Relative Optical Density Research Articles

The impact of human albumin on the activity of some anti-staphylococcal agents in an in vitro pharmacokinetics / pharmacodynamics model

The emergence of anti-staphylococcal drug resistance has significantly increased, thereby making it difficult to control the life-threatening staphylococcal infections. A validated two-compartment in vitro pharmacokinetics / pharmacody¬namics (PK/PD) model has been used in order to estimate the efficacies of some anti-staphylococcal drugs - namely, vancomycin (maximum concentration or Cmax of 3 and 5 mg/L), teicoplanin (Cmax of 5 and 10 mg/L), and minocycline (Cmax of 2 and 4 mg/L) – against a mixed staphylococcal infection (S. aureus ATCC 25923 and S. epidermidis ATCC 12228), with or without human albumin (2%). The PK profile for each drug was simulated as time-concentration de¬pending on the drug’s half-life. The minimum inhibitory concentration (MIC), the relative optical density for bacterial growth, and the exposure / effect relationship (fAUC0-24/MIC) have also been assessed in this study. Our results revealed that minocycline has the best efficacy over other antibiotics against the assessed isolates (single or mixed). Moreover, the addition of albumin exhibited a negative effect on vancomycin and a positive effect on teicoplanin in both the single and the mixed infections. In conclusion, albumin drew a different antibiotic scenario in response to different pathogens.

Assessment of antifungal drugs’ activity against some Candida albicans isolates in the presence or absence of human albumin: a study employing an in vitro pharmacokinetics / pharmacodynamics model

Invasive candidiasis associated with the dissemination of endogenous Candida species is a fatal condition linked to high rates of morbidity and mortality. Progressive drug resistance necessitates the need for prompt and effective therapy. Therefore, choosing a specific and effective treatment is crucial. A two-compartment in vitro pharmacokinetics (PK) / pharmacodynamics (PD) model has been used for this purpose, and the PD behaviours of amphotericin B (AMB; at 2.5 and 5 mg/L), voriconazole (VOR; at 1.5 and 3 mg/L), and itraconazole (ITR; at 1.5 and 3 mg/L) were assessed against two Candida albicans isolates (a sensitive and resistant one; ATCC-90028 and ATCC-10231, re¬spectively) with or without the addition of human albumin (2%). PK were simulated as time-concentration profiles, while the PD susceptibility of all drug doses has been assessed through the minimum inhibitory concentration (MIC), the relative optical density of fungal growth, and the exposure - effect relationship (fAUC0–24/MIC). A fungicidal activity without the presence of albumin was seen against both isolates of C. albicans at the highest dose of VOR, while the addition of albumin potentiated the efficacies of AMB and of VOR against both isolates, with no effect for ITR. Finally, human albumin exerted a variable and dose-dependent effect on the activities of some antifungal agents.

Vocal and tongue exercise in early to mid-stage Parkinson disease using the Pink1-/- rat

Vocal and swallowing deficits are common in Parkinson disease (PD). Because these impairments are resistant to dopamine replacement therapies, vocal and lingual exercise are the primary treatment, but not all individuals respond to exercise and neural mechanisms of treatment response are unclear. To explore putative mechanisms, we used the progressive Pink1-/- rat model of early to mid-stage PD and employed vocal and lingual exercises at 6- and 10-months of age in male Pink1-/- and wild type (WT) rats. We hypothesized that vocal and lingual exercise would improve vocal and tongue use dynamics and increase serotonin (5HT) immunoreactivity in related brainstem nuclei. Rats were tested at baseline and after 8 weeks of exercise or sham exercise. At early-stage PD (6 months), vocal exercise resulted in increased call complexity, but did not change intensity, while at mid-stage (10 months), vocal exercise no longer influenced vocalization complexity. Lingual exercise increased tongue force generation and reduced relative optical density of 5HT in the hypoglossal nucleus at both time points. The effects of vocal and lingual exercise at these time points are less robust than in prodromal stages observed in previous work, suggesting that early exercise interventions may yield greater benefit. Future work targeting optimization of exercise at later time points may facilitate clinical translation.

