Method to obtain PRP and L-PRP for the use in routine medical practice of orthopedic traumatologist

The paper presents the results of experimental study of two methods for blood processing to obtain platelet-rich plasma (PRP). Due to the widespread clinical use of platelet-rich plasma in orthopedics and traumatology, a differential approach to the use of PRP and leukocyte- and platelet-rich plasma (L-PRP) has become relevant. Most commercial kits for PRP preparation significantly increase the cost of treatment. In addition, the main attention is focused on the concentration of platelets in the final product and the marketing features of a particular kit (separating gel, tube shape, anticoagulant, activator, etc.). The role of leukocytes in the features of the regenerative effect of platelet-rich plasma stays out of focus. The blood of volunteers was processed in two different ways and the cellular composition of the obtained products was analyzed. Sample No. 1 was used as a control for the baseline blood cell composition. Sample No. 2 was centrifuged in a sterile plastic tube, platelet rich plasma was collected manually. Sample No. 3 was centrifuged in a sterile modified syringe, platelet rich plasma was collected using a closed technique by means of connectors together with a rich layer. The results of statistical analysis showed that there were obtained totally different biological products with different concentrations of platelets and leukocytes. The first sample (control) had platelet concentration of (228.69 ± 39.15) × 109/l and leukocyte concentration of (5.18 ± 1.32) × 109/l. In the second sample (tube + manual sampling), platelet concentration was (429.38 ± 79.92) × 109/l and leukocyte concentration — (0.85 ± 0.34) × 109/l. The third sample (syringe plunger + closed vacuum collection) had platelet concentration of (541.15 ± 85.49) × 109/l and leukocytes of (6.56 ± 1.92) × 109/l. The data are given without regard to –12.5% deviation on dilution with citrate dextrose. As the result of the work, orthopedic traumatologists can use the simplest non-commercial methods for obtaining PRP and L-PRP in their practice depending on nosology and clinical tasks.

Kajian Cemaran Salmonella spp pada Daging Ayam Broiler di Pasar Tradisional dan Modern Kota Bandar Lampung

Food can be a carrier for pathogenic agents that can disease in consumers (foodborne disease). Chicken meat is one of the products from livestock which has a high consumption rate, because besides being easy to obtain, the growth of chickens is fast, and the price is also more affordable compared to large livestock types. Chicken meat is an excellent medium for microbial growth and makes it a perishable food ingredient. Foodborne illness is a disease caused by pathogenic microorganisms that contaminate food, such as Salmonella spp, Staphylococcus aureus, and Escherichia coli (E.coli). Salmonella spp infection can cause salmonellosis which irritates the digestive tract and many of them result in death. This study aims to assess the presence and number of pathogenic microorganisms Salmonella spp in broiler chicken sold in traditional and modern markets in Bandar Lampung. The research was conducted by purposive sampling of 30 chicken meat samples. Broiler chicken carcass samples were taken as whole and then stored in sterile plastic, labeled and put into a cool box filled with ice. The variables observed in this study were the presence and number of Salmonella bacteria. The data obtained were analyzed descriptively. The results of Salmonella spp examination on chicken meat samples in traditional markets in Bandar Lampung City showed a positive in 1 (one) sample or 6.7%, while in modern markets Salmonella was not found. The presence of Salmonella in chicken meat is thought to be due to contamination from the water used during the process of slaughtering and preparing the carcass, contamination from the carcass slaughtering and selling environment which does not apply good sanitary hygiene and poor personal hygiene of the traders. The conclusion obtained is that the number of Salmonella spp in chicken meat in traditional markets is not in accordance with SNI 7388 of 2009 concerning the Maximum Limit of Microbial Contamination (BMCM) in fresh meat.

