Estimation of age at onset of linear enamel hypoplasia. New calculation tool, description and comparison of current methods.

Enamel Hypoplasia (EH) is known to be a useful indicator for wide range of detrimental factors in early childhood in past populations, such as nutritional disturbances, mechanical trauma, disease, metabolic, and/or genetic disorders. EH may be divided into three categories: pits, grooves, and lines, where the last two are referred to as "Linear Enamel Hypoplasia" (LEH). The regularity of enamel formation allows retrospective determination of the age of LEH formation. The current article reviews and compares the best-known methods used to estimate age at LEH formation and provides a new computational tool. Growth curves for canines and incisors were developed based on tooth growth tables by previous authors. Optimal models were selected using the Akaike Information Criterion. A Microsoft Excel spreadsheet was created to calculate age at LEH formation using the most common methods. All method results were compared with an archaeological sample (44 teeth of 18 individuals from an early modern cemetery from Wroclaw, Poland) and a theoretical model. The results of the methods were compared pairwise with Bland-Altman plots. The current article provides a quick and easy-to-use tool for analyzing LEH chronology and comparing the results of different methods. As shown by the Bland-Altman plots, most methods provide approximately consistent results for LEHs formed at around 2-3 years of age. However, LEHs formed particularly early or late are more prone to discrepancies between different methods. Comparison of the age at LEH formation obtained by different methods should be done carefully - and the new LEH calculation tool with optimized equations provided in this publication can facilitate this process.

Correlation between Periostin Expression and Pro-Angiogenic Factors in Non-Small-Cell Lung Carcinoma.

The role of periostin (POSTN) in remodeling the microenvironment surrounding solid tumors and its effect on the tumor cells in non-small-cell lung carcinoma (NSCLC) have not yet been fully understood. The aim of this study was to determine the relationship between POSTN expression (in tumor cells [NSCLC cells] and the tumor stroma) and pro-angiogenic factors (CD31, CD34, CD105, and VEGF-A) and microvascular density (MVD) in NSCLC. In addition, these associations were analyzed in individual histological subtypes of NSCLC (SCC, AC, and LCC) and their correlations with clinicopathological factors and prognosis were examined. Immunohistochemistry using tissue microarrays (TMAs) was used to assess the expression of POSTN (in tumor cells and cancer-associated fibroblasts [CAFs]) and the pro-angiogenic factors. A significant positive correlation was found between the expression of POSTN (in cancer cells/CAFs) and the expression of the analyzed pro-angiogenic factors (CD31, CD34, CD105, and VEGF-A) and MVD in the entire population of patients with NSCLC and individual histological subtypes (AC, SCC). In addition, this study found that POSTN expression (in tumor cells/CAFs) increased with tumor size (pT), histopathological grade (G), and lymph-node involvement (pN). In addition, a high expression of POSTN (in tumor cells and CAFs) was associated with shorter survival among patients with NSCLC. In conclusion, a high expression of POSTN (in cancer cells and CAFs) may be crucial for angiogenesis and NSCLC progression and can constitute an independent prognostic factor for NSCLC.

Vitrification of feline ovarian tissue: Comparison of protocols based on equilibration time and temperature.

Global contraction of biodiversity pushed most members of Felidae into threatened or endangered list except the domestic cat (Felis catus) thence preferred as the best model for conservation studies. One of the emerging conservation strategies is vitrification of ovarian tissue which is field-friendly but not yet standardized. Thus, our main goal was to establish a suitable vitrification protocol for feline ovarian tissue in field condition. Feline ovarian tissue fragments were punched with biopsy punch (1.5 mm diameter) and divided into 4 groups. Group 1 was fresh control (Fr), while the other three were exposed to 3 vitrification protocols (VIT_CT, VIT_RT1 and VIT_RT2). VIT_CT involved two step equilibrations in solutions containing dimethyl sulfoxide (DMSO) and ethylene glycol (EG) for 10 min each at 4 °C. VIT_RT1 involved three step equilibration in solutions containing DMSO, EG, polyvinylpyrrolidone and sucrose for 14 min in total at room temperature, while in VIT_RT2 all conditions remained the same as in VIT_RT1 except equilibration timing which was reduced by half. After vitrification and warming, fragments were morphologically evaluated and then cultured for six days. Subsequently, follicular morphology, cellular proliferation (expression of Ki-67, MCM-7) and apoptosis (expression of caspase-3) were evaluated, and data obtained were analysed using generalised linear mixed model and chi square tests. Proportions of intact follicles were higher in Fr (P = 0.0001) and VIT_RT2 (P = 0.0383) in comparison to the other protocols both post warming and after the six-day culture. Generally, most follicles remained at primordial state which was confirmed by the low expression of Ki-67, MCM-7 markers. In conclusion, VIT_RT2 protocol, which has lower equilibration time at room temperature has proven superior thus recommended for vitrification of feline ovarian tissue.

