Antimicrobial Peptides Epinecidin-1 and Beta-Defesin-3 Are Effective against a Broad Spectrum of Antibiotic-Resistant Bacterial Isolates and Increase Survival Rate in Experimental Sepsis

The antimicrobial peptides human Beta-defensin-3 (hBD-3) and Epinecidin-1 (Epi-1; by Epinephelus coioides) could be a promising tool to develop novel antibacterials to combat antibiotic resistance. The antibacterial activity of Epi-1 + vancomycin against methicillin-resistant Staphylococcus aureus (22 isolates) and Epi-1 + hBD-3 against carbapenem-resistant isolates of Klebsiella pneumoniae (n = 23), Klebsiella aerogenes (n = 17), Acinetobacter baumannii (n = 9), and Pseudomonas aeruginosa (n = 13) was studied in vitro. To evaluate the in vivo efficacy of hBD-3 and Epi-1, ICR (CD-1) mice were injected intraperitoneally with a lethal dose of K. pneumoniae or P. aeruginosa. The animals received a single injection of either sterile saline, hBD-3 monotherapy, meropenem monotherapy, hBD-3 + meropenem, or hBD-3 + Epi-1. Studied peptides showed antibacterial activity in vitro against all studied clinical isolates in a concentration of 2 to 32 mg/L. In both experimental models of murine sepsis, an increase in survival rate was seen with hBD-3 monotherapy, hBD-3 + meropenem, and hBD-3 + Epi-1. For K. pneumoniae-sepsis, hBD-3 was shown to be a promising option in overcoming the resistance of Klebsiella spp. to carbapenems, though more research is needed. In the P. aeruginosa-sepsis model, the addition of Epi-1 to hBD-3 was found to have a slightly reduced mortality rate compared to hBD-3 monotherapy.

A Nonrandomized Phase 2 Trial of EG-Mirotin, a Novel, First-in-class, Subcutaneously Delivered Peptide Drug for Non-Proliferative Diabetic Retinopathy

ABSTRACTBackgroundEG-Mirotin, which includes an active ingredient; EGT022, targeting Non-Proliferative Diabetic Retinopathy (NPDR), the early stage of retinopathy. EG-Mirotin is a drug that is used before capillary damage progresses to an irreversible stage. Safety and efficacy of EG-Mirotin were investigated in subjects with Type 1 or 2 diabetes and NPDR with the degree from moderate to severe.MethodsSubjects (n=10, 20 eyes) satisfying the selection criteria through the screening test were administered EG-Mirotin once a day (3 mg in 1.5 ml of sterile saline) for 5 days, 5 times in total, and were evaluated of the Ischemic index changes and safety. End-of-study (EOS) is performed approximately 8 weeks ± 1 (57 days ± 7) from the first dose.ResultsA total of 4 Treatment Emergent Adverse Events (TEAE) were observed in 2 subjects out of 10 (20.00%) who received the investigational drug. Among them, no subjects were reported experiencing a TEAE related to the investigational drug. All injections were well tolerated (3 mg in 1.5 ml of sterile saline) with no dose-limiting adverse events, deaths, serious adverse events. The overall average percent change in ischemic index at each evaluation point compared to baseline was statistically significant (Greenhouse-Geisser F=9.456, p=0.004 for the main effect of time), and a larger change was observed when the baseline ischemic index value was high (Greenhouse-Geisser F=10.946, p=0.002 for the time*group interaction).ConclusionsEG-Mirotin was well tolerated and found to reverse the ischemia and leakage of capillaries in the retina caused by diabetes.

Accelerative Effect of Cinnamon Nanoparticles as well as HAMLET on Healing of Wounds Infected with MRSA in Diabetic Rats