The Efficiency of Hand Sanitizers Against Microbes From Coins

Objective:This study aimed to examine the effectiveness of hand sanitizers against bacteria from circulating coins during COVID 19 era.Method:22-coin samples have been collected by the research group from taxi drivers in Nablus city, Then the samples were transferred in a sterile plastic bag to the Microorganism Laboratory (17B1140) at the Faculty of Medicine and Health Sciences at An-Najah National University. The collected coins were swabbed with normal saline and cultured on nutrient agar for enrichment. After their appearance, the grown colony has been sub-cultured on Blood, MacConkey, and Uri select, for identification with the assistance of microscopical examination and chemical tests for confirmation. To answer the question of the study, four different sanitizers have been applied to measure their antimicrobial activity against microbes from circulating metal coins. Result:The results showed that sanitizer A (Hand sanitizer gel) (ג'ל להיגיינת הידיים ללא צורך , בשטיפה ובמים) is the most effective one of the four sanitizers that were used, followed by sanitizer (D)(HiGeen )(JO), then sanitizer (C)(National ) (PS), while sanitizer (B)(Icona London) (UK) was the least effective one.Conclusion:The research group have conducted research on coins circulated from taxi drivers during COVID 19 era, hence the number of isolated bacteria was low, due to the current sterilization and prevention behaviors like the using of sanitizers containing 70% of alcohol, which was the best among the used types in the current study as well as it has been observed that excessive use of alcohol- based hand sanitizers can cause microorganisms resistant and or less affected to repeated use of hand sanitizer. Hence, Hand sanitizers may need to be used appropriately in combination with other procedures.

Analysis of Salivary Metabolites by Nuclear Magnetic Resonance Before and After Oral Mucosa Cleaning of Infants in the Pre-dental Period

The aim of the present study was to verify if a protocol for cleaning the oral cavity of infants in the pre-dental period can reduce extrinsic salivary metabolites observed through Nuclear Magnetic Resonance (NMR). A cross-sectional clinical study with a convenience sample was conducted, and infants were recruited at the UFRJ Pediatric Dentistry Clinic. Participants who had used antibiotics and/or antifungals up to 3 months before and whose legal guardians did not consent or sign the Informed Consent Form were excluded. An anamnesis was performed with the guardians and the participants' intraoral clinical examination. Initial collection of unstimulated total saliva was performed using an automatic pipette with sterile plastic tips in the buccal floor region, at least 1 h after the last feeding. Subsequently, the infants' oral mucosa was cleaned with gauze moistened with filtered water, and after 5 min, a new collection was performed, using the same methodology. The obtained samples were immediately transferred on ice to the laboratory, centrifuged (10,000 g), and stored at −80°C. The NMR analyses were performed using a 500-MHz spectrometer Bruker, Germany); evaluations were done via the 1H and 1H-1H TOCSY spectra for metabolite signaling. Eleven pre-dental infants were evaluated, with a mean age of 3.8 months, including six girls (55%). Of these, nine participants (82%) were exclusively breastfed. The higher presence of components such as lactose, glucose, sugars, acetate, alanine, and lactate were observed in the samples before oral mucosa cleaning. Regarding the type of diet, more lactose was observed in the saliva of patients who were exclusively breastfed than those that received mixed feeding. We conclude that the oral mucosa cleaning of infants in the pre-dental period tends to reduce the concentration of extrinsic components from the diet, such as lactose, in the salivary metabolomic profile analyzed by NMR.

The Fertility Outcome of Royal Jelly versus Intra Uterine Insemination: A Pilot Randomized Controlled Trial Study

Background: The popularity of complementary and alternative medicine is on the rise. Objectives: The current study aimed to compare the effect of vaginal royal jelly with intrauterine insemination (IUI) on sub-fertility in men. Methods: The sample size was estimated as 27 subjects, based on a study power and confidence interval of 80% and 95%, respectively. The study was conducted in Mashhad, in the Northeast of Iran, from 2015 to 2017. Men with sub-fertility (asthenozoospermia, defined as total sperm motility below 40% and sperm concentration below 20 million/mL), with healthy wives, based on salpingography, participated in this study. Patients were assigned to the royal jelly group, which received 5 grams of royal jelly, and the IUI group, which received 75 units of Follicle Stimulating Hormone (FSH) from the second day of the menstrual cycle. Semen samples were collected in sterile plastic caps after 72 hours of sexual abstinence by normal sexual intercourse. Samples were prepared using the standard swim-up technique. Results: This study showed no significant difference between groups concerning spousal age, occupation, education (husband and wife), and social class (P = 0.745). Also, no significant difference was found between groups concerning fertility success rate (P = 0.573). Conclusions: Based on similar treatment success rates of royal jelly and IUI, it seems that royal jelly can be considered as an alternative to IUI.

