Superficial Cells Research Articles

Effect of Probiotic Lactobacillus Casei on Secretory Immunoglobulin A (S-IgA) Levels in Rats Norvegicus Strain Wistar Post Ovariectomy

Estrogen promotes the growth of Lactobacillus colonies in the vagina, converting glycogen in superficial cells into lactic acid, resulting in a low vaginal pH that inhibits pathogenic bacteria. Post-menopause, Lactobacillus growth declines, leading to thinner secretions, higher vaginal pH, and a weakened immune system. This study investigates the effect of Lactobacillus casei extract on secretory immunoglobulin A (IgA) levels, vaginal acidity, and beneficial bacteria in the vaginal mucosa. An experimental post-test-only control group design was employed with 24 female rats aged 8-10 weeks, weighing 180-220 grams. Following ovariectomy and a 14-day acclimation, the rats were divided into four groups: a control group (K) with no treatment and three treatment groups (P1, P2, P3) receiving Lactobacillus casei extract at doses of 2ml, 2.25ml, and 2.5ml, respectively, administered orally for 14 days. On day 15, vaginal acidity was measured using a swab method, and IgA levels were assessed via ELISA; gram-positive staining was performed for bacterial identification. One-way ANOVA revealed that Lactobacillus casei extract significantly increased IgA levels and reduced vaginal acidity (p = 0.003 and p = 0.000). The reduction in acidity is attributed to Lactobacillus converting glycogen into lactic acid, maintaining vaginal acidity, while S-IgA on the mucosal surface acts as an opsonin and antigen-presenting cell to inhibit pathogenic growth. The study concludes that Lactobacillus casei extract can elevate secretory IgA levels and normalize vaginal pH.

Guilu Erxian glue reduces endoplasmic reticulum stress-mediated apoptosis and restores the balance of extracellular matrix synthesis and degradation in chondrocytes by inhibiting the ATF6/GRP78/CHOP signaling pathway

Knee Osteoarthritis (KOA) is characterized by phenotypic alterations, apoptosis, and the breakdown of the extracellular matrix (ECM) in the superficial articular cartilage cells. The inflammatory response activates the Endoplasmic Reticulum Stress (ERS) signaling pathway, which plays a critical role in the pathophysiology and progression of KOA. Chondrocytes stimulated by thapsigargin(TG)exhibit heightened ERS and significantly increase the expression of ERS-associated proteins. Key mediators of ERS-induced apoptosis include X-box-binding protein 1(XBP1), elevated levels of the protein transport protein Sec61 subunit (SEC61), and C/EBP homologous protein (CHOP).While the precise mechanism of action of Guilu Erxian Glue (GEG), a medication commonly used in the clinical treatment of KOA, remains to be fully elucidated, our research has shown that GEG mitigates the imbalance between ECM synthesis and degradation, as well as chondrocyte apoptosis resulting from ERS. This effect is likely achieved through the suppression of the Activating Transcription Factor 6 (ATF6)/Glucose-Regulatory Protein 78 (GRP78)/CHOP signaling pathway.In summary,our research results indicate that GEG can activate the ATF6/GRP78/CHOP signaling pathway to restore endoplasmic reticulum (ER) homeostasis in chondrocytes, thereby reducing chondrocyte apoptosis and ultimately promoting the balance between ECM synthesis and degradation.

A Novel Intravaginal Contraceptive Plug for Cats: A Preliminary Biocompatibility Assessment on Haematology and Vaginal Swab.

This preliminary study evaluated the biocompatibility of a novel degradable intravaginal plug contraceptive composed of PEG 4000 and chitosan in cats using haematological profiling and vaginal cytology. Five healthy, non-pregnant female cats were fully anaesthetised and fitted with an intravaginal plug (10 × 0.3 mm) using an applicator, following oestrogen administration 3 h prior. Blood samples were collected from the cephalic vein on days 0 (pre-insertion) and 3 and 7 (post-insertion). Vaginal cytology examinations were conducted on day 0 (pre- and post-oestrogen injection) and days 1, 3 and 7 post-insertion. Haematological parameters, including red blood cell count, haemoglobin levels, haematocrit values, total white blood cell count and differentiation, showed no significant changes after contraceptive insertion (p > 0.05). Vaginal cytology indicated an acute inflammatory response in one out of five subjects on day three post-insertion. The distribution of vaginal epithelial cells (parabasal, intermediate and superficial) remained unaffected by contraception. Oestrogen injection resulted in the dominance of superficial cells up to day 7 of observation (p < 0.05). Overall, PEG 4000 and chitosan-based intravaginal plug contraceptives demonstrated sufficient biocompatibility, indicating their potential as viable contraceptive options for feline use.

