Ml Of Sterile Water Research Articles

First Report of Bacterial Leaf Streak Disease of Rice Caused by Xanthomonas oryzae pv. oryzicola in Ivory Coast.

Bacterial Leaf Streak (BLS) of rice caused by Xanthomonas oryzae pv. oryzicola (Xoc) is considered as the third emerging infectious disease of rice in Africa. First reported in Africa in the 1980s, the disease is now present in at least eight African countries including Burundi, Burkina Faso, Kenya, Madagascar, Mali, Nigeria, Senegal and Uganda. Yield loss caused by BLS is estimated at 20 to 30% (Sileshi and Gebeyehu, 2021). To our knowledge BLS has so far not been reported in Ivory Coast. While BLS has not been described in the adjacent rice-growing countries Ghana and Liberia, Xoc strains isolated from samples collected between 2003 and 2011 in Burkina Faso and Mali have been characterized (Wonni et al., 2014). Xoc is transmitted through rice seeds which favors its spread through rice trading (Sileshi and Gebeyehu, 2021). Given the extensive rice trade between Burkina Faso, Mali and Ivory Coast, we hypothesized that BLS might also be present in this country. Field surveys were carried out across Ivory Coast in October 2018. Typical symptoms of the disease, e.g. translucent lesions in the form of yellow-brown to black streaks with sometimes visible droplets of exudates on the leaf surface, were observed in the area of Korhogo. 5cm-long leaf pieces were successively disinfected, rinsed in sterile water, and then ground using the Qiagen Tissue Lyser System (QIAGEN, Courtaboeuf, France). Leaf powder was resuspended in 1.5 ml of sterile water and incubated at room temperature for 30 minutes. Then, 10 μl of the suspension was streaked on semi-selective PSA medium and incubated at 28 ° C for 3 to 7 days. Colonies characteristic of Xoc, i.e. round, convex, mucous and straw yellow in color were purified from 6 individual samples from 2 distinct sites in Korhogo. To confirm their identity, isolated strains underwent a pathogenicity and molecular characterization test. The multiplex PCR developed for the identification of X. oryzae pathovars (Lang et al., 2010) revealed for all the isolates the characteristic PCR profile of Xoc (two amplicons of 324 and 691 base pairs). Strains of Xoc BLS256 and Xoo PXO99 were used as controls. The pathogenicity test was performed on 5 weeks-old plants of O. sativa cv. Azucena leaves by infiltration with a needleless syringe of a bacterial suspension at an optical density of 0.5. After 7 days of greenhouse incubation (27 ± 1°C with a 12-hour photoperiod), all infiltration points (2 infiltrations x 3 plants per isolate) developed water-soaked lesions identical to the one challenged with BLS256 while water-infiltrated leaves remained asymptomatic. These lesions were collected and subjected to the isolation and multiplex PCR processes described above, thus fulfilling Koch's postulate. Finally, three of the isolates were subjected to sequencing of the housekeeping gene gyrB by PCR amplification using the primers XgyrB1F and XgyrB1R (Young et al., 2008). Analysis of 780bp of the gyrB sequence of strains CI_k1-1, CI_k2-2 and CI_k3-2 revealed 100% identity with the gyrB sequence of Xoc reference strain BLS256 (Acc. No. CP003057) and 10 polymorphic nucleotides compared to the Xoo reference strain PXO99A (Acc. No. CP000967). To our knowledge, this is the first report of BLS in Ivory Coast supported by molecular characterization methods. New surveys in Ivory Coast and neighboring countries where the disease has not been reported will allow to implement collections and assess disease incidence as part of future control strategies.

First Report of Phytopythium litorale Causing Crown and Root Rot on Rhododendron pulchrum in China.

