BIO Faculty Publications

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    Organogenetic transcriptomes of the Drosophila embryo at single cell resolution
    (The Company of Biologists Ltd., 2024-01) Peng, Da; Jackson, Dorian; Palicha, Bianca; Kernfeld, Eric; Laughner, Nathaniel; Shoemaker, Ashleigh; Celniker, Susan E.; Loganathan, Rajprasad; Cahan, Patrick; Andrew, Deborah J.
    To gain insight into the transcription programs activated during the formation of Drosophila larval structures, we carried out single cell RNA sequencing during two periods of Drosophila embryogenesis: stages 10-12, when most organs are first specified and initiate morphological and physiological specialization; and stages 13-16, when organs achieve their final mature architectures and begin to function. Our data confirm previous findings with regards to functional specialization of some organs - the salivary gland and trachea - and clarify the embryonic functions of another - the plasmatocytes. We also identify two early developmental trajectories in germ cells and uncover a potential role for proteolysis during germline stem cell specialization. We identify the likely cell type of origin for key components of the Drosophila matrisome and several commonly used Drosophila embryonic cell culture lines. Finally, we compare our findings with other recent related studies and with other modalities for identifying tissue-specific gene expression patterns. These data provide a useful community resource for identifying many new players in tissue-specific morphogenesis and functional specialization of developing organs.
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    Succession of the bacterial community from a spacecraft assembly clean room when enriched in brines relevant to Mars
    (Cambridge University Press, 2023-12) Carte, Meris; Chen, Fei; Clark, Benton C.; Schneegurt, Mark A.
    Interplanetary spacecraft are built in a spacecraft assembly facility (SAF), a clean room designed to reduce microbial contamination that could confound life detection missions or influence native ecosystems. The frigid hyperarid near-surface environment of Mars has ample hygroscopic Mg and Na salts of chloride, (per)chlorate and sulphate that may deliquesce to form dense brines, liquids with low water activity, and freezing points <0°C. The current study sought to define the climax microbial community after 6 mo of enrichment of SAF floor wipe samples in salt plains medium supplemented with 50% (w/v; ~2 M; $a_{w}$ = 0.94) MgSO$_4$ or 20% (w/v; ~1.9 M; $a_{w}$ = 0.91) NaClO$_3$. After 1 wk, 4 wk and 6 mo of incubation, metagenomic DNA extracts of the enriched SAF microbial community were used for high-throughput sequencing of 16S rRNA genes and subsequent phylogenetic analyses. Additionally, dozens of bacterial strains were isolated by repetitive streak-plating from the climax community after 6 mo of enrichment. Early in the enrichment, staphylococci greatly dominated and then remained abundant members of the community. However, actinobacteria succeeded the staphylococci as the dominant taxa as the cultures matured, including Arthrobacter, Brachybacterium and Brevibacterium. A diverse assemblage of bacilli was present, with Oceanobacillus being especially abundant. The SAF culture collection included representatives of Brachybacterium conglomeratum, Brevibacterium sediminis, Oceanobacillus picturae and Staphylococcus sciuri. These were characterized with biochemical and physiological tests, revealing their high salinotolerance. Shannon diversity indices were generally near 2, reflecting modest diversity at several levels of identity and the community structures were uneven throughout. However, minor members of the community seem capable of the ecosystem functions required for biogeochemical cycling. For instance, organisms capable of all the functions of the N cycle were detected. The microbial assemblage in SAFs is the most likely to be transported by spacecraft to another world. While individual microbial populations may exhibit the qualities needed for survival at the near-surface of Mars, certainly entire communities with the capacity for complete biogeochemical cycling, would have a greater chance of survival and proliferation.