Demonstrating the staining ability of &lt;i&gt;Zingiber officinale&lt;/i&gt; and &lt;i&gt;Curcuma longa&lt;/i&gt; as replacement to Eosin Y in EA50

Nicholas Papanicolaou developed the Pap stain, a combination of three major stains used for cervical smears. Eosin-Azure (EA50), one of these stains, includes Eosin Y, Light Green SF, and Bismarck brown. This study explores the use of Curcuma longa extract as a substitute for Eosin Y in EA50. Cervical smears from premenopausal women were collected and stained using both the standard Papanicolaou stain and a modified version where Eosin Y was replaced with Curcuma longa extract. The modified stain's effectiveness was assessed based on the quality of staining of superficial cells. The modified stain (T.A- extract) successfully stained and highlighted superficial cells, while the Zingiber officinale extract (Z.A) performed poorly in staining the nuclei and cytoplasm. The staining ability of the Curcuma longa extract was found to be 100% comparable to the traditional Papanicolaou stain. Staining aims to clearly demonstrate cellular components for diagnostic purposes, and many existing stains are long-established but require modifications for better application. Using local alternatives can enhance availability and affordability. Curcuma longa is readily available and, based on our findings, can effectively replace Eosin Y in Eosin-Azure stains. Further research into the dye components of herbal products and their staining capabilities could lead to cost-effective, locally sourced alternatives to imported stains.

Superficial Conjunctival Cells from Dupilumab-Treated Patients with Atopic Dermatitis with Ocular Adverse Events Display a Transcriptomic Psoriasis Signature

Dupilumab has demonstrated efficacy in the treatment of atopic dermatitis. However, a subset of patients experiences ocular adverse events (OAEs), including conjunctivitis and dry eye syndrome, the pathological mechanisms of which are still unknown. In a bicentric study, we used DNA microarray analysis to compare the transcriptome of conjunctival cells of patients with atopic dermatitis collected by impression cytology before (M0) and 4 months after (M4) initiating dupilumab treatment. Thirty-six patients were included and divided in 2 groups according to their ophthalmological status at M4: 12 with OAEs (OAE+) and 24 without (OAE-). The analysis revealed 52 differentially expressed genes between OAE+ and OAE- patients at M0 and 113 at M4. Ingenuity Pathway Analysis enrichment revealed a psoriasis signature in OAE+ patients, both before and after OAE outcomes. In addition, we noticed the overexpression of several genes involved in keratinocyte differentiation, particularly encoding cornified envelope components. Among the 16 differentially expressed genes selected for real-time RT-PCR validation, 9 were confirmed as upregulated at M4 in OAE+ versus OAE- patients, validating the psoriasis signature, whereas MUC7 was downregulated. In conclusion, these results suggest that a conjunctival transcriptomic profile predisposes some patients with atopic dermatitis to developing OAEs upon dupilumab treatment.

Electrosurgery: heating, sparking and electrical arcs.

The translation of impedance (R), current (I), and voltage (V) into tissue effects and the understanding of the settings of electrosurgical units is not obvious if judged by the many questions during live surgery. Below 200 V, the current heats the tissue until the steam of boiling stops the current. Thus, slower heating, because of less energy or a larger contact area, results in deeper coagulation. Above 200 V and a duty cycle (per cent of time electricity is delivered) of >50% (yellow pedal), sparks become electric arcs, and the heat causes the explosion of superficial cells, i.e. cutting. With higher voltages, cutting is associated with coagulation, i.e. blended current. With even higher voltages and a duty cycle <10% preventing arching, only coagulation occurs (blue pedal; forced coagulation). Voltage being crucially important for tissue effects, newer electrosurgical units deliver a constant voltage and limit the energy output (Maximal Watts: W=I*V= joules/sec). Unfortunately, the electrosurgical units indicate the combination of voltage and duty cycles as a force of cutting (pure cutting or blended) or coagulation (soft, forced or spray) current. It is important that the surgeon understands whether electrosurgical units control voltages or output, as well as the electrical basics of the different settings and programs used.