During a survey of pathogenic oomycetes in Nanjing, China from June 2019 to October 2020, at least ten adjacent Rhododendron pulchrum plants at a Jiangjun Mountain scenic spot showed symptoms of blight, and crown and root discoloration . Symptomatic root tissues collected from three 6-year-old plants were rinsed with water, cut into 10-mm pieces, surface sterilized with 70% ethanol for 1 min, and plated onto 10% clarified V8 PARP agar (cV8A-PARP) containing pimaricin (20mg/liter), ampicillin (125mg/liter), rifampicin (10 mg/liter), and pentachloronitrobenzene (20mg/liter). Four Pythium-like isolates were recovered after three days of incubation at 26°C, and purified using hyphal-tipping. Ten agar plugs (2×2 mm2) of each isolate were grown in 10 mL of 10% clarified V8 juice (cV8) in a 10 cm plate at 26°C for 3 days to produce mycelial mats, and then the cV8 was replaced with sterile water. To stimulate sporangial production, three to five drops of soil extract solution were added to each plate. Sporangia were terminal, ovoid to globose, and the size is 24 to 45.6 (mean 34.7) (n=10.8) in length x 23.6 to 36.0 (mean 29.8) (n=6.2) in width. Gametangia were not observed in cV8A or liquid media after 30 days. For colony morphology, the isolates were sub-cultured onto three solid microbial media (cV8A-PARP, potato dextrose agar, corn meal agar) . All isolates had identical morphological features in the three media. Complete ITS and partial LSU and cox2 gene regions were amplified using primer pairs ITS1/ITS4, NL1/NL4, and FM58/FM66 , respectively. The ITS, LSU, and cox2 sequences of isolate PC-dj1 (GenBank Acc. No. MW205746, MW208002, MW208003) were 100.00% (936/936 nt), 100.00% (772/772 nt), and 99.64% (554/556 nt) identical to those of JX985743, MT042003, and GU133521, respectively. We built a maximum-likelihood tree of Phytopythium species using the concatenated dataset (ITS, LSU, cox2) to observe interspecific differences. Based on the morphological characters and sequences, isolate PC-djl was identified as Phytopythium litorale . As the four isolates (PC-dj1, PC-dj2, PC-dj3 and PC-dj4) tested had identical morphological characters and molecular marker sequences, the pathogenicity of the representative isolate, PC-dj1, was tested using two inoculation methods on ten one-year-old R. pulchrum plants. For the first inoculation method, plants were removed from the pot, and their roots were rinsed with tap water to remove the soil. Each of these plants was placed in a glass flask containing 250 mL of sterile water and 10 blocks (10 x 10 mm2) of mycelial mats harvested from a three-day-old culture of P. litorale, while the other plant was placed in sterile water as a control, and incubated at 26°C. After three days, symptoms including crown rot, root rot and blight was observed on the inoculated plants whereas the control remained asymptomatic. For the second inoculation method, ten plants were dug up to expose the root ball. Ten three-day-old cV8A plugs (5×5 mm2) from a PC-dj1 culture or sterile cV8A plugs were evenly insert into the root ball of a plant before it was planted back into the original pots. Both plants were maintained in a growth chamber set at 26°C with a 12/12 h light/dark cycle and irrigated as needed. After 14 to 21 days, the inoculated plant had symptoms resembling those in the field , while the control plant remained asymptomatic. Each inoculation method was repeated at triplicate and the outcomes were identical. Phytopythium isolates with morphological features and sequences identical to those of PC-dj1 were recovered from rotted crown and root tissues of all inoculated plants. Previously, P. litorale was found causing diseases of apple and Platanus orientalis in Turkey, fruit rot and seedling damping-off of yellow squash in southern Georgia, USA. This is the first report of this species causing crown and root rot on R. pulchrum, an important ornamental plant species in China. Additional surveys are ongoing to determine the distribution of P. litorale in the city of Nanjing.

Evaluation of Excreta/Secreta of Lucilia sericata Larvae as a New Antibacterial Candidate for Treatment of MRSA Ocular Infection

Staphylococcus aureus is the most serious cause of bacterial keratitis in most populations. Due to the resistance of this bacteria to methicillin (MRSA), the novel antimicrobial component is required for novel treatments for keratitis. Excretions/secretions (ES) from larvae of Lucilia sericata (maggot) have antibacterial activity against a number of bacteria. Around 500 larvae were washed with 1 mL of sterile water every hour for ES development. Bactericidal activity of ES was examined by time killing assay. MRSA strain was injected intrastromal into rabbit corneas. The rabbits have been split into three categories, including artificial tear, gentamicin, and maggot ES. Following, the eyes were clinically examined by a slit lamp. This evaluation included the conjunctiva, iris, and cornea. Lastly, the cornea of rabbits was collected for bacterial colony counts. After statistical analysis by ANOVA, it was found that gentamicin and ES, significantly reduced ocular infections in rabbits. Comparisons of corneal opacity scores with Mann–Whitney U-test showed a decreasing trend of therapeutic effects as shown below: gentamicin˃ES˃control. Gentamicin and ES significantly decrease CFU (number of colony-forming units) compared with the control. The mean bacterial count (log CFU/mL) from corneal culture for ES, gentamicin, and untreated groups were 6.04, 5.0, and 8.9, respectively (P≤0.05). In conclusion, the extraction of maggot larvae is useful in treating keratitis mediated by MRSA.

