Microscopic Preparations Research Articles

Russulaceae of the Pakaraima Mountains of Guyana 5. Two newly described diminutive species in a novel lineage of the crown clade of Russula (Russulaceae)

Much of our previous work documenting fungal biodiversity in Guyana has been centered in forests of Dicymbe corymbosa Spruce ex Benth., which forms monodominant stands in the white sands region of Guyana. In order to begin to assess fungal biodiversity associated with other ectomycorrhizal host trees found in Guyana, expeditions were conducted to areas containing Dicymbe altsonii and Pakaraimaea dipterocarpacea. In this paper, we describe two new species of Russula from Guyana with diminutive basidiomata: R. lilliputia and R. pakaraimaeae, found in association with these tree hosts, using a combined approach of morphological characterization and molecular analyses. Together with R. gelatinivelata, which we previously described from Guyana, R. lilliputia and R. pakaraimaeae form what appears to be a new lineage within the crown clade of Russula, sister to the subsect. Auratinae. Synapomorphies for the lineage include a pruinose to subtomentose pileus in dry conditions, suprapellis comprised of inflated cells that give rise to long cylindrical or digitate, and sinuous, thin or thick-walled pileocystidia that are strongly emergent, easily disarticulate in microscopic preparations, and have contents that are acid resistant in Basic Fuchsin, pruinose to subomentose patches on the stipe, suprahilar plage on the basidiospores, spore ornamentation of mostly isolated verrucae, and geographic distribution restricted to the Pakaraima Mountains of Guyana. Full descriptions, photos, line drawings of microscopic features, and comparisons with related species are provided.

Microbial Biocontrol Agents and Natural Products Act as Salt Stress Mitigators in Lactuca sativa L.

One of the major problems related to climate change is the increase in land area affected by higher salt concentrations and desertification. Finding economically and environmentally friendly sustainable solutions that effectively mitigate salt stress damage to plants is of great importance. In our work, some natural products and microbial biocontrol agents were evaluated for their long-term effectiveness in reducing salt stress in lettuce (Lactuca sativa L. var. romana) plants. Fourteen different treatments applied to soil pots, with and without salt stress, were analyzed using biometric (leaf and root length and width), physiological (chlorophyll and proline content), and morphological (microscopic preparations) techniques and NGS to study the microbial communities in the soil of plants subjected to different treatments. Under our long-term experimental conditions (90 days), the results showed that salt stress negatively affected plant growth. The statistical analysis showed a high variability in the responses of the different biostimulant treatments. Notably, the biocontrol agents Papiliotrema terrestris (strain PT22AV), Bacillus amyloliquefaciens (strain B07), and Rahnella aquatilis (strain 36) can act as salt stress mitigators in L. sativa. These findings suggest that both microbial biocontrol agents and certain natural products hold promise for reducing the adverse effects of salt stress on plants.

Scots pine roots (Pinus sylvestris L.) as dendrogeomorphological indicators of soil erosion on a hiking trail in the Brodnica Lakeland, Poland NE

Trampling on hiking trails leads to vegetation degradation, promoting soil erosion. Erosion removes the topsoil, causing lasting changes and exposing tree roots. Yet, little is known about the environmental factors affecting soil erosion rates determined from tree root analysis. This study, exclusively utilizing microscopic preparations, examines tree root anatomical changes on a lowland trail in Brodnica Lakeland. It aims to understand the temporal and spatial variability of soil erosion rates using dendrogeomorphological records of Scots pine (Pinus sylvestris L.) roots. Up to 64 % of samples show exposure due to continuous denudation, significantly shaping the trail’s relief. Abrupt exposures and secondary growth occur in only 22 % and 14 %, respectively. The average soil erosion rate, calculated from 76 root exposures at the Bachotek site, is 1.93 ± 0.82 mm/year, ranging from 0.71 to 3.80 mm/year, with recently exposed roots exhibiting the highest average soil erosion values on the hiking trail. 45 % of root samples exhibit above-average erosion rates. Statistical analyses reveal that erosion rates (ER) are influenced by soil compaction (COM), bulk density (BULK), root orientation (ASP), and exposure length (EXP). Higher compaction and bulk density correlate with increased erosion, while roots parallel to the trail experience more erosion. Conversely, greater root exposure length leads to decreased erosion rates. These findings suggest that soil erosion on the hiking trail primarily results from gradual geomorphological processes. Wood anatomical analyses of pine roots confirm the role of trampling in initiating and accelerating these processes, altering the trail’s subsurface soil layers, and indirectly affecting soil erosion dynamics. Scots pine roots prove valuable as indicators of the temporal and spatial variability of soil erosion along the lowland trail.

