Mean Optical Density Research Articles

An in vitro Evaluation of Periodontal Ligament Cell Viability in Honey as a Transport Medium for an Avulsed Tooth

Abstract Background Dental avulsion is the most severe form of injury to a tooth, and its prognosis is majorly affected by the type of the storage medium prior to re-implantation. Honey is a readily available household substance with anti-inflammatory properties. However, there is paucity of data regarding its use as a transport medium for avulsed teeth. Therefore, this study aims to evaluate the viability of periodontal Ligament (PDL) cells stored in honey and other recommended transport media. Materials and Methods An in vitro study involving the culture of PDL cells, harvested from premolars freshly extracted for orthodontic reasons. The cells were exposed to Dulbecco’s Minimum Essential Medium (DMEM), honey, and Hanks Balanced Salt Solution (HBSS) at room temperature, and their viability was tested with the tetrazolium salt-based colorimetric (MTT) assay after 30 min, 2 h, and 24 h. The Optical Density (OD) of the cells were assessed with a spectrophotometer. The Mean Optical Density (MOD) of the cells stored in DMEM was compared with those of cells in other media at each interval with the independent t-test at P < 0.05 Results The MOD of cells stored in DMEM, honey, and HBSS over the 24-h period ranged between 0.32 ± 0.04 −0.31 ± 0.05; 0.30 ± 0.07−0.28 ± 0.07, and 0.23 ± 0.0−0.21 ± 0.01, respectively. The mean difference between the OD of cells stored in honey and those in DMEM at the end of the study period was 0.03 (t = 1.37; P = 0.18). Conclusion There was no difference in the viability of PDL cells stored in honey and DMEM over the study period.

Investigating the Skin Health Benefits of Rosa roxburghii, Punica granatum and Rose: A Randomized Single-Blind Controlled Clinical Trial.

Recent studies underscore the beneficial impacts of oral natural plant extracts on human skin health, though clinical evidence of their efficacy and safety is limited. This study evaluates the skin health effects of a novel oral supplement containing Rosa roxburghii, Punica granatum, and rose extracts (RPR) 0.70 healthy female participants were randomly assigned to either a control group or an RPR group, with the latter ingesting 20 mL of the RPR supplement daily on an empty stomach over 8 weeks. After 8 weeks, the RPR group exhibited significant enhancements (p < 0.001) in skin hydration, glossiness, elasticity, and skin tone, with increases of 69.02%, 30.48%, 25.97%, and 7.52%, respectively. Concurrently, decreases in skin firmness and melanin levels were observed at 21.17% (p = 0.007) and 25.06% (p < 0.001), respectively. Statistical analysis confirmed that these changes were significantly greater than those in the control group. Image analysis indicated no significant changes in mean optical density of hyperpigmented spots within the RPR group (p = 0.367), but a significant reduction in the areas of hyperpigmented spots, under-eye fine lines, and crow's feet by 41.50%, 37.55%, and 29.36%, respectively (p < 0.001), whereas no significant changes were detected in the control group. Importantly, no adverse effects were observed. These findings suggest that the combined intake of Rosa roxburghii, Punica granatum, and rose extracts can improve skin health, offering a promising natural alternative for dermatological care.

Biodegradation potentials of soil-borne fungal cultures grown on haloxyfop R methyl ester herbicide medium

The heterotrophic and haloxyfop-R methyl ester utilizing fungal counts associated with two top soil samples was determined. Five (5) fungal isolates; Aspergillus niger, Candida sp., Trichoderma sp., Fusarium sp., and Alternaria sp. Were cultured from the soil samples using serial dilution as well as pour plate techniques. These isolates were sub-cultured and screened for their ability to use haloxyfop-R methyl ester as sole source of metabolic energy as well as using the turbidimeteric method. Two of the screened fungal isolates which exhibited maximal optical density (OD) difference values; A. niger and Fusarium sp. Were selected and utilized for the subsequent degradation/shake flask experiment. Growth profile parameters which included; pH, OD, dissolved CO2 and weighed dry fungal biomass were determined during the growth profiling test at a 96 hour interval for 16 days using appropriate methods and equipment. Mean soil pH values were 5.08 ± 0.02 and 4.62 ± 0.02 for samples A and B. The mean total heterotrophic fungal count was 8.7×10³ cfu/g ± 1.5 for A and 1.6 × 10⁴ cfu/g ±1.0 for B. The mean pH value observed for A. niger in the course of the growth profile test ranged from 6.0 ± 8.7 to 7.3 ± 8.0 and 7.3 ± 5.8. The mean OD values observed for A. niger and Fusarium sp. In the course of the shake flask experiment varied from 0.66 ± 7.1 to 1.96 ± 4.5 and 1.62 ± 5.4 to1.99 ± 3.6 respectively. With reference to the detection of mean dissolved CO2 values, utilized as an indirect indicator of herbicide mineralization, the axenic cultured A. niger was the most effective amongst the growth profile fungal cultures.