Objective. The aim of the present study was to investigate the effect of cinnamon nanoparticles (CNPs) on healing of wounds infected with methicillin-resistant Staphylococcus aurous with human alpha-lactalbumin made lethal to tumor cells sensitization in diabetic rats. Methods. We included fifty diabetic male rats and divided them into 5 groups. There were 10 rats in each group as follows: CONTROL group: we did not infect the CONTROL group. The wound was only covered with sterile saline 0.9% solution (0.1 mL). INFCTD group: in this group, the wounds were infected with MRSA and covered with sterile saline 0.9% solution (0.1 mL). INFCTD-HMLT group: in this group, the wounds were infected with MRSA and HAMLET (100 μg). INFCTD-CNM group: in this group, the wounds were infected with MRSA and 0.1 mL CNPs (1 mg/mL) were applied topically to wounds. INFCTD-HMLT-CNM group: in this group, the wounds were infected with MRSA, HAMLET (100 μg), and 0.1 mL CNPs (1 mg/mL). Results. Bacteriology, wound area reduction measurements, biochemistry, histomorphometrical studies, hydroxyproline levels, and reverse transcription polymerase chain reaction for caspase-3, Bcl-2, and p53 showed significant difference between rats in the INFCTD-HMT-CNM group in comparison with other groups ( P < 0.05 ). Conclusions. Accelerated healing of diabetic wounds infected with MRSA showed that local application of cinnamon nanoparticles along with HAMLET sensitization on S. aureus-infected wound could be taken into consideration.

Change of Embryotoxic Effects of Metal Citrates Depending on the Duration of Their Introduction

Given the increase in cadmium in the environment (air, soil, water) through modern industrial processes, the absorption of significant amounts from cigarette smoke is relevant to studying the effect of cadmium compounds on embryogenesis. The purpose of the study: experimental study and comparison of embryolethality and embryotoxicity of metal citrates depending on the duration of their intragastric administration (during 13 and 20 days of gestation). Materials and methods. The study was performed on 120 white adult female Wistar rats that weigh 170-200 g. They were divided into 6 groups due to the intragastric administration of solutions of the studied metals – rats treated with citrates: cadmium at a dose of 1.0 mg/kg – 1st group (nfemale = 20, nemb = 166); cadmium at a dose of 1.0 mg/kg and cerium (1.3 mg/kg) – 2nd group (nfemale = 20, nemb = 185); cadmium in a dose of 1.0 mg/kg and germanium (0.1 mg/kg) – 3rd group (nfemale = 20, nemb = 184); cadmium at a dose of 1.0 mg/kg and zinc (1.5 mg/kg) – 4th group (nfemale = 20, nemb = 179); cadmium at a dose of 1.0 mg/kg and nanocomposite (iodine + sulfur + selenium) at a dose of 2.0 mg/kg – 5th group (nfemale = 20, nemb = 180), 6th group – control (nfemale = 20, nemb = 212) – proportional volume of sterile saline in the same way. Females were divided into 2 subgroups of 10 animals each, depending on the duration of administration of test substances. The embryotoxic and embryolethal effects of the test substances were evaluated according to generally accepted criteria, which were calculated according to well-known formulas. Results and discussion. Cadmium compounds harm the embryogenesis of rats in the experimental groups and increase the rates of embryolethality. The most pronounced differences in these parameters concerning the control were found in animals with isolated exposure to cadmium citrate: the rates of total embryonic mortality increased by 4.0 times in both study periods of embryogenesis and 20% from the 13th to the 20th day; increased pre-implantation mortality by 6.0 times with an increase in post-implantation mortality by 3 times on the 13th day and by 15.0 times and 2.8 times on the 20th day of embryogenesis, respectively. Depending on the duration of introduction, the indicators of pre-implantation mortality increased by 25.0% with an increase in post-implantation mortality by 22.2% in the same period. The experimental group of exposure to cadmium citrate at a dose of 1.0 mg/kg recorded the lowest indicators of the number of live fetuses and intrauterine survival with the highest resorption rate studied of embryonic development. At the same time, in the groups of combined exposure to cadmium citrate with metal citrates, a decrease in the indicators of total embryonic mortality was 50.0% - 30.0%, pre-implantation mortality – 50.0% - 25.0%, post-implantation mortality – 60.0% - 44.4 % and increase in the number of fetuses per female – 12.7% - 25.3%. Conclusion. Analysis of the results shows a pronounced embryotoxic effect of cadmium citrate at a dose of 1.0 mg/kg on the processes of embryogenesis, which is a significant increase in overall embryonic mortality, preimplantation, and postimplantation mortality compared with the control group in all studied terms. In the groups of combined action of cadmium citrate with metal citrates, the data obtained indicate a decrease in the accumulation of cadmium under the influence of the studied citrates, which allows them to be considered as potential bioantagonists of cadmium citrate

