Antibiofilm Activity, Compound Characterization, and Acute Toxicity of Extract from a Novel Bacterial Species of Paenibacillus

The effectiveness of many antimicrobial agents is currently decreasing; therefore, it is important to search for alternative therapeutics. Our study was carried out to assess the in vitro antibiofilm activity using microtiter plate assay, to characterize the bioactive compounds using Ultra Performance Liquid Chromatography-Diode Array Detection and Liquid Chromatography-Mass Spectrometry and to test the oral acute toxicity on Sprague Dawley rats of extract derived from a novel bacterial species of Paenibacillus strain 139SI. Our results indicate that the crude extract and its three identified compounds exhibit strong antibiofilm activity against a broad range of clinically important pathogens. Three potential compounds were identified including an amino acid antibiotic C8H20N3O4P (MW 253.237), phospholipase A2 inhibitor C21H36O5 (MW 368.512), and an antibacterial agent C14H11N3O2 (MW 253.260). The acute toxicity test indicates that the mortality rate among all rats was low and that the biochemical parameters, hematological profile, and histopathology examination of liver and kidneys showed no significant differences between experimental groups P>0.05. Overall, our findings suggest that the extract and its purified compounds derived from novel Paenibacillus sp. are nontoxic exhibiting strong antibiofilm activity against Gram-positive and Gram-negative pathogens that can be useful towards new therapeutic management of biofilm-associated infections

Characteristics and racial variations of polypoidal choroidal vasculopathy in tertiary centers in the United States and United Kingdom

PURPOSE: To evaluate the characteristics and racial variations amongst patients with polypoidal choroidal vasculopathy (PCV) in the United States and the United Kingdom. METHODS: Fundus photos and indocyanine green angiography images were evaluated in a multicenter retrospective study to establish the diagnosis of PCV. Visual acuity (VA) was recorded in ETDRS letter count. RESULTS: Eighty eyes of 71 PCV patients (average age of 69.4 ± 10.4 years) were included in the analysis. Of the total 71 subjects, 46 (65%) were women, 33 (46.5%) were Blacks, 16 (22.5%) were Whites, 19 (26.8%) were Asians and 3 (4.2%) belonged to other races. The Black subgroup had vision gain of 3.5 letters. The White and Asian subgroups had vision loss of 13.1 and 3.5 letters, respectively. There was female predominance in Blacks (67%), Whites (69%), and Asians (58%). PCV was found to be a bilateral disease in 14 patients (20%). There was significant decrease of 7 letters with every decade increase in age (p = 0.005). Final VA was worse in males when compared to females (p = 0.042), and worse in Whites when compared to Blacks (p = 0.005). For every 10 letters worse in initial VA upon diagnosis with PCV, the final VA was worse by 6 letters (p < 0.001). The location of the polypoidal lesion within the macula was associated with significant decrease of 14 letters in BCVA (p = 0.02). The length of follow up was significantly associated with worse visual outcome (p = 0.012). Final VA had no significant correlation with the lens status, or the different treatment modalities. CONCLUSIONS: Based on our cohort from tertiary centers in the United States and United Kingdom, PCV is a bilateral disease in one-fifth of patients. It features a variable female predominance based on ethnicity. Increased age, worse vision upon initial presentation, longer follow up and macular location of the polyp were associated with worse visual outcome

Characterization of Choroidal Layers in Normal Aging Eyes Using Enface Swept-Source Optical Coherence Tomography

Purpose To characterize qualitative and quantitative features of the choroid in normal eyes using enface swept-source optical coherence tomography (SS-OCT). Methods Fifty-two eyes of 26 consecutive normal subjects were prospectively recruited to obtain multiple three-dimensional 12x12mm volumetric scans using a long-wavelength high-speed SS-OCT prototype. A motion-correction algorithm merged multiple SS-OCT volumes to improve signal. Retinal pigment epithelium (RPE) was segmented as the reference and enface images were extracted at varying depths every 4.13 mu m intervals. Systematic analysis of the choroid at different depths was performed to qualitatively assess the morphology of the choroid and quantify the absolute thicknesses as well as the relative thicknesses of the choroidal vascular layers including the choroidal microvasculature (choriocapillaris, terminal arterioles and venules;CC) and choroidal vessels (CV) with respect to the subfoveal total choroidal thickness (TC). Subjects were divided into two age groups: younger (= 40 years). Results Mean age of subjects was 41.92 (24-66) years. Enface images at the level of the RPE, CC, CV, and choroidal-scleral interface were used to assess specific qualitative features. In the younger age group, the mean absolute thicknesses were: TC 379.4 mu m (SD +/- 75.7 mu m),CC 81.3 mu m (SD +/- 21.2 mu m) and CV 298.1 mu m (SD +/- 63.7 mu m). In the older group, the mean absolute thicknesses were: TC 305.0 mu m (SD +/- 50.9 mu m),CC 56.4 mu m (SD +/- 12.1 mu m) and CV 248.6 mu m (SD +/- 49.7 mu m). In the younger group, the relative thicknesses of the individual choroidal layers were: CC 21.5% (SD +/- 4.0%) and CV 78.4% (SD +/- 4.0%). In the older group, the relative thicknesses were: CC 18.9% (SD +/- 4.5%) and CV 81.1% (SD +/- 4.5%). The absolute thicknesses were smaller in the older age group for all choroidal layers (TC p=0.006, CC p=0.0003, CV p=0.03) while the relative thickness was smaller only for the CC (p=0.04). Conclusions Enface SS-OCT at 1050nm enables a precise qualitative and quantitative characterization of the individual choroidal layers in normal eyes. Only the CC is relatively thinner in the older eyes. In-vivo evaluation of the choroid at variable depths may be potentially valuable in understanding the natural history of age-related posterior segment disease

