Secondary Cultures Research Articles

Establishment and Validation of an Efficient Agrobacterium Tumefaciens-Mediated Transient Transformation System for Salix Psammophila

Salix psammophila, C. Wang & Chang Y. Yang, a desert-adapted shrub, is recognized for its exceptional drought tolerance and plays a vital role in ecosystem maintenance. However, research on S. psammophila has been limited due to the lack of an efficient and reliable genetic transformation method, including gene functional studies. The Agrobacterium-mediated transient overexpression assay is a rapid and powerful tool for analyzing gene function in plant vivo. In this study, tissue culture seedlings of S. psammophila were utilized as the recipient materials, and the plant expression vector pCAMBIA1301, containing the GUS reporter gene, was transferred into the seedlings via an Agrobacterium-mediated method. To enhance the efficiency of the system, the effects of secondary culture time, Agrobacterium concentration, infection time, and co-culture duration on the transient transformation efficiency of S. psammophila were explored. The optimal combination for the instantaneous transformation of S. psammophila tissue culture seedlings mediated by Agrobacterium was determined as follows: a secondary culture time of 30 d, a value of OD600 of 0.8, an infection time of 3 h, and a co-culture duration of 48 h. Subsequently, the effectiveness of the transformation system was validated using the S. psammophila drought response gene SpPP2C80. To further confirm the accuracy of the system, SpPP2C80-overexpressing Arabidopsis was constructed and drought resistance analysis was performed. The results were consistent with the transient overexpression of SpPP2C80 in S. psammophila tissue culture seedlings, indicating that this system can be effectively employed for studying gene function in S. psammophila. These findings provide essential information for investigating gene function in non-model plants and pave the way for advancements in molecular biology research in S. psammophila.

Factors Affecting the Density of Corneal Endothelial Cells Cultured from Donor Corneas.

We investigated which specific correlation exists between the endothelial cell density (ECD) of corneal endothelial cell (CEC) cultures and the features of the donor corneas from which they originate. CEC cultures were prepared from one donor cornea or by pooling together cells of more corneas from elderly donors with ECDs lower or higher than 2000 cells/mm2. The ECDs of such primary cultures were evaluated and showed that that ECDs > 2000 cells/mm2 can be obtained only when CECs are isolated from (1) corneas of young donors; (2) at least two elderly donor corneas (if ECD > 2000 cells/mm2), or three elderly donor corneas (if ECD < 2000 cells/mm2). Secondary cultures are all characterized by ECDs < 2000 cells/mm2. Our study highlights the difficulties in obtaining cultures with ECDs > 2000 cells/mm2. Even if achievable with corneas from young donors, this becomes a challenging task when corneas from elderly donors are used (i.e., the overall majority of those collected by eye banks) and particularly when corneas from elderly donors with ECD < 2000 cells/mm2 are used. Pooling more of two corneas to obtain suitable CECs could technically overcome the problem. The above issues should be tackled appropriately before moving into clinical studies.

Morphological, structural, and ultrastructural key features of hyperhydricity in Beta vulgaris L. var. saccharifera

Abstract Hyperhydricity, as a phenomenon specific to in vitro plants, triggers a series of changes at the cell and tissue level, which modify the plants’ physiological processes. For a better understanding of this phenomenon, we found it useful to present here the fundamental research on establishing the steps of structural and ultrastructural degradation of cells and tissues, observed by light microscopy (LM) and transmission electron microscopy (TEM) analyses, in the leaves of Beta vulgaris var. saccharifera, at 30 days of secondary culture, and their correlation with viability in the third culture. Comparative with control (non-hyperhydric plants), three degradation steps of hyperhydricity were identified in plants regenerated on a culture medium with 2.5 mg·L-1 6-benzyladenine. The first step involved cell wall deformation, which lost its rigidity and became sinuous, causing the enlargement of the intercellular spaces. In the second step, these spaces formed gaps through their union, and the whole membrane system suffered: both chloroplasts and tonoplast were broken (the cytoplasm and vacuolar composition were mixed), the nuclear membrane presented undulations, just before the damage to the tonoplast, and the nucleus became pyknotic. In step 3, the cell showed the beginning of lysis, which leads to necrosis, the cell had nothing in common with a normal ultrastructure. For in vitro plants in this final step, there was no chance of surviving but in steps I and II, the viability was 55-75%. These features can be useful to producers to calculate the level of culture damage and start measures to prevent losses.

