Maximum Number Of Cells Research Articles

Improvement of the valuable compounds of fish waste through solid-state fermentation with probiotics

The nutritional value of fish waste is increased by using solid-state fermentation with probiotic strains. The highest number of viable cells were observed in the fish waste substrate by two strains, namely Lactobacillus plantarium (MT-ZH193) and Pediococcus acidilactici (CNCM I-4622 - MA 18/5 M). The temperature of 38 °C, the moisture of 42 %, and dried non-alcoholic malt beverage waste (MBW) at 5 % were the optimal values for desired variables to achieve maximum number of viable cells during the 48-hour fermentation (12.8 Log cfu/gdw). As the fermentation time increased, fat decreased by 28 %. The production of non-protein nitrogen increased as the true protein reduced during solid-state fermentation due to protease enzyme activity. The inhibition percentage of free radicals increased during the 48-hour fermentation as compared with the non-fermented sample. The first day of fermentation showed the highest level of protease enzyme activity. The results showed that fermented fish waste was a good choice as a feed additive for livestock and poultry due to its high concentration of protein and digestibility. It is also rich in non-protein nitrogen and peptides with antioxidant characteristics.

Novel paradigm of light and CO2 feeding to improve the ‎ co-production of ‎astaxanthin and polyunsaturated fatty acids in Haematococcus ‎pluvialis

This study aimed to develop an innovative paradigm of lighting and CO2 flow rate in dilution and photoinduction stages of H. pluvialis cultivation to overcome the cell number reduction and stimulate the astaxanthin and polyunsaturated fatty acids accumulation joined by carbon bio-sequestration. The appropriate paradigm of lighting to reach the maximum cell number, astaxanthin concentration, and astaxanthin content in the dilution and photoinduction stages was obtained 2-fold (300 µmol/m2/s) and 6-fold (900 µmol/m2/s) of the final light intensity in the phototrophy stage (150 µmol/m2/s), respectively. Under the inventive lighting paradigm, feeding the 225 mL/L/min CO2 recorded optimum consequences in the dilution and photoinduction stages. The cell number, astaxanthin productivity, lipid content, polyunsaturated fatty acid and CO2 bio-fixation rate at the end of the photoinduction stages by lighting/CO2 balance were 94 %, 96 %, 32 %, 61 % and 40 % higher than control, respectively. Additionally, the levels of omega-6 fatty acids and omega-3 fatty acids were 70 % and 73 % higher than the control, respectively. This substantial increase was achieved by maintaining an optimal ratio of omega-6 to omega-3 fatty acids (3.05). Furthermore, to maximize the cell growth of microalgae and enhance the accumulation of valuable metabolites through CO2 bio sequestration, it is recommended to focus on the CO2 flow rate parameter instead of the CO2 percentage. That is affected by both the CO2 percentage and the aeration rate. Our findings provide a promising cost-effective and ecofriendly approach to improve the co-production of astaxanthin and polyunsaturated fatty acids in H. pluvialis.

Optimization of preparation conditions for Salsola laricifolia protoplasts using response surface methodology and artificial neural network modeling

BackgroundSalsola laricifolia is a typical C3–C4 typical desert plant, belonging to the family Amaranthaceae. An efficient single-cell system is crucial to study the gene function of this plant. In this study, we optimized the experimental conditions by using Box-Behnken experimental design and Response Surface Methodology (RSM)-Artificial Neural Network (ANN) model based on the previous studies.ResultsAmong the 17 experiment groups designed by Box-Behnken experimental design, the maximum yield (1.566 × 106/100 mg) and the maximum number of viable cells (1.367 × 106/100 mg) were obtained in group 12, and the maximum viability (90.81%) was obtained in group 5. Based on these results, both the RSM and ANN models were employed for evaluating the impact of experimental factors. By RSM model, cellulase R-10 content was the most influential factor on protoplast yield, followed by macerozyme R-10 content and mannitol concentration. For protoplast viability, the macerozyme R-10 content had the highest influence, followed by cellulase R-10 content and mannitol concentration. The RSM model performed better than the ANN model in predicting yield and viability. However, the ANN model showed significant improvement in predicting the number of viable cells. After comprehensive evaluation of the protoplast yield, the viability and number of viable cells, the optimal results was predicted by ANN yield model and tested. The amount of protoplast yield was 1.550 × 106/100 mg, with viability of 90.65% and the number of viable cells of 1.405 × 106/100 mg. The corresponding conditions were 1.98% cellulase R-10, 1.00% macerozyme R-10, and 0.50 mol L−1 mannitol. Using the obtained protoplasts, the reference genes (18SrRNA, β-actin and EF1-α) were screened for expression, and transformed with PEG-mediated pBI121-SaNADP-ME2-GFP plasmid vector. There was no significant difference in the expression of β-actin and EF1-α before and after treatment, suggesting that they can be used as internal reference genes in protoplast experiments. And SaNADP-ME2 localized in chloroplasts.ConclusionThe current study validated and evaluated the effectiveness and results of RSM and ANN in optimizing the conditions for protoplast preparation using S. laricifolia as materials. These two methods can be used independently of experimental materials, making them suitable for isolating protoplasts from other plant materials. The selection of the number of viable cells as an evaluation index for protoplast experiments is based on its ability to consider both protoplast yield and viability. The findings of this study provide an efficient single-cell system for future genetic experiments in S. laricifolia and can serve as a reference method for preparing protoplasts from other materials.