HISTOCHEMICAL AND IMMUNOHISTOCHEMICAL FEATURES OF DIFFERENTIATED TROPHOBLAST IN CHORIONIC VILLI OF THE PLACENTA IN PRETERM LABOR

The importance of studying the histochemical and immunohistochemical features of the diff erentiated trophoblast in chorionic villi of the placenta in preterm labor is determined by the threatening nature of this condition for both mother and fetus, as well as its high prevalence and serious consequences for the health of both. The understanding of the damage to the trophoblast of chorionic villi in the placenta can be expanded by histochemical and immunohistochemical methods, which allow the assessment of the concentration of specifi c marker molecules in one way or another.Objective. To determine certain histochemical and immunohistochemical characteristics of proteins in the trophoblast of theintermediate and terminal chorionic villi of the placenta in preterm labor.Material and Methods. The obtained material (30 placentas from preterm deliveries and 30 placentas from normalpregnancies) was fi xed for 20-24 hours in 10 % neutral formalin solution buff ered in Lilly’s phosphate buff er. After tissue removal, the placental tissue was dehydrated in an ascending ethanol series and embedded in paraffi n at a temperature of approximately 58 °C. Serial histologic sections were cut at 5.0 μm thickness using an MS-2 sliding microtome. After deparaffi nization, histological sections were stained with hematoxylin and eosin, histochemical methods for total protein with bromophenol blue according to Bonhéme, and immunohistochemical techniques according to the manufacturer’s protocols (Dako, Denmark). In particular, immunohistochemical reactions were performed with monoclonal antibodies against trophoblast hormone- placental lactogen and placental alkaline phosphatase. Visualization of primary antibodies was performed using the Dako polymer visualization system with diaminobenzidine as chromogen (resulting in brown staining of the sites of studied antigens). In addition to the descriptive method of histopathologic research, computer morphometry of digital microphotographs of histologic sections was performed using a Delta Optical Evolution 100 microscope and an Olympus SP550UZ digital camera. Digital copies of the images were processed using a legitimate copy of the ImageJ v1.52f computer program developed for histometric studies (National Institutes of Health, USA). Specifi cally, the evaluation of staining intensity (optical density) was performed on digital microphotographs using the method of computer microdensitometry. For this purpose, a microprobe method was used to obtain a computer brightness value in an 8-bit analysis system with 256 gray levels – from black (0) to white (255).The obtained values were then transformed into relative optical density values (r.OD) by logarithmic transformation (naturallogarithm method). The relative optical density value ranges from 0 (absolute transparency of the object) to 1 (absolute opacity of the object). The obtained digital data were processed using statistical analysis methods. A legitimate copy of the statistical analysis computer program PAST v4.14 was used, with a preliminary check for normal distribution using the Shapiro- Wilk test. Since, according to this test, the hypothesis of normal distribution was not rejected for the statistical samples studied (at p=0.05), parametric methods of statistical analysis were applied: calculation of the mean and its standard error, Student’s t-test (two-tailed, unpaired). In addition, the non-parametric Mann- Whitney test was used for the reliability of the conclusions. The research was conducted as part of the scientifi c research project «Preservation and Restoration of Reproductive Health in Women and Girls with Obstetric and Gynecological Pathology» at the Department of Obstetrics and Gynecology of Bukovinian State Medical University (state registration number 0121U110020, duration 2021-2025).Results. Microscopic examination of histological sections stained with hematoxylin and eosin did not reveal any diff erencesin the structure of chorionic villus trophoblast in the placenta of preterm labor compared to normal pregnancy. However,histochemical and immunohistochemical methods of investigation revealed a number of features in the trophoblast of chorionic villi, where fundamental events related to substance exchange occur – intermediate immature, intermediate mature, terminal villi, including terminal «specialized villi».Conclusions. According to the obtained histochemical and immunohistochemical data, in preterm labor, compared to normalpregnancy, no changes are observed in the trophoblast of intermediate immature villi, while in intermediate mature and terminal villi, there is a decrease in histochemical staining for total protein and immunohistochemical staining for specifi c trophoblast proteins – placental lactogen hormone and placental alkaline phosphatase enzyme.

Amoxicillin/clavulanate in combination with rifampicin/clarithromycin is bactericidal against Mycobacterium ulcerans.

Buruli ulcer (BU) is a skin neglected tropical disease (NTD) caused by Mycobacterium ulcerans. WHO-recommended treatment requires 8-weeks of daily rifampicin (RIF) and clarithromycin (CLA) with wound care. Treatment compliance may be challenging due to socioeconomic determinants. Previous minimum Inhibitory Concentration and checkerboard assays showed that amoxicillin/clavulanate (AMX/CLV) combined with RIF+CLA were synergistic against M. ulcerans. However, in vitro time kill assays (TKA) are a better approach to understand the antimicrobial activity of a drug over time. Colony forming units (CFU) enumeration is the in vitro reference method to measure bacterial load, although this is a time-consuming method due to the slow growth of M. ulcerans. The aim of this study was to assess the in vitro activity of RIF, CLA and AMX/CLV combinations against M. ulcerans clinical isolates by TKA, while comparing four methodologies: CFU enumeration, luminescence by relative light unit (RLU) and optical density (at 600 nm) measurements, and 16S rRNA/IS2404 genes quantification. TKA of RIF, CLA and AMX/CLV alone and in combination were performed against different M. ulcerans clinical isolates. Bacterial loads were quantified with different methodologies after 1, 3, 7, 10, 14, 21 and 28 days of treatment. RIF+AMX/CLV and the triple RIF+CLA+AMX/CLV combinations were bactericidal and more effective in vitro than the currently used RIF+CLA combination to treat BU. All methodologies except IS2404 quantitative PCR provided similar results with a good correlation with CFU enumeration. Measuring luminescence (RLU) was the most cost-effective methodology to quantify M. ulcerans bacterial loads in in vitro TKA. Our study suggests that alternative and faster TKA methodologies can be used in BU research instead of the cumbersome CFU quantification method. These results provide an in vitro microbiological support to of the BLMs4BU clinical trial (NCT05169554, PACTR202209521256638) to shorten BU treatment.