Lacticaseibacillus casei as Anti-blowing Agents: Impact on the Evolution of Ripening and Sensory Profile of Montasio Cheese

AbstractRecently, the Lacticaseibacillus casei group strains have been gaining growing interest due to their potential to be used as secondary adjunct cultures in cheese. This work aimed to test autochthonous Lb. casei strains as anti-blowing agents and to evaluate their impact on the evolution of the ripening and the sensory profile of Montasio cheese. The cheesemaking trial included a control production without lysozyme (C1), a control with lysozyme (C2), and four experimental productions, each containing a different pool of autochthonous Lb. casei strains (EX1-4). Samples were taken during ripening, and physicochemical, microbiological, and sensory analyses were carried out. Lb. casei counts indicate that the selected strains survived the cheesemaking and maintained their viability of about 9 log cfu g−1 at the end of the ripening. Only EX3 showed a significant slowdown of the proteolytic index compared to controls over time. Furthermore, from the principal component analysis, it emerged that at the end of the 120-day-long ripening, C2 and the experimental samples were characterized by similar profiles of volatile compounds. The late-blowing defect (LBD) was observed exclusively in C1, whereas it was not detected in the control sample with lysozyme or in any experimental samples. These findings not only confirmed the efficacy of lysozyme in preventing LBD but also supported the effectiveness of the selected Lb. casei strains as anti-blowing agents with the ability to contribute to the final volatile profile without compromising the typicality of the product.

Bafilomycin 1A Affects p62/SQSTM1 Autophagy Marker Protein Level and Autophagosome Puncta Formation Oppositely under Various Inflammatory Conditions in Cultured Rat Microglial Cells.

Regulation of autophagy through the 62 kDa ubiquitin-binding protein/autophagosome cargo protein sequestosome 1 (p62/SQSTM1), whose level is generally inversely proportional to autophagy, is crucial in microglial functions. Since autophagy is involved in inflammatory mechanisms, we investigated the actions of pro-inflammatory lipopolysaccharide (LPS) and anti-inflammatory rosuvastatin (RST) in secondary microglial cultures with or without bafilomycin A1 (BAF) pretreatment, an antibiotic that potently inhibits autophagosome fusion with lysosomes. The levels of the microglia marker protein Iba1 and the autophagosome marker protein p62/SQSTM1 were quantified by Western blots, while the number of p62/SQSTM1 immunoreactive puncta was quantitatively analyzed using fluorescent immunocytochemistry. BAF pretreatment hampered microglial survival and decreased Iba1 protein level under all culturing conditions. Cytoplasmic p62/SQSTM1 level was increased in cultures treated with LPS+RST but reversed markedly when BAF+LPS+RST were applied together. Furthermore, the number of p62/SQSTM1 immunoreactive autophagosome puncta was significantly reduced when RST was used but increased significantly in BAF+RST-treated cultures, indicating a modulation of autophagic flux through reduction in p62/SQSTM1 degradation. These findings collectively indicate that the cytoplasmic level of p62/SQSTM1 protein and autophagocytotic flux are differentially regulated, regardless of pro- or anti-inflammatory state, and provide context for understanding the role of autophagy in microglial function in various inflammatory settings.

Comparison of Scenedesmus obliquus in CO2 Capture, Biolipid Production and Nutrient Removal

The cultivation of microalgae from municipal wastewater, while simultaneously removing nutrients from the water column, has the potential to aid biodiesel production and carbon dioxide fixation, thereby alleviating the pressure of energy shortages. In this research, different ratios of sodium bicarbonate and glucose were used to prepare simulated municipal wastewater. The results demonstrated that microalgae were most effectively treated under one-stage direct treatment conditions. During direct culture, the most effective treatment was observed for IAA-3, which exhibited a dry weight of 1.4363 g/L and a lipid content of 25.05% after stimulation with 0.0005 M NaHCO3. In contrast, NaHCO3-2 demonstrated optimal performance during the secondary culture, with a dry weight of 1.6844 g/L and a lipid content of 18.05%. Finally, the economic, social and environmental benefits of direct treatment (IAA-3) and secondary treatment NaHCO3-2 were analyzed. The benefits of direct treatment were found to be USD 0.50989/L, while those of secondary treatment were USD 0.43172/L. For each tonne of municipal wastewater treated, the carbon sequestration benefits of IAA-3 during direct treatment and NaHCO3-2 during secondary treatment were USD 0.45645 and USD 0.85725, respectively.