Roudneff’s Conjecture in Dimension 4

J.-P. Roudneff conjectured in 1991 that every arrangement of n ≥ 2 d + 1 ≥ 5 pseudohyperplanes in the real projective space P d has at most ∑ i = 0 d − 2 ( n − 1 i ) complete cells (i.e., cells bounded by each hyperplane). The conjecture is true for d = 2, 3 and for arrangements arising from Lawrence oriented matroids. Our main contribution is to show the validity of Roudneff’s conjecture for d = 4. Moreover, based on computational data we show that for d ≤ 4 and n ≤ 2 d + 1 the maximum number of complete cells is only obtained (up to isomorphism) by cyclic arrangements.

Development of chitosan/carrageenan macrobeads for encapsulation of Paenibacillus polymyxa and its biocontrol efficiency against clubroot disease in Brassica crops

Clubroot, caused by the obligate parasite Plasmodiophora brassicae, is one of the most important diseases of brassicas. The antagonistic bacterium Paenibacillus polymyxa ZF129 can suppress clubroot while its effectiveness is often unstable. To control clubroot more effectively, the macrobeads for controlled release of ZF129 were prepared using microencapsulation technology. Macrobeads with various ratios of chitosan (2 % w/w): carrageenan (0.3 % w/v) were prepared by an ionotropic gelation method and the bacteria ZF129 was loaded into macrobeads. The 1:1 chitosan: carrageenan showed the maximum swelling ratio (634 %), and the maximum survival rate (61.52 ± 1.12 %) after freeze-drying. Fourier transform infrared revealed the electrostatic interactions between chitosan and carrageenan. The macrobeads can efficiently release ZF129 strains into phosphate buffer solution and reach equilibrium in 48 h. The maximum number of bacteria cells to be released in the soil was observed after 25–30 days. The control efficacy of ZF129 macrobeads (chitosan: carrageenan, 1:1) and ZF129 culture against clubroot disease was 76.33 ± 3.65 % and 59.76 ± 4.43 % in greenhouse experiments, respectively and the control efficacy was calculated as 60.74 ± 5.00 % for ZF129 macrobeads and 40.94 ± 4.05 % for ZF129 culture under field experiments, respectively. The ZF129 macrobeads had significant growth-promoting effects on pak choi and Chinese cabbage. The encapsulation method described in this study is a prudent approach toward efficient biopesticides utilization with reduced environmental implications.

Histologic Manifestations of Gastrointestinal Adenovirus Infection After Stem Cell Transplant.

Adenovirus can cause severe disease in hematopoietic stem cell transplant (HSCT) patients. Histopathologic features of this infection in gastrointestinal biopsies and their distinction from graft-versus-host disease (GVHD) have been incompletely studied. We retrospectively identified patients with gastrointestinal adenovirus infection. H&E-stained sections were reviewed and the histologic features were recorded. The extent of immunostaining was determined using a semiquantitative scale and a maximum number of positive cells per high-power field. Information regarding the clinical course and endoscopic findings were obtained from the electronic medical records. The study group included 32 HSCT patients. Most (81%) presented with diarrhea and detectable virus in the serum. Twenty patients had multiorgan involvement in the gastrointestinal tract, mostly in the duodenum (62%) and colon (56%). Characteristic features included apoptotic epithelial cells with nuclear disarray (84%) and tufted aggregates of degenerating epithelial cells (69%), the latter of which was more commonly seen in the study population more than a control group of HSCT patients with GI involvement by GVHD. Viral inclusions were limited to the superficial epithelium in 59% of samples, and the density of viral inclusions within biopsies was variable (grade 1: 40%, grade 2: 38%, and grade 3: 22%). Following therapy, 10 patients (30%) improved and 14 (42%) had progressive disease. Patients with disease progression were often older (64 vs. 36 years, P =0.01) with higher serologic viral loads, prior history of GVHD, multifocal involvement, and increased number and density of immunoreactive nuclei. Adenovirus infection elicits a spectrum of histologic changes that can simulate or occur in combination with gastrointestinal GVHD. Patients with progressive disease are more likely to have high viral loads and more extensive infection of the gastrointestinal tract.