Combined underwater endoscopic and microscopic surgery for tympanic paraganglioma: A case report.

The resection of middle ear paragangliomas can be challenging given their vascular nature and the small volume of the tympanic cavity, particularly when the tumor in the hypotympanum is close or attached to the internal carotid artery (ICA). We performed combined underwater endoscopic and microscopic surgery for a Class B1 middle ear paraganglioma according to the modified Fisch classification. The suspicious bone in the hypotympanum and around the petrous ICA was drilled with underwater endoscopy. The feeding arteries, the caroticotympanic and inferior tympanic arteries, were suctioned and cauterized under microscopy. To the best of our knowledge, no case of middle ear paraganglioma treated with underwater endoscopy has been reported. Underwater endoscopy, providing a clear operative field with blood and bone dust irrigation, is a good indication for middle ear paragangliomas. In contrast, microscopic preparation for unexpected bleeding is important, particularly when the tumor closely extends to vital structures, such as the ICA or the jugular bulb.

Geology and characteristics of petrographic rocks in the region of Trepça, Kosovo

Purpose. The purpose of this paper is to accomplish a thorough identification of rock types occurring within the Mitrovica region by describing in detail all encountered varieties. The authors aim to determine the discontinuity or continuity of all inter-formational boundaries, crucial for accurate delineation on the ground and complete reflection on a 1:25000 scale map. Additionally, the objective is to identify the nature of contact between rock types and provide a detailed their description. Methods. Field exploration in the Mitrovica region was conducted for several months. Rock identification involved detailed sampling and petrographic analysis, including thin section preparation of magmatic rocks. The method included sample preparation, polarized light source interactions with minerals and observation of optical properties. Key properties observed are birefringence, interference colors, extinction angles, identification and analysis. Findings. Based on the study of stratigraphic units and geological descriptions of mineral outcrop areas, various types of rocks through petrographic microscope preparation, as well as chemical and geochemical analyses, have been differentiated. The Mitrovica area encompasses the following lithostratigraphic units: harzburgite, gabbro, diabase, metasandstone, sandstone, quartzite and grainstone with calcium (Aeolisaccus sp.; Bioclastic grainstone with dacycladal algae and small milliolides; Salpin-goporella sp.). Originality. The originality of the research lies in employing an optical microscope for the precise identification of rocks. Through the re-search carried out in the study area, we obtained a comprehensive petrographic description of mineral composition, texture, and mineralization, facilitating the assessment of the area exploitation potential. Practical implications. The petrographic exploration yielded the conclusion that the Pb-Zn mineralization is present in the study area, which is a sig-nificant finding for the advancement of the mining sector and the local community, provided that environmental preservation measures are upheld and responsible methods of area exploitation are implemented.

Potential of Hibiscus (Hibiscus rosa-sinensis L.) Extract as a Substitute for Eosin in Hematoxylin-Eosin Histological Staining

Staining is an important stage in preparing microscopic preparations so that the histological structure can be observed clearly. Hematoxylin-Eosin is the most widely used dye. Unfortunately, eosin is a synthetic chemical compound that is carcinogenic. To reduce the health and environmental impacts of eosin compounds, it is important to look for alternative dyes from natural ingredients. Hibiscus (Hibiscus rosa-sinensis L.) is a plant known to contain anthocyanins which can be used as eosin substitutes. This study aims to determine the potential of hibiscus extract as a substitute for eosin in histological staining. In this study, unstained liver tissue preparations from mice (Mus musculus) were used. Hibiscus extract dye was compared with eosin as a control. The color quality of the tissue preparations was used as an experimental parameter. The results of the microscopic assessment showed that liver tissue stained with Hibiscus extract at concentrations of 60% and 80% was less good than eosin. However, because tissue can stain reddish, hibiscus extract has the potential to be used as an eosin substitute. The right extraction method may be able to produce intact anthocyanin so that it can color the tissue optimally.