Screening of lactic acid bacteria with the ability to reduce goaty flavor related fatty acids in goat milk

In this study, we conducted sensory evaluation and gas chromatography–mass spectrometry analysis on fermented goat milk samples prepared by 12 strains of lactic acid bacteria (LAB) isolated from goat milk to screen for strains with the ability to reduce the goaty flavor. The bacterial counts of fermented goat milk was 7.07–9.01 log CFU/mL. The electronic nose distinguished fresh goat milk (FGM) and fermented goat milk, and the electronic tongue results showed that Leuconostoc citreum 1, 4, 20, 22, 32, and 57, Latilactobacillus curvatus 144 and 147 imparted fermented goat milk a taste different from FGM. Overall, Leuconostoc citreum 57, Leuconostoc citreum 126, Latilactobacillus curvatus 142, Latilactobacillus curvatus 143, and Latilactobacillus curvatus 147 were screened with the ability to improve the flavor of goat milk. They gave fermented goat milk a goat flavor score lower than or equal to FGM. And the fermented goat milk samples 57, 126, 142, 143, and 147 contained 25, 22, 15, 24, and 17 volatile flavor compounds, respectively, with a greater variety and content of ketones and aldehydes and lower levels of hexanoic acid, octanoic acid, and decanoic acid than FGM. However, the pH and WHC results indicated that the application of these strains as secondary cultures is necessary. Our finding provides basic research data to improve the flavor of goat milk products.

Selection and characterization of spontaneous phage-resistant mutant of Limosilactobacillus fermentum

Phage infection remains a major cause of fermentation failures in the dairy industry. The development of phage-resistant mutants of important fermentation strains is an effective measure used to address phage-related issues. This study employed the secondary culture method to screen for spontaneous phage-resistant mutants from the phage sensitive strain Limosilactobacillus fermentum IMAU32646 (L. fermentum IMAU32646). The phenotypic characteristics, technological attributes, probiotic characterization, adsorption characteristics and mutant genes were investigated. The results showed that the mutant strain displayed a high degree of phage-resistance and stability. The mutant strain produced more lactic acid during fermentation than the sensitive strain, while maintaining identical cell structure and morphologies. The mutant strain exhibited superior tolerance to acid and bile salts compared to the sensitive strain. Furthermore, the adsorption rate of phage LFP01 on the mutant strain was significantly lower than that of the sensitive strain. Following genome re-sequencing analysis showed that adsorption interference and blocked DNA injection were responsible for its phage-resistance. These results may provide a new strategy for avoiding phage contamination and industrial application of phage-resistant strains with good characteristics.

Finding a Place for Third Culture Kids in Culture: A Preliminary Study Exploring Negative and Positive Impacts on Their Experiences Living in a Muslim Country During the COVID-19 Pandemic

Third culture is basically a new set of cultures developed by an individual, being a child or an adult, who goes abroad outside of his/her country of domicile. This terminology emerges when the individual confronts a new host country’s secondary culture as against the first culture of the home country, with the third culture referring to the fusion of these two distinct cultures. Adjustment process becomes even more challenging during the present COVID-19 pandemic since a significant number of their daily activities are restricted owing to the movement restriction orders (MCOs) imposed nationwide. The MCOs will definitely impede movement and their interaction process with the local community. Third Culture Kid (TCK) denotes a child who grows up in a culture distinct from the one their parents grew up in, and in the process experiences a wide array of challenges, barriers, and impediments while assimilating in the host country. Among the challenges they face, include communication and general barriers, such as food, weather, transportation system, clothing, and lifestyles that are different from their home country. This research was conducted to obtain preliminary data on both positive and negative impacts of life experiences among TCKs in Muslim country. Primary Objectives of this research are (1) To study why TCKs come to Malaysia and (2) To explore what are the positive and negative impacts on their life experiences when living in Malaysia? A qualitative approach utilising in-depth online interviews with 5 children-respondents from different countries and with varied intercultural experiences was employed in order to explore the above objectives. Data extracted were analysed using Atlas.ti software. The results of the study show that the main reason why TCKs come to Malaysia has been mainly due to the need to accompany their parents who are pursuing further studies in Malaysia and the sub-theme of the factors that their parents choose Malaysia as a destination to continue their studies are (a) Common culture with the home country, (b) Low cost of living, and (c) Exploring a new culture. Additionally, the results suggest that the positive impacts on their life experiences while in Malaysia are (a) Learning a new language and (b) Meeting new people and friends within the new cultural norms. The negative impacts are (a) Communications barrier and (b) Feeling of Homesickness. This study provides contributions to the literature review on this topic and preliminary information to parents who might wish to bring along their children in pursuing studies abroad in the future.