Determination of microbial quality of tsire offered for sale at Hadejia Local Government Area, Jigawa state

Microorganisms grow on meat causing visual, textural and organoleptic changes when they release metabolites. A lot of factors affect the growth of microorganisms on meat, which include temperature, pH, water availability, presence of nutrients, gaseous requirement and atmosphere of storage. Microbial quality of raw meat and tsire sold at retail outlets from Yayari, Yankoli and Matsaro of Hadejia metropolis, Jigawa State were evaluated. Three samples from each outlet were collected fortnightly into sterile plastic bags, stored at 4°C in ice chest filled with ice and transported immediately to the laboratory for Total Viable Count (TVC), Total Coliform Count (TCC), Total Staphylococcus Count (TCS) and Total Fungal Count (TFC) analyses using Nutrient Agar, Mac Conkey Agar, Mannitol Salt Agar and Potato Dextrose Agar, respectively. They were incubated at 37°C for 24 hours except for the detection of fungi, which was incubated at 25°C for 5 days. Tsire samples collected from Matsaro had the highest TVC (6.0 x 104) while that from Yankoli had the highest TCS (4.0x102). The study concluded that tsire sold at Hadejia Local Government Area was safe for consumption, due to its less microbial load.

Microbial Assessment of Fresh and Frozen (For Four Days) Marine Najil Fish (Plectropomus pessuliferus)

The present study was conducted at Directorate General of Preventing Medicine and PHC, Epidemiology Dept, Ports and Quarantine Health Unit, Port Sudan, endeavors for Microbial evaluation (bacteriological load) of marine Najil fish (Plectropomus pessuliferus). And to determine the occurrence of some contaminated bacteria. 20 samples of Najil fish (P. Pessuliferus) were collected from Port Sudan fish market (in sterile plastic bags) and pre chilled with ice in thermostatic container. All the samples were tested immediately, and instantly froze for four days. Then the studied samples were tested microbiologically and results showed:For fresh fish total bacterial count (TBC) was (5.68×105) CFU/gm, and for frozen samples was (4.38×105) CFU/gm, respectively.The study concluded that, Najil fish (P. Pessuliferus) showed the highest level of contamination between fresh fish. While in frozen fish freeze seem have the higher effect (less number). And the results showed highly significant differences between fresh and frozen fish.The samples were also tested for contaminant bacteria and the result indicated the presence of Staphylococcus and E. coli, while the salmonella and vibrio were absent at each of it.

Molecular detection of Acanthamoeba spp. in Seven Crater Lakes of Laguna, Philippines

Acanthamoeba spp. are ubiquitous free-living amoeba with genotypes that cause severe pathology of the eyes, central nervous systems, and rare reports of cutaneous infections. The Seven Crater Lakes are freshwater water resources in Laguna, Philippines primarily used for aquaculture and tourism. A total of 16 surface water samples were collected from different sampling areas per Crater Lake and placed in sterile plastic containers. Samples were filtered using 1.2 μm pore size, glass microfiber filter. Filtered sediments were placed on non-nutrient agar lawned with Escherichia coli and incubated aerobically at 35 °C for 14 days. Six out of 16 water samples exhibited amoebic growth. Cystic stages revealed circular to stellate morphology under light microscopy which were initially classified as Acanthamoeba spp. DNA from positive isolates were made to react with polymerase chain reaction using Acanthamoeba specific primers JDP1 5′-GGCCCAGATCGTTTACCGTGAA-3′ and JDP2 5′-TCTCACAAGCTGCTAGGGAGTCA-3′confirmed the presence of several Acanthamoeba species. Phylogenetic analysis revealed the presence of seven isolates belonging to Acanthamoeba genotypes T4, T5, and T9. The presence of potentially pathogenic Acanthamoeba genotypes in the Seven Crater Lakes of Laguna signifies risk to human health which necessitates the development of programs, policies, and guidelines on the understanding, prevention, and management of potential human infections.