A Novel Technique for Corneal Transepithelial Electrical Resistance Measurement in Mice.

We developed a technique that can measure corneal transepithelial electrical resistance (TER) in mice, which was used for evaluating corneal toxicity induced by ophthalmic drugs. We used a tissue culture well and its insert to mount the mouse globe and separated the cornea from the rest of the globe to enable corneal TER measurements to be taken. The explanted mouse eyes were divided into groups, and the corneal epithelia were exposed to different concentrations of BAC. Half of these eyes were fixed for transmission electron microscopy (TEM) examination and the other for ZO-1 immunohistochemical (IHC) evaluation. After exposure to control, 0.1%, 0.2%, and 0.5% BAC, the TER was 100 ± 0%, 91 ± 14%, 83 ± 13%, and 34 ± 12% of the pre-exposure TER at 1 min, respectively, with a statistically significant decrease in the 0.5% group. After 3 min, the TER showed a statistically significant decrease in the 0.2% and 0.5% groups. The TEM examinations showed a loss of epithelial tight junctions between superficial cells in the 0.2% and 0.5% groups. The IHC examination showed decreased ZO-1 staining of the corneal epithelium of the same groups as compared to the control. To the best of our knowledge, we succeeded in developing an innovative technique for corneal TER measurement in mice.

Efficacy and Safety of Intravaginal Estrogen in the Treatment of Atrophic Vaginitis: A Systematic Review and Meta-Analysis.

Postmenopausal females often experience genitourinary symptoms like vulvovaginal dryness due to estrogen decline. Hormone replacement therapy is effective in alleviating vaginal atrophy and genitourinary syndrome in this population. Evaluate local estrogen's safety and effectiveness for alleviating postmenopausal vaginal symptoms, including endometrial thickness, dyspareunia, vaginal pH, and dryness. We searched Google Scholar, Cochrane Library, ClinicalTrial.Gov, PubMed, and ScienceDirect databases until July 2023. All randomized controlled trials (RCTs) linking intravaginal estrogen supplementation to vaginal atrophy or vaginitis were included. The risk of bias was evaluated with RoB 2, and publication bias was assessed using Egger and Beggs analysis. All evidence pertains to females. Eighteen studies (n = 4,723) compared estrogen with placebo. Patients using estrogen showed a significant increase in superficial cells (mean differences [MD]: 19.28; 95% confidence intervals [CI]: 13.40 to 25.16; I² = 90%; P < 0.00001) and a decrease in parabasal cells (MD: -24.85; 95% CI: -32.96 to -16.73; I² = 92%; P < 0.00001). Vaginal pH and dyspareunia significantly reduced in estrogen users (MD: -0.94; 95% CI: -1.05 to -0.84; I² = 96%) and (MD: -0.52; 95% CI: -0.63 to -0.41; I² = 99%), respectively. Estrogen did not significantly affect vaginal dryness (MD: -0.04; 95% CI: -0.18 to 0.11; I² = 88%). Adverse events like vulvovaginal pruritis, mycotic infection, and urinary tract infection were reported, but the association was insignificant (risk ratio: 0.95; 95% CI: 0.88 to 1.02; I² = 0%). Our meta-analysis of 18 RCTs suggests promising potential for intravaginal estrogen therapy in alleviating vaginal atrophy and vaginitis in postmenopausal females.

Associations among the largest follicle, preovulatory estradiol concentrations, and predominant vaginal epithelial cells at the completion of hormonal ovarian stimulation for fixed-time artificial insemination in goats.

