.haemolyticus were identified as 2 S.epidermidis, and 2 S.hominis; 1 S.warneri was identified as S.capitis by BD Phoenix system. According to the previous studies Maldi-tof MS seemed to be a powerful method in ID of CoNS with up to 99.3% correct results. The next step of this study will be tuf sequencing to find the accuracy rates of these two methods. Mustafa Guney, J Microb Biochem Technol 2013, 5:4 http://dx.doi.org/10.4172/1948-5948.S1.010V species are the leading cause of bacterial illness (V. parahaemolyticus) and mortality (V. vulnificus) associated with seafood consumptionin the United States. Pathogenicity of these organisms is not fully understood, likely due to the involvement of multiple factors. Some virulence determinants have been identified for these organisms; however, no one virulence determinant, or set of determinants, has been shown to be required to cause illness. Additionally, isolates from patients with disease can lack any known virulence markers. In other instances, areas of the country with the greatest incidence of illness have lower levels of strains with identified virulence determinants in environmental samples. Taken together, these data highlight the necessity for identification of more robust virulence determinants in V. parahaemolyticus and V. vulnificus. Our laboratory has undertaken the quest for identification of new virulence biomarkers in these two pathogens using well characterized sets of clinical (presumed virulent) and environmental (presumed mostly avirulent) isolates. Using these strain panels, multiple approaches are being used to identify distinguishing features of the virulent (clinical) strains. Methods currently being employed include molecular subtyping, proteomics, and whole genome sequencing. A brief history of the understanding of vibrio virulence will be presented, followed by the detailed approach for identification of new biomarkers undertaken in our laboratory and resultant findings. Jessica L. Jones, J Microb Biochem Technol 2013, 5:4 http://dx.doi.org/10.4172/1948-5948.S1.010

24 Laser atherectomy in the treatment of peripheral arterial occlusive disease

Background: Peripheral occlusive arterial disease (PAD) represents a highly prevalent chronic disorder among the population.Clinically, we can divide it into four stages: I-IV according to Fountaine. In the critical stage of limb ischemia the preferred methods of first choice minimally invasive revascularisation procedures - PTA with or without stenting. Atherectomy represent an adjunctive technique that allows to directly remove the intravascular atheromatous plaques. We used the pulse laser beam delivered in a fine, flexible fiberoptic catheter that removes the atheromatous plaque in the artery wall and allows the blood stream to oxygenate the ischemic tissue- the excisional, rotational atherectomy technique based on mechanical removing of the plaque by rotating knife located in a thin catheter Material: In the year 2011, 346 patients with PAD in clinical stage IIb-IV were treated in the Hospital of Sonneberg using endovascular PTA. Out of this number 12 patients undewent atherectomy. Atherectomy was combined with balloon angioplasty in all the 12 cases. Excisional (rotational) atherectomy (1.6%) was performed six times and laser atherectomy (1.6%) was exhibited another six times. Clinical examination, duplex ultrasound and ankle-brachial index were performed in a standard way: before the treatment, during the first few days after the procedure and at six-month follow-up.In the year 2011, six patients (5 men, 1 woman) in the average age of 73 were treated with laser. After each atherectomy, PTA was performed. Excisional atherectomy was performed using the Turbohawk Plaque Excision System. In the year 2011, six patients (4 men, 2 women) in the average age of 69 were treated with excisional atherectomy. In all the cases, the primary crossing technique with the guiding wire insertion was used and PTA was additionally performed. Conclusion: The recent mechanical interventional endovascular systems allow in majority of the cases good interventional results. Also, the corresponding rapidity of the treatment is an advantage.In the femoropopliteal area, the mechanical (rotational) atherectomy allows similar results as the laser angioplasty, however, below the knee, , the laser angioplasty offers more interventional capabilities (due to the small diameter of the vessel and the impossibility of guidewire insertion).We suppose that the laser-assisted angioplasty is a useful technique in cases where the used mechanical methods represent higher risk of penetration or dissection as they do in popliteal area or by hard plaques by the dense calcinosis. A limitation of this study is given by the small sample size and short follow-up time. The laser assisted angioplasty represents a competent therapy option for the selected patients constituing an addition to the conventional, minimal invasive techniques.