O226: OTD is cytotoxic to A-375 melanoma cells without compromising fibroblast migratory wound healing function – a potential role in perioperative melanoma surgery

Abstract Introduction Melanoma incidence is increasing with 325,000 new cases reported in 2020 worldwide and 57,000 deaths. High recurrence rates are associated with prolonged inflammation and subsequent immunosuppression post-operatively. Surgical adjuvants may play a role in mitigating these adverse immune responses while increasing the cytotoxicity of cancer cells. Our previous research demonstrated anti-inflammatory effects of 1,4,5-oxathiazinane-4,4-dioxide (OTD) on Human Dermal Fibroblasts (HDFs) by the reduction of cytokine production. This study evaluates its cytotoxicity on melanoma cells. Methods OTD treatments of 0.25mM, 0.5mM, 0.75mM, 1mM and 1.25mM were applied to A-375 primary melanoma cells in triplicate for 24h and an MTT assay assessed cell viability. The absorbance was measured at 570nm wavelength. Cell viability was expressed as a percentage of the control. Results were compared using one-way analysis of variance (ANOVA) with GraphPad Prism software. Results Application of OTD treatments demonstrated a cytotoxic effect on melanoma cells at all concentrations. Relative optical density (OD) value was compared for each concentration against the control. A significant difference in cytotoxicity, relative to untreated control, was observed for each concentration; 56.8% at 0.25mM OTD (p*** = 0.001) and 68 to 71.6% at 0.5mM to 1.25mM OTD (p****<0.001). Conclusion OTD induces cytotoxicity in primary melanoma cells at all concentrations. Whereas our previous research demonstrated at 0.75mM it reduced inflammatory cytokine production in HDFs without adversely affecting migration. Therefore, OTD as a surgical adjuvant has the potential to be cytotoxic towards primary melanoma cells, reduce HDF inflammation and preserve migratory wound healing function.

Macular pigment profile characteristics and stability over periods of up to 25 years

This study compares the foveal and total retinal macular pigment (MP) amount in healthy subjects and examines the stability of MP spatial distribution profiles, measured almost annually for periods of up to 25 years.Measurements of MP spatial distribution profiles were made by 51 subjects using the psychophysical technique of minimum motion photometry, utilizing a moving bichromatic grating (test 460 nm, comparison 580 nm). Two foveal fields (0.9° and 2.2° diameter) and 11 annular segments (eccentricities from 0.8° to 7.5°) were used. The total amount of MP within the central 15 degrees was estimated by numerical integration and compared with foveal values. Four subjects with widely different MP profiles underwent almost annual testing for periods of 19–25 years. Eleven of the same subjects and 19 others underwent 2‐wavelength autofluorescence (AF) assessment of MP using a scanning laser ophthalmoscope (incident radiation 488 nm and 514 nm), allowing quantification of the peak relative optical density, distribution and total complement of MP over the central 21 degrees.Detailed examinations revealed widely different MP distribution profiles using motion photometry and/or 2‐wavelength fundus autofluorescence. There was a lack of correlation between peak MP optical density at the fovea and assessments of the total complement of MP within the central 15 or 21 degrees. Serial psychophysical assessments over periods of 19–25 years in 4 cases demonstrated a high degree of stability at all macular locations including the fovea; plots of peak MP optical density against time showed minimal positive and negative gradients (<0.0003).The total amount of macular pigment cannot be accurately estimated from foveal or central values. Widely different MP distribution profiles in healthy subjects demonstrate a high degree of long‐term stability, relevant to investigations that aim to monitor MP or fundus autofluorescence in disease or in studies that aim to modify MP through dietary supplementation.