The objective of the present study was to investigate the association among the largest follicle (LF), preovulatory estradiol (E2), and predominant vaginal epithelial cell at the completion of hormonal ovarian stimulation for fixed-time artificial insemination (FTAI) in goats. Thirty-seven crossbred Boer does received gonadotropin-releasing hormone (GnRH) and intravaginal progesterone (P4)-releasing devices (day 0). On day 5, P4 devices were removed and does received prostaglandin F2α and equine chorionic gonadotrophin. On day 7, does received GnRH, and FTAI was undertaken. On day 7, does were divided into three groups, i.e. small-sized (3-3.9 mm; n = 5), medium-sized (4-4.9 mm; n = 8), and large-sized (≥5 mm; n = 24) according to the diameter of the ovarian LF; follicular characteristics (number and diameter) were identified, and blood samples and vaginal smears were collected. The average diameters of total antral follicles and LF and the percentage of superficial cell were greatest in large-sized LF does (p < .01). The average diameters of total antral follicle (r = .68) and LF (r = .71), number of preovulatory follicle (r = .58), and plasma E2 concentrations (r = .61) were positively correlated with the percentage of superficial cells (p < .01). The likelihood of a pregnancy outcome after the FTAI increased by 13.71 times in does with a greater average diameter of antral follicle, 14.18 times with emergence of a large preovulatory follicle, and 36.83 times with a higher percentage of vaginal superficial cells (p < .01). It was concluded that there is a relationship between the cell types of the vaginal epithelium, the diameters of the largest ovarian follicles, and the concentration of E2 in goats subjected to FTAI protocols.

Chondrogenic potential of superficial versus cartilage layer cells of the temporomandibular joint condyle in photopolymerizable gelatin-based hydrogels.

The objectives of this study were to compare the chondrogenic potential of cells derived from different layers of Mandibular condyle cartilage and to gain further understanding of the impact of chondrogenic cues when embedded into a novel hydrogel scaffold (PGH, a polymer blend of poly (ethylene glycol), gelatin, and heparin) compared to a gelatin hydrogel scaffold (GEL). Cartilage layer cells (CLCs) and fibroblastic superficial layer cells (SLCs) were harvested from the mandibular condyle of boer goats obtained from a local abattoir. After expansion, cells were seeded into PGH and GEL hydrogels and cultured in chondrogenic media for 3 weeks. Scaffolds were harvested at 0, 1, and 3 week(s) and processed for gross appearance, histochemical, biochemical, and mechanical assays. In terms of chondrogenesis, major differences were observed between scaffold materials, but not cell types. Glycosaminoglycan (GAG) staining showed GEL scaffolds deposited GAG during the 3 week period, which was also confirmed with the biochemical testing. Moreover, GEL scaffolds had significantly higher compressive modulus and peak stress than PGH scaffolds at all time points with the largest difference seen in week 3. It can be concluded that GEL outperformed PGH in chondrogenesis. It can also be concluded that materials play a more important role in the process of chondrogenesis than the tested cell populations. Fibroblastic SLCs were shown to have similar chondrogenic potential as CLCs cells, suggesting a rich pool of progenitor cells in the superficial fibroblastic layer capable of undergoing chondrogenesis given appropriate physical and chemical cues.

Establishment of urinary catheter materials antibacterial efficacy evaluation platform in pigs