Renal cell carcinoma metastasis to the ciliary body responds to proton beam radiotherapy: a case report

<p>Abstract</p> <p>Introduction</p> <p>We report an unexpected presentation of metastatic renal cell carcinoma (RCC) to the ciliary body and an interesting response to proton beam radiotherapy.</p> <p>Case presentation</p> <p>We encountered a case of angle-closure glaucoma as the initial presentation of ocular metastasis to the ciliary body in a 65-year-old Caucasian man who had undergone right radical nephrectomy for RCC 15 years earlier. He underwent YAG (yttrium aluminium garnet) laser peripheral iridotomy while further metastatic workup took place. His condition was eventually diagnosed as stage IV metastatic RCC of the clear cell type and involved multiple sites, including the ciliary body, brain, lungs, liver, and pancreas. The progression of RCC metastasis to the ciliary body was studied for 16 months. The ciliary body mass continued to grow despite systemic treatment with temsirolimus and interleukin-2 and intravitreal injections of bevacizumab. The tumor size peaked at 6.11 × 6.06 mm before the start of proton therapy, which reduced the tumor size to 5.07 × 4.39 mm.</p> <p>Conclusions</p> <p>RCC can produce metastases involving unusual sites many years after resection of the primary tumor. Proton therapy was found to be effective in treating RCC metastasis to the ciliary body in settings in which other treatment modalities failed.</p

Antibiofilm activity from novel soil bacterial species of Paenibacillus Haemolyticus strain 139si towards new therapeutic management of chronic and recurrent tonsillitis / Saad Musbah Naji Alasil

A biofilm is a layer of microbial cells that is firmly attached to surfaces and enclosed in a matrix of polysaccharide material. Biofilms pose a problem to the environment, industry and the medical field causing a variety of chronic infections particularly in the ear, nose and throat like chronic and recurrent tonsillitis. Therefore, development of new therapeutic strategies against biofilm-forming pathogens is essential in the management of biofilm-associated tonsillar diseases. One hundred and forty (140) palatine tonsils were collected from 70 patients undergoing tonsillectomy at University Malaya Medical Centre (UMMC) and the type of microorganisms and their antimicrobial susceptibility of both swab and biopsy tonsillar specimens were identified. The presence of bacterial biofilms in the excised tonsils was detected via Scanning Electron Microscopy and Confocal Laser Scanning Microscopy, and the ability of tonsillar bacterial isolates to form biofilms was determined via Congo Red Agar method and Microtiter Plate assay. A culture filtrate from novel soil bacterial species of Paenibacillus haemolyticus strain 139SI was tested for its acute toxicity in rats and evaluated for its antibiofilm activity both in vitro via Microtiter Plate assay and in vivo using a rat model of chronic lung infection. The soil bacterial filtrate was purified via High Performance Liquid Chromatography, and the selected compounds were tested against clinical isolates for their antibiofilm activity. The most potential antibiofilm compounds were further analysed via Liquid Chromatography-Mass Spectrometry. Two main groups of clinical cases were classified as infected tonsils represented by 49 (70%) cases of recurrent tonsillitis, 9 (12.85%) cases of chronic tonsillitis in addition to non-infected (hypertrophied) tonsils represented by 12 (17.14%) cases of obstructive sleep apnea. The presence of bacterial biofilms was detected in 42 (60%) patients. iii A total number of 464 bacterial isolates were recovered from tonsillar specimens with 184 (39.65%) Staphylococcus aureus isolates as the most common followed by 86 (18.53%) of Haemophilus influenzae isolates. Three susceptibility patterns were found among S. aureus isolates with 89.4% being susceptible to all tested antimicrobial agents, 10.6% resistant to fusidic acid and 0.5% resistant to both methicillin and fusidic acid. The antimicrobial agent co-trimoxazole showed the highest rate of resistance among all bacterial isolates including 55 (98.2%) isolates of Group B Streptococci and 11 (78.5%) isolates of Group A Beta-Haemolytic Streptococci, 14 (56%) isolates of Group G Streptococci, 10 (32.2%) isolates of Haemophilus pararinfluenzae and 27 (31.3%) isolates of Haemophilus influenzae. The soil bacterial filtrate of Paenibacillus haemolyticus strain 139SI showed no signs of toxicity and a significant antibiofilm activity was detected in the lungs of chronically-infected rats. Four potential antibiofilm compounds of the bacterial filtrate namely FR4, FR5, FR8 and FR13 were identified. The compound FR5 with a molecular weight of 253.237 and a molecular formula of C8H20N3O4P exhibited the strongest antibiofilm activity. In conclusion, discovery of an antibiofilm activity from a novel soil bacterial species of Paenibacillus haemolyticus strain 139SI adds an important dimension in the search for new potent compounds against biofilm infections