Preliminary field evaluation of indirect ELISA test using the recombinant antigen rLicNTPDase-2 for serodiagnosis of canine leishmaniasis in Colombia

Preliminary field evaluation of indirect ELISA test using the recombinant antigen rLicNTPDase-2 for serodiagnosis of canine leishmaniasis in Colombia

A Histological Study on the Effect of Different Methods of Mobile Phone Exposure on the Hippocampus in Adult Male Albino Rat

Abstract Introduction Using mobile phones has dramatically increased in the last few years. Mobile phones are carried mainly near the head during talking. Electromagnetic fields (EMF) emitted from cell phones may exert detrimental effects on human organs especially hippocampus which is an important organ concerned with memory and cognitive function. Aim To study the effect of different methods of mobile phone EMF exposure on the structure of hippocampus in adult male rats. Materials and Methods 28 adult male albino Wistar rats were equally divided into four groups: Group I (Control): were left without any interference. Group II (silent non-vibrating): exposed to continuous 900-1800 megahertz cell phone- emitted EMF for 60 minutes/day, for four weeks. Mobile phones were kept in silent, non-vibrating mode. Group III (silent vibrating): exposed to EMF as in group II, but with mobile phones kept in vibrating mode. Group IV (ringing non- vibrating): exposed to EMF as in group II, but with ringtone turned on and adjusted at 80 decibels. After four weeks of exposure, hippocampi were harvested form all rats. They were subjected to hematoxylin and eosin, toluidine blue, Caspase-3, and glial fibrillary acidic protein (GFAP) immunohistochemical stain as well as transmission electron microscopic examination. Morphometric and statistical studies were also done. Results pyramidal cells in CA3 area of Group IV (ringing non-vibrating) showed condensed chromatin clumps, distorted mitochondria, and clumped neurofilaments. A significant decrease in the mean thickness of the pyramidal cell layer of the CA3 area was noticed compared to other groups. While a significant increase in the mean number of degenerated pyramidal cells, number of caspase-3 positive cells as well as number of astrocytes was noticed in CA3 area of group IV compared to other groups. A significant increase in the mean optical density of caspase-3 and GFAP was also noticed in CA3 of group IV compared to other groups. Conclusions Exposure to EMF caused significant changes in the structure of hippocampus especially CA3 area. There was no significant difference between silent and vibrating mode. The worst changes were detected in ringing mode.

The Macular Pigment Optical Density (MPOD) Decrease in Chinese Primary Angle Closure Glaucoma Using the One-Wavelength Reflectometry Method

Purpose The objective of this study was to observe the macular pigment optical density (MPOD) and the relationship between MPOD and retinal thickness in Chinese primary angle-closure glaucoma (PACG) patients by the one-wavelength reflectometry method. Methods This study was a prospective comparative observational study, including 39 eyes from 39 PACG patients (15 men and 24 women, mean age 61.89 ± 12.30) and 41 eyes from 41 controls (20 men and 21 women, mean age 63.24 ± 14.02). We measured the MPOD 7-degree area by the one-wavelength reflectometry method and analyzed both the max and mean optical density (OD). The central retinal thickness (CRT) and the total thickness of the macular ganglion cell layer (GCL), and inner plexiform layer (IPL)were measured by spectral-domain-optical coherence tomography (SD-OCT). Statistical methods such as Shapiro–Wilk test, Fisher’s exact test, chi-square test, two independent samples test and Spearman’s correlation coefficient were used to observe the differences in the MPOD between normal subjects and PACG patients and the correlation between the MPOD and retinal thickness. Results The max optical density (Max OD) (PACG group: 0.302 ± 0.067d.u, control group: 0.372 ± 0.059d.u., p < .001) and mean optical density (Mean OD) (PACG group: 0.124 ± 0.035d.u., control group: 0.141 ± 0.028d.u., p < 0.05) were significantly reduced in PACG patients compared with control subjects. Significant decreases in GCL + IPL thickness (PACG group: 74.71 ± 39.56 μm, control group:113.61 ± 8.14 μm, p < 0.001) and CRT (PACG group: 254.49 ± 41.47 μm, control group:329.10 ± 18.57 μm, p < 0.001) were also observed in PACG eyes. There was no statistically significant correlation between the MPOD and GCL + IPL thickness (p = .639, p = .828). Conclusions MPOD was significantly lower in Chinese PACG patients than in the control group, potentially due to thinning of the GCL + IPL thickness. This study provides insights for the pathophysiology, assessment of PACG and potential guidance for lifestyle modifications.