Urinary catheters are one of the common invasive medical catheters, mostly composed of high-polymer materials such as polyurethanes. During clinical use, the high friction between the catheter material surface and tissues can lead to tissue damage, patient discomfort, and pain during insertion or removal. Moreover, due to the hydrophobic nature of the polymer material surface, there is a risk of urinary tract infections (CTIs) caused by problems such as protein adsorption and bacterial infection upon insertion into the body. Prolonged indwelling time of urinary catheters in the body increases the probability of other complications, potentially leading to life-threatening consequences and imposing medical burdens. Therefore, the purpose of this study was focused on the insertion of the Escherichia coli (E. coli, 1 × 105 CFU/mL)-coated urinary catheter to implant into a female pig’s urethra for establishing the urinary catheter materials antibacterial efficacy evaluation platform in pigs. Results were shown that during the experimental period, the week 1 and week 2 average pig’ body temperature were 39.4 ± 0.6°C and 39.1 ± 0.2°C, respectively. The average pig’s body temperature was normal and there was no significant differences in body temperature at week 1 and week 2 urinary catheter implanted-experiment period. Moreover, the clinical observation of the pigs showed that their vitality, appetite, and excretion were all normal, and they survived until the end of the experiment without presenting any abnormal clinical symptoms. In addition, E. coli in urine culture respectively showed 0.95 ± 0.03 × 105 CFU/mL, 1.23 ± 0.03 × 105 CFU/mL, 1.43 ± 0.04 × 105 CFU/mL, 1.75 ± 0.08 × 105 CFU/mL, and 1.91 ± 0.09 × 105 CFU/mL on day 0, 3, 7, 10, and 14 of urinary catheter implantation. As time progressed after urinary catheter implantation, there was an increasing trend in E. coli counts. Staining with crystal violet for urinary catheter showed that the urinary catheter wall was positive for crystal violet staining. E. coli isolation and counts from biofilm on inside and outside of urinary catheter respectively showed 5.10 ± 0.10 × 105 CFU/mL and 7.50 ± 1.08 × 105 CFU/mL on day 14 of urinary catheter implantation. As time progressed after urinary catheter implantation, there was an increasing trend in E. coli counts. Finally, the pathological interpretation of the urinary tract indicates that in the tissue morphology of the bladder, there is observed a localized mild shedding of superficial and intermediate layer cells in the transitional epithelium of the ureteral stent group, with only basal cells remaining connected to its submucosa. There are no significant pathological changes observed in the tissue morphology of the urethra and kidneys. According to the results of this study, a urinary catheter materials antibacterial efficacy evaluation platform in pigs has successfully established, which can be provided for R&amp;D of urinary catheter antibacterial materials.

Hierarchical Gradients of Encoded Spatial and Sensory Information in the Neocortex Are Attenuated by Dorsal Hippocampal Lesions.

During navigation, the neocortex actively integrates learned spatial context with current sensory experience to guide behaviors. However, the relative encoding of spatial and sensorimotor information among cortical cells, and whether hippocampal feedback continues to modify these properties after learning, remains poorly understood. Thus, two-photon microscopy of male and female Thy1-GCaMP6s mice was used to longitudinally image neurons spanning superficial retrosplenial cortex and layers II-Va of primary and secondary motor cortices before and after bilateral dorsal hippocampal lesions. During behavior on a familiar cued treadmill, the locations of two obstacles were interchanged to decouple place-tuning from cue-tuning among position-correlated cells with fields at those locations. Subpopulations of place and cue cells each formed interareal gradients such that higher-level cortical regions exhibited higher fractions of place cells, whereas lower-level regions exhibited higher fractions of cue cells. Position-correlated cells in the motor cortex also formed translaminar gradients; more superficial cells were more likely to exhibit fields and were more sparsely and precisely tuned than deeper cells. After dorsal hippocampal lesions, a neural representation of the learned environment persisted, but retrosplenial cortex exhibited significantly increased cue-tuning, and, in motor cortices, both position-correlated cell recruitment and population activity at the unstable obstacle locations became more homogeneously elevated across laminae. Altogether, these results support that the hippocampus continues to modulate cortical responses in familiar environments, and the relative impact of descending feedback obeys hierarchical interareal and interlaminar gradients opposite to the flow of ascending sensory inputs.

Molecular anatomy of emerging Xenopus left-right organizer at successive developmental stages.

Vertebrate left-right symmetry breaking is preceded by formation of left-right organizer. In Amphibian, this structure is formed by gastrocoel roof plate, which emerges from superficial suprablastoporal cells. GRP is subdivided into medial area, which generates leftward flow by rotating monocilia and lateral Nodal1 expressing areas, which are involved in sensing of the flow. After successful symmetry breaking, medial cells are incorporated into a deep layer where they contribute to the axial mesoderm, while lateral domains join somitic mesoderm. Here, we performed detailed analysis of spatial and temporal gene expression of important markers and the corresponding morphology of emerging GRP. Endodermal marker Sox17 and markers of superficial mesoderm display complementary patterns at all studied stages. At early stages, GRP forms Tekt2 positive epithelial domain clearly separated from underlying deep layers, while at later stages, this separation disappears. Marker of early somitic mesoderm MyoD1 was absent in emerging GRP and was induced together with Nodal1 during early neurulation. Decreasing morphological separation is accompanied by lateral to medial covering of GRP by endoderm. Our data supports continuous link between superficial mesoderm at the start of gastrulation and mature GRP and suggests late induction of somitic fate in lateral GRP.