Induction, Proliferation and Differentiation of Callus in Paris polyphylla Sm. through Leaf Culture

Paris polyphylla Sm. is a vulnerable medicinal plant employed in the treatment of various ailments. This study seeks to establish a protocol for callus induction, proliferation, and differentiation of P. polyphylla. Immature leaf explants were cultured on MS medium with varying concentrations of plant growth regulators (PGRs), including 2,4-dichlorophenoxyacetic acid (2,4-D), kinetin (Kn), 6-benzylaminopurine (BAP), Thidiazuron (TDZ), α-Naphthalene acetic acid (NAA), and Gibberellic acid (GA3), along with 10% coconut water. After 12 weeks of primary culture, the optimal callus induction was observed in MS medium supplemented with 0.25 mg/l 2,4-D + 0.5 mg/l Kn. In the secondary culture at 8 weeks, the best callus proliferation, as determined by callus weight or growth index, occurred in MS medium supplemented with 2.0 mg/l BAP alone, 2.0 mg/l Kn alone, 1.0 mg/l TDZ alone, combinations of 2.0 mg/l Kn + 1.0 mg/l BAP + 2.0 mg/l GA3, and combinations of 0.5 mg/l NAA + 2.0 mg/l BAP + 2.0 mg/l GA3, as well as 10% coconut water. Furthermore, callus differentiation into mini rhizomes with root primordia was successfully achieved in MS media containing 2.5 mg/l Kn and 10% coconut water. This study reports, for the first time, the formation and differentiation of callus from leaf explants in P. polyphylla. Large-scale callus generation from leaf explants has the potential to enhance the production of bioactive secondary metabolites for therapeutic purposes and facilitate the development of plantlets through organogenesis.

Pronuclear transfer rescues poor embryo development of in vitro-grown secondary mouse follicles.

Pronuclear transfer rescues poor embryo development of in vitro-grown secondary mouse follicles.

THE RELATIONSHIP BETWEEN ORGANIZATIONAL SILENCE EDUCATION EXPERIENCED IN SECONDARY EDUCATION INSTITUTIONS AND SCHOOL CULTURE

