Sterile Physiological Solution Research Articles

Effect of a water-soluble form of dihydroquercetin on age-dependent LPS-induced gliovascular remodeling of the substantia nigra in rats

Background. The molecular and cellular mechanisms of action of the water-soluble form of dihydroquercetin (DHA-VF) on neurogliovascular units, age-related disturbances of intrasystem connections that may underlie neuroinflammatory and neurodegenerative diseases, remain unclear.The aim: to study structural changes of microcirculatory vessels and functional responses of micro- and astroglial cells in the substantia nigra of young and old rats in response to intranigral injection of lipopolysaccharide (LPS) and subsequent oral administration of DHA-VF.Methods. Young (250-320 g) and old (390-450 g) Wistar rats were injected with 2 μL LPS solution at a concentration of 0.01 μL/mL (experimental group; n=24) or 2 μL sterile physiological solution (control group; n=12) into the substantia nigra area of the brain using a stereotaxic device. Half of the animals in the experimental groups (6 animals of each age group) were given 2 ml of a solution containing DHA-VF at a concentration of 3 mg/mL daily by gavage with a probe. After 8 weeks, the animals were decapitated and cryostat sections were obtained for histochemical (FITC-labeled tomato lectin) and immunohistochemical (antibodies against GFAP and CD-11β) staining of vascular endothelium and glial cells, respectively. Results. 8 weeks after LPS administration to old rats in the CS, a significant excess of areas occupied by cell bodies and processes of microglial and astroglial cells as well as the number of vessels at standard sites was found compared to both young animals exposed to similar exposure and old control animals. Oral administration of DHA-VF to rats significantly reduced LPS-induced glial activation in young and old animals. In addition, administration of DHA-VF to old animals reduced the intensity of LPS-induced microvascular remodeling of the CS. Conclusions. LPS administration in the rat CS induces neuroinflammation and vascular angiogenesis, which are maximally expressed in old animals. Administration of DHA-VF for 8 weeks significantly reduced these LPS-induced changes.

A Study of the Effect of Specific Antibacterial Therapy on the Course of Tuberculous Spondylitis in an Experiment

A Study of the Effect of Specific Antibacterial Therapy on the Course of Tuberculous Spondylitis in an Experiment

Transabdominal self-expandable metal stent placement as a rescue therapy for drainage of walled-off necrosis.

Video 1CT scan of a walled-off necrosis in the right pararenal space extended to the iliac area.

Retrieving the in vivo Scopoletin Fluorescence Excitation Band Allows the Non-invasive Investigation of the Plant-Pathogen Early Events in Tobacco Leaves.

In this study, we developed and applied a new spectroscopic fluorescence method for the in vivo detection of the early events in the interaction between tobacco (Nicotiana tabacum L.) plants and pathogenic bacteria. The leaf disks were infiltrated with a bacterial suspension in sterile physiological solution (SPS), or with SPS alone as control. The virulent Pseudomonas syringae pv. tabaci strain ATCC 11528, its non-pathogenic ΔhrpA mutant, and the avirulent P. syringae pv. tomato strain DC3000 were used. At different post-infiltration time–points, the in vivo fluorescence spectra on leaf disks were acquired by a fiber bundle-spectrofluorimeter. The excitation spectra of the leaf blue emission at 460 nm, which is mainly due to the accumulation of coumarins following a bacterial infiltration, were processed by using a two-bands Gaussian fitting that enabled us to isolate the scopoletin (SCT) contribution. The pH-dependent fluorescence of SCT and scopolin (SCL), as determined by in vitro data and their intracellular localization, as determined by confocal microscopy, suggested the use of the longer wavelength excitation band at 385 nm of 460 nm emission (F385_460) to follow the metabolic evolution of SCT during the plant–bacteria interaction. It was found to be directly correlated (R2 = 0.84) to the leaf SCT content, but not to that of SCL, determined by HPLC analysis. The technique applied to the time-course monitoring of the bacteria–plant interaction clearly showed that the amount and the timing of SCT accumulation, estimated by F385_460, was correlated with the resistance to the pathogen. As expected, this host defense response was delayed after P. syringae pv. tabaci ATCC 11528 infiltration, in comparison to P. syringae pv. tomato DC3000. Furthermore, no significant increase of F385_460 (SCT) was observed when using the non-pathogenic ΔhrpA mutant of P. syringae pv. tabaci ATCC 11528, which lacks a functional Type Three Secretion System (TTSS). Our study showed the reliability of the developed fluorimetric method for a rapid and non-invasive monitoring of bacteria-induced first events related to the metabolite-based defense response in tobacco leaves. This technique could allow a fast selection of pathogen-resistant cultivars, as well as the on-site early diagnosis of tobacco plant diseases by using suitable fluorescence sensors.