Morphofunctional Characteristics of Dorsal Root Ganglion Neurons in the Dynamics of Healing of a Purulent Wound

Abstract. between the morphological and functional state of spinal ganglia neurons and the dynamics of healing of a purulent wound in an experimental animal during its natural course and complex therapy with hydropulse sanitation (HIS) and the addition of platelet-rich blood plasma (PRP). Material and methods. In an experiment on 100 white outbred rats, a purulent wound on the lateral surface of the thigh was modeled by introducing a daily culture of Staphyllococcus aureus. Two groups were formed: natural healing and healing after therapy with hydropulse sanitation of the wound with an isotonic NaCl solution followed by the addition of platelet-enriched autoplasma. The material for the study was taken on the 1st, 3rd, 5th, 7th, 14th, and 21st days. The area of the wound and the dynamics of its healing were assessed using the planimetric method. For the study, lumbar spinal ganglia (SG) of segments LIII–LV. were taken. At the light-optical level, after staining with cresyl violet according to Nissl and azure B according to the S. Shea method, the area of nerve cells was measured, photometry of RNA content was carried out, and the number of satellite gliocytes was counted. The obtained data were processed using statistical analysis methods. Results. The healing process of a purulent skin wound ended with the formation of a scar on days 7–14 for both experimental groups. The use of GIS and PRP demonstrated better dynamics of defect closure - a reduction in wound area by 59.35±3.4% (p=0.018) in the period from 3 to 5 days, compared with 40.05±2.9% (p=0.009) with natural healing. The response of SG neurons during healing was characterized by pronounced polymorphism. The maximum number of cells with reactive changes during natural healing was observed on the 5th day of the experiment (17.2±3.2% for A-neurons and 36.7±4.3% for B-neurons). Complex therapy led to a decrease in the proportion of both reactively altered and destructive neurons. The cell area, RNA content, and the number of satellite glia increased on average by the 7th day of the experiment. Separate close correlations were identified between the dynamics of changes in the wound area and the studied quantitative characteristics. Conclusion. A purulent wound causes a complex of nonspecific reactions in the neurons of the SG. In the dynamics of wound healing, neurons with reactive changes more often demonstrated biphasic dynamics, more pronounced for B-type cells with a primary reaction on days 1–3 and a further transition to regenerative hypertrophy or destruction. Regenerative changes in SG neurons had a close correlation with the processes of regeneration of the wound defect.

Testing Adaptive Therapy Protocols Using Gemcitabine and Capecitabine in a Preclinical Model of Endocrine-Resistant Breast Cancer.

Adaptive therapy, an ecologically inspired approach to cancer treatment, aims to overcome resistance and reduce toxicity by leveraging competitive interactions between drug-sensitive and drug-resistant subclones, prioritizing patient survival and quality of life instead of killing the maximum number of cancer cells. In preparation for a clinical trial, we used endocrine-resistant MCF7 breast cancer to stimulate second-line therapy and tested adaptive therapy using capecitabine, gemcitabine, or their combination in a mouse xenograft model. Dose modulation adaptive therapy with capecitabine alone increased survival time relative to MTD but not statistically significantly (HR = 0.22, 95% CI = 0.043-1.1, p = 0.065). However, when we alternated the drugs in both dose modulation (HR = 0.11, 95% CI = 0.024-0.55, p = 0.007) and intermittent adaptive therapies, the survival time was significantly increased compared to high-dose combination therapy (HR = 0.07, 95% CI = 0.013-0.42, p = 0.003). Overall, the survival time increased with reduced dose for both single drugs (p < 0.01) and combined drugs (p < 0.001), resulting in tumors with fewer proliferation cells (p = 0.0026) and more apoptotic cells (p = 0.045) compared to high-dose therapy. Adaptive therapy favors slower-growing tumors and shows promise in two-drug alternating regimens instead of being combined.