Effects of dexmedetomidine on depression-like behaviour in chronic restraint stress mice: Involvement of specific brain regions

Effects of dexmedetomidine on depression-like behaviour in chronic restraint stress mice: Involvement of specific brain regions

Expression of angiotensin converting enzyme 2 in patients with oral squamous cell carcinoma

Expression of angiotensin converting enzyme 2 in patients with oral squamous cell carcinoma

Abstract PO4-26-01: Computational pathology: Revolutionizing diagnostics and clearing the way for precision medicine

Abstract While targeted cancer therapies often rely on subjective and semi-quantitative visual assessment of protein biomarkers by pathologists through immunohistochemically stained tissue, the transformative force of computational pathology is reshaping healthcare by unleashing unprecedented diagnostic accuracy and unlocking personalized treatments. In recent years, we have established a large integrated computational pathology unit to foster collaboration between interdisciplinary teams of computer scientists, pathologists, molecular biologists, and data scientists. This integration of cutting-edge technologies enabled us to develop a computational pathology approach called Quantitative Continuous Scoring (QCS). QCS deploys the power of Deep Learning (DL) to provide objective and continuous expression data of biomarkers in digitized IHC whole slide images (WSI), particularly of proteins expressed at low levels. While manual scoring of IHC WSIs is limited by subjectivity and semi-quantitative assessment of protein expression, QCS overcomes these limitations with an unprecedented accuracy. QCS utilizes two DL-based algorithms, which we developed fully supervised by using pathologist input as the reference standard. These algorithms identify invasive tumour areas and segment each tumour cell across the WSI into cell nuclei, cytoplasm and membrane. Based on an accurate subcellular segmentation, we can compute biomarker expression, on a continuous scale, as mean Optical Density (OD) in each subcellular compartment based on the Hue-Saturation-Density (HSD) model. Therefore, this approach enables precise detection of the low biomarker expression range with single-cell resolution. Importantly, it also allows the computation of the spatial distribution of tumour cells across the WSI. We have successfully used QCS to drive the selection of antibody clones for IHC assays and to delineate the mode of action and PK/PD mechanisms. Of note, the combination of assessing continuous target expression and capturing the spatial distribution of tumor cells has provided surrogate markers to predict potential bystander activity of antibody drug conjugates (ADCs). This approach outperformed traditional pathologist scoring in identifying patient populations having maximum treatment benefit through retrospective analysis of multiple clinical trials. At present, all computational pathology approaches are developed based on conventional IHC assays that have been optimized for manual scoring. Importantly, we draw a vision in which the assay serves as a critical catalyst for unleashing the full potential of computational pathology by providing high-quality, standardized data inputs. We suggest an approach utilizing orthogonal methods as a reference standard to develop highly sensitive IHC assays, capable of detecting even subtle molecular and cellular changes with precision and exhibiting exceptional specificity for accurately identifying and distinguishing target biomarkers from background noise. In summary, we here describe and discuss a computational pathology-based approach for precise biomarker quantification and superior patient selection with broad applicability and the potential to transform the very fabric of how we diagnose and treat cancer. Citation Format: Mark Gustavson, Markus Schick, Ansh Kapil, Anatoliy Shumilov, Carl Barrett, Hadassah Sade. Computational pathology: Revolutionizing diagnostics and clearing the way for precision medicine [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO4-26-01.

Small ubiquitin-like modifier-tag and modified protein purification significantly increase the quality and quantity of recombinant African swine fever virus p30 protein.

