Sterile Laminar Flow Research Articles

IMPACT OF A GEL COMPOSITION CONTAINING VARIOUS SUBSTANCES ON INDICES OF IMMUNITY AND TISSUE INFLAMMATION IN PALATINE TONSILS OF PATIENTS WITH CATARRHAL GINGIVITIS DURING CHRONIC TONSILLITIS

The study of pharmacotherapeutic (immunomodulating) action in conditions of in vitro cell cultures is currently considered appropriate and justified for determining the action of drugs at the preclinical level. It is also important to address the pathogenetic nature in using such agents, especially fro local application. This study investigated the efficacy of a multicomponent gel containing decamethoxine and hyaluronic acid in treating chronic catarrhal gingivitis among patients with chronic tonsillitis. The purpose of the work is to assess the efficacy of a gel containing decamethoxine and hyaluronic acid on the tissue of the palatine tonsils in the therapy of chronic catarrhal gingivitis in patients with chronic tonsillitis.
 Materials and methods. To investigate the effects of the main active components in the gel, we followed the recommendations of I.P. Kaidashev et al. for testing pharmacological preparations in in vitro cultures. Tissue samples from the palatine tonsils were obtained from 20 patients with chronic tonsillitis and chronic catarrhal gingivitis undergoing tonsillectomy. All procedures adhered to the guidelines set by the Laboratory of Pathophysiology and Immunology at Prof. O.S. Kolomiichenko Institute of Otolaryngology, National Academy of Sciences of Ukraine. Palatine tonsil tissue was incubated in 199 medium (Serva, Germany) containing gentamicin sulfate (Darnytsa, Ukraine) at a concentration of 120 μg/ml for 20 minutes at 4-8°C. After that, the hemorrhagic parts were removed from the whole tonsil and placed into Eagle's medium (Sigma, USA) and rinsed three to four times. Subsequently, the prepared tissue was placed in sterile vials containing enriched Eagle's medium supplemented with L-glutamine, sodium bicarbonate, 100x vitamin concentrate, 5% fetal calf serum, and 60 μg/mL gentamicin. Tonsil tissue was dissected using scissors and then washed repeatedly (5-7 times) with fresh medium to remove loosely attached cells, then, the resulting cell suspension was filtered. All procedures were conducted within a sterile laminar flow cabinet. Cell viability was assessed using light microscopy (Olympus-23, Japan) and a hemocytometer. A final cell suspension containing 2.5 million lymphocytes per milliliter of medium was prepared. Suspensions with a viability of less than 90% (blue, "dead" cells) were excluded from further experiments.
 Results. Treatment of in vitro palatine tonsil cells from patients with chronic catarrhal gingivitis and recurrent tonsillitis with the main components of the developed gel (decamethoxine and hyaluronic acid) did not induce significant changes in the concentration of α and γ-interferons. Our findings suggest that the gel composition reduces the levels of pro-inflammatory factors: interleukin-1β and immune complexes. Additionally, it appears to stimulate the production of the antimicrobial factor antistreptolysin-O by tonsil cells, potentially leading to increased antibody production against hemolytic streptococcus antigens.
 Conclusion. Based on the in vitro findings, the investigated gel composition containing decamethoxine and hyaluronic acid demonstrates potential anti-inflammatory and antimicrobial properties that stimulate the production of the antimicrobial factor antistreptolysin-O by tonsil cells, which might lead to increased antibody production against hemolytic streptococcus antigens.

Effect of Agar Weight Loss with Passive Continuous Air Monitoring According GMP Annex 1 2023

The GMP Annex 1 2023 requests that microbial environmental monitoring by active and passive microbial air samplers is actuated by continuous air monitoring. A prolonged sampling produces a loss of moisture in the agar of the Petri dish and therefore it is necessary that during the “continuous time” the dryness of the nutrient media is under control to guarantee a regular growing of the cfu. The purpose of this paper is to evaluate if the agar of settle plate (which volume was reduced with a treatment under sterile laminar flow before aspiration) has still the correct nutrient characteristics during the incubation. The results of the study are summarised in Table 1 and 2. The average loss of humidity after 4 hours in the first part of test under laminar flow was 14,7%. In the second part of test, the Petri dishes aspirated the air in a normal open environment and the average growth after incubation (indicated in cfu/ plate), in comparation with control plates with the original volume of agar, was 87%. The acceptance criteria using the US Pharmacopeia Validation Recommendation for microbiological examination (chapter ) is that the test plates had to recover < 70% of the challenge.