Physiological and Biochemical State of Green Microalgae (Chlorophyta) under Different Nutrients' Content in the Cultural Medium

The physiological and biochemical state of green microalgae &lt;i&gt;Tetradesmus dimorphus&lt;/i&gt; (Turpin) M.J. Wynne and &lt;i&gt;Desmodesmus brasiliensis&lt;/i&gt; (Bohlin) E. Hegew at different growth phases depending on the nutrients' content in the cultural medium was investigated. Maximal cell numbers, specific growth rate and biomass increment in &lt;i&gt;T. dimorphus&lt;/i&gt; were recorded at the exponential stage under high concentrations of nitrate nitrogen and phosphorus of phosphates, and in &lt;i&gt;D. brasiliensis&lt;/i&gt; - under low. The increase of the cells' division frequency resulted in their lesser size and increase of length/width ratio. Maximal content of lipids and carbohydrates in the cells of &lt;i&gt;T. dimorphus &lt;/i&gt;was recorded at the end of the exponential stage over cultivation in the medium with low initial concentrations of nitrate nitrogen and phosphorus of phosphates, and in &lt;i&gt;D. brasiliensis&lt;/i&gt; -with high content. At this is no difference in the proteins' content in their cells was revealed.

Evaluation of cytotoxicity of 3.8 % SDF and BioAKT solutions on the viability of dental pulp stem cells

IntroductionNonsurgical endodontic therapies have evolved from classic endodontic therapies to regenerative endodontic treatments (RETs) in recent years. In context of the cytotoxic activity of the most commonly used endodontic irrigant, NaOCl, newer endodontic irrigating solutions should be tested for its effective use in RETs. The aim of this trial was to examine and assess the cytotoxic response of 3.8 % SDF and BioAKT irrigating solutions on the viability of DPSCs. MethodsThe viability of DPSCs cultivated in 5.25 % NaOCl, 3.8 % SDF & BioAKT at dilutions of 1:100,1:20 &1:10 were evaluated through MTT assay after 10 min, 60 min and 24 h incubation, detection of apoptosis and ALP activity after 7,14 & 21-days incubation. A two-way analysis of variance (ANOVA) with post hoc Turkey HSD was performed to determine significant differences between the specimens tested. ResultsWhen compared to the control at all time periods, all test specimens at varied dilutions (1:100, 1:20, and 1:10) caused no cytotoxic effects. The maximum number of live cells and ALP activity was observed with DPSCs cultivated in BioAKT followed by 3.8 % SDF and 5.25 % NaOCl at all time intervals. ConclusionDifferent doses of 3.8 % SDF and BioAKT solution revealed encouraging outcomes when compared to 5.25 % NaOCl in terms of viability, proliferation and long-term ALP functioning potential when cultivated in DPSCs.

Calcium alginate elastic capsules for microalgal cultivation.

Calcium alginate elastic capsules with a core-shell structure are versatile spherical solid beads that can be produced in large quantities using various techniques. This type of capsule is a promising platform for cell culture applications, owing to its mechanical elasticity and transparency. This paper reports the production of calcium alginate capsules with high consistency, and for the first time, demonstrates the feasibility of the capsules for microalgal cultivation. Cell growth analysis reveals that the vibrationally-shaken calcium alginate elastic capsule platform yielded a higher maximum cell number (4.86 × 108 cells per mL) during the cultivation period than the control solution platforms. Aquafeed and food supplements for humans are the targeted applications of this novel platform.

Effect of fertilizers on the growth performance of Azolla pinnata: A potential feed additive for fish culture in Bangladesh

Azolla is among the most popular, low-cost, and high-nutrient-content fish feeds. Azolla pinnata's growth performance was evaluated on the effects of fertilizers containing phosphorus and nitrogen. The study used a completely randomized design (CRD) over thirty-five days in 150 L water containers. Two sets of sub-experiments were established, each with three replications: one for the combined application of phosphorus and nitrogen and another for the sole application of phosphorus. The results showed that the combined application of phosphorus (46% P) and nitrogen (46% N) was more effective than the application of phosphorus alone. The highest cell number and protein content of combined P and N applications were (70.01±4.73 g and 33.71±2.3%) in the T4 treatment. On the other hand, the maximum cell number and protein content of the sole P application were 58.76±3.8 and 33.06±2.25%, respectively, in the T4 treatment. A Significant variation was observed in terms of the mean cell number, weight, and protein content among the experimental treatments. This study's primary focus was to demonstrate the effects of inorganic fertilizers on Azolla growth. The findings suggest that the combined application of phosphorus and nitrogen fertilizer can significantly enhance Azolla growth compared to the sole application of phosphorus.