Analysis of the acute neuroinflammatory response and endogenous markers in the amygdala of animals submitted to status epilepticus by intrahippocampal pilocarpine application

Introduction: Experimental evidence and clinical evidence indicate that the inflammatory process is a crucial mechanism in the pathophysiology of epileptic seizures and temporal lobe epilepsy. The amygdala when involved in an atypical processing is associated with multiple moods such as depression and anxiety disorder and psychiatric disorders such as schizophrenia. Objective: This study investigated the acute inflammatory process and modulation of the endogenous proteins’ galectins and AnxA1 in the amygdala of animals submitted to an experimental model of temporal lobe epilepsy. Methods: The experimental procedures were approved by the Ethics Committee on the Use of Animals at UNIFESP (CEUA nº2958050814). The experiments performed in this study used data and materials that were obtained from the project “Neuroprotective and anti-inflammatory role of the mimetic peptide ac2-26 of the annexin a1 protein in intrahippocampal pilocarpine-induced status epilepticus” conducted by the advisor. The experimental model used male Wistar rats that were divided into 3 experimental groups (NAIVE; SHAM, Status Epilepticus or SE - n = 5 animals/group). Once acclimated, the animals in the SHAM and SE groups underwent stereotaxic surgery for implantation of the intracerebral cannula in the right hippocampus. The SHAM animals received sterile saline in all procedures and the NAIVE group only manipulated. The animals were monitored throughout the period and after 24 hours of experiment all animals were euthanized by overdose of thiopental to remove the brain and performed histological and immunohistochemical analysis. Results and Conclusion: Initial results demonstrate that SE and the acute inflammatory process cause damage to the amygdala, and there is also modulation of inflammatory markers in this structure. However, further studies are needed to better understand the mechanism of action in neuroinflammation in status epilepticus.

Microbial Analysis of Contact Lenses

Introduction: Contact lenses are small, thin lenses which are worn directly on the surface of the eyes. They can be worn aesthetically or to correct vision. Contact lens related eye infections can lead to serious complications such as blindness, and are associated with several risk factors such as sleeping with lenses, exposure to water, not adhering to replacement schedules, and reusing disinfecting solutions, among others. The severity of the infection may vary with the degree of pathogenicity of the microorganism. Hygiene and handling of contact lenses play a very important role. The main aim of this study is to assess the microbial analysis of contact lenses. Materials and Methods: A total of 15 lenses were collected in 2 ml sterile saline solution individually and manually agitated for five minutes. The lens was then removed using a sterile toothpick from the container. The sterile container was stored at 4°C till it was processed. 50 microliter of the sample was transferred using a pipette and inoculated on nutrient agar, blood agar and sabouraud dextrose agar. The microorganism (fungus or bacteria) were identified by standard protocol. Results and Discussion: Bacillus, Staphylococcus aureus and CONS [Coagulase Negative Staphylococci] were found in blood agar. No fungal growth was found among the samples. More importance could be given to contact lenses handling and hygiene to avoid eye related bacterial and fungal infections. Conclusion: In the present study, the total CFU (Colony Forming Unit) was found to be confluent in all the participants who wore contact lenses for 4 months compared to those who have worn contact lenses from 15 and 28 days. Only bacterial growth was seen in the culture plate. There was no fungal growth seen from the samples collected.

Novel Antimicrobial Peptides Designed Using Recurrent Neural Network Reduce Mortality in Experimental Sepsis

The amino acid sequences of 198 novel peptides were obtained using a generative long short-term memory recurrent neural network (LTSM RNN). To assess their antimicrobial effect, I synthesized 5 out of 198 generated peptides. The PEP-38 and PEP-137 peptides were active in vitro against carbapenem-resistant isolates of Klebsiella aerogenes (n=12) and K. pneumoniae (n=18). PEP-137 was also active against Pseudomonas aeruginosa (n=17). The remaining three peptides (PEP-36, PEP-136 and PEP-174) showed no antibacterial effect. Then I investigated the effect of PEP-38 and PEP-137 (a single intraperitoneal administration of a 100 µg dose 30 min after infection) on animal survival in an experimental murine model of K. pneumoniae-induced sepsis. As a control, I used two groups of mice: one received sterile saline, and the other received inactive in vitro PEP-36 (a single 100 µg dose). The PEP-36 peptide was shown to provide the highest survival rate (66.7%). PEP-137 showed a survival rate of 50%. PEP-38 was found to be ineffective. The data obtained can be used to develop new antibacterial peptide drugs to combat antibiotic resistance.