African swine fever (ASF) is a highly virulent and contagious viral disease caused by the ASF virus (ASFV). It has a significant impact on swine production throughout the world, while existing vaccines and specific treatments remain ineffective. ASFV p30 is a potent antigenic protein that induces protective antibodies immediately after infection; however, most recombinant p30 is insoluble. This study aimed to improve the solubility, yield, and purity of recombinant p30 by tagging it with a small ubiquitin-like modifier (SUMO) and modifying the protein purification process. SUMO fused with ASFV p30 (SUMO-p30) and p30 alone were cloned and expressed in Escherichia coli. SUMO-p30 and p30 solubility and expression levels were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Protein purification was modified by combining ammonium sulfate precipitation method with affinity chromatography. In addition, large-scale production of all versions of p30 were compared using SDS-PAGE and western blotting, and the purified p30 was used to develop the indirect enzyme-linked immunosorbent assay (ELISA). The solubility and expression levels of SUMO-p30 were dramatically enhanced compared with that of p30. Modification of the purification process significantly increased purified and soluble SUMO-p30 and p30 yields by 6.59 and 1.02 μg/mL, respectively. Large-scale production confirmed that this procedure increased the quantity of recombinant p30 while maintaining protein purity and immunogenicity. The p30-based indirect ELISA was able to discriminate between positive and negative serum samples with statistically significant differences in mean optical density 450 values (p < 0.001). This study demonstrates the enhancement of solubility, purity, and yield of ASFV p30 expressed in E.coli by SUMO fusion tagging and combining ammonium sulfate precipitation with affinity chromatography for protein purification. These positive effects were sustained in large-scale production. Cleavage and removal of hexahistidine-SUMO tag from the fusion protein by protease may not be suitable when handling a large amount of the protein. However, the SUMO-fused p30 retained strong immunoreactivity to convalescent swine serum, indicating its application in immunization and diagnostic purposes. The expression and purification procedures in this study could be applied to increase solubility, quality, and quantity of other recombinant proteins as well.

Determination of the antibiofilm property of aqueous extract of the Amorphophallus paeoniifolius on some early and late colonizers in an artificially synthesized dental biofilm - An in vitro study.

Mechanical therapy along with adjunctive therapy, using agents like chlorhexidine digluconate mouthwash helps to disrupt the plaque biofilm. Recently, herbs with medicinal value have been tested for their antimicrobial properties. The present study was designed to assess the anti-biofilm activity of Amorphophallus paeoniifolius against some periodontal pathogens in an artificially synthesized dental biofilm. The aqueous extract of A. paeoniifolius was constituted and its minimum inhibitory concentration (MIC) against standard strains of some periodontal pathogens was determined. A total of 21 biofilm samples were synthesized on extracted teeth and microtiter plates, and these were divided into two groups of 10 samples each. One group was treated with the predetermined MIC values of A. paeoniifolius, while the other group was treated with chlorhexidine. The anti-biofilm activity of both compounds was assessed by calculating colony-forming units (CFUs) for the extracted teeth and optical density (OD) values for the microtiter plates. The mean CFU at baseline was 55,000/μl while posttreatment with chlorhexidine digluconate and aqueous extract of A. paeoniifolius was 23,280 ± 5274.00 and 28,560 ± 4509.545/μl, respectively. The mean OD value (at 595 nm) posttreatment with chlorhexidine digluconate was 0.9876 ± 0.49179 and A. paeoniifolius was 1.4990 ± 0.37851. Results indicate that the aqueous extract of A. paeoniifolius showed an inhibitory effect on biofilm obtained on microtiter plates and the one constituted on extracted teeth. Anti-biofilm activity of aqueous extract of A. paeoniifolius was appreciable and also comparable to that of chlorhexidine digluconate, both on extracted teeth and microtiter plates.

Photoactivated disinfection procedure for denture stomatitis in diabetic rats.