Diagnostic yield and the number of tumor cells of ultrathin bronchoscopy for peripheral lung lesions: A comparison with thin bronchoscopy.

Ultrathin bronchoscopy has been reported to have a higher diagnostic yield than thin bronchoscopy for small peripheral lung lesions in transbronchial biopsy under radial endobronchial ultrasonography (EBUS). However, data comparing the number of tumor cells in non-small cell lung cancer (NSCLC) are limited. We retrospectively compared the number of NSCLC tumor cells in peripheral lung lesions obtained using an ultrathin bronchoscope and a thin bronchoscope with radial EBUS between April 2020 and October 2021. In all patients, we used virtual bronchoscopic navigation (VBN) software, and guide sheaths were used in thin bronchoscopy cases. A total of 175 patients were enrolled in this study. Ultrathin bronchoscopy cases (n = 69) had lesions with a smaller diameter that are more peripherally located compared to thin bronchoscopy cases (n = 106) (median, 25.0 vs. 26.5 mm, mean bronchial generations accessed by bronchoscopy; 4.4±1.2 vs. 3.8±1.0, respectively; p<0.010). There were no significant differences in the overall diagnostic yield (ultrathin vs. thin bronchoscopy cases, 68.1% vs. 72.6%, p = 0.610) or diagnostic yield in only lung cancer cases (78.6% vs. 78.5%, p = 1.000). In histologically NSCLC cases (n = 102), the maximum number of tumor cells per slide as the primary endpoint was similar (average, 307.6±246.7 vs. 328.7±314.9, p = 0.710). The success rate of the Oncomine™ analysis did not differ significantly (80.0% vs. 55.6%, p = 0.247). The yield of NSCLC tumor cells was not different between the samples obtained by the ultrathin bronchoscope and those obtained by the thin bronchoscope.

E72 Distribution of cells and extracellular matrix in the marginal synovium during the postnatal period

Abstract Introduction The marginal transitional zone of the joint is of fundamental importance in joint pathology. The normal marginal synovium takes part in articular cartilage repair (Adachi et al. 2012). The inflamed marginal synovium, in turn, promotes irreversible damage to the adjacent cartilage and bone (Hitchon &amp; El-Gabalawy, 2011). Early erosive damage in rheumatoid arthritis and osteophyte formation in osteoarthritis occurs primarily here (Allard et al. 1990). Objectives The morphology of the marginal synovium from birth till the 90th day was studied using the hip joints of Wistar rats. Methods The collection of joints was carried out on the first, seventh, 14th, 30th, 45th, 60th and 90th day (six rats in each group, 42 in total). Joints were fixed, decalcified and dehydrated. Histologic paraffine sections were stained with Mallory's trichrome and Hart's elastin staining, analyzed at the acetabular labrum by light microscopy (χ100), morphometrical and statistical (P &amp;lt; 0.05) methods with extrapolation to humans (Sengupta, 2013). The obtained data were presented as percentages. Results In the marginal synovium, cells and components of the extracellular matrix, which are typical for fibrous connective tissue, could be seen. The synovial lining cells are embedded in the modified basement membrane which includes collagen and elastic fibers. They form a three-dimensional reticulum in it and adjacent synovial subintima. Vessels of the microcirculatory bed are also widely represented in the marginal synovium. The overall quantity of collagen fibers increased gradually, exceeding on the 90th day (approximately corresponds to 18–19 human years) that of newborns by one and a half times (48.92 ± 3.39 vs 72.17 ± 7.77; P = 0.023). The number of arranged collagen fibers increased gradually, exceeding on the 90th day that of newborns almost twofold (37.25 ± 3.21 vs 60.08 ± 5.99; P = 0.008). The area of distribution of disarranged collagen fibers in newborns was almost four and a half times less than the overall quantity of collagen fibers (11.67 ± 1.03 vs 48.92 ± 3.39; P &amp;lt; 0.001) and did not change substantially throughout the experiment. The number of elastic fibers increased till the 30th day (0.63 ± 0.11 vs 18.3 ± 1.25; P &amp;lt; 0.001) and did not undergo significant changes later. In newborns, the area of distribution of ground substance was not considerably higher than the ratio of disarranged collagen fibers (10.42 ± 1.8 vs 11.67 ± 1.03; P = 0.562) and did not undergo substantial changes later. The distribution density of vessels increased till the 30th-45th day (4.5 ± 0.67 vs 7.5 ± 0.82 and 7.7 ± 0.54; P &amp;lt; 0.001; approximately corresponds to the 9th and 14th human years respectively), followed by a gradual decrease from the 60th day (7.05 ± 0.72; roughly corresponds to the 16th human years). The maximum number of cells was in newborns. Subsequently, it gradually decreased and was almost three times less on the 90th day than that of newborns (38.01 ± 3.06 vs 12.58 ± 1.53; P &amp;lt; 0.001). Conclusion The morphogenesis of the marginal synovium during the postnatal period is accompanied by a gradual decrease in the number of cells, an increase in the area occupied by elastic and arranged collagen fibers with a relatively constant distribution density of disarranged collagen fibers and ground substance together with an undulatory pattern in the distribution of vessels. These peculiarities emphasize the morphological uniqueness of the marginal transitional zone, which forms the innate nonspecific barrier between the joint capsule and the articular cartilage, and thereby provides the necessary anatomical and physiological integrity of the joint as a whole organ. Knowledge about the patterns of morphogenesis of the marginal synovium will help in the future to analyze the modeled pathological or boundary states in it. Ethics The study was approved by the ethics committee of the Zaporizhzhia State Medical University.