Bacterial minicells to the rescue: cyto-Immunotherapy for the treatment of late stage cancers with minimal to no toxicity.

Bacterial minicells to the rescue: cyto-Immunotherapy for the treatment of late stage cancers with minimal to no toxicity.

Intraureteral Indocyanine Green (ICG) in Benign Gynecologic Surgery

<h3>Study Objective</h3> To demonstrate the use of intraureteral indocyanine green (ICG) in gynecologic surgery <h3>Design</h3> Educational video highlighting a surgical technique. <h3>Setting</h3> Academic medical center. <h3>Patients or Participants</h3> Women undergoing laparoscopic pelvic surgery for benign gynecologic indications including both conventional and robotic approaches. We use this in our practice in cases where significant distortion of anatomy from presenting pathology is anticipated or in the setting of patient factors including multiple prior surgeries, radiation, active inflammation, or congenital anomalies. <h3>Interventions</h3> At the start of the procedure, rigid cystoscopy is performed using a 22 French rigid cystoscope. We combine 25 mg of ICG with 10 mL of sterile water (concentration of 2.5 mg/mL). A 5 French ureteral catheter is passed into the ureter up to 20 cm and 5 mL of ICG is gradually injected as the catheter is slowly withdrawn. This process is repeated in the contralateral ureter. Using specialized equipment, near infrared light or laser penetrates the ureter causing the ICG molecules fluoresce and emit near infrared light that is captured and electronically converted to give the ureter a green color. <h3>Measurements and Main Results</h3> The fluorescence of the ICG allows for quick identification of the ureter. The catheter used may have a smallerdiameter than stents, potentially reducing risk of ureteral edema, injury, and obstruction. Indwelling stents also have greater likelihood of thinning the ureteral wall. ICG allows for detection of small full thickness injury that may be missed by stents as extravasation will present in the surrounding tissues. There are fewer externalized foreign bodies intraoperatively for a shorter duration of time, potentially reducing risk of infection or inadvertent injury. Limitations include the need for specialized equipment and possible washout effect with ICG used intravenously or intracervically. <h3>Conclusion</h3> Intraureteral ICG helps identify the ureter intraoperatively during benign gynecologic surgery with low risk of complications and potential benefits in comparison to stents.

Earrings: An Obscured Source of Cross- Contamination in Dental Clinics of India

Purpose: This study aimed to check for the bacterial contamination of earrings worn by female dentists involved in direct patient care.&#x0D; Method: This cross-sectional microbiological study was carried out among female dentists working in clinical departments of a tertiary care dental hospital. A questionnaire concerning awareness regarding the donning of ornaments as possible source of contamination was administered. It also included questions on measures taken to prevent cross contamination through earrings and methods for disinfection of earrings used if any. Two swabs from each participant were taken, one from the earring and other from the underlying skin site using a sterile microbial swab moistened in saline. The samples were then dispensed into 1 ml of sterile water by vortexing for 45s. The vortexed sample was then inoculated onto Blood Agar. The inoculated plates were incubated for 24 hours at 37˚C aerobically. Further examination was done for the count of bacteria, type of bacteria and antibiotic sensitivity.&#x0D; Result: In total 90 swabs, 45 each from earring and underlying skin were cultured for microorganisms. All the (100%) samples were positive for bacterial contamination. The swabs obtained from underlying skin sites were found to have significantly higher microbial load. (p-value = 0.008). Sixty two percent (62%) of the participants were aware of earrings being a possible source of contamination and 55% had the knowledge that ornaments are supposed to be removed while working in patient clinics. Only 5% practiced removing the ornaments which included bangles/bracelets, rings and watches. None reported to have removed earrings while working in clinics.&#x0D; Conclusion: The results of this study show that earrings are potential source of cross contamination in a dental setting. This can act as nidus for colonization of bacteria and possible source for nosocomial infections and pathogenic transmission.