Modulating the Antifungal Activity of Antimycotic Drugs with Farnesol

Introduction. Clinical strains of microorganisms, including opportunistic yeast-like fungi (YLF) of the genus Candida, are resistant to currently used antifungal drugs. In this regard, the search for alternative ways to potentiate the activity of antimicrobial agents in relation to the infectious agent is an important and relevant area of research. The study of combinations of existing antimycotic drugs and a medicinal extract of plant origin – farnesol – is one of the promising approaches in the fight against resistant strains of YLF genus Candida. In our previous studies, farnesol has been shown to exhibit relative activity against YLF Candida albicans biofilms. In this study, we used 6 clinical isolates and one museum strain YLF C. albicans to study the effect of farnesol on the antifungal activity of antimycotic drugs.Aim. To prove that farnesol can increase the antifungal activity of certain antimycotics.Materials and methods. To determine the sensitivity of 7 strains of YLF C. albicans to the antimycotic drugs "Nystatin" (NYS 50 µg), "Ketoconazole" (KET 10 µg), "Clotrimazole" (CTR 10 µg), "Amphotericin B" (AMB 10 µg), "Voriconazole" (VRC 10 µg) disk diffusion test was used. A solution of farnesol in concentrations of 100, 50 and 25 µM in a volume of 25 µl was applied to the disk with the antimycotic drug. Sterile physiological (PhS) solution was used as a control (pH 7.0; V = 25 µl).Results and discussion. In 34.3 % of of experiments we can talk about the modulating effect of farnesol solutions on the antifungal activity of antimycotic drugs. In all these cases, the sensitivity of YLF C. albicans to the antimycotic drug increases.Conclusion. The results of this study provide useful information for understanding the mechanism of QS-molecules action with antifungal activity, as well as they are the basis for the practical application of some QS-molecules in the treatment of infectious diseases caused by YLF of the genus Candida. The study demonstrates that farnesol can be recommended as an active substance that improves the sensitivity of YLF Candida to antimycotic drugs, especially in the case of multi-resistant strains Candida.

Експериментальне моделювання туберкульозного спондиліту

Актуальність. Туберкульозний спондиліт (ТС) в структурі кістково­суглобового туберкульозу у дорослих займає провідне місце і досягає 40–61,5 %. Метою даної публікації є висвітлення процесу моделювання туберкульозного спондиліту і його особливостей у морських свинок для подальшого використання результатів експерименту у лікувальній та науковій практиці. Матеріали та методи. Експеримент ставився на 40 статевозрілих морських свинках. Моделювання туберкульозного спондиліту проводилось на основі розробленого нами способу (патент № 112423 (UА) Україна). Усі тварини були розподілені на чотири рівні групи: 1­ша — 3­тя групи (основні) — виконувалась ін’єкція 0,5 мл суспензії М.tuberculosis (0,1 мг сухої маси в 1 мл) в тіло хребця згідно з методикою; 4­та група — контрольна. Тваринам робилась ін’єкція стерильного фізіологічного розчину (0,9% — 0,5 мл) в тіло хребця. Група 1 (10 свинок) — лікування специфічними антибактеріальними препаратами (АБП) першого ряду (ізоніазид, стрептоміцин, рифампіцин). Група 2 (10 свинок) — лікування специфічними антибактеріальними препаратами другого ряду (амікацин, рифабутин, офлоксацин). Група 3 (10 свинок) — не проводилося лікування. За всіма тваринами проводилося динамічне спостереження з клінічними, рентгенологічними, патоморфологічними і лабораторними дослідженнями. Виведення ­експериментальних тварин з досліду проходило за раніше розробленим графіком — при виявленні ознак досліджуваних стадій туберкульозного процесу. Всім виведеним з експерименту тваринам проводилося патоморфологічне дослідження. При цьому основна увага приділялася вивченню патоморфології уражених специфічним процесом хребців. Результати. У результаті морфологічного дослідження встановлено наявність активного туберкульозного процесу в тілах хребців і паравертебральних тканинах у тварин із модельованим туберкульозом і лікуванням специфічним АБП першого ряду, а також у тварин, які не приймали специфічного лікування. Важливо зазначити, що ступінь вираженості деструктивних змін у вражених хребцях у тварин нелікованих і тих, які приймали АБП першого ряду, практично однаковий. У тварин, яким моделювали туберкульоз і лікували специфічним АБП другого ряду, виявлено пригнічення патологічного процесу з утворенням молодої кісткової та сполучної тканини різного ступеня зрілості та наявністю зони, що відмежовує вогнище запалення від здорової тканини в ранні терміни захворювання (один місяць). В результаті виконаної роботи вдалося простежити стадійність розвитку туберкульозного спондиліту у морської свинки та сучасні особливості клінічного, рентгенологічного та патоморфологічного перебігу. Виявлена ідентичність моделі основних клінічних форм туберкульозного спондиліту у морської свинки та людини. Висновки. Дане дослідження показало, що проведення сучасної інтенсивної специфічної антибактеріальної терапії в умовах експерименту дозволяє досягти відмежування деструктивного процесу в порівняно ранні терміни розвитку захворювання (4–5 тижнів). Отримані нові знання про патоморфологічні особливості перебігу туберкульозного спондиліту дозволяють проводити радикальні оперативні втручання на хребті без ризику генералізації туберкульозного процесу в більш ранні терміни.

