Differential Protection from Tobramycin by Extracellular Polymeric Substances from Acinetobacter baumannii and Staphylococcus aureus Biofilms

We investigated biofilms of two pathogens, Acinetobacter baumannii and Staphylococcus aureus, to characterize mechanisms by which the extracellular polymeric substance (EPS) found in biofilms can protect bacteria against tobramycin exposure. To do so, it is critical to study EPS-antibiotic interactions in a homogeneous environment without mass transfer limitations. Consequently, we developed a method to grow biofilms, harvest EPS, and then augment planktonic cultures with isolated EPS and tobramycin. We demonstrated that planktonic cultures respond differently to being treated with different types of EPS (A. baumannii versus S. aureus) in the presence of tobramycin. By harvesting EPS from the biofilms, we found that A. baumannii EPS acts as a “universal protector” by inhibiting tobramycin activity against bacterial cells regardless of species; S. aureus EPS did not show any protective ability in cell cultures. Adding Mg²⁺ or Ca²⁺ reduced the protective effect of A. baumannii EPS. Finally, when we selectively digested the proteins or DNA of the EPS, we found that the protective ability did not change, suggesting that neither has a significant role in protection. To the best of our knowledge, this is the first study that demonstrates how EPS protects pathogens against antibiotics in a homogeneous system without mass transfer limitations. Our results suggest that EPS protects biofilm communities, in part, by adsorbing antibiotics near the surface. This may limit antibiotic diffusion to the bottom of the biofilms but is not likely to be the only mechanism of protection.     

Mechanoreceptors of the ligaments and tendons around the knee

Proprioceptive inputs from the joints and limbs arise from mechanoreceptors in the muscles, ligaments and tendons. The knee joint has a wide range of movements, and proper neuroanatomical organization is critical for knee stability. Four ligaments (the anterior (ACL) and posterior (PCL) cruciate ligaments and the medial (MCL) and lateral (LCL) collateral ligaments) and four tendons (the semitendinosus (STT), gracilis (GT), popliteal (PoT), and patellar (PaT) tendons) from eight fresh frozen cadavers were harvested. Each harvested tissue was divided into its bone insertion side and its tendinous part for immunohistochemical examination using S100 staining. Freeman–Wyke's classification was used to identify the mechanoreceptors. The mechanoreceptors were usually located close to the bone insertion. Free nerve endings followed by Ruffini endings were the most common mechanoreceptors overall. No Pacini corpuscles were observed; free nerve endings and Golgi‐like endings were most frequent in the PCL (PCL‐PaT: P = 0.0.1, PCL‐STT: P = 0.00), and Ruffini endings in the popliteal tendon (PoT‐PaT: P = 0.00, Pot‐STT: P = 0.00, PoT‐LCL: P = 0.00, PoT‐GT: P = 0.00, PoT‐ACL: P = 0.09). The cruciate ligaments had more mechanoreceptors than the medial structures (MS) or the patellar tendon (CR‐Pat: P = 0.000, CR‐MS: P = 0.01). The differences in mechanoreceptor distributions between the ligaments and tendons could reflect the different roles of these structures in the dynamic coordination of knee motion. Clin. Anat. 29:789–795, 2016. © 2016 Wiley Periodicals, Inc.     

Effects of different surface treatments on shear bond strength in two different ceramic systems