The BENAR RELAXATION THERAPY (BRT) DECREASING PSYCHOLOGIC SYMPTOMS IN DIABETES TYPE II IN CANDIBINANGUN DISTRIC

Diabetes prevalence in Yogyakarta is the second highest in Indonesia, and Diabetes sufferers are prone to experiencing physical and psychological disorders. Psychological disorders such as depression, stress, and anxiety are more at risk in Diabetes patients. This condition will be worsen the prognosis and outcome, so managing psychological disorders in Diabetes patients is essential. One of the efforts to manage mental disorders in Diabetes patients is non-pharmacological therapy such as Benar Relaxation Therapy (BRT). It combining between aromatherapy and Benson relaxation techniques which is effective for mild and moderate anxiety therapy. In this study, the BRT try to test the effectivity stress, anxiety, until depression. This therapy using three sickles of therapy and the dose of the aromatherapy was three drops in 20 ml of sterile water which formulated by the researcher. The aim study is evaluate the effect of Benar Relaxation Therapy (BRT) in reducing psychological symptoms in diabetes patients. This study did Quasi-experimental research with a pre-posttest without control group design. 14 adults Diabetes patients who ADL independently and have cellphones with video features in Candibinangun district purposively taken to this study. For three consecutive days, patients received therapy for 15 minutes. Their psychological symptoms were measured using the DASS 21 and PSQI questionnaires. Meanwhile, the measurement of physiological symptoms is to measure blood pressure, pulse, and blood glucose. The data processed using the T-test and Wilcoxon test according to the results of data normality. The results showed that Benar Relaxation Therapy effectively reduced psychological symptoms in the form of depression (p = 0.038) and improved sleep quality (p = 0.042). Physiologically, this therapy was also effective in lowering blood pressure (p = 0.00). However, this therapy is not significant in reducing psychological and physiological symptoms in stress, anxiety, pulse, and blood glucose levels. The conclusions is the Benar Relaxation Therapy in this study was effective in lowering depression scores and blood pressure. This therapy also improves the quality of sleep in type 2 Diabetes patients. However, this therapy significantly increases the patient's stress and anxiety and has no significant effect on pulse and blood glucose levels.&#x0D; Keywords: Benar Relaxation Therapy; Psychologic Symptoms

Nursery evaluation of potential endophytic Trichoderma spp. from North Sumatra, Indonesia as a biocontrol agent against Ganoderma boninense

Basal stem rot (BSR) caused by the fungus Ganoderma boninense is regarded as the most destructive disease of oil palm (Elaeis guineensis Jaqc.) causes significant economic losses in the oil palm industry. An early study on the endophytic Trichoderma spp. showed they were promising potential in suppessing and preventing BSR. However, investigation on the effectiveness of endophytic Trichoderma from North Sumatra, Indonesia in preventing BSR disease in oil palms still scarce. The aim of this study was to evaluate the effectiveness of endophytic Trichoderma from North Sumatra, Indonesia in preventing BSR disease in the nursery. The active Trichoderma mixture (ATM) was prepared by first grown in PDA for six days before mixed and homogenized in 3 ml of sterile water. The mixture was later inoculated with a ratio of 1 ml to 150 g of mixture of cooked and sterilized broken corn and rice (3:4) medium. ATM was further dried and mixed homogeneously with zeolite (1:10) for application in the nursery. The potential endophytic Trichoderma spp. in suppressing BSR was monitored in the nursery for ten months. The external and internal symptoms of disease development were observed and disease incidence (DI) was also recorded. Total phenolic content (TPC) was analyzed using Folin-Ciocalteu method to determine the effect of endophytic Trichoderma in inducing oil palm seedling resistance to BSR disease. The result showed that application of endophytic Trichoderma spp. was found to be effective in suppressing G. boninense infection, where endophytic T. reesei ET501 to be the most effective in preventing disease incidence (DI) with only 13.4 % infection after ten months of observation followed by endophytic T. asperellum ET537 (48.4 %) and T. asperellum ET523 (71.8 %), respectively. Application of endophytic Trichoderma spp. in the nursery also exhibited the potential in inducing TPC amount and it helps in developing resistance in oil palm seedlings, thus protecting the seedlings from G. boninense infection.

First report of Fusarium fujikuroi causing bulb rot on Lilium lancifolium in China.

First report of Fusarium fujikuroi causing bulb rot on Lilium lancifolium in China.

Can topical epinephrine application to the papilla prevent pancreatitis after endoscopic retrograde cholangiopancreatography? Results from a double blind, multicentre, placebo controlled, randomised clinical trial

Background and study aimsPost-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis (PEP) is a complication associated with important morbidity, occasional mortality and high costs. Preventive strategies are suboptimal as PEP continues to affect...