Effects of Green Alga, Chaetomorpha aerea Extract on Non-Specific Immune Responses and Disease Resistance against Edwardsiella tarda Infection in Labeo rohita

The current study evaluated the effects of a methanol extract from Chaetomorpha aerea (a green alga) on non-specific immune responses and resistance against Edwardsiella tarda infection in Labeo rohita. Different doses of the extract (5, 50 and 500 mg/kg of body weight) were injected into the fish intraperitoneally while a control group was injected with 0.2 mL of sterile physiological salt solution. Variations in several immunostimulatory parameters (i.e., neutrophil, serum lysozyme, myeloperoxidase, serum antiprotease, and ceruloplasmin activity), reactive oxygen species (ROS) and reactive nitrogen species (RNS) were assessed after 7, 14, 21, and 28 days of post stimulation. E. tarda culture was injected into the fish after 28 days of post stimulation to induce infection to monitor fish mortality within 14 days. Interestingly, all doses of methanolic extract enhanced neutrophil, lysozyme, and myeloperoxidase activity, ROS and RNS, while a dose of 50 mg/kg was the most effective. Fish injected with this optimal dose were also protected against infection with virulent strain of E. tarda. The results of the study suggest that C. aerea extract is a potential prophylactic agent against bacterial infections in finfish.

Third Molar Extraction: Irrigation and Cooling with Water or Sterile Physiological Solution: A Double-Blind Randomized Study.

Background: The present study aimed to ascertain whether any significant reduction in patients’ postoperative pain and inflammation could be achieved by using sterile physiological solution instead of normal water to irrigate the surgical field and cool the dental bur during third molar extractions. Methods: The study concerned 22 patients (11 females and 11 males) in good general health, who were referred to the Dental Clinic at Padova University hospital for lower third molar extractions. They were randomly assigned to two groups. Only the fluid used to irrigate the surgical field and cool the dental bur differed between the two study groups, being sterile physiological solution for group A, and mains water for group B. Postoperative pain, swelling, trismus and inflammation with high sensitivity CRP where measured and statistically evaluated. The numerosity of our sample was calculated on the grounds of an endpoint based on data in the literature. Results: Eighteen patients needed bilateral extractions, and 4 required only one extraction, so a total of 40 third molars were extracted. A sterile physiological solution was used to irrigate the surgical field in 20 extractions, while water was used in the other 20 cases. Data analysis with Wilcoxon test show no differences between the two groups (p < 0.05). Conclusions: no differences between groups for any of the parameters considered, after third molar extraction procedures undertaken using either sterile physiological solution or water for irrigation and cooling purposes.