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    Hydrophilic and Antibacterial Electrospun Nanofibers from Monofilament Fishing Lines
    (Korean Fiber Society, 2023-12) Ijaola, Ahmed O.; Mohammed, Qamar Saberi; Obi, Mmasi; Akamo, Damilola O.; Ajiboye, Emmanuel Gboyega; Twomey, Janet M.; Yang, Shang-You; Asmatulu, Eylem
    Microplastics such as monofilament fishing lines (MFLs) are major pollutants in the marine environment and affect marine life and water quality. To solve this global menace, many researchers have been working on several ways to recycle these wastes and convert them into value-added products such as nanofibers. In this study, we produced novel nanofibers through the electrospinning of a polymeric solution consisting of MFL, hydroxyapatite (HAP), and silver nanoparticles (AgNPs). These fabricated nanofibers were further characterized to study their wettability, surface morphology, surface chemistry, thermal degradation, and antibacterial capability. Results from the incorporation of HAP and AgNPs showed increased fiber diameter and scattered fiber orientation. The addition of 0.4 and 1.5 wt% AgNPs to the nanofibers improved their thermal stabilities for temperatures above 350°C. The MFL + HAP + 1.5 wt% AgNPs nanofibers showed the best antibacterial performance against Escherichia coli (gram-negative) and Staphylococcus aureus (gram-positive), with bactericidal efficiencies of 70.5% and 68.6%, respectively. Also, increasing the size of the nanofiber aids cell proliferation. These fabricated nanofibers could be used for biomedical and water purification applications.
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    Oocyte quality is enhanced by hypoglycosylated FSH through increased cell-to-cell interaction during mouse follicle development
    (The Company of Biologists Ltd., 2023-11) Converse, Aubrey; Liu, Zhenghui; Patel, Jai C.; Shakyawar, Sushil K.; Guda, Chittibabu; Bousfield, George R.; Kumar, T. Rajendra; Duncan, Francesca E.
    Macroheterogeneity in follicle-stimulating hormone (FSH) β-subunit N-glycosylation results in distinct FSH glycoforms. Hypoglycosylated FSH$^{21}$ is the abundant and more bioactive form in pituitaries of females under 35 years of age, whereas fully glycosylated FSH$^{24}$ is less bioactive and increases with age. To investigate whether the shift in FSH glycoform abundance contributes to the age-dependent decline in oocyte quality, the direct effects of FSH glycoforms on folliculogenesis and oocyte quality were determined using an encapsulated in vitro mouse follicle growth system. Long-term culture (10-12 days) with FSH$^{21}$ (10 ng/ml) enhanced follicle growth, estradiol secretion and oocyte quality compared with FSH$^{24}$ (10 ng/ml) treatment. FSH$^{21}$ enhanced establishment of transzonal projections, gap junctions and cell-to-cell communication within 24 h in culture. Transient inhibition of FSH$^{21}$-mediated bidirectional communication abrogated the positive effects of FSH$^{21}$ on follicle growth, estradiol secretion and oocyte quality. Our data indicate that FSH$^{21}$ promotes folliculogenesis and oocyte quality in vitro by increasing cell-to-cell communication early in folliculogenesis, and that the shift in in vivo abundance from FSH$^{21}$ to FSH$^{24}$ with reproductive aging may contribute to the age-dependent decline in oocyte quality.
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    Fabrication, Characterization, and In Vitro Cytotoxicity Assessment of Tri-Layered Multifunctional Scaffold for Effective Chronic Wound Healing
    (Multidisciplinary Digital Publishing Institute (MDPI), 2023-09) Ijaola, Ahmed Olanrewaju; Subeshan, Balakrishnan; Pham, Anh; Uddin, Md. Nizam; Yang, Shang-You; Asmatulu, Eylem
    Chronic wounds have been a global health risk that demands intensive exploration. A tri-layered biomaterial scaffold has been developed for skin wounds. The top layer of the scaffold is superhydrophobic, and the bottom layer is hydrophilic, both of which were electrospun using recycled expanded polystyrene (EPS) and monofilament fishing line (MFL), respectively. The intermediate layer of the scaffold comprised hydrogel by cross-linking chitosan (CS) with polyethylene glycol. The surface morphology, surface chemistry, thermal degradation, and wettability characteristics of each layer of the scaffold were examined. Also, the antibacterial activity and in vitro cytotoxicity study on the combined tri-layered scaffold were assessed against $Escherichia coli (E. coli)$ and $Staphylococcus aureus (S. aureus)$. Data revealed exceptional water repellency of the heat-treated electrospun top superhydrophobic layer (TSL) with a high-water contact angle (WCA) of 172.44°. A TSL with 15 wt% of micro-/nano-inclusions had the best thermal stability above 400 °C. The bottom hydrophilic layer (BHL) displayed a WCA of 9.91°. Therapeutically, the synergistic effect of the combined tri-layered scaffold significantly inhibited bacteria growth by 70.5% for $E. coli$ and 68.6% for $S. aureus$. Furthermore, cell viability is enhanced when PEG is included as part of the intermediate CS hydrogel layer (ICHL) composition.