Morphological and morphometric changes of bone tissue in patients with osteoporosis and osteomalation

Bone tissue was studied in 56 postmenopausal women (mean age 62.30 ± 2.74 years), of which 46 patients who worked in unfavorable working conditions had a decreased bone mineral density (BMD) (osteoporosis (OP) — in 31 women, osteomalacia (OM) — in 13); 10 women had no metabolic changes in bone tissue (BT). A BT scan fragment was obtained during surgery for a fracture of the femoral neck. Non-decalcified QD sections were prepared, the functional activity of the QD cell nuclei was determined using the method of differential staining of nuclei with different functional activity. Morphological changes in OP and OM have both common features and differences. The common is the thinning of the bone rods, the expansion of the canals of osteons, the presence of cell-free areas, and cell-free lacunae. In contrast to OP, OM presents with the thickness and area of the osteoid increase, a less pronounced decrease in oxyphyllin matrix, a higher functional activity of BT cells. A decrease in BMD and the occurrence of low-energy fractures may result not only from OP but also OM. When prescribing treatment, it is necessary to carry out diffe-rential diagnostic measures that determine the cause of the decrease in bone mass.

2D morphometric analysis of Arabidopsis thaliana nuclei reveals characteristic profiles of different cell types and accessions

Functional changes of cells upon developmental switches and in response to environmental cues are often reflected in nuclear phenotypes, showing distinctive chromatin states corresponding to transcriptional changes. Such characteristic nuclear shapes have been microscopically monitored and can be quantified after differential staining of euchromatin and heterochromatin domains. Here, we examined several nuclear parameters (size, DNA content, DNA density, chromatin compaction, relative heterochromatin fraction (RHF), and number of chromocenters) in relation to spatial distribution of genes and transposon elements (TEs), using standard 2D fluorescence microscopy. We provide nuclear profiles for different cell types and different accessions of Arabidopsis thaliana. A variable, yet significant, fraction of TEs was found outside chromocenters in all cell types, except for guard cells. The latter cell type features nuclei with the highest level of chromatin compaction, while their chromocenters seem to contain gene-rich regions. The highest number of parameter correlations was found in the accession Cvi, whereas Ler showed only few correlations. This may point at differences in phenotype robustness between accessions. The significantly high association of NOR chromocenters in accessions Ws and Cvi corresponds to their low RHF level.

A STUDY ON ADENOSINE DEAMINASE AND C-REACTIVE PROTEIN IN CEREBROSPINAL FLUID OF MENINGITIS PATIENTS OF DIFFERENT TYPES.

Introduction: The three commonest types of Meningitis are Bacterial, Viral and Tubercular; they differ in their pathophysiology and management. That is why early differentiation between them is important. Bacterial meningitis have heavy fatality rates whereas Tubercular meningitis inicts severe neurological sequalae. Aims & Objectives: To determine whether ADA and CRP can be used successfully to predict the type of Meningitis affection Methodology: This is a Hospital based cross-sectional observation study which was undertaken in Biochemistry dept. Medical College, Kolkata. Eighty cases of meningitis were taken. ADA and CRP was measured along with Protein, Sugar and Differential staining cell counting. The cases were divided into three groups (Bacterial, Viral, and Tubercular) on the basis of microscopic and Biochemical Examination. Results & Analysis: ANNOVA test was done along with Bon Ferronis Test which revealed that there was signicant difference in mean of ADA & CRP in the three different groups.ROC curve for CRP between Reactive and Non-reactive meningitis revealed that if CRP values are more than 11.0, 95% of the cases were reactive whereas ROC curve for ADA between Tubercular and Non Tubercular forms showed when the concentration was more then 9.0, 95% of the cases were of Tubercular origin. Conclusion: When taken together ADA and CRP successfully predict the type of Meningitis. It is also helpful in diagnosing those cases when the Cell count and type was equivocal.