Downregulation of the GABAA receptor β2 subunit in a rat model of autism

Introduction: Gamma-aminobutyric acid (GABA) is the primary inhibitory neurotransmitter in the brain, and GABA type A receptor (GABAA) activation mediates fast inhibitory actions. Numerous studies have shown that individual with autism spectrum disorder (ASD) exhibit abnormalities in the expression of GABAA receptors in various brain areas. Additionally, animal models of ASD have suggested alterations in GABAergic neurotransmission and a dysregulation in the balance between inhibitory and excitatory systems. Objective: We investigated the immunolabeling of the GABAA receptor β2 subunit (GARB2) in the hippocampus, the amygdala, and thalamus of infant rats prenatally exposed to valproic acid (VPA) as an ASD model. Methods: Pregnant females were injected with VPA (600mg/Kg, i.p.) during the twelfth embryonic day; control rats were injected with saline. On the fourteen-postnatal-day, rats from both experimental groups were anesthetized, transcardially perfused with 0.9% NaCl and 4% paraformaldehyde, and sequential coronal brain sections (40μm thickness) were obtained. Immunohistochemistry was performed to detect GARB2 and the relative optical density (OD) of expression was analyzed. Results: Our data showed a statistically significant downregulation of GARB2 in the lateral amygdaloid nucleus, as well as in the ventral and lateral thalamic nuclei when compared to control rats. No statistically significant differences were detected in the hippocampus. Discussion: Our findings suggest that prenatal exposure to VPA reduces GARB2 expression in limbic brain regions involved in social-emotional behaviors, like previous reports in individuals with ASD. Conclusion These results support for the involvement of the GABAergic system in the pathogenesis of ASD.

NEUN EXPRESSION IN SPINAL NEURONS ORGANIZING PROJECTIONS TO THE CEREBELLUM

We analyzed the peculiarities of the NeuN immunostaining of the cat spinal cord neurons located in four structures organizing projections to the cerebellum: the Clarke’s nucleus and border cells – in the L4 segment, the central cervical nucleus – in the C3 segment, and the Stilling’s nucleus – in the S2 segment. Morphometric and densitometric studies were carried out. It was shown that all neurons of interest have a striking feature: an extremely weak level of the cytoplasmic NeuN-staining, while maintaining a high level of the nuclear NeuN-staining. The soma size of neurons of interest was 1000–1850 mkm2, which is comparable to the size of another type of large neurons at slices – motoneurons (1140–1660 mkm2). Thus, we used a motoneuronal population of the corresponding segments to compare the values of optical density. The relative optical density of neurons of interest was several times lower than for the motoneurons (0.060 ± 0.030 vs 0.330 ± 0.127). There were no significant differences in optical density between different structures of interest. Given the morphological uniqueness and similarity of these four cell populations, we believe that the feature of NeuN protein expression can be used as a simple tool for the visualization of cells organizing projections to the cerebellum. It can be valuable both for targeted morphological examination and for histological control after a physiological experiment.

THU117 Lipocalin-2 As A Mediator Of Negative Affect After Sepsis

Abstract Disclosure: S.E. Yang: None. C. Johnston: None. K. Laborc: None. S. Kounelis-Wuillaume: None. W. Schwieterman: None. A.R. Hill: None. J.L. Spencer-Segal: None. Sepsis is characterized by a life-threatening response to severe infection including systemic inflammation. Many sepsis survivors suffer long-term from a post-sepsis syndrome including prominent neuropsychiatric symptoms, but the mechanisms underlying these symptoms remain unknown. In mice, cecal ligation and puncture (CLP) is used to induce sepsis, which is then treated with antibiotics and fluids. Although mice successfully recover within a week, we observe long-lasting anxiety-like behaviors in C57BL/6J mice of both sexes up to 3 weeks post-CLP as shown by decreased time spent in the open areas of an elevated zero maze compared to SHAM mice (52.24±24.60 vs. 74.32±20.25 seconds; p<0.05; n=12 CLP, n=18 SHAM), reminiscent of the symptoms seen in human sepsis survivors. To elucidate the mechanisms underlying anxiety-like behavior in CLP survivors, we performed bulk RNA-sequencing in hippocampal tissues 3 weeks after SHAM or CLP surgeries. We found that lipocalin-2 (LCN2), a key component of the innate immune response, was one of the most upregulated gene transcripts in the hippocampus after CLP (24.3-fold increase relative to SHAM). We also observed a significant increase in systemic LCN2 protein levels in plasma samples 3 weeks after CLP compared to SHAM (1246±1902.62 vs. 77.53±59.09 ng/mL; p<0.001; n=30 CLP, n=39 SHAM). To identify the cellular source of lcn2 mRNA upregulation, we performed RNAscope in hippocampal tissues and found that lcn2 mRNA expression is similar between SHAM and CLP mice in CA1 hippocampal neurons, and highly abundant in TEK+ endothelial cells after CLP (0.43±0.18 vs. 0.25±0.05 relative optical density; p<0.05; n=8 CLP, n=20 SHAM) suggesting that endothelial cells are the major source of LCN2 upregulation after CLP. To investigate the role of LCN2 as a hormonal mediator of negative affect in sepsis, we administered recombinant LCN2 (rLCN2) once a day for 7 days via subcutaneous injections in both male and female mice and assessed affect and memory using the open field test, the elevated zero maze and the novel objection recognition test. While vehicle and rLCN2-injected female mice showed no significant differences in behavior, rLCN2-injected male mice exhibited anxiety-like behaviors as indicated by a significant decrease in the time spent in the center of an open field compared to vehicle-injected controls (48.25±18.62 vs. 65.87±14.76 seconds; p<0.05; n=9 per group). This difference was not caused by locomotion deficits as the mean total distance travelled was similar between groups. Overall, our results demonstrate that LCN2 is a hormone produced in excess by endothelial cells in sepsis survivors, and that chronic rLCN2 administration was sufficient to induce anxiety-like behaviors in male mice but not female mice. Taken together, our data suggest that LCN2 is a novel endocrine mediator of post-sepsis syndrome. Presentation: Thursday, June 15, 2023