Organizations can be thought of as small social examples created by people. In order for the organization to make progress in the rapidly changing and developing world, it may be necessary for the members of the organization to be in effective communication and to express their opinions without hesitation on every issue that concerns the organization. Silence is one of the most important factors preventing the development and change of the organization. Organizational silence may occur in organizations where there are members who cannot express their opinions and where effective communication is not dominant. However, just as different societies around the world have different cultures, each organization has its own unique culture. Culture is the most important organizational element that enables the organization to survive and maintain its existence against various changes in its environment. The cultures of organizations whose ultimate aim is to survive and adapt to every development can be affected by situations such as organizational silence and this effect can be reflected in the development of the organization. The purpose of this study is to determine the relationship between organizational silence in secondary education institutions and school culture according to teachers' opinions. The relationship between organizational silence and school culture was examined according to the variables of gender, type of school they work and professional seniority of teachers working in secondary education institutions. The population of the research consists of 859 teachers working in secondary education institutions in the Central district of Düzce, and the sample consists of 402 teachers working in 23 public schools in the Central district of Düzce. All of the general high schools in the central district of Düzce were included in the study, and the teachers to whom the measurement tool was applied were determined by easy sampling method. In the study, Organizational Silence scale and School Culture scale were used to collect data. The collected data were analyzed in the SPSS program. The role and relationship of organizational silence on school culture; independent samples t-test, one-way analysis of variance and kruskal wallis tests were applied to determine whether teachers differ according to gender, type of school and professional seniority. Pearson Correlation analysis was conducted to determine whether there is a significant relationship between organizational silence and school culture. The results obtained as a result of the research can be summarized as follows: According to the opinions of teachers working in secondary education institutions in the central district of Düzce, a low-level negative significant relationship was found between organizational silence and school culture. There is no significant difference between the views of teachers on organizational silence and its sub-dimensions and their gender. There was no significant difference between organizational silence and its sub-dimensions and school type. A significant relationship was found between organizational silence sub-dimensions, silence for the benefit of the organization and professional seniority. There is no significant difference between the views of teachers on school culture and all its sub-dimensions, and their gender. A significant difference was found between all sub-dimensions of school culture and the school type variable. A significant difference was determined between the other sub-dimensions of the school culture variable except the support culture sub-dimension and the professional seniority variable. Organizational silence and sub-dimensions were found to have a moderately significant negative relationship with success culture and support culture. A low-level significant positive correlation was found between organizational silence and sub-dimensions and bureaucratic culture. Organizational silence is not a significant predictor of task culture.

Strengthening Public School Culture: Integration of Local Wisdom, Religious Values, and Universal Values

This study focuses on integrating local wisdom, religious values, and universal values. This study aims at revealing various secondary school cultures in public schools. This study employed a qualitative method with a case study design at Junior High School SMPN 7 Muaro Jambi. Data was collected using observation, in-depth interviews, and document study techniques. The collected data were analyzed using an interactive model, including data condensation, presentation, and verification. Triangulation techniques, extended observations, and member checks are employed to increase the validity of the data. The study revealed that five school cultures still characterize school managerial life: Gotong Royong (cooperation), Musyawarah (deliberation), Islamic Ritual, Clean and Healthy, and Discipline. These five cultures are manifestations of adherence to local values (indigenousity), religious values, and universal values. Combining these three integrated sources of values is a distinctive pattern of school culture that becomes the foundation for developing school quality at public schools at SMPN 7 Muaro Jambi. This research implies that integrating local, religious, and universal values in school culture enriches learning and prepares students for a globally complex world.

RELATING THE ORGANIZATIONAL CULTURE ASSESSMENT INSTRUMENT TO ADHERENCE IN A PRAGMATIC TRIAL OF A MUSIC INTERVENTION

Abstract Embedded pragmatic trials encourage the translation of evidence-based interventions to “real-world” settings. Most pragmatic trials of behavioral interventions for people with dementia suffer from low adherence. Understanding how organizational values and structure may increase adherence is important. We report findings from an embedded, pragmatic trial (ePCT) of a personalized music intervention for managing behaviors in residents with dementia, conducted in 54 nursing homes (NHs) from four corporations between June 2019 and February 2020. Before the trial began, the administrator and a nursing staff member from each NH completed the Organizational Culture Assessment Instrument (OCAI). Using the OCAI, respondents rated their organizational culture by allocating a total of 100 points across four competing domains: Clan, Adhocracy, Hierarchy, and Market. Results were aggregated to understand how differences in culture impacted corporate-level adoption of the intervention. All four corporations allocated the majority of their points to Clan culture, which is focused on collaboration and staff engagement. However, corporations differed in their scoring of the secondary culture type. The two corporations that rated Hierarchical culture, which prioritizes consistency and efficiency, highly were more likely to adhere to the intervention protocols. The corporation with Market highly rated had the lowest adherence to the protocols. After controlling for other corporate characteristics, including for-profit status, size, and overall quality, hierarchical culture was associated with greater numbers of exposed residents and a higher dose of the music, compared to other culture types. Understanding the role of organizational culture on pragmatic implementation is an understudied area for research.

Influence Of Principals’ Leadership Practices on Collaborative School Culture in Public Secondary Schools in Tiaty East and Tiaty West Sub-Counties, Kenya.