Le tecniche e le soluzioni utilizzate nella detersione delle ulcere: una revisione della letteratura

L’ulcera è la perdita di continuità dell’epitelio, dell’epidermide o della mucosa causata dall’espulsione del tessuto necrotico infiammato. È l’espressione di processi degenerativi provocati da fenomeni infiammatori, infettivi, da disturbi circolatori o da danneggiamento tissutale per cause chimiche e fisiche. Il trattamento di una lesione cutanea cronica deve essere multidisciplinare e richiedere la collaborazione di numerosi specialisti, del personale infermieristico, del paziente e/o del nucleo familiare. Lo scopo della revisione è quello di verificare gli effetti della detersione sulle ferite considerando sia gli approcci adottati, sia le soluzioni utilizzate, valutando i relativi tassi di guarigione e di infezione delle ulcere. La ricerca della letteratura è stata eseguita nelle banche dati di Pubmed e Cochrane Library. Sono stati considerati studi clinici randomizzati e revisioni di letteratura dal 2003 al 2018. 276 lavori scientifici sono stati individuati. Di questi, sono 15 studi sono stati considerati potenzialmente eleggibili. Successivamente alla lettura completa di ciascun lavoro sono stati inclusi infine 6 articoli. La gestione delle ulcere coinvolge molte figure professionali esperte in wound care che dovrebbero collaborare tra loro per guarire le ferite. Specificamente la detersione viene considerata un intervento di routine che viene effettuato con soluzione fisiologica sterile attraverso il metodo del tamponamento, questo perché la letteratura non fornisce ricerche soddisfacenti nell’ambito dell’utilizzo dell’acqua di rubinetto.

Combined effect of glyphosate, saccharin and sodium benzoate on the gut microbiota of rats

Glyphosate is the main component of many broadly used herbicides due to its safety for humans and animals. It is known that the remains of glyphosate are present in allowable doses in fodders and food products, and, consumrd in low doses, it is found in insignificant amounts in milk, eggs and even in the internal organs (liver, kidneys) of animals. For determining combined impact of glyphosate and the commonest food additives on the composition of microbiota of animals, four groups of laboratory male rats were formed, which during 42 days consumed pure water without any restrictions; 1% aqueous solution of glyphosate; 1% solution of glyphosate in combination with 1% solution of sodium benzoate; 1% solution of glyphosate with 1% solution of saccharin. After killing the animals, 1 g of feces were collected and by serial dilutions with 10–1 to 10–9 sterile physiologic solution, a microbiological analysis was undertaken. Out of each dilution an inoculation of the studied material to the elective growth media was performed, by 0.1 cm3, then the material was incubated in a thermostat (24–72 hours, temperature 37 °С), the results were recorded after 24–72 h. The microorganisms were identified by studying morphological parameters, tinctorial, cultural and enzymic properties. Results are provided in CFU/g (colony-forming unit per gramm) of feces. The impact of glyphosate and glyphosate with food additives led to no changes in the number of Escherichia coli and emergence of this species of microorganism with changed enzymic activity. Also no changes occurred in the number of microorganisms of Bifidobactrium and Lactobacillus spp. Addition of glyphosate, and also glyphosate in combination with saccharin to the diet contributes to broader reproduction of microorganisms of Klebsiella, Citrobacter, Enterobacter and Pseudomonas genera. Mixtures of glyphosate and food additives allow conditionally pathogenic yeast-like Candida fungi (Candida glabrata and C. albicans) to spread more widely in the intestine. Significant fluctuations in the number of Enterococcus spp. bacteria genus were observed: by 80 times within range of each of the three experimental groups of rats with addition of herbicide with benzoate and saccharin to the diet.

Evaluation of the cleansing and disinfection protocol of pig-breeding premises contaminated by the porcine epidemic diarrhea virus