Correlation Between I/T Ratio, S100B, Interleukin-10 Serum, and NEWS2 Score in Sepsis

Immature/total neutrophils (I/T) ratio is a simple and inexpensive method used as an early sign of bacterial infection. Meanwhile, the S100B protein is found in glial cells and functions as a neuroprotection. Then, the interleukin-10 cytokine acts as an anti-inflammatory cytokine during infection. In addition, the NEWS2 score is used to see the early and fast changes in patients with sepsis. The purpose of the study was to prove the correlation between the I/T ratio, S100B, IL-10, and NEWS2 score in patients with sepsis. A cross-sectional study was conducted on 34 patients with sepsis from September 2020 to February 2021. I/T ratio was calculated in 100 leukocytes from microscopic blood smear preparation. Levels of S100B and IL-10 serum were measured using the ELISA method. Pearson correlation test was used for normally distributed data, and the Spearman correlation test was utilized for abnormally distributed data. Correlation test between I/T ratio, S100B, and IL-10 with NEWS2 score respectively showed values of r=0.58; P=0.01, r=0.36; P=0.03, and r=0.39; P=0.02, in which P<0.05. There was a moderate positive correlation between I/T ratio and NEWS2 score, a weak positive correlation between S100B and NEWS2 score, and a weak positive correlation between IL-10 and NEWS2 score.

An alternative method for SARS-CoV-2 detection with use modified fluorescent in situ hybridization

The real-time reverse-transcriptase polymerase-chain-reaction (rRT-PCR) tests are the gold standard in detecting SARS-CoV-2 virus infection. However, despite high sensitivity and specificity, they have limitations that in some cases may result in false negative results. Therefore, it is reasonable to search for additional tools that could support microbiological diagnosis of SARS-CoV-2. The aim of the study was to develop a highly specific molecular test capable of detecting and visualizing SARS-CoV-2 infection. A universal probe and a set of 18 specific oligonucleotides with a FLAP sequence attached to them on both sides were designed to visualize SARS-CoV-2 virus infection based on the fluorescence in situ hybridization method (FISH). FISH conditions using the developed kit were standardized on the Vero CCL-81 cell line infected by SARS-CoV-2 virus. The method was tested on 290 nasopharyngeal swabs (collected in a doublet) from patients with clinical symptoms of SARS-CoV-2. Each one swab from the doublet was subjected to RNA isolation and amplification by rRT-PCR. From the second swab, a microscopic preparation was performed for FISH. The use of the rRT-PCR allowed obtaining 200 positive and 90 negative results, while our FISH method allowed for 220 positive results and 70 negative results. The differences obtained using both methods were statistically significant (p = 0.008). The obtained results support the use of FISH as an additional method in microbiological diagnostics of SARS-CoV-2.

The Influence of Environmental Factors on the Incidence of Malaria on the Coast of Malacca Regency

Environmental conditions that support malaria transmission include the physical environment such as temperature, air humidity, rainfall, wind speed, altitude of a place or location and sunlight. The biological environment which is the environment of flora and fauna such as: mangroves, mosses and algae can affect the life of mosquito larvae. The presence of plants can protect larvae from sunlight and attacks from other living things. The mosquito population in an area is also determined by the presence of various types of larva-eating fish such as tin head fish, tilapia and tilapia. The presence of large livestock such as cows and buffaloes can reduce the number of mosquito bites in humans if the cage is located close to the house. The purpose of this study is to determine and analyze the Influence of Environmental Factors on the Incidence of Malaria on the Coast of Malacca Regency. The type of research used is quantitative research with Case Control design. The population in the study was all people living in coastal areas covering the working areas of the Weoe and Lamea health centers who carried out microscopic and RDT thick drops blood preparation examinations based on malaria register data at the Puskesmas in 2020. The case sampling technique in this study was to use a total sampling technique saturated with a total of 73 local positive malaria cases (indigenus) recorded in 2020 at the Weoe and Alkani health centers, while the control sample was set at a ratio of 1: 1, so that a large sample of 73 people was obtained. So the total sample number is 146 people. The data was then analyzed using a simple logistic regression test. The results showed that there was an influence of mosquito breeding sites, mosquito breeding sites on the incidence of malaria on the coast of Malacca Regency with a p.value value<α (0.05). While the variables Temperature, Humidity, Large livestock ownership had no influence on the incidence of Malaria (p > 0.05).