Histological analysis of forearm superficial veins structure.

BACKGROUND: The connection between the basilic and cephalic veins of the forearm shows considerable interindividual variation. Depending on its form, the most common types of venous connections are M-, N- or Y-shaped. This study aims to compare the metric traits of the basilic and cephalic veins and the relative content of smooth muscle/collagen fibers/elastic fibers in their walls and to determine the differences between the forearm venous systems. MATERIALS AND METHODS: The study was performed on 42 veins collected from 26 deceased individuals between the ages of 19 and 50 years. Vein sections were fixed, embedded in paraffin blocks and used to prepare histological slides, stained according to pentachrome Movat's method. Venous metrics were assessed and the percentage of muscle, elastic and collagen fibers was determined using the Trainable Weka segmentation. Statistical analysis compared the M-type vein with the Y- and N-types, which were combined into one category. RESULTS AND CONCLUSIONS: Analysis showed a greater tunica media thickness in the M-type vein, with a greater lumen circumference in the Y/N types. Correlation analysis showed a correlation of vein metrics with elastic fibre content and a weak inverse correlation with the tunica media thickness. It can be hypothesized that the increased performance of N- and Y-types may be related to elastic fibers content.

Ultrastructural changes in feline oocytes during ovary storage for 24- and 48-hours.

Despite established microscopic markers of feline oocyte quality, little is known about their ultrastructural traits. To the best of our knowledge, there is no published report analysing the effect of 24 and 48 h ovarian storage time on the domestic cat oocytes characteristics at the ultrastructural level. Oocytes (n = 30) were classified using the light microscopy as good or bad quality and then proceeded for TEM observations. The location, shape, size and distribution of each organelle was noted in each examined oocyte. In in good quality oocytes the cytoplasmic organelles were generally easier to identify, and furthermore its distribution pattern was more obvious to spot than in bad quality ones. Whereas bad quality oocytes were typically characterised by the lower visibility of the cellular structures and cytoplasmic architecture was less apparent and often arranged without a predictable pattern. In good quality oocytes obtained from fresh ovaries cytoplasmic vacuoles (CVs) occupied a significantly larger area (0,72 vs. 0.18 CVs/µm2, respectively) than in bad quality ones, whereas in bad quality and stored oocytes more cytoplasm was occupied by lipid droplets (LDs) than in fresh good oocytes (0,22 ± 0,09 vs. 0,09 ± 0,05 respectively). It can be concluded that ultrastructure changes in feline oocytes during 24 and 48 h ovarian storage cannot be assessed in light microscopy. The ultrastructure of oocytes was seriously disturbed after 48 h of ovary storage, despite being classified as good quality. However, further investigations utilizing more cells are necessary to confirm reported traits of ultrastructure changes in stored and non-stored oocytes of good and bad quality.

Nutritional reconstruction in an early modern population: Searching for a relationship between dental microwear and bone element composition.