Hirudotherapy in Wound Healing

Leeches are hermaphrodite, bloodsucking parasitic worms usually found in places with fresh water. Leech therapy existed 3000 years, and it is being used at a different scope. Several species of leeches have been used in medicine, and the most common species used is Hirudo medicinalis. Leeches suck the excess blood, reduce the swelling in the tissues, and promote healing by allowing fresh oxygenated blood to reach the area until normal circulation can be restored. Pain relief from leech therapy is rapid, effective, and long-lasting in many conditions. The aim of this study was to evaluate the efficacy and duration of healing utilizing sterile medicinal leeches, Hirudinaria manillensis, in the management of pain and wound healing. Leech was taken out from its sterile tube by using a pair of non-tooth sterile plastic forceps and gloved hands. Each leech was left in place for as long as it was feeding. Leeches were removed only after they became detached from the patient. The specimen jars containing the used leeches were sealed in either a biohazard bag or in a small yellow clinical waste bin liner securely fastened with a cable tie. The leech was killed by using 70% alcohol prior to disposal into a yellow hazard bin, which undergoes incineration. All 3 patients had improvements in their condition, especially in terms of reduction in the pain and improvement in their sense of balance. All the wounds healed well. Therefore, leech therapy is effective in reducing pain and increasing perfusion to allow the wounds to heal quickly. However, a more robust trial is needed to show significance as the sample size is small.

86 Impact of steeping corn DDGS over 72 hours without or with fiber degrading enzymes and protease alone or in combination on concentration of sugars and organic acids and pH in the liquid medium

We investigated the impact of steeping corn DDGS over 72 h with or without fiber degrading enzymes (FDE) and protease (PRO) on concentration of sugar, acetic and lactic acid, and pH. The concentration of crude fat, CP, NDF and ADF in DDGS sample was 8.6, 30.8, 36.1, 12.2% DM, respectively. Treatments were: 1) DDGS steeped without enzymes (Control), 2) DDGS steeped with FDE (FDE), 3) DDGS steeped with protease (PRO), 4) DDGS steeped with combination (FDEPRO). Enzymes were added at 1% of DDGS. Briefly, 50g of DDGS was mixed with 500-mL water with or without enzymes in sterile plastic bottles and steeped for 0, 12, 24, 48 and 72-h at 37℃ with continuous agitation. Samples were aliquoted to individual bottle for each timepoint. At each time point, bottles were pulled out from the incubator for pH measurements and liquid medium supernatant for sugars and acids. Highest arabinose, xylose and glucose was observed at 12 and 24 h, followed by decrease at 48 and 72 h. At 12 h, arabinose was higher (P < 0.05) for FDE and FDEPRO than control. The highest xylose and glucose was seen at 12 h for PRO, FDE and FDEPRO while control was highest at 24 h. Acetic acid and lactic acid increased over time. At 72 h, FEDPRO had highest acetic acid compared to control and PRO, while lactic acid was highest for FDE and FDPRO compared with (P < 0.0001) control and PRO. The pH decreased (P < 0.05) over time for all treatments. At 72 h, FDE had lowest pH followed by FDEPRO, PRO and control (P < .0001). Overall, arabinose, xylose and glucose increased to a certain timepoint and decreased, while lactic and acetic acid increased over time, subsequently leading to lowest pH at 72 h. In conclusion, FDE increased concentration of sugars and acids in steeped DDGS whereas PRO had no effect.