E71 Distribution of cells and extracellular matrix in the normal marginal synovium of the hip joint capsule during the postnatal period

Abstract Introduction The marginal transitional zone is of fundamental importance in joint pathology. It includes the marginal cartilage and marginal synovium. The normal marginal synovium takes part in articular cartilage repair (Adachi et al. 2012). The inflamed marginal synovium, in turn, promotes irreversible damage to the adjacent cartilage and bone (Hitchon &amp; El-Gabalawy, 2011). Objectives The morphology of the normal marginal synovium from birth till the 90th day was studied using the hip joints of Wistar rats. Methods The collection of joints was carried out on the first, seventh, 14th, 30th, 45th, 60th and 90th day (six rats in each group, 42 in total). Joints were fixed, decalcified and dehydrated. Histologic paraffine sections were stained with Mallory's trichrome and Hart's elastin staining, analyzed at the acetabular labrum by light microscopy (χ100), morphometrical and statistical (P &amp;lt; 0.05) methods with extrapolation to humans (Sengupta, 2013). The obtained data were presented as percentages. Results In the marginal synovium, cells and components of the extracellular matrix, which are typical for fibrous connective tissue, could be distinguished. The synovial lining cells are embedded in the modified basement membrane which is a meshwork of collagen and elastic fibers. They form a three-dimensional reticulum in it and adjacent synovial subintima. Vessels of the microcirculatory bed are also widely represented in the marginal synovium. The overall quantity of collagen fibers increased gradually, exceeding on the 90th day (∼18–19 human years) that of newborns by one and a half times (48.92 ± 3.39 vs 72.17 ± 7.77; P = 0.023). The number of arranged collagen fibers increased gradually, exceeding on the 90th day that of newborns almost twofold (37.25 ± 3.21 vs 60.08 ± 5.99; P = 0.008). The area of distribution of disarranged collagen fibers in newborns was almost four and a half times less than the overall quantity of collagen fibers (11.67 ± 1.03 vs 48.92 ± 3.39; P &amp;lt; 0.001) and did not change substantially throughout the experiment. The number of elastic fibers increased till the 30th day (0.63 ± 0.11 vs 18.3 ± 1.25; P &amp;lt; 0.001) and did not undergo significant changes later. In newborns, the area of distribution of ground substance was not considerably higher than the ratio of disarranged collagen fibers (10.42 ± 1.8 vs 11.67 ± 1.03; P = 0.562) and did not undergo substantial changes later. The distribution density of vessels increased till the 30th-45th day (4.5 ± 0.67 vs 7.5 ± 0.82 and 7.7 ± 0.54; P &amp;lt; 0.001; ∼9-14th human years), followed by a gradual decrease from the 60th day (7.05 ± 0.72; ∼16 human years). The maximum number of cells was in newborns. Subsequently, it gradually decreased and was almost three times less on the 90th day than that of newborns (38.01 ± 3.06 vs 12.58 ± 1.53; P &amp;lt; 0.001). Conclusion The morphogenesis of the marginal synovium during the postnatal period is accompanied by a gradual decrease in the number of cells, an increase in the area occupied by elastic and arranged collagen fibers with a relatively constant distribution density of disarranged collagen fibers and ground substance together with an undulatory pattern in the distribution of vessels. These peculiarities emphasize the morphological uniqueness of the marginal transitional zone, which forms the innate nonspecific barrier between the joint capsule and the articular cartilage, and thereby provides the necessary anatomical and physiological integrity of the joint as a whole organ. Knowledge about the patterns of morphogenesis of the normal marginal synovium will help to understand joint pathology. Ethics The study was approved by the ethics committee of the Zaporizhzhia State Medical University.