An ideal collaborative school culture in the two sub-counties should entail teacher’s collaboration, teacher’s involvements in decision-making practices, effective open communication and high teacher motivation. The current situation in Tiaty East and Tiaty West sub counties is that the area is plagued by poverty and frequent banditry, this has made the school environment in the areas to be hostile and the safety of both the learner and the teacher is not assured. A hostile school culture of collaboration can cause the problem of poor performance to persist since there are no harmonious relations among teachers, and teachers work in isolation. As a result, the principals’ task is to create an enabling school climate towards the creation of a collaborative culture of the school. The purpose of this study was to find out influence of school principals’ leadership practices on collaborative school culture in public secondary schools in Tiaty East and Tiaty west sub-counties. The objectives of this study were : to determine influence of principals’ mentorship practices on collaborative school culture and to establish how principals’ communication practices influence collaborative school culture. The research approach was the qualitative research and a cross-cultural study design with two cases and multiple units of analysis within each case. This study was grounded on the perspective that valuable insights about the issue of principal leadership practices and collaborative school culture can be obtained from a target population of 77 comprising of principals (1), teachers (156), support staff (276) and student leaders (328). A sample of 161 respondents; 1 principals, 31 teachers, 56 support staff and 64 student leaders was selected using purposeful sampling. The main instrument for primary data collection was use of interview schedules for principals, teachers and support staff and focus group discussion guides for student leaders. Data analysis was done using Computer Assisted Qualitative Data Analysis Software (CAQDA); ATLAS. Ti 9. The study found that the principals invited mentors for staff development to improve collaboration, ultimately positively impacting on student outcomes to promote a collaborative secondary school culture. Moreover, on principals communication practices, school meetings and social media especially Whatsapp groups are used to engage the school community in communication.The study concluded that principal's mentorship practices, and principal's communication practices are integral and symbiotic in creating a positive and collaborative school culture. On building positive collaborativecultures, the study is beneficial to future researchers in the field as they can expound more on other principals’ leadership practices and it will also contribute to existing research data. The study recommended that principals should involve teachers and other stakeholders in mentorship process in the schools. Key words: Collaboration, decision-making, motivation, mentorship, communication.

Changing Strategies for the Detection of Bacteria in Platelet Components in Ireland: From Primary and Secondary Culture (2010-2020) to Large Volume Delayed Sampling (2020-2023).

Bacterial contamination of platelet components (PC) poses the greatest microbial risk to recipients, as bacteria can multiply over the course of PC storage at room temperature. Between 2010 and 2020, the Irish Blood Transfusion Service (IBTS) screened over 170,000 buffy coat-derived pooled (BCDP) and single-donor apheresis platelets (SDAPs) with the BACT/ALERT 3D microbial detection system (Biomerieux, L'Etoile, France), using a two-step screening protocol which incorporated primary and secondary cultures. Although the protocol was successful in averting septic transfusion reactions (STRs), testing large sample volumes at later time points was reported to improve detection of bacterial contamination. A modified large-volume delayed sampling (LVDS)-type protocol was adopted in 2020, which in the case of SDAP was applied to collections rather than individual splits (2020-2023, 44,642 PC screened). Rates of bacterial contamination for BCDP were 0.125% on Day-2, 0.043% on Day-4 vs. 0.191% in the post-LVDS period. SDAP contamination rates in the pre-LVDS period were 0.065% on Day-1, 0.017% on Day-4 vs. 0.072% in the post-LVDS period. Confirmed STRs were absent, and the interdiction rate for possibly contaminated SDAP was over 70%. In the post-LVDS period, BCDPs had a higher total positivity rate than SDAPs, 0.191% (1:525) versus 0.072% (1:1385), respectively, (chi-squared 12.124, 1 df, p = 0.0005). The majority of organisms detected were skin-flora-type, low pathogenicity organisms, including coagulase-negative staphylococci and Cutibacterium acnes, with little change in the frequency of clinically significant organisms identified over time. Both protocols prevented the issue of potentially harmful components contaminated (rarely) with a range of pathogenic bacteria, including Escherichia coli, Serratia marcesens, Staphylococcus aureus, and streptococci. Culture positivity of outdates post-LVDS whereby 100% of expired platelets are retested provides a residual risk estimate of 0.06% (95% CI 0.016-0.150). However, bacterial contamination rates in expired platelets did not demonstrate a statistically significant difference between the pre-LVDS 0.100% (CI 0.033-0.234) and post-LVDS 0.059% (0.016-0.150) periods (chi-squared = 0.651, 1 df, p = 0.42).