Echocardiographic comparison between oral acetaminophen versus ibuprofen in management of patent ductus arteriosus in neonates

BackgroundPatent ductus arteriosus (PDA) is a common complication in preterm neonates with the incidence of 30–67% in extremely preterm neonates, which results in increased pulmonary blood flow and increased incidence of further comorbidities such as chronic lung disease.ObjectiveTo evaluate the effect of oral acetaminophen versus ibuprofen in the management of PDA in premature infants by echocardiography.Patients and methodThe study was carried out on 50 preterm neonates diagnosed by echocardiography to have PDA in the neonatal intensive care unit in Menoufia University Hospital from March 2019 to September 2019. Patients were classified into two groups: group 1 included neonates who received oral acetaminophen at a dose of 15 mg/kg every 6 h for 3 days and group 2 included neonates who received oral ibuprofen at the initial dose of 20 mg/kg followed by 10 mg/kg for 3 days. Both acetaminophen and ibuprofen were administrated via an orogastric tube, which was flushed with 1–2 ml of sterile water to ensure delivery of the drug. Both groups were evaluated by echocardiography. All patients were subjected to complete history taking, thorough clinical examination, complete blood count, and kidney and liver function tests.ResultsAfter the first course of treatment, PDA closed in 21 (84%) patients who received oral acetaminophen versus 21 (84%) of those given oral ibuprofen (P = 0.766). After the second course of treatment, PDA closed in 12% of oral acetaminophen group and 8% of oral ibuprofen group.ConclusionThere was no significant difference between the treatment of PDA with either oral acetaminophen or oral ibuprofen in preterm neonates, but acetaminophen has minimal adverse effects.

1303. Characterization of Isavuconazole Serum Concentrations with Various Administration Routes in a Hospitalized Cohort

BackgroundPatients with invasive fungal infections are often critically ill and immunosuppressed with multiple comorbidities that may impact drug absorption and exposure. This study sought to characterize isavuconazole serum concentrations (ISCs) in a cohort of real-world hospitalized patients when administered by intravenous solution (IV), enteral as intact capsules, or tube as opened capsule contents.MethodsThis retrospective cohort analysis included all hospitalized patients who received isavuconazole as prophylaxis or treatment between September 2017 and September 2018 and had therapeutic drug monitoring performed. For patients receiving isavuconazole by tube, the capsules were opened and contents were diluted with 10-30 mL of sterile water. Administration was per package insert for intact capsules and IV solution. ISCs were obtained as part of routine care and were quantified by high-performance liquid chromatography. An appropriate trough was defined as within 4 hours of the next scheduled dose. Currently, there is a lack of correlation between isavuconazole exposure and efficacy or toxicity; thus, ISCs were compared between administration routes.Results93 ISCs were obtained during 65 encounters from 55 unique patients. The majority of patients were post-transplant (69.1%) and death occurred during 12 (18.5%) encounters. ISCs based on different characteristics of the cohort are shown in Table 1. All ISC assessments were detectable, median 2.3 mg/dL (Q1: 1.5 mg/dL, Q3: 3.3 mg/dL). Administration via tube achieved similar ISCs compared with IV therapy (1.6 mg/dL vs. 1.9 mg/dL, respectively). However, administration of intact capsules resulted in higher median ISCs, 3 mg/dL (Q1: 1.9 mg/dL, Q3: 4.1 mg/dL). All 14 patients with administration via tube were post-transplant, which was not shown to have a significant impact on ISCs (median, transplant 2.2 mg/dL vs. non-transplant 2.7 mg/dL).Table 1. Characterization of Isavuconazole Concentrations ConclusionISCs were detectable in all patients regardless of transplant status or location at the time of assessment. Administration of isavuconazole via an enteral feeding tube achieved comparable serum concentrations compared with FDA-approved routes of administration and may represent an important alternative for select patients.Disclosures All Authors: No reported disclosures

First report of root mat disease in a hydroponic tomato production system caused by rhizogenic Agrobacterium biovar 1 in South Korea.