The purpose of this study was to evaluate the influence of different surface treatments (sandblasting, acid etching, and laser irradiation) on the shear bond strength of lithium disilicate-based core (IPS Empress 2) and feldspathic ceramics (VITA VM 9). One hundred ceramic discs were divided into two groups of 50 discs each for two ceramic systems: IPS Empress 2 (group I) and VITA VM 9 (group II). Each of the two groups was further divided into five surface treatment groups (ten each) as follows: group SB, sandblasting with alumina particles (50 μm); group HF, 5 % hydrofluoric acid etching; group L, Er:YAG laser irradiation (distance, 1 mm; 500 mJ; 20 Hz; 10 W; manually, noncontact R14 handpiece); group SB–L, sandblasting + Er:YAG laser; and group HF–L, 5 % hydrofluoric acid + Er:YAG laser. Luting cement (Panavia 2.0) was bonded to the ceramic specimens using Teflon tubes. After 24 h of water storage, a shear bond strength test was performed using a universal testing machine at a crosshead speed of 0.5 mm/min. The data were analyzed with a two-way analysis of variance (ANOVA) and Tukey's honestly significant difference tests (α?=?0.05). The two-way ANOVA indicated that the shear bond strength was significantly affected by the surface treatment methods (p?<?0.05), but there was no significant interaction between the ceramic systems. Group SB–L had the highest mean values for each ceramic system. Sandblasting, followed by Er:YAG laser irradiation, enhanced the bond strength, indicating its potential use as an alternative method. The atomic force microscopic evaluation revealed that group SB had the most distinct sharp peaks among the groups.     

The Effect of K-ATP Channel Blockage During Erythropoietin Treatment in Renal Ischemia-Reperfusion Injury

ATP dependent K channels (K-ATP) take part in the Erythropoietin (EPO) induced cardioprotection but these channel activations have role in cytoprotective role of EPO in the renal ischemia reperfusion (IR) damage is still unknown. For this purpose rats were pretreated with EPO (500 IU/kg) and/or K-ATP channel blocker glibenclamide (40mM/kg) i.p. before bilateral renal IR damage. Renal tissues were used for histological examination and measurement of caspase-3 and TNF-α levels. Renal functions were evaluated by glomerular filtration rate (GFR) fractional excretion of sodium (FENa) and potassium (FEK). Renal TNF-α and caspase-3 levels were decreased in both glibenclamide and EPO-treated IR rats compared to untreated rats. The protection afforded by the pretreatment with EPO alone was greater than that of administering glibenclamide alone. Application of glibenclamide at the same time partly abolished the cytoprotective effect of EPO treatment. K-ATP mediated cytoprotection is not the main mechanism of protective effect of EPO.     

Effect of cardiac resynchronization therapy on left atrial reverse remodeling and spontaneous echo contrast

Recent studies revealed reverse remodeling in left ventricle with cardiac resynchronization therapy (CRT). However, effects on left atrial remodeling, left atrial total emptying fraction and left atrial spontaneous echo contrast (SEC) have not been adequately evaluated. The aim of this study was to investigate the long-term changes in SEC, left atrial reverse remodeling, and left atrial total emptying fraction after CRT. Twenty patients with systolic heart failure and complete left bundle-branch block underwent implantation of biventricular pacemaker devices. Transthoracic and transesophageal echocardiography were performed one week before and one and six months after pacemaker implantation. After biventricular pacemaker implantation, significant clinical improvement was observed in all patients. Left atrial maximal and minimal volumes showed a significant progressive decline after CRT (reverse remodeling). Left atrial total emptying ejection fraction (LATEF) was 33 +/- 19% at baseline and increased to 37 +/- 10% and 41 +/- 11% at the 1st and 6th months respectively (p = 0.01 and p = 0.04). SEC was detected in 18 of 20 patients (90%) at the beginning of the study. After six months SEC disappeared in 5 patients and frequency of SEC reduced to 45%. Decrease in the intensity of the SEC was also statistically significant (at the 1st and 6th months; p = 0.001 and p < 0.001 respectively). Long-term CRT results in atrial reverse remodeling, increases LATEF, and reduces both frequency and intensity of atrial SEC.     

Clinical outcome of rhabdomyosarcoma in adolescent and adult patients: single center experience from Turkey

