Folinic Acid Potentiates the Liver Regeneration Process after Selective Portal Vein Ligation in Rats

Liver resection remains the gold standard for hepatic metastases. The future liver remnant (FLR) and its functional status are two key points to consider before performing major liver resections, since patients with less than 25% FLR or a Child–Pugh B or C grade are not eligible for this procedure. Folinic acid (FA) is an essential agent in cell replication processes. Herein, we analyze the effect of FA as an enhancer of liver regeneration after selective portal vein ligation (PVL). Sixty-four male WAG/RijHsd rats were randomly distributed into eight groups: a control group and seven subjected to 50% PVL, by ligation of left portal branch. The treated animals received FA (2.5 m/kg), while the rest were given saline. After 36 h, 3 days or 7 days, liver tissue and blood samples were obtained. FA slightly but significantly increased FLR percentage (FLR%) on the 7th day (91.88 ± 0.61%) compared to control or saline-treated groups (86.72 ± 2.5 vs. 87 ± 3.33%; p < 0.01). The hepatocyte nuclear area was also increased both at 36 h and 7days with FA (61.55 ± 16.09 µm2, and 49.91 ± 15.38 µm2; p < 0.001). Finally, FA also improved liver function. In conclusion, FA has boosted liver regeneration assessed by FLR%, nuclear area size and restoration of liver function after PVL.

Correlation of Nuclear Morphometry with Clinicopathologic Parameters in Malignant Breast Aspirates

Background Nuclear size, shape, chromatin pattern, and nucleolar size and number have all been reported to change in breast cancer. Aim The aim of the study was to quantify nuclear changes on malignant breast aspirates using morphometry and to correlate the morphometric parameters with clinicopathologic features such as cytologic grade, tumor size, lymph node status, mitotic index, and histopathologic grade. Materials and Methods Forty-five cases of carcinoma breast diagnosed on cytology were included in this study. Cytologic grading was performed as per the Robinson’s cytologic grading system. Nuclear morphometry was done on Papanicolaou stained smears. One hundred nonoverlapping cells per case were evaluated. Both geometrical and textural parameters were evaluated. Results Comparison of cytologic grades with most morphometric features (nuclear area, perimeter, shape, long axis, short axis, intensity, total run length, and TI homogeneity) was highly significant on statistical analysis. Correlation with tumor size yielded significant results for nuclear area, perimeter, long and short axes, and intensity with p < 0.05. The study of lymph node status and morphometry showed a highly significant statistical association with all the parameters. Mitotic count was significantly associated with all the geometric parameters and one textural parameter (total run length). On correlation of ductal carcinoma in situ and histopathological Grades 1 to 3 with morphometry, it was found that all the parameters except long–run emphasis were highly significant with p < 0.001. Conclusion Morphometry as a technique holds immense promise in prognostication in breast carcinoma.

Canine mast cell tumors: utility of stereologic tools in cytology

Quantitative morphologic parameters assessed in cytologic samples of canine cutaneous mast cell tumors (ccMCTs) may assist with surgical planning and prognostication. Robust cutoffs can be defined, with high reproducibility, for parameters such as the nuclear area (NA). The NA may be determined by morphometry (image analysis, NAI) or by stereology, such as the 2D-nucleator method (NAN); stereologic techniques have not been applied to cytologic specimens of ccMCT, to our knowledge. We retrospectively selected routine cytology smears from 51 ccMCT cases and screened them to determine the percentage of neoplastic mast cells with indistinct nuclear borders; this was repeated after the slides were restained with H&E. The NAI and the NAN were estimated in 100 mast cells per animal in H&E-stained slides. All nuclei were visible in H&E smears, and unbiased quantification was feasible. The NAN was similar to NAI, but less time-consuming. Both the NAN and NAI determined by cytology differed in histologic low- and high-grade ccMCTs, and in histologic grade I plus II versus grade III ccMCTs. Stereologic parameters such as the NAN could be considered as complementary techniques for the cytologic evaluation of ccMCTs.

Relationship between Ag-NORs and T Lymphocyte Subsets in Children with Epilepsy

With the acceleration of the pace of life, the number of children with epilepsy is increasing. This study was to investigate the relationship between Ag-NORs and T lymphocyte subsets in children with epilepsy. In the morning, 3ml of elbow vein blood was collected from epileptic children on an empty stomach. After standing at room temperature for 0.3-1 hour, the serum was centrifuged (2000 rpm, 4 minutes). 3 ml of mixed venous blood was collected in EDTA-K2 vacuum container. The sample was taken out within 5 hours after staining. 30 n I of Itcite reagentand 30 n I of anticoagulant were put into TruCount tube and mixed evenly. Then flow cytometry was used for detection, and facscomp software was used for automatic inspection and calibration. In the same way, the automatic analysis software muitiset was used to obtain 20000 white blood cells, count 8000-10000 lymphocytes automatically, and calculate CD4 + T lymphocytes, CD8 + T lymphocytes and CD4 + / CD8 + ratio. Ag-NORs detection: under aseptic conditions, 0.5ml anticoagulant blood was added into the medium flask containing rpm-1640 and incubated in the incubator at 37°C for 72 hours. After mixing the cell suspension at room temperature, put it into a 10ml glass tube, heat it and dry it. Wait until the temperature of the water solution tank rises to 80-90 °C, and then put aluminum plate on it. The staining was placed under the microscope on the stage, and the image was adjusted to make the image analysis program effective. The ratio of nuclear area to nuclear silver staining area of 30 lymphocytes was counted. This ratio reflects the content of Ag-NORs in the nucleolar forming region of T lymphocytes. The ratio of Ag-NORs area to nuclear area was 0.32 ± 0.03, 0.38 ± 0.03 and 0.46 ± 0.03, respectively. This study is helpful to provide guidance for the treatment of epilepsy in children.