In June 2019, root mat disease was observed in hydroponically cultivated tomatoes in Jinju, South Korea, which occurred in at least 30% of the plants in the greenhouse. To isolate the causal bacteria, 10 g of infested tomato root mat sample was ground with 50 mL of sterile water. A 100-µL aliquot of the homogenate was serially diluted and spread on Mannitol-Glutamate (MG) medium amended with 0.1% yeast extract (MGY) and incubated at 28°C for 48 hours. Fifteen dominant colonies that formed on the MGY medium were purified and subjected to diagnostic polymerase chain reaction (PCR) based on the virD2-ipt gene loci. Because Ti-plasmid harbors both virD2 and ipt genes, Ri-plasmid-borne Agrobacterium species with only virD2 can be differentiated using virD2-ipt PCR. To amplify virD2, the primers 5'-ATG CCC GAT CGA GCT CAA GT-3' and 5'-TCG TCT GGC TGA CTT TCG TCA TAA-3' were used; for ipt amplification, the primers 5'-GAT CG(G/C) GTC CAA TG(C/T) TGT-3' and 5'-GAT ATC CAT CGA TC(T/C) CTT-3' were used. Amplification involved an initial 94°C for 1 min and then 40 cycles at 94°C, 50°C, and 72°C for 1 min at each temperature, with a final 5-min extension at 72°C. For three strains (GNIY2, J10, and J11), virD2 PCR products, but no ipt PCR products, were identified, indicating that three strains are Ri-plasmid-borne Agrobacterium species. A pathogenicity test was performed on 2-week-old tomato plants. Bacteria isolates (GNIY2, J10, and J11) cultured overnight in LB were made into a bacterial suspension (107 cfu/mL) in 50 mM phosphate buffer. Five tomato seedling roots were cut with sterilized scissors and soaked in each bacterial suspension for 1 hour. Phosphate buffer was used as a negative control. The inoculated tomato seedlings were transplanted in new pots and placed in a greenhouse with 25°C day and 20°C night temperature set points in natural light. After 9 weeks, all inoculated tomato plants produced overgrown roots, while the negative control plants had no symptoms. To satisfy Koch's postulates, re-isolation was performed from the diseased tomato and was the re-isolated bacteria were subject to partial 16S rDNA sequencing. Biovar tests performed as previously described revealed that all three isolates were biovar 1. A representative strain (GNIY2) was deposited in the Korean Agricultural Culture Collection (KACC 21759). To confirm the identity, four housekeeping genes of KACC 21759 were sequenced (16S rRNA, trpE, rpoB, and recA) and deposited in GenBank (accession nos. MT071560, MT444428, MT444429, and MT444430). Multilocus sequence analysis performed as previously described showed that the KACC 21759 strain was grouped in Agrobacterium genomospecies 4. This is the first report on mat root disease caused by Agrobacterium biovar 1 in hydroponic tomatoes in South Korea.

Immediate Use After Reconstitution of a Biostimulatory Poly-L-Lactic Acid Injectable Implant.

Poly-L-lactic acid (PLLA) is a biodegradable, synthetic polymer that stimulates collagen production and can improve skin quality, volume, and thickness. The current reconstitution procedure for Sculptra, a PLLA-containing injectable device involves 2 hours standing time before use. To evaluate and validate an immediate-use procedure for reconstituting a PLLA-containing injectable device. Three batches of the product were shaken for 1 minute immediately after reconstitution with 8 mL of sterile water. Different physicochemical tests including viscosity, concentration of excipients (sodium carboxymethylcellulose and mannitol), pH, and particle size distribution were performed for standing times 0, 2, 24, and 72 hours after immediate shaking, and compared with the standard 2 hours standing time before shaking. The recovery and stability of optional addition of 1 mL of 2% lidocaine hydrochloride was also assessed. All physiochemical parameters evaluated were equivalent, regardless of reconstitution procedure, showing that shaking vigorously for 1 minute dissolves the excipients of the product properly without a required standing time and with no impact to the PLLA particles. There were no differences in lidocaine hydrochloride content of suspensions after 0 and 72 hours. The PLLA-containing product can be used immediately after reconstitution including vigorous shaking, as shown from physicochemical analyses. Optional addition of lidocaine hydrochloride is feasible. J Drugs Dermatol. 2020;19(12): doi:10.36849/JDD.2020.5228.

COVID-19 and Brain Pathology

Experiments on 33 Wistar rats were performed to test two interrelated hypotheses. Firstly, is the migration of fluorescein isothiocyanate (FITC) labeled endotoxin (Escherichia coli lipopolysaccharide, LPS) (5.0 μg in 5.0 μl of sterile water) via olfactory nerve into the brain after single submucous injection of LPS real? Secondly, in case such migration is feasible, is it possible to explain a number of clinical signs in patients infected with COVID-19 using the obtained facts? We are talking namely about the symptoms of damage to the terminals of olfactory and trigeminal nerves and corresponding brain neural networks due to COVID-19. Positive answers were received to both questions as a result of the experimental analysis.