The results of the determination of the efficiency of the cleaning and disinfection protocol of pig farms contaminated by the porcine epidemic diarrhea virus are presented in this study. The research was conducted in the Scientific Research Center of Biosafety and Environmental Control AIC of the Dnipro State Agrarian and Economic University. The experimental part was carried out on the basis of a permanently infected pig breeding farm of the southern region of Ukraine. In order to determine the critical points, a selection of samples of washings/scraps from the surfaces of the pig farm’s premises and the objects of the environment was carried out before cleaning and disinfection and after 1 and 2 premises’ sanitation. A total of 204 tests were analyzed. Disinfection of the territory of pig farms and livestock premises was carried out in accordance with generally accepted methods. Before disinfection, a thorough mechanical cleaning of livestock premises was carried out, with subsequent irrigation of all surfaces with cold tap water. Disinfection of premises and the territory of the pig complex was carried out by the wet cleaning of surfaces with a solution of disinfectant based on glutaraldehyde after preparatory operations in the absence of animals. After disinfection the premises were additionally processed with Ca(OH)2 solution. Washes were obtained with a sterile swab from a surface of 10.0 cm2 by inserting a swab into a test tube with a sterile physiological solution. Scraps from the surfaces were obtained with a disposable sterile blades from the scalpel. The obtained material was placed in disposable sterile plastic 1.5–2.0 cm3 test tubes. The study of the presence of the PEDV was performed using a real-time polymerase chain reaction with the previous reverse transcription of RNA. It was established that the causative agent of PED in a permanently infected farm contaminates most of the production and auxiliary premises and objects of the environment. The one-time cleaning and disinfection protocol does not allow free the farm from the PEDV completely. Only two-time sanitation of premises and objects of the environment with the laboratory control of the efficiency of cleaning and disinfection contributes to the effective eradication of the PEDV.

The Use of Platelet-Rich Fibrin for Immediate Closure of Acute Oroantral Communications: An Alternative Approach

Currently, the buccal advancement flap, palatal rotational flap, and buccal fat pad techniques are frequently used to close an oroantral communication (OAC). In this study, platelet-rich fibrin (PRF) clots were used for immediate closure of acute OACs after extraction of posterior maxillary molars. Acute oroantral perforations larger than 3mm in diameter in 21 patients were treated uneventfully using PRF clots. None of the patients had systemic diseases or symptoms of sinus disease. After tooth extraction, the diagnosis of acute OAC was examined by the Valsalva maneuver (compressing the nostrils with the fingers and then blowing out air). After detection of an air leak, a modified ball burnisher instrument 3mm in diameter was used to determine whether the perforation was larger than 3mm. The postextraction socket of the tooth was cleaned with sterile physiologic saline solution and the cavity was filled with PRF. PRF clots were sutured to the gingiva to prevent them from migrating to the sinus and for stabilization. On the seventh day of follow-up, healthy granulation tissue was observed; during the third week of follow-up, epithelialized oral mucosa was detected at the extraction site in all cases. The use of PRF enables the closure of OACs without primary flap closure or any other surgical interventions. Thus, the use of PRF for immediate closure of acute OACs will make the treatment of OACs less traumatic and easier and will eliminate the need for special surgical expertise.

Antimicrobial Activity of Chlorhexidine against Multi-Species Biofilm Formation

In the present work, the efficacy of the Ti–7.5Mo alloy nanotube and Ti–7.5Mo alloy nanotube with chlorhexidine against bacterial biofilm formation was evaluated. Nanotubes were processed using anodization in 0.25% NH4F electrolyte solution. Biofilms were cultured in discs immersed in sterile brain heart infusion broth (BHI) containing 5% sucrose, inoculated with microbial suspension (106 cells/ml) and incubated for 5 days. Next, the discs were placed in tubes with sterile physiological solution 0.9% sodium chloride (NaCl) and sonicated to disperse the biofilms. Tenfold serial dilutions were carried and aliquots seeded in selective agar, which were then incubated for 48 h. Then, the numbers CFU/ml (log 10) were counted and analyzed statistically. Scanning electron microscopy (SEM) on discs with biofilms groups and contact angle was carried out. The results show that there is no difference in bacterial adhesion between of the Ti–7.5Mo alloy nanotube and Ti–7.5Mo alloy nanotube with chlorhexidine.

