MO319BEDSIDE URINE SEDIMENT EXAMINATION IN IMMUNOGLOBULIN A NEPHROPATHY PATIENTS PERFORMED BY NEPHROLOGISTS

Background and Aims Urine sediment microscopy is mostly abandoned by nephrologists nowadays, however it is an important diagnostic tool in kidney and urinary tract diseases. The aim of this study is to emphasize the benefits of urine microscopy performed by a nephrologist. Method A prospective cohort study at Pauls Stradins Clinical University Hospital Nephrology center included patients with histologically confirmed diagnosis of IgA nephropathy from 1st January 2020 till December 2020. Appropriately collected urine samples were examined using manual microscopy within an hour after sample collection and by automated urinalysis. Samples were centrifuged at 4000 rpm for 4 minutes, the supernatant urine was carefully decanted, 1 - 1,5 ml of the left urine was mixed by gentle agitation and placed on a standard glass slide with a cover slip. Sample examination was performed using low (magnification x10) and high power (magnification x40) using brightfield microscopy with a minimum of 10 fields. Results A total of 37 patients (24 men, mean age 42.7 ± 10.9 years) were included in the study. 59.5 % of patients (n = 22) had hematuria based on automated urine sediment analysis and 62.2 % (n = 23) of patients had hematuria based on manual urine microscopy. 45.9 % of patients (n=17) had dysmorphic erythrocytes, 13.5 % of patients (n = 5) had isomorphic red blood cells (RBC) and 40.5 % of patients (n = 15) did not have RBC in urine samples by manual urine microscopy. 54.2 % (13/24) of men and 30.8 % (4/13) of women had dysmorphic RBC in urine. Conclusion Manual urine sediment examination was more sensitive than automated analysis. Majority of IgA nephropathy patients have active urine sediment with hematuria and dysmorphic RBC. Manual microscopy remains an effective and reliable method that can be easily and quickly performed by nephrologists.

A REVIEW ON CURRENT STATUS OF SELF-EMULSIFYING DRUG DELIVERY SYSTEMS

The modern drug delivery system of hydrophobic drugs presents a main challenge because of the poor aqueous solubility of such compounds. Self emulsifying drug delivery systems (SEDDS) are usually used to enhance the bioavailability of hydrophobic drugs. SEDDS can be administered orally in soft or hard gelatin capsules and form fine relatively stable oil in water (o/w) emulsions upon aqueous dilution due to the gentle agitation of the gastrointestinal fluids. From time to time so many workers have claimed different rational applications of Self-emulsifying formulation for increcing bioavailability and site-specific targeting of highly lipophilic drugs. Significant improvement in the oral bioavailability of these drug compounds has been demonstrated for each case. The fact that almost 40% of the new drug compounds are hydrophobic in nature implies that studies with SEDDS will continue, and more drug compounds formulated as SEDDS will reach the pharmaceutical market in the future. The present article gives an overview of the Composition, mechanism, advantages, disadvantages, characterization, recent advancements, patents related information of SEEDS and commercial products approved for oral transmucosal administration.

Abstract 147: Plasma Factor XIII Binding to Cold-stored Platelets Results in Increased Fibrin Crosslinking and Clot Strength

Currently, platelets (PLTs) stored at room temperature (RT) for 5-7 days with gentle agitation are exclusively used for transfusion although FDA recently clarified that apheresis PLTs stored at 4°C for up to 72 hours may be used for treating active hemorrhage. We have demonstrated that cold (4C) storage of PLT is an attractive alternative to RT storage since it better preserves the PLT metabolic reserves, in vitro responses to agonists of activation, aggregation and physiologic inhibitors, as well as adhesion to thrombogenic surfaces. In this study, we tested the hypothesis that 4C-stored PLT will form clots with mechanical strength superior to those from RT-stored PLT due to higher hemostatic potential. From rheological measurements, we observed that the clots formed from 5 day 4C-stored PLTs are significantly stiffer (elastic modulus) and stronger (critical stress) than those formed from RT-stored PLT but comparable to fresh PLT (Fig. A). We also observed from ultrastructural microscopy that the fibrin fibers in clots from cold-stored PLT were thinner with more branch points than those from RT-stored PLTs, indicating the presence of increased crosslinks (Fig. B, C). Finally, molecular analysis revealed an increase FXIII transglutaminase activity due to the binding of plasma FXIII/fibrinogen to the surface of 4C-stored PLTs (Fig D).In conclusion, we have shown that cold-induced plasma FXIII binding to PLT surface result in increased fibrin crosslinking and enhanced clot strength. Our data, together with the benefit of reduced risk of late thrombosis due to their rapid clearance in vivo , underscores the consideration of 4C-stored PLT for acute response to hemorrhage. Figure 1 (A) RT-stored platelets form clots with less stiffness (n=5); (B) Representative SEM images of clots (n=3) (C) Clots from 4C-stored platelets have higher cross-linking density (n=5) (D) In FXIII-deficient plasma, 4C-stored platelets form clots with higher stiffness than fresh platelets (n=4)

One size doesn’t fit all: Should we reconsider the introduction of cold-stored platelets in blood bank inventories?