EBV antibody and gastric cancer risk: a population-based nested case-control study in southern China

BackgroundWe aim to clarify the controversial associations between EBV-related antibodies and gastric cancer risk.MethodsWe analysed the associations between serological Epstein-Barr nuclear antigen 1 immunoglobulin A (EBNA1-IgA) and viral capsid antigen immunoglobulin A (VCA-IgA) by enzyme-linked immunosorbent assay and the risk of gastric cancer in a nested case-control study originated from a population-based nasopharyngeal carcinoma (NPC) screening cohort in Zhongshan, a city of southern China, including 18 gastric cancer cases and 444 controls. Conditional logistic regression was used to calculate the odds ratios (ORs) and corresponding 95% confidence intervals (CIs).ResultsAll the sera of cases were sampled before diagnosis and the median time interval was 3.04 (range: 0.04, 7.59) years. Both increased relative optical density (rOD) values of EBNA1-IgA and VCA-IgA were associated with higher risks of gastric cancer with age adjusted ORs of 1.99 (95%CI: 1.07, 3.70) and 2.64 (95%CI: 1.33, 5.23), respectively. Each participant was further classified as high or medium/low risk based on a combination of two anti-EBV antibody levels. Participants in the high-risk group had substantially higher odds of developing gastric cancer than that in the medium/low risk group with an age adjusted OR of 6.53 (95%CI: 1.69, 25.26).ConclusionsOur research reveals positive associations between EBNA1-IgA and VCA-IgA and gastric cancer risk in southern China. We thus postulate that EBNA1-IgA and VCA-IgA might appear to be potential biomarkers for gastric cancer. More research to further validate the results among diverse populations and investigate its underlying biological mechanism is needed.

Effect of a Tetraethoxysilane Hydrolysis Reaction Catalyst on the Precipitation of Hydrolysis Products in the Pores of a Polyimide Track Membrane

This paper presents the results of obtaining a composite film based on polyimide track membranes filled with a silica filler, although the issue of the deposition of this filler in the pores of the given membranes remained unexplored. The filler was obtained by hydrolysis of tetraethoxysilane using an alkaline and acid catalyst. This paper presents the results of the effect of the tetraethoxysilane hydrolysis reaction catalyst on the precipitation of hydrolysis products in the pores of the polyimide track membrane. The factors influencing the formation of silicon oxide nanofibers within the matrix template (polyimide track membrane) are determined. It was found that the use of an acid catalyst provides the highest rates of filling, while when using an alkaline catalyst, the filling is practically not observed, and only single pores are filled. The properties of the composite film obtained were investigated. SEM images of the surface and chip of the composite while using alkaline and acid catalyst are presented. The spatial structure of composite films based on track membranes was investigated by FTIR spectroscopy. The hydrolysis of tetraethoxysilane in an acid medium significantly decreases the optical density index of the membranes and simultaneously increases their light transmission index. The greatest changes are observed in the range of 500–1000 nm, and there are no detectable changes in the range of 340–500 nm. When using an alkaline catalyst, there is not the same significant decrease in the relative optical density index D.