Spontaneous globe subluxation: a case report and review of the literature

Background Spontaneous globe subluxation (SGS) is an atraumatic anterior dislocation of the eyeball. It is exceedingly rare. Understanding SGS predisposing factors may help uncover its etiology and undertake vision-saving management. Case presentation A 48-year-old female presented to the ED with her right eye out of its socket. She reported blurry vision, photophobia, and pain in the affected eye. She was unable to close her right eyelid and was in obvious distress. On arrival, her blood pressure was elevated. Her medical history was notable for hypertension and obesity. On physical examination, extraocular eye movements were not intact, and the globe appeared whole and round. She was also unable to count fingers with the affected eye. There was no visible trauma to the face. Multiple wet gauzes with sterile saline were placed over the displaced eyeball. Direct and even pressure was applied on the globe. Within 30 s, the globe was reduced back in. The patient was able to close her eyelids and reports substantial pain relief with reduction. A CT scan of the orbits was then obtained, demonstrating mild bilateral proptosis. The globes were normal and symmetric. No intraconal or extraconal abscess or infection was seen. There were no intraconal or extraconal masses. There was no acute orbital traumatic injury, no avulsion of the optic nerve, ocular rupture, or retrobulbar hematoma. After reviewing the case with an ophthalmologist, a follow-up appointment with the ophthalmologist was arranged. The patient was discharged on erythromycin ointment. Post-discharge investigation of the CT imaging revealed dilated optic nerve sheaths, tortuosity of the optic nerve, and empty sella. Conclusions In addition to causing distress and severe anxiety, SGS poses numerous immediate as well as long-term complications. Traction of the optic nerve and retinal vasculature may potentially cause retinal venous congestion and loss of visual acuity with potential vision loss. In the absence of known risk factors or disease processes, orbital imaging and serological studies for thyroid ophthalmopathy should be considered.

EVALUATION OF CUMULATIVE PROPERTIES OF THE COMPLEX TOOL Z-88

This present work is devoted to the study of cumulative properties of the complex means Z-88. The studies were carried out on intact mongrel laboratory rats that had previously passed quarantine measures for 12 days. Two equivalent groups were created. The animals of the experimental group were injected intramuscularly in the thigh area with Z-88, and the control group was similarly injected with 0.9% sterile sodium chloride solution. On the first day of the experiment, the rats were injected with test drugs at a dose of 0.5 ml per animal, which was 1/10 of the maximum volume used in experiments to determine the parameters of acute toxicity. Both Z-88 and sterile saline solution were administered once a day throughout the experiment. Z-88 was administered once a day throughout the experiment. Starting from the fifth day, in every subsequent four days, the dose of the drug was increased by 1.5 times. The duration of the experiment was 24 days. During the experimental period of visible changes on the part of the studied indicators.

Comparison of Iodoform and Paraminobenzoate for The Managment of Dry Socket

Objective: The aim of present study is to compare effectiveness of two different modalities Iodoform and Para-Aminobenzoate for the management of Dry socket. Material And Methods: Patients reported with dry socket at the OPD of Oral and Maxillofacial Surgery, Institute of Dentistry, Liaquat University of Medical and Health Sciences, after mandibular molar tooth extraction were included After thorough irrigation with sterile saline and followed up for three alternative days by replacing dressing and then findings were recorded in the designated proforma of the patients. Pain was measured by Visual Analogue Scale (VAS). Results: The mean age of group A was 26.18+4.41 years and mean age in group B was observed 26.0+3.92 years. Males were predominantly reported in both groups. In this study 3rd molar extraction was commonest as 81.1% in group A. Similarly 3rd molar extraction was 85.5% in group B. Most of the cases of both groups underwent surgical extraction. In our study mostly onset symptoms were seen at 72 hours in both study groups. On day 3 and 4 pain was markedly decrease in patients of group B as compared to group A, p-values were quite insignificant (p=0.001). Conclusion: Para-Aminobenzoate showed better effectiveness in decreasing the pain from day 2nd. Keywords: Dry socket, Iodoform, Para-Aminobenzoate, Pain