Postinfantile Giant Cell Hepatitis in Native and Allograft Livers: A Multi-Institutional Clinicopathologic Study of 70 Cases

Postinfantile giant cell hepatitis (PIGCH) is a rare hepatitis pattern in adults with variable etiologies and clinical outcomes. We conducted a multi-institutional retrospective study to define the clinicopathologic characteristics of patients with PIGCH. A total of 70 PIGCH cases were identified and reviewed for pathological features, including fibrosis, cholestasis, inflammation, steatosis, necrosis, and apoptosis, as well as the distribution of giant cells and the maximum number of giant cells per high-power field. Demographic and clinical data, including age, sex, laboratory results, etiologies, and follow-up results, were recorded. Among the 70 cases, 40% (28/70) were associated with autoimmune liver diseases, followed by 9 (13%) with unknown etiology, 8 (11%) with viral infection, 5 (7%) with medications, 5 with combined etiologies, and 4 (6%) with malignancies (mostly chronic lymphocytic leukemia). Notably, another 16% were de novo PIGCH in liver allografts, most of which occurred after a rejection event. During follow-up, 26 (37%) patients died of the disease and 44 (63%) were alive. Deceased patients were characterized by older age (mean age, 54.9 vs 45.5 years; P = .02), higher alkaline phosphatase level (mean value, 253.3U/L vs 166.3 U/L; P = .03), higher fibrosis stage (stage 3-4 vs stage 0-2, 57.7% vs 29.6%; P = .03), being more likely to have de novo PIGCH after transplantation (23.1% vs 11.4%; P = .04), and being less likely to have primary autoimmune liver disease etiology (26.9% vs 47.7%; P = .04). These results indicate that PIGCH is a rare pattern of liver injury associated with different etiologies and variable clinical outcomes. Autoimmune liver disease with PIGCH is associated with better survival, whereas de novo PIGCH in allografts is associated with poorer survival. Older age, higher alkaline phosphatase level, and advanced fibrosis are adverse prognostic factors.

The Effect of Optimum Photoperiod from Blue LED Light on Growth of Chlorella Vulgaris in Photobioreactor Tank

Abstract Study for investigated the optimum photoperiod from blue LED light for the growth of Chlorella vulgaris in culture tank. Photoperiods from blue LED Light (Light/Dark) were set up at 24:0, 16:8 and 12:12 h. The initial number of C. vulgaris cells was 1.6 x 105 ± 0.12 cells mL-1. Culture medium was using commercially available fertilizers. After 17 days of cultivation, the results showed that the 24:0 h photoperiod had better growth of algae than 16:8 and 12:12 h, with maximum number of cells of 1.64 x 107 ± 0.23 cells mL-1 specific growth rate (SGR) was 0.17 ± 0.05, -0.06 ± 0.11 and -0.19 ± 0.1% day-1 with statistically significant differences (P &lt; 0.05) in day 5 of cultivation. The stationary phase continued up to day 3 of cultivation, between the treatment. The optimum temperature for growth ranged from 35.5–38.5°C. The results showed that blue LED lighting at 24:0 h is the optimum photoperiod for cultivation of C. vulgaris. Keywords: Photobioreactor tank, Photoperiod, Blue LED light, Chlorella vulgaris, Growth rate

The Utilisation of Antarctic Microalgae Isolated from Paradise Bay (Antarctic Peninsula) in the Bioremediation of Diesel.