Pre-diagnostic anti-EBV antibodies and primary liver cancer risk: a population-based nested case-control study in southern China

BackgroundWe aimed to investigate associations between pre-diagnostic anti-Epstein-Barr virus (EBV) antibodies, including interactions with hepatitis B virus (HBV), and risk of primary liver cancer in southern China.MethodsIn a population-based nested case-control study, we measured pre-diagnostic immunoglobulin A (IgA) against EBV nuclear antigen 1 (EBNA1) and viral capsid antigen (VCA) in 125 primary liver cancer cases and 2077 matched controls. We also explored the interaction between HBV surface antigen (HBsAg) and anti-EBV antibodies.ResultsParticipants with positive EBNA1-IgA, positive VCA-IgA or single-positive anti-EBV antibodies had two-fold odds of developing liver cancer, compared with seronegative subjects. The odds ratios (ORs) between the relative optical density of EBNA1-IgA and VCA-IgA and primary cancer, controlling for age and HBsAg, were 1.59 (95% confidence interval (CI): 1.17, 2.14) and 1.60 (95% CI: 1.07, 2.41), respectively. Subjects with both HBsAg and anti-EBV antibody seropositivity were at 50-fold increased risk compared with those negative for both biomarkers (OR: 50.67, 95% CI: 18.28, 140.46), yielding a relative excess risk due to interaction of 30.81 (95% CI: 3.42, 114.93).ConclusionPre-diagnostic seropositivity for EBNA1-IgA and/or VCA-IgA was positively associated with primary liver cancer risk, especially in combination with HBsAg positivity. EBV may interact with HBV in the development of primary liver cancer, and anti-EBV antibodies might be potential biomarkers for primary liver cancer in this high-risk population.

Evidence for the Hydration of Some Organic Compounds during Reverse-Phase HPLC Analysis.

Some polar analytes (X) can reversibly form hydrates in water-containing eluents under the conditions of reversed-phase HPLC analysis, X + H2O ⇄ X × H2O. One of the methods to detect their formation is the recurrent approximation of the net retention times of such analytes, tR(C + ΔC) = atR(C) + b, where ΔC = const is the constant step in the variation of the organic modifier content of an eluent. These dependencies are linear if hydrates are not formed, but in the case of hydrate formation, they deviate from linearity under high water content. It has been shown that UV spectroscopic parameters, namely, relative optical densities: Arel = A(λ1)/A(λ2), depend on eluent composition for some organic compounds, but their variations cannot be used as indicators for hydrate formation. The coefficients that characterize the dependence of the analyte retention indices on the organic component concentration of an eluent, dRI/dC, appeared to be the most informative additional criterion for hydration. The values of these coefficients for most polar analytes are largely negative (dRI/dC < 0), whereas, for nonpolar compounds, they are largely positive (dRI/dC > 0).

Effects of Long-Term Dietary Zinc Oxide Nanoparticle on Liver Function, Deposition, and Absorption of Trace Minerals in Intrauterine Growth Retardation Pigs.

To investigate the long-term effects of dietary zinc oxide nanoparticle (Nano-ZnO, 20-40 nm) on the relative organ weight, liver function, deposition, and absorption of trace minerals in intrauterine growth retardation (IUGR) pigs, piglets were allocated to NBW (6 normal birth weight piglets fed basal diets), IUGR (6 IUGR piglets fed basal diets) and IUGR+NZ (6 IUGR piglets fed basal diets + 600 mg Zn/kg from Nano-ZnO) groups at weaning (21 days of age), which were sampled at 163 days of age. There were no noteworthy changes in the relative weight of organs, hepatic histomorphology, serum alkaline phosphatase, glutamic pyruvic transaminase and glutamic oxalacetic transaminase activities, and Mn, Cu, and Fe concentrations in leg muscle, the liver, the tibia, and feces among the IUGR, NBW, and IUGR+NZ groups (P>0.05), and no intact Nano-ZnO in the jejunum, liver, and muscle was observed, while dietary Nano-ZnO increased the Zn concentrations in the tibia, the liver, serum, and feces (P<0.05) and mRNA expression of metallothionein (MT) 1A, MT2A, solute carrier family 39 member (ZIP) 4, ZIP14, ZIP8, divalent metal transporter 1, solute carrier family 30 member (ZnT) 1, ZnT4 and metal regulatory transcription factor 1, and ZIP8 protein expression in jejunal mucosa (P<0.05). Immunohistochemistry showed that dietary Nano-ZnO increased the relative optical density of ZIP8 (mainly expressed in cells of brush border) and MT2A (mainly expressed in villus lamina propria and gland/crypt) (P<0.05). In conclusion, long-term dietary Nano-ZnO showed no obvious side effects on the development of the major organs, liver function, and metabolism of Cu, Fe, and Mn in IUGR pigs, while it increased the Zn absorption and deposition via enhancing the expression of transporters (MT, ZIP, and ZnT families) in the jejunum, rather than via endocytosis as the form of intact nanoparticles.