Microbiological evaluation of dental stone casts after immersion in sodium hypochlorite and peracetic acid

Objective: the purpose of this study was to evaluate the efficacy of disinfection of type III dental stone by immersion in 1% sodium hypochlorite and 0.25% peracetic acid at different periods of time (1, 5 and 10 min). MaterialandMethods: silicon dies were previously infected with strains of Bacillus subtilis for 15 min. Then, type III gypsum stone (Herodent, Vigodent COLTÈNE SA, Rio de Janeiro, Brazil) was inserted into the cavities to obtain contaminated specimens. A sterile silicone die was used to obtain uncontaminated specimens. The specimens were separated into positive and negative control groups, and further divided into the following groups: blocks immersed in sterile physiologic solution for 1, 5 or 10 min; blocks immersed in 1% sodium hypochlorite for 1, 5 or 10 min; and blocks immersed in 0.25% peracetic acid for 1, 5 or 10 min. All the groups were double-plated and incubated at 37 ?C for 24 h. Results: the results were expressed in colony forming units (CFU/ml) and the data were submitted to the Kruskal-Wallis test followed by Dunn’s test. The results showed that immersion in 1% sodium hypochlorite and 0.25% peracetic acid resulted in complete disinfection of the test specimens at all test periods (p &lt; 0.01), whereas immersion in saline did not provide effective disinfection. Conclusion: it can be concluded that both 1% sodium hypochlorite and 0.25% peracetic acid provided effective disinfection in dental stone specimens immersed in the solutions described above, at different periods of time.KeywordsDisinfection; Peracetic acid; Sodium hypochlorite; Stone casts.

172 ANTIVIRAL ACTION OF PROPOLIS AQUEOUS EXTRACT DURING MATURATION OF INFECTED BOVINE OOCYTES

The objective of this study was to evaluate the decrease of virus replication in BoHV-1 (Colorado strain, 106.5 TCID50/mL) after the treatment using propolis aqueous extract (PAE) during in vitro maturation of infected bovine oocytes (24 h). Cow ovaries were obtained from a local slaughterhouse (Nelore breed) and transported to the laboratory. Cumulus-oocyte complexes (COC) were aspirated from follicles and separated into 4 groups (number of replicates for all groups = 6), which were exposed to 20 μL of sterile physiological solution (SPS), 100 μL of the in vitro maturation (IVM) medium [G1 (control), n = 609]; 10 μL of BoHV-1 (106.5 TCID50/mL) virus, 100 μL IVM medium, and 10 μL of SPS (G2, n = 786); 10 μL of PAE in 0.001% in SPS, 100 μL IVM medium, and 10 μL of SPS (G3, n = 819); 10 μL of PAE extract in 0.001% in SPS, 10 μL of BoHV-1 (106.5 TCID50/mL) virus and 100 μL of the IVM medium (G4, n = 734). All groups were kept for 24 h at 38.5°C, 5% CO2 in air. After the IVM, we analysed COC expansion and the presence of a polar body by optical microscope as well as viral replication by titration (Reed and Muench test) after 72 h of co-culture with Madin-Darby bovine kidney (MDBK) cells. The G1, G3, and G4 showed steady expansion of the cumulus cells and ooplasm with uniform appearance. The G2 did not have expansion of the cumulus cells. In contrast, the cytoplasm showed degenerative appearance and an absence of maturity in numerous oocytes. The maturation rates were as follows: G1 = 79% (482/609), G2 = 51% (407/786), G3 = 80% (662/819), and G4 = 76% (565/734). The differences among groups in maturation rates were compared using the chi-squared test (α = 5%) and the average titrations using the Mann–Whitney test (α = 5%). There was a significant difference (P &lt; 0.01) among G1 and G2 evincing the interference of the virus maturation. The extract did not affect maturation as there was no difference among G1 and G3 (P = 0.43). The main step was finding no significant difference between the groups G1 and G4, (P = 17) proving that the extract interferes with viral replication. The titration after co-culturing the oocytes in MDBK demonstrated that G4 (average titrations = 1.63 × 103 titration) showed a lower rate of viral replication, the Mann-Whitney test, when compared to group G2 (average titrations = 6.04 × 107) which has not been subjected to treatment with PAE (P = 0.02). These results indicate that the propolis aqueous extract reduces the rate of viral replication without interfering with the maturation of oocytes and, therefore, it can be a conclusion that the analysis of the action of the molecules of this extract (by proteomics, for example) and future studies should be directed towards identifying the effect of extract on antiviral activity during the assessment of oocyte competence and embryo development.

Isolation of Bacillus sphaericus from Lombok Island, Indonesia, and Their Toxicity against Anopheles aconitus.