Effects of E. coli bivalent vaccine and of host genetic susceptibility to E. coli on the growth performance and faecal microbial profile of weaned pigs

F4- and F18-positive enterotoxigenic Escherichia coli (ETEC) are well-known pathogens able to cause a severe swine disease with a worldwide economically importance in pig production. Vaccination of piglets with an oral live bivalent F4/F18 Escherichia coli vaccine has shown some promise and efficacy results in preventing ETEC post weaning diarrhoea (PWD); however, currently, there is a lack of knowledge on its effect on the intestinal microbial community. Furthermore, is not known if the effect of vaccination on gut eubiosis can vary according to pig genetic susceptibility to ETEC F4 and F18. The aim of this study is to evaluate the effect of an E. coli bivalent vaccine and the potential influence of the host genetic susceptibility to ETEC growth performance and faecal microbial profile of healthy piglets around weaning. A total of 288 healthy piglets at 26 days of age (d0; weaning) were divided into two different groups balanced for body weight: i) vaccine (VAX: 6.33 kg) and control (CO: 6.45 kg). At d0 VAX group was orally inoculated with 2 mL of live non-pathogenic E. coli O8:K87 (F4ac-positive) and O141:K94 (F18ac positive) vaccine (Coliprotec F4/F18; Prevtec Microbia an Elanco Company), while CO was orally inoculated with 2 mL of sterile water. At d0 and 18 days later (d18), piglets were individually weighed, and a faecal sample was collected for microbial profile of V3-V4 regions of the 16S rRNA gene using MiSeq Illumina platform. Bristles were collected to determine the genetic ETEC susceptibility by genotyping analysis of MUC4 and FUT1 polymorphisms. No effect of vaccination and genetic susceptibility was observed on growth performance. The vaccination modulated the faecal microbial composition, reducing the alpha diversity indices (P<0.05) and affecting the beta diversity (R2=0.02; P = 0.05). VAX was discriminated from CO by beneficial bacteria genera including Dialister, Prevotella, Blautia, Ruminiclostridium, Parabacteroides and Faecalibacterium. No interaction between vaccination and animal genetic susceptibility to ETEC was observed. F4 resistant genotype showed a higher alpha diversity index than the susceptible ones pre- and post-weaning (P< 0.05). These findings contributed to the body of knowledge regarding the beneficial effect of oral live bivalent F4/F18 E. coli vaccine on piglets’ gut homeostasis resulting in improved gut eubiosis.

Stability of extemporaneously compounded amiloride nasal spray.

Anxiety disorders (AD) are the most common mental conditions affecting an estimated 40 million adults in the United States. Amiloride, a diuretic agent, has shown efficacy in reducing anxious responses in preclinical models by inhibiting the acid-sensing ion channels (ASIC). By delivering amiloride via nasal route, rapid onset of action can be achieved due to direct “nose-to-brain” access. Therefore, this study reports the formulation, physical, chemical, and microbiological stability of an extemporaneously prepared amiloride 2 mg/mL nasal spray. The amiloride nasal spray was prepared by adding 100 mg of amiloride hydrochloride to 50 mL of sterile water for injection in a sterile reagent bottle. A stability-indicating high-performance liquid chromatography (HPLC) method was developed and validated. Forced-degradation studies were performed to confirm the ability of the HPLC method to identify the degradation products from amiloride distinctively. The physical stability of the amiloride nasal spray was assessed by pH, clarity, and viscosity assessments. For chemical stability studies, samples of nasal sprays stored at room temperature were collected at time-points 0, 3 hr., 24 hr., and 7 days and were assayed in triplicate using the stability-indicating HPLC method. Microbiological stability of the nasal spray solution was evaluated for up to 7 days based on the sterility test outlined in United States Pharmacopoeia (USP) chapter 71. The stability-indicating HPLC method identified the degradation products of amiloride without interference from amiloride. All tested solutions retained over 90% of the initial amiloride concentration for the 7-day study period. There were no changes in color, pH, and viscosity in any sample. The nasal spray solutions were sterile for up to 7 days in all samples tested. An extemporaneously prepared nasal spray solution of amiloride hydrochloride (2 mg/mL) was physically, chemically, and microbiologically stable for 7 days when stored at room temperature.

Antimicrobial properties of antibiotic-loaded implants.