To study the efficacy of PADTM Plus-based photoactivated disinfection (PAD) for treating denture stomatitis (DS) in diabetic rats by establishing a diabetic rat DS model. The diabetic rat DS model was developed by randomly selecting 2-month-old male Sprague-Dawley rats and dividing them into four groups. The palate and denture surfaces of rats in the PAD groups were incubated with 1 mg/mL toluidine blue O for 1 min each, followed by a 1-min exposure to 750-mW light-emitting diode light. The PAD-1 group received one radiation treatment, and the PAD-2 group received three radiation treatments over 5 days with a 1-day interval. The nystatin (NYS) group received treatment for 5 days with a suspension of NYS of 100,000 IU. The infection group did not receive any treatment. In each group, assessments included an inflammation score of the palate, tests for fungal load, histological evaluation, and immunohistochemical detection of interleukin-17 (IL-17) and tumor necrosis factor (TNF-α) conducted 1 and 7 days following the conclusion of treatment. One day after treatment, the fungal load on the palate and dentures, as well as the mean optical density values of IL-17 and TNF-α, were found to be greater in the infection group than in the other three treatment groups (P < 0.05). On the 7th day after treatment, these values were significantly higher in the infection group than in the PAD-2 and NYS groups (P < 0.05). Importantly, there were no differences between the infection and PAD-1 groups nor between the PAD-2 and NYS groups (P > 0.05). PAD effectively reduced the fungal load and the expressions of IL-17 and TNF-α in the palate and denture of diabetic DS rats. The efficacy of multiple-light treatments was superior to that of single-light treatments and similar to that of NYS.

Bacillus subtilis alleviates excessive apoptosis of intestinal epithelial cells in intrauterine growth restriction suckling piglets via the members of Bcl-2 and caspase families.

The intestine is a barrier resisting various stress responses. Intrauterine growth restriction (IUGR) can cause damage to the intestinal barrier via destroying the balance of intestinal epithelial cells' proliferation and apoptosis. Bacillus subtilis has been reported to regulate intestinal epithelial cells' proliferation and apoptosis. Thus, the purpose of this study was to determine if B. subtilis could regulate intestinal epithelial cells' proliferation and apoptosis in intrauterine growth restriction suckling piglets. Compared with the normal birth weight group, the IUGR group showed greater mean optical density values of Ki-67-positive cells in the ileal crypt (P < 0.05). IUGR resulted in higher ability of proliferation and apoptosis of intestinal epithelial cells, by upregulation of the messenger RNA (mRNA) or proteins expression of leucine rich repeat containing G protein coupled receptor 5, Caspase-3, Caspase-7, β-catenin, cyclinD1, B-cell lymphoma-2 associated agonist of cell death, and BCL2 associated X (P < 0.05), and downregulation of the mRNA or protein expression of B-cell lymphoma-2 and B-cell lymphoma-2-like 1 (P < 0.05). However, B. subtilis supplementation decreased the mRNA or proteins expression of leucine rich repeat containing G protein coupled receptor 5, SPARC related modular calcium binding 2, tumor necrosis factor receptor superfamily member 19, cyclinD1, Caspase-7, β-catenin, B-cell lymphoma-2 associated agonist of cell death, and Caspase-3 (P < 0.05), and increased the mRNA expression of B-cell lymphoma-2 (P < 0.05). IUGR led to excessive apoptosis of intestinal epithelial cells, which induced compensatory proliferation. However, B. subtilis treatment prevented intestinal epithelial cells of IUGR suckling piglets from excessive apoptosis. © 2024 Society of Chemical Industry.

Improving Access to Anti-HDV Testing: Development and Validation of an Affordable In-House ELISA Assay.

In certain low-income nations, the hepatitis Delta virus and hepatitis B virus (HBV) pose a serious medical burden, where the prevalence of hepatitis B surface antigen (HBsAg) is greater than 8%. Especially in rural places, irregular diagnostic exams are the main restriction and reason for underestimation. Utilizing serum samples from a Pakistani isolate, an internal ELISA for the quick identification of anti-HDV was created, and the effectiveness of the test was compared to a commercial diagnostic kit. HDV-positive serum samples were collected, and a highly antigenic domain of HDAg antigen was derived from them. This antigenic HDAg was expressed in a bacterial expression system, purified by Ni-chromatography, and confirmed by SDS-PAGE and Western blot analysis. The purified antigen was utilized to develop an in-house ELISA assay for anti-HDV antibody detection of the patient's serum samples at very low cost. Purified antigens and positive and negative controls can detect anti-HDV (antibodies) in ELISA plates. The in-house developed kit's efficiency was compared with that of a commercial kit (Witech Inc., USA) by the mean optical density values of both kits. No significant difference was observed (a P value of 0.576) by applying statistical analysis. The newly developed in-house ELISA is equally efficient compared to commercial kits, and these may be useful in regular diagnostic laboratories, especially for analyzing local isolates.