Porphyromonas somerae Invasion of Endometrial Cancer Cells

Recent evidence suggests an association between endometrial cancer and the understudied bacterial species Porphyromonas somerae. This association was demonstrated in previous work that indicated a significantly enriched abundance of P. somerae in the uterine microbiome of endometrial cancer patients. Given the known associations of the Porphyromonas genus and oral cancer, we hypothesized that P. somerae may play a similar pathogenic role in endometrial cancer via intracellular activity. Before testing our hypothesis, we first characterized P. somerae biology, as current background data is limited. These novel characterizations include growth curves in liquid medium and susceptibility tests to antibiotics. We tested our hypothesis by examining growth changes in response to 17β-estradiol, a known risk factor for endometrial cancer, followed by metabolomic profiling in the presence and absence of 17β-estradiol. We found that P. somerae exhibits increased growth in the presence of 17β-estradiol of various concentrations. However, we did not find significant changes in metabolite levels in response to 17β-estradiol. To study direct host-microbe interactions, we used in vitro invasion assays under hypoxic conditions and found evidence for intracellular invasion of P. somerae in endometrial adenocarcinoma cells. We also examined these interactions in the presence of 17β-estradiol but did not observe changes in invasion frequency. Invasion was shown using three lines of evidence including visualization via differential staining and brightfield microscopy, increased frequency of bacterial recovery after co-culturing, and in silico methods to detail relevant genomic and transcriptomic components. These results underscore potential intracellular phenotypes of P. somerae within the uterine microbiome. Furthermore, these results raise new questions pertaining to the role of P. somerae in the progression of endometrial cancer.

Modified staining protocol with Safranin O and Astra Blue for the plant histology

Many staining protocols are widely applied in botanical microtechniques and serve specific histological purposes. In particular, some dyes are used simultaneously to receive contrasting colorations of different chemical structures, e.g., lignin and cellulose. One of the most popular differential staining protocols is based on the Safranin O / Astra Blue dyes combination. Safranin O is a water-soluble basic dye that stains lignin in red. Astra Blue is also a water-soluble dye but having an acidic reaction, which stains cellulose in blue. Usually, a 1–2 % solution of Safranin O in distilled water or 50–70 % ethanol is applied in combination with the 0.5–1 % water solution of Astra Blue to detect lignified structures and obtain contrasting pictures convenient for the light microscopy. For a long time, Astra Blue was used exclusively with water solutions, and such recommendation without additional options is indicated on producers’ web sites. However, in 2002 it was proposed to use 1 % Astra Blue solution in 95 % ethanol to identify the lignified tissues. Later, such an ethanol solution of Astra Blue was also successfully applied by other researchers for different experimental purposes.We tested the modified staining protocol with the application of both 1 % Safranin O and Astra Blue solutions in a slightly lower concentration of ethanol (70 %) on the flower buds of Gagea lutea (Liliaceae) and found it working well. We believe that such a modified protocol with the solutions of these two dyes in 70 % ethanol allows simplifying the procedure of the plant material staining due to application of the same concentrations of dissolvent and reducing the difference in solvent concentration between two following contrasting staining solutions. Such differential staining can be effectively applied for plant histology purposes, especially where there is a need to distinguish lignified structures and secretory tissues.

Revisiting the Karyotypes of Alligators and Caimans (Crocodylia, Alligatoridae) after a Half-Century Delay: Bridging the Gap in the Chromosomal Evolution of Reptiles

Although crocodilians have attracted enormous attention in other research fields, from the cytogenetic point of view, this group remains understudied. Here, we analyzed the karyotypes of eight species formally described from the Alligatoridae family using differential staining, fluorescence in situ hybridization with rDNA and repetitive motifs as a probe, whole chromosome painting (WCP), and comparative genome hybridization. All Caimaninae species have a diploid chromosome number (2n) 42 and karyotypes dominated by acrocentric chromosomes, in contrast to both species of Alligatorinae, which have 2n = 32 and karyotypes that are predominantly metacentric, suggesting fusion/fission rearrangements. Our WCP results supported this scenario by revealing the homeology of the largest metacentric pair present in both Alligator spp. with two smaller pairs of acrocentrics in Caimaninae species. The clusters of 18S rDNA were found on one chromosome pair in all species, except for Paleosuchus spp., which possessed three chromosome pairs bearing these sites. Similarly, comparative genomic hybridization demonstrated an advanced stage of sequence divergence among the caiman genomes, with Paleosuchus standing out as the most divergent. Thus, although Alligatoridae exhibited rather low species diversity and some level of karyotype stasis, their genomic content indicates that they are not as conserved as previously thought. These new data deepen the discussion of cytotaxonomy in this family.