Cytological and Cytometric Analysis of Epithelial Cell Changes Under the Surface of Acrylate Prosthesis in Diabetic Patients

Background: The aim of this study was to assess cytological alterations of the squamous epithelium of the alveolar ridge mucosal surface under the acrylate prosthesis in T2D patients. Methods and Results: The subjects of interest were patients in whom the total acrylate prosthesis had been applied 3-5 years or more prior to the examination. The subjects were divided into two groups: 30 adult subjects with T2D (T2D group) and 30 adult subjects without T2D (control group). Both groups were over 49 years of age. Cytological smears were obtained by cytobrush from the mucosal surface of the gingival crest underlying the acrylate prosthesis. The following parameters were assessed: type of cells (basal, intermediate, superficial, superficial without nucleus, and parakeratotic), proportions of the types of cells, cytoplasmic diameter, nuclear diameter, and nucleus-to-cytoplasm ratio. An independent sample t-test was used to compare the two study groups. The percentage of superficial cells was significantly lower in the T2D group than in the control group (P=0.001). The T2D group tended to have a higher rate of parakeratotic cells (13.0±11.12 vs. 8.07±7.92, P=0.052). Superficial cells in the T2D group had a significantly lower mean cytoplasmic diameter than the control group (32.7947±8.61929 µ vs. 36.6383±4.32228 µ, P=0.03). Additionally, the nucleus-to-cytoplasm ratio of intermediate and superficial cells in the T2D group was significantly higher than in the control group (0.2407±0.07206 vs. 0.2000±o.03291, P=0.007 and 0.2573±0.06330 vs. 0.2280±0.03178, P=0.027). Conclusion: The results of our study show that total acrylate prostheses in diabetic patients are responsible for the disrupted maturation of squamous epithelial cells. This is reflected in smaller superficial cells, increased parakeratosis, and the higher nucleus-to-cytoplasm ratio of intermediate and superficial cells.

Cholecystokinin-expressing superficial tufted cells modulate odour representation in the olfactory bulb and olfactory behaviours.

In mammals, odour information within the olfactory bulb (OB) is processed by complex neural circuits before being ultimately represented in the action potential activity of mitral/tufted cells (M/Ts). Cholecystokinin-expressing (CCK+) superficial tufted cells (sTCs) are a subset of tufted cells that potentially contribute to olfactory processing in the OB by orchestrating M/T activity. However, the exact role of CCK+ sTCs in modulating odour processing and olfactory function in vivo is largely unknown. Here, we demonstrate that manipulating CCK+ sTCs can generate perception and induce place avoidance. Optogenetic activation/inactivation of CCK+ sTCs exerted strong but differing effects on spontaneous and odour-evoked M/T firing. Furthermore, inactivation of CCK+ sTCs disrupted M/T odour encoding and impaired olfactory detection and odour discrimination. These results establish the role of CCK+ sTCs in odour representation and olfactory behaviours. KEY POINTS: Mice could perceive the activity of CCK+ sTCs and show place avoidance to CCK+ sTC inactivation. Optical activation of CCK+ sTCs increased the percentage of cells with odour response but reduced the odour-evoked response in M/Ts in awake mice. Optical inactivation of CCK+ sTCs greatly decreased spontaneous firing and odour-evoked response in M/Ts. Inactivation of CCK+ sTCs impairs the odour decoding performance of M/Ts and disrupts odour detection and discrimination behaviours in mice. These results indicate that CCK+ sTCs participate in modulating the odour representation and maintaining normal olfactory-related behaviours.