Research has confirmed that the utilisation of Antarctic microorganisms, such as bacteria, yeasts and fungi, in the bioremediation of diesel may provide practical alternative approaches. However, to date there has been very little attention towards Antarctic microalgae as potential hydrocarbon degraders. Therefore, this study focused on the utilisation of an Antarctic microalga in the bioremediation of diesel. The studied microalgal strain was originally obtained from a freshwater ecosystem in Paradise Bay, western Antarctic Peninsula. When analysed in systems with and without aeration, this microalgal strain achieved a higher growth rate under aeration. To maintain the growth of this microalga optimally, a conventional one-factor-at a-time (OFAT) analysis was also conducted. Based on the optimized parameters, algal growth and diesel degradation performance was highest at pH 7.5 with 0.5 mg/L NaCl concentration and 0.5 g/L of NaNO3 as a nitrogen source. This currently unidentified microalga flourished in the presence of diesel, with maximum algal cell numbers on day 7 of incubation in the presence of 1% v/v diesel. Chlorophyll a, b and carotenoid contents of the culture were greatest on day 9 of incubation. The diesel degradation achieved was 64.5% of the original concentration after 9 days. Gas chromatography analysis showed the complete mineralisation of C7-C13 hydrocarbon chains. Fourier transform infrared spectroscopy analysis confirmed that strain WCY_AQ5_3 fully degraded the hydrocarbon with bioabsorption of the products. Morphological and molecular analyses suggested that this spherical, single-celled green microalga was a member of the genus Micractinium. The data obtained confirm that this microalga is a suitable candidate for further research into the degradation of diesel in Antarctica.

Optimized operation of a controlled stirred tank reactor system for the production of mesenchymal stromal cells and their extracellular vesicles.

The therapeutic effects of human mesenchymal stromal cells (MSC) have been attributed mostly to their paracrine activity, exerted through small-secreted extracellular vesicles (EVs) rather than their engraftment into injured tissues. Currently, the production of MSC-derived EVs (MSC-EVs) is performed in laborious static culture systems with limited manufacturing capacity using serum-containing media. In this work, a serum-/xenogeneic-free microcarrier-based culture system was successfully established for bone marrow-derived MSC cultivation and MSC-EV production using a 2 l-scale controlled stirred tank reactor (STR) operated under fed-batch (FB) or fed-batch combined with continuous perfusion (FB/CP). Overall, maximal cell numbers of (3.0 ± 0.12) × 108 and (5.3 ± 0.32) × 108 were attained at Days 8 and 12 for FB and FB/CP cultures, respectively, and MSC(M) expanded under both conditions retained their immunophenotype. MSC-EVs were identified in the conditioned medium collected from all STR cultures by transmission electron microscopy, and EV protein markers were successfully identified by Western blot analysis. Overall, no significant differences were observed between EVs isolated from MSC expanded in STR operated under the two feeding approaches. EV mean sizes of 163 ± 5.27 nm and 162 ± 4.44 nm (p > 0.05) and concentrations of (2.4 ± 0.35) × 1011 EVs/mL and (3.0 ± 0.48) × 1011 EVs/mL (p > 0.05) were estimated by nanoparticle tracking analysis for FB and FB/CP cultures, respectively. The STR-based platform optimized herein represents a major contribution towardthe development of human MSC- and MSC-EV-based products as promising therapeutic agents for Regenerative Medicine settings.

Effect of Simulated Transport Conditions on Microbiological Properties of Bottled Natural Mineral Water

Bottled mineral water is distributed globally through complex supply chains, making it available far beyond its bottling plants. In low-viscosity food matrices, invisible changes may occur due to shaking. The primary purpose of this research was to investigate the potential correlation between the intensity of mechanical agitation and the number of detectable microorganisms in bottled mineral water. The simulation of dynamic mechanical vibration was conducted using both time-accelerated and real-time tests. Freshly bottled natural mineral water and commercially available mineral water brands from different bottling locations and times were subjected to random vibration at three intensities as specified by the ASTM D-4169-16 standard, which simulates road transport on semi-trailer trucks. The study investigated the specific growth rate, the generation time, and the maximum cell numbers of microorganisms. The quantitative PCR (qPCR) technique was used to determine and compare the concentrations of microbes. Dynamic mechanical vibration affected the microbiome of mineral waters, influencing growth rates and generation times. In the case of waters from different bottling locations and times, the specific growth rate varied significantly for each water and for each intensity. This finding demonstrates that the microbiome composition of the water source and the interaction between microbes influence the response to mechanical impact. The time-accelerated test was shown to be suitable for analyzing the reaction of the microbiome of the tested matrix to the intensity and duration of vibration. The applied test protocol enabled the monitoring of changes in cell numbers by qPCR. All three intensities of the time-accelerated method were effective in testing the effects of real-time mechanical agitation on the microbiome.