Rhabdomyosarcoma (RMS) is rare disease in adults (age >or= 16 years). The data from randomized prospective trials are scarce; the clinical outcome of these patients seems poor with the currently available treatment strategies. In this study, we report a single institution's experience in the treatment of adult RMS. We reviewed the medical records of patients with RMS who were >or= 16 years and have been treated in our institution between 1988 and 2003 retrospectively. We analyzed the survival outcome of these patients and the prognostic impact of clinical/pathological factors on their survival. In total, 23 patients with RMS were identified. Median age was 26 years (range, 16-72 years). Majority of patients were male (n: 17, 73.9%), and had large tumors (>or= 5 cm, n: 13, 56.5%), localized disease (N0, M0, n: 12, 52.2%), and embryonal histology (n: 10, 43.5%). Median overall survival was 31.3 months, and the 3-year progression-free survival and overall survival rates were 19.9% and 34.94%, respectively. Patients with smaller tumors (< 5 cm) (p < 0.04), local disease (p < 0.01), and normal lactic dehydrogenase (LDH) level (p < 0.01) at the time of diagnosis were found to have better survival outcome. The tumor size, serum LDH level, and metastatic disease at the time of diagnosis are potential predictors of outcome in patients with adult RMS. Adult RMS is an aggressive disease with poor survival despite treatment. The data from prospective, randomized multicenter trials are necessary in order to improve the clinical outcome of adult RMS patients.     

Susceptibility patterns of enterococci causing infections

Enterococci are among the common organisms associated with hospital-acquired infections. We examined in vitro activities of different antibiotics to 103 enterococcal isolates. Minimal inhibitory concentrations (MICs) of penicillin G, ampicillin, gentamicin, ciprofloxacin, ofloxacin, levofloxacin, grepafloxacin, trovafloxacin and gemifloxacin were determined by broth microdilution testing method. Among the isolates 71 (69%) were identified as E. faecalis and 32 (31%) as E. faecium. While over 75% of E. faecium isolates were resistant to penicillin and ampicillin, approximately 25% of E. faecalis isolates were resistant to penicillin and ampicillin. None of the E. faecalis and E. faecium isolates were resistant to vancomycin. While 17 (52%) of E. faecium isolates exhibited high-level gentamicin resistance (HLGR), high level streptomycin resistance (HLSR) was detected in 24 (74%) of the isolates. In contrast, HLGR and HLSR rates for E. faecalis were 14 (20%) and 22 (31%), respectively. Both HLGR and HLSR were detected with higher frequency in ampicillin resistant isolates. Among fluoroquinolones, gemifloxacin and trovafloxacin were the most potent antibiotics tested. There was no increase in MIC90 values of the fluoroquinolones in ampicillin resistant isolates in comparison with ampicillin susceptible isolates. Our data suggest newer fluoroquinolones would be good alternative agents to use especially for combination drug therapy where enterococci with ampicillin resistance and HLAR are prevalent.     

Computed tomographic evaluation of lumbar spinal structures during traction

In the previous studies, it is reported that traction diminishes the compressive load on intervertebral discs, reduces herniation, stretches lumbar spinal muscle and ligaments, decreases muscle spasm, and widens intervertebral foramina. The aim of this study was to evaluate the effects of horizontal motorized static traction on spinal anatomic structures (herniated area, spinal canal area, intervertebral disc heights, neural foraminal diameter, and m.psoas diameter) by quantitative measures in patients with lumbar disc herniation (LDH). At the same time the effect of traction in different localizations (median and posterolateral herniation) and at different levels (L4-L5 and L5-S1) was assessed. Thirty two patients with acute LDH participated in the study. A special traction system was used to apply horizontally-motorized static lumbar traction. Before and during traction a CT- scan was made to observe the changes in the area of spinal canal and herniated disc material, in the width of neural foramina, intervertebral disc heights, and in the thickness of psoas muscle. During traction, the area of protruded disc area, and the thickness of psoas muscle decreased 24.5% (p = 0.0001), and 5.7% (p = 0.0001), respectively. The area of the spinal canal and the width of the neural foramen increased 21.6% (p = 0.0001) and 26.7% (p = 0.0001), respectively. The anterior intervertebral disc height remained unchanged with traction however the posterior intervertebral disc height was significantly expanded. This study is the first to evaluated in detail and quantitatively the effect of motorized horizontal lumbar spinal traction on spinal structures and herniated area. According to detailed measures it was concluded that during traction of individuals with acute LDH there was a reduction of the size of the herniation, increased space within the spinal canal, widening of the neural foramina, and decreased thickness of the psoas muscle.