A Venn Diagram of Secondary-Postsecondary Teaching and Learning: The Transformative Power of Concurrent Enrollment Partnerships

For decades, researchers have explored the reasons why students show up for college unprepared. In an effort to explain this phenomenon, scholars have gravitated toward a “gap” metaphor to explain the liminal space between what high school students are capable of and what colleges expect of them. The pandemic has exacerbated this situation leaving many students further behind in the academic and personal growth needed to succeed in college. Conventional wisdom says that college is “harder” than high school and students simply must adapt. And while college may, indeed, be “harder,” another perspective is that college is different from high school: in design, in intent, in pedagogical strategies, and more. Still, systems that perpetuate these educational differences place much of the responsibility of transitioning from one educational level to the other solely on the students. Enter Concurrent Enrollment (CE). In the CE space, secondary and post-secondary cultures are superimposed resulting in a Venn Diagram of teaching and learning that has the potentially for bringing into focus the best elements of each. With respect, intention, and support, high school and college instructors can utilize CE spaces to learn from and with one another to better support the continuum of student learning—regardless of where that learning takes place.

P‐TS‐39 | Impact to Put in Place Secondary Bacterial Culture in Order to Reduce Bacterial Contamination of Platelet Transfusions

P‐TS‐39 | Impact to Put in Place Secondary Bacterial Culture in Order to Reduce Bacterial Contamination of Platelet Transfusions

Impact of media culture, freeze-drying and storage conditions on preservation of Lacticaseibacillus paracasei 90: Viability and metabolic potential as a secondary culture in semi-hard cheese

Freeze-drying is the main process used to preserve lactic bacterial cultures at an industrial level; however, their viability and metabolic activity can be affected not only by the process per se but also by storage conditions. This study investigated the effect of freeze-drying and long-term storage (14 months) on the viability and metabolic activity of Lacticaseibacillus paracasei 90 (L90) grown in three culture media. The freeze-drying process did not influence its culturability and viability, regardless of the culture media used. The storage of the freeze-dried cultures at room temperature had a negative impact on viability, culturability and its ability to grow and acidify milk. However, L90 was able to withstand a long period of storage at low temperatures (14 months, 4 °C) and keep the technological features desirable of a ripening culture. Overall, the results demonstrate the robustness of this strain as a secondary culture in the food industry.

The kynurenic acid analog SZR104 induces cytomorphological changes associated with the anti-inflammatory phenotype in cultured microglia

We previously showed the anti-inflammatory effects of kynurenic acid (KYNA) and its brain-penetrable analog N-(2-(dimethylamino)ethyl)-3-(morpholinomethyl)-4-hydroxyquinoline-2-carboxamide (SZR104) both in vivo and in vitro. Here, we identified the cytomorphological effects of KYNA and SZR104 in secondary microglial cultures established from newborn rat forebrains. We quantitatively analyzed selected morphological aspects of microglia in control (unchallenged), lipopolysaccharide (LPS)-treated (challenged), KYNA- or SZR104-treated, and LPS + KYNA or LPS + SZR104-treated cultures. Multicolor immunofluorescence labeling followed by morphometric analysis (area, perimeter, transformation index, lacunarity, density, span ratio, maximum span across the convex hull, hull circularity, hull area, hull perimeter, max/min radii, mean radius, diameter of bounding circle, fractal dimension, roughness, circularity) on binary (digital) silhouettes of the microglia revealed their morphological plasticity under experimental conditions. SZR104 and, to a lesser degree, KYNA inhibited proinflammatory phenotypic changes. For example, SZR104 treatment resulted in hypertrophied microglia characterized by a swollen cell body, enlarged perimeter, increased transformation index/decreased circularity, increased convex hull values (area, perimeter, mean radius, maximum span, diameter of the bounding circle and hull circularity), altered box-counting parameters (such as fractal dimension), and increased roughness/decreased density. Taken together, analysis of cytomorphological features could contribute to the characterization of the anti-inflammatory activity of SZR104 on cultured microglia.