Estrus detection on different thoroughbred crossbreed horses with vaginal smear and vaginal acidity method

Abstract Breeding manipulation season in racehorses is required to ensure the birth at the specific period (August-December), which will maximize the foal’s age when participating in their first race (July). Therefore, it is important to time the mating of mares using highly accurate estrus detection methods. This research was conducted to identify the differences data using vaginal smear and pH vagina of Thorougbred crossbreed mares in estrus and anestrus phases. Fifteen multiparous mares were conducted with a period of estrus of 19-22d. The result data showed significant differences (P&lt;0.05) in the vaginal cell distribution patterns at each phase of the estrus cycle. The superficial cells predominate (92.43±5.14%) in the estrus phase while significant parabasal cells (87.89±3.09%) predominate in the anestrus phase. The results also showed a significant difference (P&lt;0.05) in vaginal pH which was much more alkaline (8.71±0.25) in the estrus phase while more significantly acidic-normal (6.02±0.26) in the anestrus phase. However, the results showed no difference both in vaginal smear and pH vagina based on pedigree group. Finally, the pattern of superficial cells being more common and vaginal pH levels being higher during estrus may offer a new way to find out when an equine is in estrus.

Keratin 5 basal cells are temporally regulated developmental and tissue repair progenitors in bladder urothelium.

Urothelium forms a distensible yet impermeable barrier, senses and transduces stimuli, and defends the urinary tract from mechanical, chemical, and bacterial injuries. Biochemical and genetic labeling studies support the existence of one or more progenitor populations with the capacity to rapidly regenerate the urothelium following injury, but slow turnover, a low mitotic index, and inconsistent methodologies obscure progenitor identity. The progenitor properties of basal keratin 5 urothelial cells (K5-UCs) have been previously investigated, but those studies focused on embryonic or adult bladder urothelium. Urothelium undergoes desquamation and apoptosis after birth, which requires postnatal proliferation and restoration. Therefore, we mapped the fate of bladder K5-UCs across postnatal development/maturation and following administration of cyclophosphamide to measure homeostatic and reparative progenitor capacities, respectively. In vivo studies demonstrate that basal K5-UCs are age-restricted progenitors in neonates and juveniles, but not in adult mice. Neonatal K5-UCs retain a superior progenitor capacity in vitro, forming larger and more differentiated urothelial organoids than adult K5-UCs. Accordingly, K5-UC transcriptomes are temporally distinct, with enrichment of transcripts associated with cell proliferation and differentiation in neonates. Induction of urothelial proliferation is sufficient to restore adult K5-UC progenitor capacity. Our findings advance the understanding of urothelial progenitors and support a linear model of urothelial formation and regeneration, which may have significant impact on therapeutic development or tissue engineering strategies.NEW & NOTEWORTHY Fate mapping reveals an important linear relationship, whereby bladder basal urothelial cells give rise to intermediate and superficial cells in an age-restricted manner and contribute to tissue repair. Neonatal basal cells reprise their role as superior progenitors in vitro and display distinct transcriptional signatures, which suggest progenitor function is at least partially cell intrinsic. However, the urothelium progenitor niche cannot be overlooked, since FGF7 rescues adult basal cell progenitor function.