Evaluation of Cytotoxic Effects of Purified Mercury in Human Gingival Fibroblasts-In vitro Study.

Since the introduction of amalgam for tooth fillings, there have been concerns that mercury toxicity could pose unacceptable health risks. Rasa shastra is an ancient medical discipline that focuses on the utilization of metals and minerals for the treatment of diseases. Nevertheless, these minerals cannot be directly administered to the human body in their natural state due to their potential adverse effects. Hence, for medicinal purposes, these metals and minerals need to undergo purification (Shodhana) to eliminate impurities and modify their physical, chemical, and biological characteristics. Human gingival fibroblasts (HGF) were exposed to commercially available mercury (CA-Hg) and ayurvedically purified mercury (AP-Hg) at concentrations of 6.25 μM, 12.5 μM, 25 μM and 50 μM. The unexposed HGF cultured in basal media was considered a control. All the samples were cultured for 24 hours and 48 hours, and the cytotoxicity was analyzed by MTT assay. Cell viability between the control and experimental groups varied at 24 hours, however, the results were not statistically significant (p>0.05). At 48 hours, cell viability was higher in the AP-Hg group as compared to the CA-Hg group at the concentration of 6.25 μM, and the difference was statistically significant (p<0.05). The cell proliferation assay results demonstrated a statistically significant difference in the mean optical density values (p<0.05) between CA-Hg and AP-Hg at 12.50 μM, 25 μM, and 50, μM concentrations observed at 24 hours. At 48 hours, a statistically significant difference in the mean OD values (p<0.05) between CA-Hg and AP-Hg at all four concentrations was observed. Conclusion: AP-Hg at a concentration of 6.25 μM demonstrated higher cell viability at 48 hours. Further, the cell proliferation rate was also higher for AP-Hg at all concentrations at 24 and 48 hours. These results indicated a less cytotoxic effect of AP-Hg than CA-Hg in HGF and hence could be employed for dental amalgam preparations.

EXAMINATION OF GALACTOMANNAN LEVELS IN INTRAOCULAR FLUID TO ASSIST THE DIAGNOSIS OF ASPERGILLUS ENDOPHTHALMITIS.

To evaluate the utility of galactomannan testing of intraocular fluid in the diagnosis of Aspergillus endophthalmitis (AE). This retrospective study enrolled three groups of patients, including those with 17 eyes with AE; 20 eyes with intraocular infection of bacteria, viruses, or other fungi; and 19 eyes with cataract. Intraocular fluid from all these patients was collected for galactomannan testing. In addition, the receiver operating characteristic curves and diagnostic significance were analyzed. The mean optical density index (ODI) of galactomannan was 5.77 ± 1.73 in the AE group, which was significantly higher than that in the non- Aspergillus intraocular infection group (0.19 ± 0.11, P < 0.001) and the negative control group (0.29 ± 0.27, P < 0.001). The area under the receiver operating characteristic curve (area under the curve) was 1.00 (95% confidence interval, 1.00-1.00; P < 0.001) in the AE group and the other two groups. At a cutoff optical density index of 1.88, the sensitivity and specificity were 100.0% and 100.0%, respectively, and the Youden index reached its highest value of 1.00. Galactomannan testing of intraocular fluid indicated good sensitivity and specificity for the diagnosis of AE, thereby promising a rapid diagnostic modality for AE.

Comparative evaluation of the antibacterial efficacy of two experimental calcium silicate-based intracanal medicaments: An in-vitro study.

Success of endodontic treatment relies on minimizing microbial load by chemo-mechanical preparation and intra-canal medication(ICM). Calcium hydroxide based ICMs have known disadvantages. Calcium silicate-based cements(CSC) exhibit antibacterial activity, thus promoting researchers to experiment with their formulations to use them as ICMs. Evaluation and comparison of the antimicrobial efficacy of two experimental CSC (MTA & Biodentine + 2%chlorhexidine) and Bio-C Temp against E.faecalis. Test materials were divided into four groups namely Group1-Bio-C Temp, Group2-UltraCAL XS, Group3-Biodentine+2%CHX and Group4-MTA+2%CHX. Direct contact test was done by placing a standardized suspension of E.faecalis on test materials and bacterial growth was assessed spectrophotometrically using ELISA at one, three and seven days. Data was analysed using one-way ANOVA, Tukey's multiple post hoc test and paired-t test. Results: Intragroup comparison revealed decreased mean optical density(OD) in groups 1, 2, and 4; no significant difference in group 3. Intergroup comparison showed statistical differences in mean OD values between groups (3 and 4); groups (1 and 2) at days one(p-0.018) and three(p-0.035), but no difference individually. Group 4 showed the highest antimicrobial efficacy on day seven. MTA+2%CHX & Biodentine+2%CHX showed better antimicrobial efficacy and hence could be used as potential ICMs.