Cryopreserved amniotic membrane in the treatment of limb skin defects of aplasia cutis congenita: a case study

To report the efficacy and long-term outcomes of treating the skin defects of aplasia cutis congenita (ACC) with cryopreserved amniotic membrane (AM). Human amnion was obtained from the caesarean delivery of a full-term healthy pregnancy and processed in a sterile laminar flow hood, and cryopreserved in liquid nitrogen. The structure of the AM was investigated histologically and the viability of the epithelial cells was assessed after cryopreservation and compared with fresh AM and with AM preserved in phosphate-buffered saline (PBS) at 4°C. The cryopreserved AM was applied onto the lower limb skin defects of a one-month old baby with ACC. Timely AM changes were performed as necessary until the wounds healed. The structure of the cryopreserved AM was intact, with little visible difference compared with fresh AM. The viability of the epithelial cells was partially lost but still much better retained than in those preserved in PBS at 4°C. The limb skin defects were gradually re-epithelialised upon application of the AM and were completely healed after one month. The 4-month and 2-year follow-ups presented good skin texture and colour, without hypertrophic scar formation. In this case study, cryopreservation of AM presented a well preserved stromal compartment and viable epithelial layer. It also offered features such as pain relief, good attachment and adhesiveness, improved wound healing and suppressed scar formation in the treatment of ACC skin defects.

Clinical Nursing during CD4-Specific CAR T Cells Treatment for One Mycosis Fungoides Patient Improves Generalized Skin Lesion

Objective:To summarize the clinical nursing approach to CART treatment in one mycosis fungoides patient with generalized skin lesions, and to provide experience for reference in clinical nursing for such patients undergoing CART treatment.Method:Case Presentation: The patient, female, aged 43 years, was pathologically diagnosed to mycosis fungoides. Clinical state was tumor stage, as well TNMB stage was T4N3M0B1. The patient was admitted to the hospital with diffuse crimson papules, nodules, tumor, associated with brown, black pigmentation, complained of pruritus. The tumor surface ulcerated and oozing fluid at the surface of the head, bilateral cheeks, bilateral axillae, abdomen, right groin appeared infiltrated with flower like growth, the large of which was about 6x7 cm, and microorganism culture of secretory showed acinetobacter baumannii and corynebacterium striatum. Human derived CD4-CART treatment was given after chemotherapy pretreatment. The patient occurred severe granulocytic deficiency, fever, skin infection, gastrointestinal bleeding, respiratory failure, cardiac insufficiency, secondary hemophagocytic syndrome.Nursing strategy: 1. Strengthen basic care and application of sterile laminar flow beds. 2. Closely monitor physical signs, and discover early high fever, hypotension, low blood oxygen and other CRS reactions. 3. Special professional person conducted sterile dressing change to avoid cross infection; Perform weekly microorganism culture of secretory and adjust antibiotics symptomatically; Mechanical debridement and the use of debridement glue are given to the wound accompanied by yellow excoriation, black scab; Prevention of infection using fusidic acid cream, silver pyrimidine sulfate dressings, and hydrophilic silver containing dressings; Lesional wounds are covered with a lipidic hydrogel foam dressing to achieve wet healing. 4. Reinforce nutritional support and psychological care.Result:After the fusion of CAR T cells, the patient developed grade 4 cytokine release syndrome (CRS), grade 0 ICANS. Fever appeared on the day 0 of fusion, the body temperature was up to 40.1℃. The fever sustained 16 days. No septic shock occurred; Hypoxemia was noted on day 4 after fusion to a minimum of 69% with invasive ventilator assisted breathing and life support given; On day 4 after fusion, the capillary leak syndrome occurred, and the systemic tumor and skin lesion were swollen compared with before, resulting in marked wound redness and oozing fluid. After active treatment, careful nursing with aseptic dressing change, the patient successfully passed through the CRS reaction period, and the lesional skin wound was completely repaired after 3 months.Conclusions:T cells are absent after CD4-CAR T cell therapy, and patients present with a state of immunodeficiency that predisposes to opportunistic infections. Severe patients with mycosis fungoides associated with systemic skin lesions treated with CART are prone to severe cytokine release syndrome, which can manifest as hyperthermia, severe agranulocytosis, thrombocytopenia, and capillary leak syndrome, and may easily lead to more precancerous wounds. At this time, close observation of the changes in the life condition, active control of the CRS response, while supplemented with sterile debridement replacement, systemic symptomatic use of antibiotics, can reduce the incidence of infection, improve the safety of treatment, and ultimately promote the repair of tumor skin lesions wounds. DisclosuresNo relevant conflicts of interest to declare.

Influence of dehydration on cryopreservation of Musa spp. germplasm

Cryopreservation is an important technique for the long-term storage of economically important plant germplasm. In this study, an efficient protocol was developed for the long-term conservation of seven economically important Musa taxa: M. acuminata Colla ssp. burmannica N.W. Simmonds, M. acuminata Colla ssp. zebrina (Van Houtte) R.E. Nasution, M. balbisiana Colla, M. basjoo Sieb., M. ornata W. Roxburgh (St. Lavender), M. velutina H. Wendl. et Drude (Velvet Pink Banana), and M. acuminata’ balbisiana. The seeds were dehydrated in a sterile laminar flow cabinet for different exposure times and then they were directly immersed in liquid nitrogen. The critical point was to support the initial germination of cryopreserved seeds and this was achieved by the excision of zygotic embryos after liquid nitrogen treatment that allowed the seed germination. The best moisture content for tolerance to cryopreservation ranged from 15.8% (M. acuminata ssp. zebrina) to 17.1% (M. ornata) and the maximum post-cryopreservation germination rates varied from 86.4% (M. velutina) to 55.0% (M. ornata). All seedlings derived from seeds germinated after cryopreservation were easily rooted and acclimated to greenhouse conditions.