Inhibitory Effects of Alendronate on Adhesion and Viability of Preosteoblast Cells on Titanium Discs

Objective The study aimed to investigate the effects of alendronate (ALN; a bisphosphonate) on adhesion and viability of preosteoblasts using different cell passages on sandblasted and acid-etched (SLA) Ti surfaces. Materials and Methods Preosteoblast, MC3T3, cells (passage 42; P42 and passage 62; P62) were cultured with ALN (1 and 5 µM) on cell culture plate for 7 days. Cells were lifted, counted, and seeded on SLA Ti surfaces. Cells were incubated on the discs for 6 hours to examine cell adhesion by using confocal microscopy and for 24 hours to determine cell viability by using MTT assay. Results ALN interfered with cell adhesion on Ti surfaces by reducing the cell number in both cell passages. Nuclei of untreated cells showed oval shape, whereas some nuclei of ALN-treated cells demonstrated crescent and condensed appearance. ALN at 1 and 5 µM significantly decreased nuclear area and perimeter in P42, while ALN at 5 µM reduced nuclear area and perimeter in P62. After 24 hours, cells (P42) grown on Ti surfaces showed decreased cell viability when culturing with 5 µM ALN. Conclusion ALN reduced cell adhesion and viability of preosteoblasts on Ti surfaces. ALN treatment seemed to exert higher inhibitory effects on nuclear shape and size as well as cell viability in lower cell passage. This led to the reduction in cell to implant surface interaction after encountering bisphosphonate treatment.

NUCLEAR AREA OF SPEECH GENRE SPACE OF THE RUSSIAN LEGISLATIVE DISCOURSE

Обсуждается речежанровая модель российского законодательного дискурса: предлагаются жанровые и дискурсивные параметры моделирования; в соответствии с выделенными параметрами определяются речевые жанры ядерной зоны российского законодательного дискурса первого и второго порядка; привлекаются материалы компонентного анализа лексем, называющих рассматриваемые речевые жанры. The paper discusses the speech genre model of the Russian legislative discourse. We attempt to offer modelling genre and discourse parameters according to which we determine the first-order and second-order speech genres of the nuclear area of the Russian legislative discourse. The undertaken componential analysis highlights essential differences between the lexemes naming the speech genres in question.

Comparing World Health Organization/International Society of Urological Pathology Grading and Fuhrman Grading with the Prognostic Value of Nuclear Area in Patients with Renal Cell Carcinoma

This study was undertaken to compare Fuhrman grading with World Health Organization/International Society of Urological Pathology (WHO/ISUP) grading and stereologically measured nuclear area in patients with Clear Cell Renal Cell Carcinoma (ccRCC) or Papillary Renal Cell Carcinoma (PRCC) and to evaluate the independent predictive value of Fuhrman, WHO/ISUP and stereologically measured nuclear area combined with necrosis in a series of patients with ccRCC in relation to cancer-specific survival. In all, 124 cases of ccRCC and PRCC were included. All slides were blindly scored by two trained pathologists according to the Fuhrman and WHO/ISUP grading systems. Nuclear measurements were performed on digitally scanned slides in Visiopharm® and correlated to survival. Analysis of ccRCC and PRCC cases showed that application of WHO/ISUP grading resulted in a significant downgrading of cases from G2 to G1, when comparing with Fuhrman grading. Neither of these patients experienced progression. Cancer specific survival estimates in 101 ccRCC patients showed that WHO/ISUP grading was slightly superior in predicting cancer-specific survival. Novel models included WHO/ISUP grading and mean nuclear area (MNA) each of which combined with necrosis. Both demonstrated an increased ability to predict cancer-specific survival. The study demonstrates that WHO/ISUP grading provides superior prognostic information compared to Fuhrman grading and stereologically measured nuclear area. Necrosis in combination with either WHO/ISUP grading or MNA adds additional prognostic information.

Experiments in the EpiDerm 3D Skin In Vitro Model and Minipigs In Vivo Indicate Comparatively Lower In Vivo Skin Sensitivity of Topically Applied Aneugenic Compounds

Risk management of in vitro aneugens for topically applied compounds is not clearly defined because there is no validated methodology to accurately measure compound concentration in proliferating stratum basale keratinocytes of the skin. Here, we experimentally tested several known aneugens in the EpiDerm reconstructed human skin in vitro micronucleus assay and compared the results to flow cytometric mechanistic biomarkers (phospho-H3; MPM2, DNA content). We then evaluated similar biomarkers (Ki-67, nuclear area) using immunohistochemistry in skin sections of minipigs following topical exposure the potent aneugens, colchicine, and hesperadin. Data from the EpiDerm model showed positive micronucleus responses for all aneugens tested following topical or direct media dosing with similar sensitivity when adjusted for applied dose. Quantitative benchmark dose-response analysis exhibited increases in the mitotic index biomarkers phospho-H3 and MPM2 for tubulin binders and polyploidy for aurora kinase inhibitors are at least as sensitive as the micronucleus endpoint. By comparison, the aneugens tested did not induce histopathological changes, increases in Ki-67 immunolabeling or nuclear area in skin sections from the in vivo minipig study at doses in significant excess of those eliciting a response in vitro. Results indicate the EpiDerm in vitro micronucleus assay is suitable for the hazard identification of aneugens. The lack of response in the minipig studies indicates that the barrier function of the minipig skin, which is comparable to human skin, protects from the effects of aneugens in vivo. These results provide a basis for conducting additional studies in the future to further refine this understanding.