BACKGROUND: Observing the microwear patterns of the dental crowns enamel surface can provide information on the ingredients and structure of the food consumed, but also on eating habits and lifestyle. Major role in reconstructing the diet and lifestyle of past populations is played by the analysis of alkaline earth metals, such as strontium, barium, zinc and calcium. Ba and Sr are indicators of the consumption of vegetables, plants (cereals and legumes) and marine organisms. This study aims to assess dietary diversity and identify its components based on microscopic techniques and chemical analyses of material from early modern archaeological sites in Wroclaw, Poland. METHODS: The material consisted of 36 permanent molars and the intrasternal parts of 122 first ribs, collected from 6 Wroclaw early modern cemeteries. Tooth microwear was evaluated on Scanning electron microscopy images, with Microwear 4.02 software. Bone chemical composition (Ca, P, Ba, Sr content) was evaluated with mass spectrometry. RESULTS: Most lines were present on the teeth from St. Mary Magdalene Cemetery, with the lowest average number of lines observed on the teeth from St. Barbara Cemetery. The Ca/P ratios calculated for different sites formed two clusters that allows to distinguish two groups of archaeological sites with different bone preservation status. Number of differences in Ba/Sr, Sr/Ca, Ba/Ca ratios was found between disctinct archaeological sites. A number of correlations were found between the concentration of the chemical elements, but no statistically significant correlation was found between the microwear characteristics and the proportion of the elements studied. Some strong correlations were found between microwear features and the Ca/P ratio. CONCLUSIONS: the different values of Sr/Ca, Ba/Ca and Ba/Sr ratios indicate different diets in the different communities. The data obtained indicate a mixed diet, with a relatively high proportion of animal products throughout Wroclaw. Conclusions should be treated with caution due to secondary diagenesis. The lack of interdependence between microwear and chemical composition characteristics suggests that the two methods should be considered complementary and not overlapping, as they provide different insights into the diets of past populations. The comparison of microwear between different sites should always take into account secondary diagenesis and the burial environment, as these influence the characteristics of microwear.

Antlerogenic stem cells extract accelerate chronic wound healing: a preliminary study.

BACKGROUND: Chronic wounds constitute a significant medical and social problem. Chronic wound treatment may be supported by various techniques, such as negative pressure therapy, phototherapy or stem cells therapy, yet most of those supporting therapies need more evidence to be used for standard wound care. Current study covers the use of sonicated Antlerogenic Stem Cells (ASC) extract on chronic wounds. METHODS: Study was performed on 20 dermatological patients with venous leg ulcers, divided into two groups - treated with and without ASC extract respectively. The area and circumference of the wounds during the follow-up visits were measured on the wound imprint. Dynamics of wound healing was determined and compared between control and study group; statistics includes changes in absolute values (wound area, circumference), as well as relative (percentage of wound decrease, circumference/area ratio) and their change in time. For the purpose of Ki-67 immunohistochemical staining, sections were sampled from the wound edge at distinct check-points during therapy. Results of both groups were compared with Student test or Mann-Whitney test, depending on results distribution. RESULTS: Besides Ki-67 expression, all tested wound healing parameters (including relative and absolute wound decrease and changes in circumference/area ratio) were statistically significant more favorable in experimental group. CONCLUSION: ASC extract significantly supported standard chronic wound treatment. Due to small population of study the results should be considered preliminary, yet promising for further research.

A Case of Syphilis with High Bone Arsenic Concentration from Early Modern Cemetery (Wroclaw, Poland).

Venereal syphilis is a sexually transmitted disease caused by Treponema pallidum - Gram-negative, slowly growing bacteria. The spread of the disease in the Old World was due to increased birth rate, urban population growth, migration and lack of knowledge concerning the epidemiology. In the past, the treatment was mainly symptomatic and included application of mercury compounds. The goal of the study was to present the case of advanced venereal syphilis found in early modern (16th-18thc) graveyard localized in Wroclaw, Poland. The object of the study is a cranium of a male whose age at death has been estimated to be over 55. In order to observe the morphological and paleopathological characteristics of the examined material, anthropometrics, computed tomography, spectrometry and microscopic methods were incorporated. Microscopic analysis revealed the presence of the extensive inflammatory lesions. Analyses indicate tertiary stage of venereal syphilis as the most probable cause of the observed lesions. Concentration of arsenic (16.17±0.58 µg/g) in examined bone samples was about hundred times bigger than average arsenic concentration in bones reported in other studies. Advanced stage of observed lesions along with high arsenic level may suggest long-lasting palliative care and usage of arsenic compound in therapeutic treatment of this chronic disease.