Photoperiod-regulated mitophagy in the germ cells of Brandt's voles (Lasiopodomys brandtii).

Photoperiod is a pivotal factor in affecting testicular function and spermatogenesis in seasonal-breeding animals. Mitophagy is essential for spermatogenesis, but its association with seasonal photoperiods has not been studied extensively. To explore this, we exposed male Brandt's voles (Lasiopodomys brandtii) to long-photoperiod (LP, 16 h/day) and short-photoperiod (SP, 8 h/day) conditions from their embryonic stages. Our results indicated that testis weight, volume, and relative testes weight were all significantly increased in LP compared to SP. Additionally, blood testosterone levels were markedly higher in LP than SP. Histological examination revealed that seminiferous diameter and epithelium thickness were greater in LP, with an increased abundance of germ cell types and cell numbers compared to SP. RT-qPCR analysis showed that mitophagy-promoting genes, such as Pink1, Prkn, Tomm7, Mnf2, Lc3, Optn, Gabarap, and Nbr1 were all upregulated in LP. Fluorescence in situ hybridization indicated that Pink1 expression was present in spermatogonia in SP, while in LP, Pink1 expression extended to almost all germ cell types with significantly higher mean optical density. Prkn expression was found in all germ cell types in both LP and SP, with a significantly higher mean optical density of 10-week-old LP males. Transmission electron microscopy showed normal mitochondrial morphology with clear membranes in SP, while the LP group had reduced cristae in mitochondria and damaged mitochondria undergoing autophagy. This study suggests that mitophagy may be involved in the photoperiodic spermatogenesis in Brandt's voles, providing insights into the role of photoperiod in seasonal reproduction in wild animals.

Differential contributions of the C5b-9 and C5a/C5aR pathways to microvascular and macrovascular thrombosis in complement-mediated thrombotic microangiopathy patients

Differential contributions of the C5b-9 and C5a/C5aR pathways to microvascular and macrovascular thrombosis in complement-mediated thrombotic microangiopathy patients

CYTOTOXICITY TEST OF BOVINE DEMINERALIZED BONE MATRIX ON HUMAN MESENCHYMAL STEM CELLS USING THE MTT ASSAY METHOD

The use of bone grafts in Indonesia continues to increase each year. Although Autograft is considered the gold standard in bone grafting, its use is often confronted with various challenges, similar to allograft. To address this issue, Bovine Demineralized Bone Matrix (DBM) can be considered as a substitute for bone grafts with the advantages of unlimited availability and more affordable costs. Currently, the Tissue Bank of Dr. Soetomo Hospital is developing bovine DBM, although there is no research yet on its potential toxicity.This study aims to evaluate whether bovine DBM has cytotoxic effects on human mesenchymal stem cells. In this experimental study, a total of 48 samples were involved, including a control group and two treatment groups (50% and 25%), each consisting of 16 samples. Mesenchymal stem cells were cultured and then treated with the addition of 50% and 25% DBM. Subsequently, cell viability was measured using the MTT Assay method.The collected data were processed by conducting normality and homogeneity tests and then analyzed using comparative tests with an independent t-test. The criteria for declaring cell toxicity were set at a viability of not less than 60% compared to the control group.The results of the MTT assay measurements showed that the mean Optical Density (OD) in the control group was 0.656 ± 0.021 (range 0.620-0.696), while in the treatment groups, it was 0.565 ± 0.022 (range 0.529-0.614) and 0.520 ± 0.022 (range 0.461-0.552), respectively. Statistically, the differences in OD between the control group and both treatment groups (50% and 25%) were significant (p&lt;0.05). The average cell viability in both treatment groups was found to be more than 60%, indicating that Bovine Demineralized Bone Matrix is not toxic to human mesenchymal stem cells.