Toxicological Characterization of Ten Medicinal Plants of the Beninese Flora Used in the Traditional Treatment of Diarrheal Diseases

The use of medicinal plants in traditional medicine is a common practice in developing countries. However, this unregulated or poorly rational use may present a dose-dependent risk of toxicity to humans. This study aimed to explore the phytochemical and toxicological characteristics of ten (10) plant species used in the traditional treatment of infectious diarrhea in Benin. The acute toxicity of aqueous and hydroethanolic extracts of Khaya senegalensis, Daniellia oliveri, Rauvolfia vomitoria, Vernonia amygdalina, Manihot esculenta, Ocimum gratissimum, Senna italica, Diospyros mespiliformis, Pterocarpus erinaceus, and Anacardium occidentale was evaluated following the OECD 423 protocol at a single dose of 2000 mg/kg. This safety test was complemented by a larval cytotoxicity test. Hematological and biochemical examinations, as well as a histological study of the liver and kidneys, were performed. Larval cytotoxicity was assessed by the sensitivity of Artemia salina larvae to different concentrations of the plant extracts studied. Testing for chemical compounds was performed on the basis of differential staining and precipitation reactions. The mean lethal concentration (LC50) was determined by the probit method. The qualitative phytochemical screening of the plants studied revealed the presence of catechic tannins, gallic tannins, flavonoids, anthocyanins and sterol-terpenes, alkaloids, saponosides, and reducing compounds. This composition varied according to the plants studied. Acute toxicity data indicated that there was no mortality and no structural and functional alterations of the liver and kidneys of treated animals. Larval cytotoxicity data suggest that the plants studied are not cytotoxic (LC50 ≥ 0.1 mg/mL). These observations reflect the safety of these plants and justify their use in traditional medicine in the treatment of many diseases including diarrheal diseases.

Vascular Connections Into the Grape Berry: The Link of Structural Investment to Seededness

Vascular bundles in the grape pedicel and berry contain the conduits, phloem and xylem, for transport of water, sugar, nutrients and signals into and through the grape berry and play a critical role in berry growth and composition. Here, we assess the vascular anatomy within the proximal region of the berry. Guided using a 3D berry model generated by micro-CT, differential staining of transverse sections of berries and receptacles was followed by fluorescent microscopy. Morphometric and vascular characteristics were analyzed within the central proximal region (brush zone, a fibrous extension from the pedicel vascular system into the berry) of the seeded cultivars Shiraz and Sauvignon Blanc, as well as the stenospermocarpic cultivars Ruby Seedless and Flame Seedless. Observations revealed a change in vascular arrangement from the receptacle into the berry brush zone and differences in xylem element size as well as xylem and phloem area relationships. Xylem anatomical and derived hydraulic parameters, as well as total tissue area of xylem and phloem varied between cultivars and in receptacle and berry components. Variation in vascular growth between grape pedicels and berries was independent of seededness. Differences in receptacle xylem vessel size and distribution could contribute to cultivar-dependent xylem backflow constraint.

KEMAMPUAN BAKTERI ASAM LAKTAT DALAM MENGHAMBAT Salmonella thypii

Lactic acid bacteria are known to have the ability to produce antimicrobial compounds known as bacteriocins. This study aims to determine the ability of lactic acid bacteria to inhibit the growth of Salmonella thypii bacteria. In this study, the lactic acid bacteria used were a collection of the Biology Laboratory of the University of Medan Area. Eight types of lactic acid bacterial isolates were used, including IN01, IN02, SF01, SF02, NN01, NN02 SPU01 and SPU04. A confirmation test was carried out on the growth of lactic acid bacteria and Salmonella thypii by visual and microcopic confirmation through differential staining. The results showed that all lactic acid bacteria were a group of gram-positive bacteria in the form of bacilli. The survival of lactic acid bacterial isolates in the environment with variations in pH and salt levels was indicated by the number of colonies that grew after the incubation period. The antagonistic ability of lactic acid bacteria in suppressing pathogenic bacteria was proven by the presence of an inhibition zone in the form of a clean area around the pathogenic colony. It is known that the SPU01 and SF01 bacterial isolates have the highest inhibition zone values, namely 29.79 mm and 28.13 mm.

Cytogenetic studies of three cultivated hill cotton (Gossypium arboreum L.) varieties from Bangladesh

Three hill cotton (Gossypium arboreum L.) varieties viz., HC-1, HC-2 and HC-3, released by Bangladesh Cotton Development Board were investigated through orcein, CMAand DAPI-banding for cytogenetical characterization and to elucidate the karyotypic diversity among these varieties. All these three varieties were found to possess 2n = 26 metacentric chromosomes with ‘1A’ karyotype. Based on TF%, AsK% and Syi index, HC-3 was little advanced over HC-1 and HC-2. These three varieties showed differential Chromomycin A3 (CMA)- and 4Ê-6 Diamidino-2-Phenyl Indole (DAPI)-banding patterns and a tendency of acumulation of repetitive sequences at the terminal regions was observed. Despite possessing same somatic chromosome number these three hill cotton varieties could be characterized by diversified karyotypic parameters through differential staining.