Abstract PR001: TRIM29 is required for basal bladder cancer invasive progression

Abstract TRIM29 (Tripartite Motif 29) also known ATDC, is an E3 ubiquitin ligase frequently upregulated in muscle-invasive bladder cancer (MIBC). We have previously shown TRIM29 global overexpression in a genetically engineered mouse model (GEMM) is sufficient to ` bladder cancer development and progression to MIBC. Other studies strongly suggest cytokeratin-5/14 (Krt5+/Krt14+) expressing basal-stem progenitor cells in the urothelium are the cells of origin for MIBC in mice. We therefore hypothesized that Trim29 expression in Krt5+ basal urothelial cells is required for development of MIBC in mice. To test this, we generated a GEMM harboring a Trim29 conditional KO allele and tamoxifen responsive Cre under control of the bovine Krt5promoter. Exposure of Krt5-CreERT2 Trim29flox/flox mice to tamoxifen resulted in activation of Cre and loss of Trim29 expression only in the Krt5+ basal urothelium. To induce bladder tumorigenesis, we utilized the urothelial specific carcinogen N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) in drinking water at 0.05%. After 6 months, tumor invasion was staged by a genitourinary pathologist based on depth of invasion: T1 (stromal invasive), T2 (muscle invasive) and T3 (deep muscle/peri-vesicular invasion). In agreement with previous studies, BBN exposure resulted in loss of Krt5-negative intermediate and superficial cells, and expansion of Krt5+/Krt14+ basal cells across the urothelium. In combination with a tamoxifen treatment this led to total Trim29 KO in the urothelium. All male Trim29 WT mice exposed to BBN developed Trim29+ bladder tumors, &amp;gt;50% of which demonstrated extensive progression to muscle-invasion (57% n=23). In contrast, only 16% of mice with Trim29 loss in Krt5+ basal cells developed MIBC (n=18). As an E3 ubiquitin ligase TRIM29 is known to target immune adaptor molecules, notably the stimulator of interferon genes (STING), for proteasomal degradation. We hypothesized TRIM29 targets STING during invasive progression and supports the establishment of an immune-privileged niche at the bladder cancer invasive front. We find TRIM29 expression in human bladder tissues is upregulated in the urothelium under stress and in MIBC tumors. In mice, BBN exposure upregulated Trim29 expression in the bladder urothelium which correlated with low urothelial Sting. In contrast, Krt5+ Trim29 loss resulted in significantly elevated Sting in the bladder urothelium. Multiplex immunofluorescence imaging and immunotyping of Trim29 KO mouse tissues revealed peritumoral enrichment of CD4+ and CD8+ T cells, CD206+ immunosuppressive macrophages, and CD4+FoxP3+ regulatory T cells. Taken together, these results suggest that TRIM29-mediated downregulation of STING in the bladder urothelium results in decreased immune surveillance at the tumor-stromal interface which expedites invasion and tumor burden. Citation Format: Alan Kelleher, Luke Broses, Yin Wang, Marian Henderson, Mark Day, Aaron Udager, Phillip Palmbos. TRIM29 is required for basal bladder cancer invasive progression [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2024 May 17-20; Charlotte, NC. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(10_Suppl):Abstract nr PR001.

Observations on the Germination of Spores of &lt;i&gt;Padina tetrastromatica&lt;/i&gt; Hauck

The germination of the spores of Padina tetrastromatica Hauck from Mohaballipuram, South of Madras is described in detail for the first time from India for any member of the Dictyotaceae. The tetrasporangium, instead of forming the usual tetraspores, liberates its entire contents which germinate immediately on settling down on the thallus of Padina tetrastromatica, Hauck. The liberated body is spherical in shape and on germination it produces at first a protuberance which on growing for some length as a tubular structure is cut off by a transverse wall from a spherical upper portion. The lower tubular portion, by a few divisions, forms a filamentous structure, the end cell of which produces a sort of a branched haptera on coming into contact with any foreign body. In the upper spherical portion cell divisions occur to result in a somewhat nodular structure. One of the superficial cells of this nodule grows out and by a transverse wall in this, an apical cell in cut off. By the activity of this apical cell new shoots of Padina are developed at first as cylindrical structures. By the growth and division in all cells, later a flattened structure and then a fan shaped thallus is formed characteristic of the Padina plant. In the sequence of divisions and the formation of the rhizoid like structures, Padina tetrastromatica Hauck shows some differences from the other members of the Dictyotaceae reported earlier from abroad.

Cellular profile of the skin and ocular surface in healthy shih-tzu dogs

Cytology involves analyzing individual cells and is crucial for diagnosing skin and ocular surface diseases. The skin acts as a barrier against various threats and communicates with the immune system via receptors. The conjunctiva also provides protection to the eyes. This study aimed to describe the cytological profiles of healthy Shih-Tzu dogs' skin and ocular surfaces, assessing how age impacts these profiles. Materials for cytological analysis were collected using a nylon microbrush and evaluated with the Schirmer test. Results showed lymphocytes were predominant in ocular cytology (RE - 47%, LE - 41.93%), while polymorphonuclear cells dominated in nasal fold (62%) and palmar pad cytologies (100%). Superficial cells were prevalent in all regions (RE 83.11%, LE 90.52%, palmar pads 100%, nasal fold 96.91%). Malassezia spp was detected in all areas. This research underscores the cellular variations across different anatomical sites and provides insights into normal cytological patterns in these dogs.