Malaria is endemic to Lombok Island, Indonesia. One approach to suppress malaria spread is to eliminate anopheline larvae in their habitat and the environmentally safe agent is bacteria, that is, Bacillus sphaericus. However, there is no information regarding local isolate of B. sphaericus that is toxic to mosquito larvae from Lombok. The aims of the study were to isolate B. sphaericus from soil in areas close to beach surrounding Lombok Island and to test their toxicity against 3rd instar Anopheles aconitus larvae. Soil samples were collected from 20 different sampling locations from Lombok Island and homogenized with sterile physiological salt solution. Suspension was heat-shocked at 80°C for 30 minutes and then spread onto antibiotic-supplemented NYSM solid medium. Colonies grown were characterized and subjected to initial toxicity test against anopheline larvae. Isolates with more than 50% killing percentage were subjected to bioassay testing against anopheline larvae. From 20 locations, 1 isolate showed mild toxicity (namely, isolate MNT) and 2 isolates showed high toxicity (namely, isolates SLG and TJL2) against An. aconitus. Those 3 isolates were potentially useful isolates, as they killed almost all larvae in 24 hours. The discovery of toxic indigenous isolates of B. sphaericus from Lombok Island opens opportunity to develop a biopesticide from local resources.

PCR identification of endodontic pathogens and DNA quantification in samples from teeth with posttreatment apical periodontitis

Aim: The aim of this clinical study was to quantify the concentration of DNA and to detect selected bacterial species from samples of infected root-fi lled teeth with post-treatment apical periodontitis after removal of gutta-percha (S1), after chemo-mechanical preparation at the fi rst appointment (S2), 5 days after the canal was fi lled with sterile physiological solution (S3), after reinstrumentation at the second appointment (S4), and 14 days after an intracanal dressing was placed at the third appointment (S5). Methods: Fifteen root-fi lled teeth were selected. Removal of gutta-percha was performed using the crown-down technique. Chemo-mechanical preparation was performed with hand fi les associated with 2% chlorhexidine gel. An intracanal dressing based on Ca(OH)2 was used. DNA was extracted from the samples and 14 endodontic 16S rDNA species-specifi c primers were tested. The concentration of DNA was quantifi ed using a NanoDropTM 2000 spectrophotometer. Results: Bacteria were present in all cases at all sampling times, as revealed by a universal primer. DNA was isolated from all samples, with an average concentration of 4.24 ± 2.9 ng/µL (S1), 3.39 ± 1.54 ng/µL (S2), 4.0 ± 1.94 ng/µL (S3), 2.66 ± 0.98 ng/µL (S4) and 3.97 ± 2.32 ng/µL (S5). Parvimonas micra and Enterococcus faecalis (S1), P. micra (S2), Porphyromonas endodontalis and E. faecalis (S3), E. faecalis and Prevotella nigrescens (S4/S5) were the species most frequently deteced. DNA concentration reductions were detected between S3 and S4 (p = 0.0256), whereas an increase was found between S4 and S5. Conclusion: A wide variety of bacterial species was detected in root-fi lled teeth with post-treatment apical periodontitis. Moreover, the use of an intracanal dressing was unable to further reduce the concentration of bacterial DNA.

Whole Pelvic Radiation Therapy Versus Prostate Only in Patients With Intermediate-, High-, and Very High-Risk Prostate Cancer According NCCN Criteria Treated With Radical Intention: Recap Data Base