The aim of this study was to compare the ability of tantalum, 3D porous titanium, antibiotic-loaded bone cement, and smooth titanium alloy to inhibit staphylococci in an in vitro environment, based on the evaluation of the zone of inhibition (ZOI). The hypothesis was that there would be no significant difference in the inhibition of methicillin-sensitive or methicillin-resistant Staphylococcus aureus (MSSA/MRSA) between the two groups. A total of 30 beads made of three different materials (tantalum/3D porous titanium and smooth titanium alloy) were bathed for one hour in a solution of 1 g vancomycin in 20 ml of sterile water for injection (bath concentration: 50 mg/mL). Ten 1 cm3 cylinders of antibiotic-loaded cement were also created by mixing standard surgical cement with 1 g of vancomycin in standardized sterile moulds. The cylinders were then placed on agar plates inoculated with MSSA and MRSA. The ZOIs were measured each day and the cylinders were transferred onto a new inoculated plate. For MSSA and MRSA, no inhibitory effect was found in the control group, and antibiotic-loaded smooth titanium alloy beads showed a short inhibitory effect until day 2. For MSSA, both tantalum and 3D porous titanium beads showed significantly larger mean ZOIs than cement beads (all p < 0.01) each day until day 7 for tantalum and until day 3 for 3D porous titanium. After six days, antibiotic-loaded cement had significantly larger mean ZOIs than the 3D porous titanium (p = 0.027), but no significant difference was found with tantalum (p = 0.082). For MRSA, both tantalum and 3D porous titanium beads had significantly larger mean ZOIs than antibiotic-loaded cement each day until day 6 for tantalum (all p < 0.01) and until day 3 for 3D porous titanium (all p < 0.04). Antibiotic-loaded cement had significantly larger mean ZOIs than tantalum and 3D porous titanium from day 7 to 9 (all p < 0.042). These results show that porous metal implants can deliver local antibiotics over slightly varying time frames based on in vitro analysis. Cite this article: Bone Joint J 2020;102-B(6 Supple A):158-162.

Oral nystatin prophylaxis to prevent systemic fungal infection in very low birth weight preterm infants: a randomized controlled trial

BackgroundSystemic fungal infection (SFI) is one of leading causes of morbidity and mortality in very low birth weight (VLBW) preterm infants. Because early diagnosis of SFI is challenging due to nonspecific manifestations, prophylaxis becomes crucial. This study aimed to assess effectiveness of oral nystatin as an antifungal prophylaxis to prevent SFI in VLBW preterm infants.MethodsA prospective, open-labelled, randomized controlled trial was performed in a neonatal intensive care unit (NICU) of an academic hospital in Indonesia. Infants with a gestational age ≤ 32 weeks and/or birth weight of ≤ 1500 g with risk factors for fungal infection were assessed for eligibility and randomized to either an intervention group (nystatin) or control group. The intervention group received 1 ml of oral nystatin three times a day, and the control group received a dose of 1 ml of sterile water three times a day. The incidence of fungal colonization and SFI were observed and evaluated during the six-week study period. Overall mortality rates and nystatin-related adverse drug reactions during the study period were also documented.ResultsA total of 95 patients were enrolled. The incidence of fungal colonization was lower among infants in nystatin group compared to those in control group (29.8 and 56.3%, respectively; relative risk 0.559; 95% confidence interval 0.357–0.899; p-value = 0.009). There were five cases of SFI, all of which were found in the control group (p-value = 0.056). There was no difference in overall mortality between the two groups. No adverse drug reactions were noted during the study period.ConclusionsNystatin is effective and safe as an antifungal prophylactic medication in reducing colonization rates in the study population. Whilst the use of nystatin showed a potential protective effect against SFI among VLBW preterm infants, there was no statistical significant difference in SFI rates between groups.Trial registrationNCT03390374. Registered 4 January 2018 - Retrospectively registered.

Diversity of microbes in organic and non-organic vegetable ecosystem

This study aims to determine microorganisms in organically managed land. The study was conducted in the organic land and non-organic farmer’s land in Makassar and continued at the Disease Sciences laboratory, Department of Pests and Plant Diseases, Faculty of Agriculture, Universitas Hasanuddin. The stages of the study consisted of soil sampling and 10 grams were taken to be diluted into 90 ml of sterile water, then a 10-3 dilution was carried out. The results of the dilution are taken 1ml each and then poured into the PDA media. The fungus that grows is purified on PDA media while bacteria on NA media, then identified. The results showed that in organic vegetable fields more fungi isolates were found (6 isolates): Gliocladium sp, Rhizoctonia sp, Aspergillus sp and two other isolates were unknown and gram-positive bacteria were found while non-organic was found five fungus isolates, Scopulariopsis sp., Verticillium sp., Fusarium sp., Gliocladium sp, Percentage of inhibition of fungi from organic vegetable fields ranged from 33.3 to 100% while non-organic lands 16.7 to 66.7%.