Brain Tumour Lessons From Alzheimer’s Disease: Beta-Amyloid and SorLA Expression in Glioblastoma

Abstract AIMS Exploring how two proteins strongly associated with Alzheimer’s disease are expressed in glioma. Amyloid beta (Aβ) is a peptide cleaved from amyloid precursor protein (APP) that is shuttled around the cell by sorting-related receptor with A-type repeats (SorLA). Low expression of SorLA has been linked to increased risk of Alzheimer’s disease as APP processing shifts towards production of the plaque-forming Aβ42 rather than the more benign Aβ40. METHOD Expression of Aβ40, Aβ42 and SorLA was determined in immortalised cell lines (grade 4 glioblastoma U87MG, grade 2 astrocytoma 1321N1, foetal astrocytes SVGp12), glioblastoma patient-derived cells (PD301, PD304) and normal human astrocytes through western blotting or immunocytochemistry. RESULTS SorLA was present in all cells. Relative optical density was more than 60-fold higher for U87MG glioblastoma cells and approximately 10-fold higher for 1321N1 astrocytoma cells than SVGp12 cells. Similarly, SorLA had a significantly higher expression (p&amp;lt;0.01) in patient derived cells than normal human astrocytes. High expression of SorLA was accompanied by lower Aβ42 in patient derived cells compared to normal human astrocytes. Expression of Aβ40 was similar across patient derived cells and normal human astrocytes. CONCLUSION These data suggest that SorLA is retained in glioblastoma and remains functional to drive APP processing away from Aβ42 production. Future work will determine how high SorLA expression contributes to the increased motility and invasiveness seen in glioblastoma cells by transfecting siRNA to knock down SorLA.

Chemokine (C–C motif) receptor 2 is associated with the pathological grade and inflammatory response in IgAN children

BackgroundChemokine (C–C motif) receptor 2 (CCR2) is involved in important physiological and pathological processes, such as inflammation and autoimmune diseases. Abnormal immune and inflammatory responses play a critical role in the development and progression of IgA nephritis (IgAN). However, the role of CCR2 in IgAN is unknown.MethodsFifteen IgAN children who were diagnosed by kidney biopsy provided kidney biopsy tissue, blood and urine samples, and age-matched healthy control subjects (blood donators n = 12; tissue donators n = 8) were included. Immunohistochemical analysis was used to detect the expression of CCR2, MCP-1, IL-6, IL-17, and TNF-α in the kidney tissues. Relative optical density (OD) was calculated by Image J software, and the correlation between CCR2 expression and pathological grade in IgAN children was analyzed.ResultsThe expression of CCR2 significantly increased in mesangial cells of children with IgAN compared to that in control group (P < 0.001), especially in IgAN patients with Lee’s grade III to IV (P < 0.001). Interestingly, CCR2 expression was positively correlated with Lee’s grade (r = 0.9152, P = 0.0001) in IgAN children. The expression levels of inflammatory factors were markedly increased in IgAN children, and importantly CCR2 expression was positively correlated with it’s expression level.ConclusionsThe results suggest that CCR2 signaling might be involved in pathological process and inflammatory responses of children IgAN, and could potentially be an intervention target in children IgAN.

Optical Response of Expired EBT3 Film for Absorbed Dose Measurement in X-ray and Electron Beam Range

The purpose of this study was to investigate the optical response of an expired External Beam Therapy (EBT3) film, which expired in 2018, using X-rays and electron beam doses. The film’s optical responses were evaluated for its usability in measuring different radiation sources, energy, and absorbed doses ranging up to 5 Gy. Pieces of the expired EBT3 film were irradiated with 90 kVp, 6 MV X-ray photons, and 6 MeV electron beam. The analysis was performed using the Jaz visible spectrometer and EPSON Perfection V370 Photo scanner to obtain the absorbance and the net relative optical density (ROD) of the film samples respectively. The results showed that spectroscopic measurements of the exposed expired EBT3 films under these radiation sources were able to produce primary secondary peaks at λ = 633.52 nm and λ = 582.3 nm respectively. The best wavelength subsets that presented the best MLR regression fitting for all experiments were 541.48 nm, 561.11 nm, and 600.28 nm. While, for the 6 MV photon and the 6 MeV electron beam they were 600.28 nm, 650.79 nm and 654.10 nm. In case of the irradiation with the 6 MV photon and the 6 MeV electron beam, expired EBT3 film showed no significant differences, which made it suitable for dosimetry in various sources of radiation. The individual calibration of radiation dose produces very high measurement accuracy with coefficient of determination, R2 above 0.99 and root mean square of error, RMSE of 0.038 Gy, 0.113 Gy, and 0.115 Gy for films irradiated with 90 kVp X-rays, 6 MV photon beam, and 6 MeV electron beam respectively. Hence, from the results, the expired EBT3 film used in this study showed promising usability of expired EBT3 films beyond their prescribed expiry dates.