rectal dose and serving as a fiducial marker in patients undergoing radiation therapy for prostate cancer. Materials/Methods: Biodegradable balloon was inserted transperineally, between the anterior rectum and the prostate, through guidance of transrectal ultrasound. Following insertion, the balloon was inflated with sterile physiological solution and sealed. The procedure was performed under mild sedation and in an outpatient clinic. A week later the patients underwent CT simulation and three weeks later they started the radiation course. The radiation protocol consisted of 2.5 Gy once a day, 5 times a week for 28 fractions, to a total dose of 70 Gy in 5 pts. The treatment was delivered through volumetric modulated arc therapy and accuracy was evaluated through cone beam CT (CBCT). Results: Six patients, so far, underwent implantation. Their characteristics include mean age 71 years (range, 61-79 years), PSA 9.3 (range, 5.9-17.3), Gleason score 3+3 in 5 pts and 4+3 in one, and stage T1c (5 pts) and T2 (1 pt). The dimension of the balloon were 4 x 2 x 1.5 cm (CC, lateral and AP). One patient developed urinary retention (the urine catheter was removed a day later) and another had mild bleeding (spontaneously stopped, stable parameters). Following treatment planning the mean V70 of the rectum was 0 (range, 0-0.3) V60 7.1 (range, 2-16) and the V50 was 23 (range, 9-39). The V60 of the bladder was 21 (range, 6-41). The balloon was demonstrated in the CBCT reconstruction and served as a reference point for daily radiation delivery. No immediate side effects were noted during the treatment course. Along a mean F/U of 8 months (range, 6-10 months) 1 out of 6 pts developed grade 1 rectal toxicity and urinary toxicity. No other complications were noted. Conclusions: The biodegradable perineal balloon has a good safety profile, and is efficient in reducing the dose to the rectal wall and in serving as a fiducial marker. Higher daily radiation dose and possible injection of contrast material into the balloon are currently being considered. Author Disclosure: R. Ben-Yosef: L. Stock Options; yes. T. Heres: None. A. Berniger: None. R. Carmi: None. I. lvovich: None. Y. Genesin: None.

Towards in vivo biosensors for low‐cost protein sensing

In vivo biosensing requires stable transistor operation in high-salt concentration bodily fluids while exhibiting impermeability to mobile alkali ions that would otherwise render the metal-oxide-semiconductor (MOS) threshold voltage to drift. Metal oxide semiconductor capacitor structures using Al 2 O 3 as the gate dielectric were soaked in a sterile physiological buffer solution (PBS) up to 24 hours and for thicknesses from 100 to 10 nm. The triangular voltage sweep technique characterised alkali ion penetration, and measured no detectable alkali ions for the Al 2 O 3 capacitors. By contrast, the dose of alkali ions in silicon dioxide MOS capacitors steadily increased with increasing soak times in the PBS solution.

152 EVALUATION OF POMEGRANATE (PUNICA GRANATUM) AS REPLICATION INHIBITOR OF BoHV-1 COLORADO STRAIN DURING IN VITRO MATURATION OF INFECTED BOVINE OOCYTES

The widespread use of biotechnologies in animal breeding in Brazil has enabled greater control over the transmission of pathogens. Nevertheless, disease transmission continues to be a significant concern and justifies the search for better control agents. The objective of this study was to evaluate whether the detrimental effects of the experimental infection of oocytes with BoHV-1 (Colorado strain, 108 TCID 50 mL–1) during in vitro maturation (24 h) could be reduced by the viral inhibitor ethanolic extract of Punica granatum (PG). Cattle ovaries were obtained from a local slaughterhouse (unknown breed), and transported to the laboratory. Cumulus–oocyte complexes (COC) were aspirated from follicles and allocated into four groups, which were exposed to: 10 µL of sterile physiological solution (G1 [control], n = 125); 10 µL of PG extract in 0.01% sterile saline solution (G2, n = 149); 10 µL of BoHV-1 virus (G3, n = 151); or 10 µL of PG extract in 0.01% and 10 µL of BoHV-1 virus (G4, n = 144). All groups were matured in 100 mL of IVM for 24 h at 37.5°C, 5% CO2 in air. After in vitro maturation, we evaluated COC expansion and presence of a polar body by optical microscope, and viral replication by titration (Reed and Muench test) after 72 h co-culture with Madin-Darby bovine kidney (MDBK) cells. Differences among groups in maturation rates were compared by chi-square test, and in titration by t-test. The G1, G2, and G4 showed steady expansion of the cumulus cells and ooplasm with uniform appearance. The G3 did not show expansion of the cumulus cells and ooplasma showed degenerative appearance. The maturation rates were as follows: G1 52% (65/125); G2 45.63% (68/149); G3 27.81% (42/151); G4 41.66% (60/144). We used the χ2 test (P ≤ 0.05) for the rate of maturation and the t-test (P ≤ 0.05) for the titration data. A reduction (P ≤ 0.05) in maturation rate was observed in G3, when compared to the G1 [control]. Oocytes exposed to BoHV-1 virus and matured in the presence of PG extract (G4) had lower (P ≤ 0.05) rates of viral replication than those matured in the absence of PG (G3). These results support the conclusion that PG extract reduces the viral rate of replication without interfering in oocyte maturation, and may be an alternative to sanitary control protocols.