Platelet concentrates are universally prepared with a standard method and stored for 5 days at room temperature (20–24°C) in gentle agitation. Currently, there is a renewed interest in the possibility of storing platelet concentrates below the standard temperatures. In fact, cold platelets might be more effective in bleeding patients and have a lower risk of bacterial transmission. Inventories including platelets at different temperatures may favour patient-centred strategies for prophylactic or therapeutic transfusions.

Nanostructured Aluminum Oxide Black Coating: Electrochemical, Mechanical, and Optical Characterizations

Nanoporous anodized aluminum oxide (AAO) template is continuously investigated since it is vital for producing a variety of promised nanomaterial. In this study, two steps anodization of aluminum working electrode was carried out in H2SO4 under control of temperature at 17 °C and revolution rate 150 rpm using thermostatic cooling system and revolution control mechanical stirrer, respectively. Different times 15 mins, 30 mins, and 45 mins of first anodization were used and followed by electrolytic detachment for 1 min. Then, second anodization was applied for 10 mins, 20 mins, 30 mins, and 40 mins. Each sample of the prepared nanoporous AAO was used as working electrode into the electrolytic coloring cell containing coloring salt solution of 40 g/l CuSO4. Copper ions were deposited into the porous layer by AC current with gentle agitation using magnetic stirrer. The surface was characterized by field-emission scanning electron microscope device (FESEM), atomic force microscope (AFM), microhardness, corrosion resistance, and optical characteristics. The optical characteristics and reflectivity measurements revealed that the prepared selective coating is promised for solar heating systems and solar water heaters (SWH).

Modified AOAC Three Step Method (Official Method 2008.05): Consolidation of Fractions B and C

The AOAC Quantitative Three Step Method (TSM; AOAC Official MethodSM 2008.05) is validated for testing the efficacy of liquid sporicides against spores of Bacillus subtilis and Bacillus anthracis on selected hard, nonporous, and porous surfaces. The TSM uses 5 × 5 × 1 mm inoculated coupons (carriers), which are placed in 400 μL liquid sporicidal agent contained in a microcentrifuge tube. Following exposure of inoculated carriers to the test chemical and subsequent neutralization, viable spores are recovered in three fractions: A (gentle tapping), B (sonication), and C (gentle agitation). The spores in suspension are serially diluted and plated on a recovery medium for enumeration. The plate counts are summed over the three fractions to provide the number of viable spores per carrier, which is log10-transformed to generate a mean log density (LD) value across carriers. As a measure of product efficacy, a log reduction (LR) value is calculated by subtracting the mean LD for treated carriers from the mean LD for control carriers. This paper reports on the comparative evaluation of the current and modified versions of the TSM in order to support a modification to simplify the procedure. The proposed modified TSM (mTSM) consolidates fractions B and C in the same tube. Thus, the sonication (fraction B) and gentle agitation (fraction C) steps are carried out in the same tube, thereby reducing the number of tubes and associated resources and time necessary to complete the test. Glass, steel, pine wood, and ceramic tile carriers were included in the comparative study. Inoculated carriers were evaluated against two preparations of sodium hypochlorite to generate two presumed levels of efficacy (intermediate and high); the control LD and LR values associated with testing each carrier type for the TSM and the mTSM were compared. For control carriers, the mean log densities per carrier (for each carrier material) were not significantly different based on the TSM compared to the mTSM. Furthermore, the treated carrier data showed comparable LR values for the TSM and mTSM. The data provided in this report demonstrate equivalency between the TSM and mTSM and support the proposed procedural modification to consolidate fractions B and C.

Acrosomal status and mitochondrial activity of human spermatozoa vitrified with sucrose