A Potential Research Target for Scleral Remodeling: Effect of MiR-29a on Scleral Fibroblasts

Introduction: The purpose of this study was to determine whether miR-29a regulates cell survival and apoptosis and the expression of phosphatase and tensin homolog deleted on chromosome 10 (PTEN), MMP-2, and collagen I in scleral fibroblasts. Methods: We transfected scleral fibroblasts with the miR-29a mimic and inhibitor. The effects of miR-29a on cell proliferation and apoptosis were determined using the CCK-8 assay and flow cytometry, respectively. Quantitative polymerase chain reaction (qPCR) was used to determine whether miR-29a regulates the mRNA levels of PTEN, MMP-2, and collagen I. The protein expression of PTEN, MMP-2, and collagen I was also assessed by western blot analysis. Results: The results of CCK-8 showed that, at 0, 24, 48, and 72 h after transfection, the relative optical density values in the mimic group were 0.233 ± 0.005, 0.380 ± 0.008, 0.650 ± 0.040, and 0.906 ± 0.032, and in the inhibitor group were 0.272 ± 0.011, 0.393 ± 0.029, 0.597 ± 0.059, and 0.950 ± 0.101, respectively. The flow cytometry results showed that the apoptosis rates of each group were as follows: the mimic group (0.043 ± 0.007), the NC group (0.040 ± 0.006), the inhibitor group (0.032 ± 0.003), the inhibitor NC group (0.027 ± 0.010), the lipofectamine group (0.027 ± 0.005), and the blank group (0.031 ± 0.009). The qPCR results indicated that in the mimic group, PTEN (0.795 ± 0.182, p = 0.2783), MMP-2 (0.621 ± 0.105, p = 0.0033), and COL1A1 (0.271 ± 0.100, p = 0.0002) expression decreased, whereas in the inhibitor group, PTEN (1.211 ± 0.100, p = 0.2614), MMP-2 (1.161 ± 0.053, p = 0.1190), and COL1A1 (1.7040 ± 0.093, p = 0.0003) increased. Western blot analysis showed that in the mimic group, the expression of PTEN (0.392 ± 0.039, p < 0.0001), MMP-2 (0.577 ± 0.017, p < 0.0001), and COL1A1 (0.072 ± 0.006, p < 0.0001) protein decreased, whereas in the inhibitor group, PTEN (1.043 ± 0.042, p = 0.9413), MMP-2 (1.397 ± 0.075, p = 0.0002), and COL1A1 (1.935 ± 0.081, p < 0.0001) expression increased. Conclusion: MiR-29a inhibits the expression of PTEN, MMP-2, and collagen I on scleral fibroblasts, which may provide a basis studies in sclera.

Low-Level Laser Irradiation Promotes Proliferation and Differentiation on Apical Papilla Stem Cells.

Introduction: Low-level laser therapy (LLLT) has been reported to improve cell proliferation and differentiation. The stem cells derived from dental apical papilla (SCAPs) are a promising therapy because they are easily obtained from immature human teeth. The effect of LLLT over SCAPs is still unknown. This study aimed to evaluate the proliferation and osteogenic potential of the SCAPs stimulated with LLLT. Methods: SCAPs were isolated from the third molars of a healthy donor and characterized according to the minimum established criteria. SCAPs were cultured for 24 hours before being exposed to LLLT. Cells were exposed to different doses, energy, and wavelengths for selecting the irradiation parameters. SCAPs proliferation was evaluated with the MTT assay at 24 hours and 7-day post-laser exposure. VEGF and TGFβ2 expression were assessed with a specific enzyme-linked immunosorbent assay (ELISA). The osteogenic differentiation potential was analyzed with alizarin red staining, and the nodule quantification was performed by the relative optical density (ROD) analysis using ImageJ software. Results: The cells isolated from the apical papilla showed phenotype and stem cell properties. SCAPs irradiated with one dose at 6 J/m2 and 650 nm exhibited significantly higher proliferation (P>0.05) than the controls nonirradiated. LLLT stimulated SCAPs' expression of factors VEGF and TGFβ2. Also, SCAPs irradiated showed higher osteogenic activity (P<0.05). Conclusion: LLLT promotes proliferation, osteogenic differentiation, and VEGF and TGFβ2 expression on SCAPs. LLLT is a practical approach for the preconditioning of SCAPs in vitro for future regenerative therapies. More studies are needed to determine the underlying molecular processes that determine the mechanism of the LLLT.