PM 7/128 (1) Xanthomonas axonopodis pv. allii

Specific scopeThis Standard describes a diagnostic protocol for Xanthomonas axonopodis pv. allii.This Standard should be used in conjunction with PM 7/76 Use of EPPO diagnostic protocols.Specific approval and amendmentFirst approved in 2016‐09.

Sterilization and preservation influence the biophysical properties of human amnion grafts

The introduction of amniotic membrane (AM) transplantation in ophthalmic surgery holds great promise and in many clinical situations it offers an alternative to existing management options. The purpose of this study was to examine the influence of established sterilization and preservation procedures on biophysical and histological properties of AM grafts. Amnion was sterilized by peracetic acid/ethanol sterilization [PES] and preserved by air-drying (sterile laminar flow) [AD] or in glycerol [GLYC]. Unsterilized AM were preserved at −80 °C [−80 °C] and served as an experimental control. Amnion allografts were characterized by the determination of their thickness, moisture vapour permeability (MVP), oxygen permeability (OPERM), tensile strength and sulphur content. Immunostaining for tissue-specific and basement membrane-related proteins was performed. Differences in biophysical parameters were found between the unsterilized allografts and the sterilized, air-dried or glycerol-preserved allografts. [PES/AD] showed the highest MVP and OPERM, the highest tensile strength and the lowest sulphur content and thickness. [PES/GLYC] exhibited the lowest OPERM and the highest thickness compared to [−80 °C] and [PES/AD]. Collagen types V and VII were preserved the best in the control group. Sterilization and preservation affect biophysical properties important for the use of AM as allogenic grafts. It has to be determined if any change, as noted, has a clinical impact.

Extended Stability of Oxytocin in Ringer's Lactate Solution at 4° and 25°C

Purpose Since IV admixtures of oxytocin in the final working concentrations are not commercially available, oxytocin admixtures must be prepared immediately before use or compounded in a sterile laminar flow hood and stored until needed. Although published reports indicate short-term stability of oxytocin in Ringer's lactate solution and Ringer's lactate-dextrose 5%, longer-term stability data at various temperatures are lacking. Methods A stability assay was developed, validated, and executed with oxytocin at two different concentrations in Ringer's lactate solution. Stability was assessed at 4 and 25°C over 31 days. Results This study shows that commonly used dilutions of oxytocin in Ringer's lactate solution retain greater than 90% of initial concentrations, when stored at room temperature or refrigerated for 31 days. These data show that oxytocin admixed with Ringer's lactate solution has extended chemical stability after preparation. Conclusions Demonstration of extended stability of oxytocin in Ringer's lactate solution will allow hospitals to give longer dating to oxytocin IV admixtures, provided they are prepared in a sterile environment and monitored in compliance with USP <797> guidelines. This should help minimize the potentially dangerous practice of admixing this IV solution at the bedside.

Laser biostimulation of cartilage: in vitro evaluation

An in vitro study was performed to evaluate the laser biostimulation effect on cartilage using a new gallium-aluminium-arsenic diode laser. Chondrocyte cultures were derived from rabbit and human cartilage. These cells were exposed to laser treatment for 5 days, using the following parameters: 300 joules, 1 watt, 100 (treatment A) or 300 (treatment B) hertz, pulsating emission for 10 minutes, under a sterile laminar flow. Control cultures (no treatment) received the same treatment with the laser device off. Cell viability was measured by MTT assay at the end of the laser treatment and then after 5 days. Neither rabbit nor human cultured chondrocytes showed any damage under a light microscope and immunostaining control following laser treatment. The MTT test results indicated a positive biostimulation effect on cell proliferation with respect to the control group. The increase in viability of irradiated chondrocytes was maintained for five days following the end of the laser treatment. The results obtained with the Ga-Al-As diode laser using the above tested parameters for in vitro biostimulation of cartilage tissues provide a basis for a rational approach to the experimental and clinical use of this device.

Growth of human normal and neoplastic mammary tissues in the cleared mammary fat pad of the nude mouse.

Dysplastic and malignant human breast tissues were grown successfully in the cleared mammary fat pads (CFP) of nude mice. The mammary fat pads were cleared while the mice were in a germfree isolator. Prepared mice were removed fron the germfree enviornment to facilitate transplantation of the human mammary tissue into their CFP and subsequently were maintained in sterile laminar flow racks.