This study investigates the ability of sucrose to protect spermatozoa against mitochondrial damage, artificial cryoinduction of capacitation, and acrosome reaction. Spermatozoa were isolated using the swim-up procedure performed using three different media: (a) human tubal fluid (HTF, control) medium; (b) HTF with 1% human serum albumin (HSA); and (c) HTF with 1% HSA and 0.25 M sucrose. From each group, 30 μl suspensions of cells were dropped directly into liquid nitrogen and stored for at least 24 h. Cells were thawed by quickly submerging the spheres in HTF with 1% HSA at 37 °C with gentle agitation. Sperm motility, viability, mitochondrial membrane potential integrity, spontaneous capacitation, and acrosome reaction were investigated. Sperm viability, acrosome reaction, and capacitation were detected using the double fluorescence chlortetracycline-Hoechst 33258 staining technique. Mitochondrial function was evaluated using a unique fluorescent cationic dye, 5,5′,6,6′-tetrachloro-1-1′,3,3′-tetraethyl-benzamidazolocarbocyanin iodide, commonly known as JC-1. The number of progressively motile spermatozoa was significantly higher in the sucrose-supplemented medium group (57.1±3.2%,P<0.05) when compared with controls (19.4±1.9%). The combination of HSA and sucrose (65.2±2.6%) has a stronger cryoprotective effect on the integrity of mitochondrial membrane potential (P<0.05) compared with HSA alone (32.6±4.7%). In conclusion, vitrification of human spermatozoa with non-permeable cryoprotectants such as HSA and sucrose can effectively cryopreserve the cells without significant loss of important physiological parameters.

Gelled Calcium Polyphosphate Matrices Delay Antibiotic Release

Introducing a gelling step during antibiotic incorporation has previously been found to delay vancomycin delivery from a calcium polyphosphate matrix intended for local treatment of bone infections. This study examined the general applicability of this approach using cefuroxime, a lower-molecular-weight antibiotic with different charge characteristics compared with those of vancomycin. A calcium polyphosphate/cefuroxime paste was “gelled” in disk form in a humid environment for 5 or 24 hours prior to drying. Antibiotic release in Tris-buffered saline under gentle agitation was monitored over a seven-day period. While non-gelled samples clearly exhibited a burst release, the gelling process significantly retarded early antibiotic release from five- and 24-hour gelled matrices, yielding a constant release rate over the first four days. Cefuroxime incorporation did not appear to alter matrix structure or degradation. Overall, this non-aggressive process effectively trapped cefuroxime and reduced its release rate, suggesting its potential applicability with molecularly diverse therapeutic agents.

Hyperconcentrated (Dry) Versus Standard Platelet Apheresis: An In Vitro Quality Study.

Introduction: Platelet concentrates (PC) collected by apheresis are effective in supporting deeply thrombocytopenic patients. The reduced risk of multiple allogeneic exposure and transmissible infectious diseases together with the high WBC depletion and diminished transfusion reactions are the main advantages offered by PC transfusion. At moment, the availability of several synthetic solutions for platelets storage permits to prepare hyperconcentrate(dry) apheresis platelets with the advantage of reducing febrile non-hemolytic transfusion reactions and, in low body weight patients the citrate toxicity, without the necessity of further manipulations. The aim of this study was to test the quality of 20 dry platelets (DP) in comparison to 20 standard plateletpheresis (SP) concentrates. Materials and methods: A total of 40 apheresis procedures were performed by the single-needle Cobe Trima separation device (Gambro BCT, Lakewood, CO, USA) collecting either DP or SP concentrates. Within 1h after collection, the bag containing DP was added with the appropriate amount (70% of DP final volume) of synthetic solution for platelets storage (SSP, MacoPharma). Both DP and SP concentrates were stored at room temperature with gentle agitation for 4 days. For both concentrates, platelet yield was calculated and in vitro studies of membrane glycoproteins expression and aggregation at day +1 and day +4 were carried out. Results: The comparison between 20 DP and 20 SP concentrates in terms of ability to aggregate in vitro and membrane glycoproteins expression at day +1 and day +4 of storage is reported in table A and B respectively. Conclusions: The in vitro tests documented a major activation of dry platelets. In particular, the ability to aggregate was reduced in the 20 DP concentrates analised and this phenomenon was more evident at day +4 of storage. The alteration of membrane glycoproteins expression (markers of storage lesion) confirms the lower in vitro quality of DP concentrates. The effectiveness of this new blood component in vivo should be evaluated in a controlled clinical trial. Table A. At collection Day +1 Day +4 SP DP SP DP SP DP Collagen μg/ml 4 93 88 97 82 80 53 ADP 10 μM 32 11 24 16 16 5 Ristocetin 1.5 mg/ml 91 97 77 81 71 52 Collagen 10 μg/ml + Adrenaline 10 μM 98 95 93 94 93 80 ADP 10 μM + Adrenaline 10 μM 87 72 76 61 57 30 Table B. At collection Day +1 Day +4 SP DP SP DP SP DP GPIb alfa (MFI) 5.06 5.75 6.31 6.13 5.26 4.54 GPIIb-IIIa (MFI) 35.47 36.71 34.61 37.7 31.76 40.9 GP IV (MFI) 11.49 11.4 11.67 10.28 11.65 11.38 GP 53 24.23 27.11 21.74 25.91 21.46 31.84 GMP 140 21.79 29.29 22.65 30.38 20.58 34.89