Distribution and FTIR-based fingerprint of secondary metabolites in different organs of ant-plant (Myrmecodia tuberosa)

Abstract. Lestari NB, Sulistyaningsih YC, Umar AH, Ratnadewi D. 2024. Distribution and FTIR-based fingerprint of secondary metabolites in different organs of ant-plant (Myrmecodia tuberosa). Biodiversitas 25: 1104-1115. Myrmecodia tuberosa Jack, or ant-plant, is widely used as a traditional medicine. The plant has the potential to be antimicrobial, anticancer, and antioxidant. Its domatium is believed to contain valuable substances, rendering people to collect the organ that devastates the whole plant. Therefore, the distribution and nature of the major substances in the leaf, petiole, and domatium must be investigated. This study aimed to determine the secondary metabolites and their accumulation sites through anatomy and histochemistry. Fourier-transform infrared (FTIR) and chemometrics supported the determination. Microscopic preparations made observations of the anatomy and secretory structures of transverse and paradermal sections. Histochemical studies were performed with specific reagents. M. tuberosa has a hypostomatic leaf blade with parasitic stomata. The main secretory structure is the idioblast. Idioblasts are scattered over various tissues, such as the epidermis, hypodermis, palisade, spongy mesophyll, and cuticle of the leaf blade, in the epidermal tissue, chlorenchyma cortex, and cuticle of petiole and the periderm, cortex, and pith of domatium. Terpenoids, phenols, lipophilic compounds, essential oils, and flavonoids were found in the leaf blade, petiole, and domatium, whereas alkaloids were present only in the petioles. FTIR demonstrated the main functional groups in the three organs are similar, while the principal component analysis distinguished domatium from leaf blade and petiole.

Long-term organogenic callus cultivation of Ranunculus illyricus L.: a blueprint for sustainable ex situ conservation of the species in urban greenery

The growing trend of introducing wild plant species into urban environments necessitates the identification of novel species adapted to prevailing conditions. A promising reservoir of such species may be xerothermic communities where Ranunculus illyricus occurs. This study aimed to establish a micropropagation protocol for R. illyricus using indirect organogenesis. The protocol includes initiation of culture from various explants, callus proliferation, shoot regeneration, multiplication, and concurrent rooting. Callus appeared on most types of vegetative explants tested, but stolons were considered the best due to their good availability, high disinfection (85%), and robust callus production (maximum increase − 363.1%). The growth rate of the callus fresh matter (CFM) obtained from stolons was calculated. Greater CFM was obtained on the medium with the supplemented picloram 8.0 mg L− 1 with kinetin 5.0 mg L− 1 and in second part of experiment on medium with the addition of 2,4-D (2,4-dichlorophenoxyacetic acid) 2.0 mg L− 1 alone or picloram 6.0 mg L− 1 with kinetin 8.0 mg L− 1. Shoot organogenesis was observed on macronutrients B5 (Gamborg medium), micronutrients MS (Murashige and Skoog) medium with the addition of 2.0 mg L− 1 IBA (indole-3-butyric acid) and 4.0 mg L− 1 BAP (6-benzylaminopurine). To document the process of callus differentiation, microscopic preparations were prepared. Subsequently, the regenerated plants underwent acclimatisation and their growth in an ex situ collection was monitored over three growing seasons. In particular, in vitro-origin plants exhibited developmental patterns similar to those of their seed-origin counterparts. The incorporation of R. illyricus into urban landscapes not only increases aesthetic appeal, but also ensures the preservation of valuable genetic resources for this rare species, potentially contributing to effective ex situ conservation in the future. This marks the first scientific report on in vitro cultures of R. illyricus.

Histology of the digestive tract of the endangered Salmo obtusirostris (Teleostei: Salmonidae) from the Buna river (Neretva river basin, Bosnia and Herzegovina)

Morphological and histological studies are very useful for the characterization of the digestive tract, which provides important information for understanding the feeding habits and digestive physiology of fish. This paper presents the results of research of the histological structure of the digestive system of the endemic fish Salmo obtusirostris (Heckel, 1851) from the Buna River (Neretva River basin, Bosnia and Herzegovina). According to IUCN Red List, S. obtusirostris is considered Endangered B2ab (v) ver. 3.1, with decreasing population trend throughout its distribution area. A specimen (TL = 265 mm, SL = 245 mm, m = 251.52 g) of the species S. obtusirostris was caught in the Buna River in May 2019. As the basic material, digestive tract of the above-mentioned species was used, divided into esophagus, stomach, pyloric appendages, intestine, as well as the digestive glands, the liver and the pancreas. By using basic histological techniques, microscopic preparations were made from the tissues. When studying the microscopic structure of the organs of the digestive system of S. obtusirostris, it was established that the majority basically consists of mucosa (tunica mucosa), submucosa (tunica submucosa), muscular layer (tunica muscularis) and outer layer (tunica serosa). The histological structure of the digestive system of S. obtusirostris Heckel, 1851 is consistent with its diet.

Assessment of Proteinase and Phospholipases Enzymes Isolated from Pathogenic Candida species from Women Attending Antenatal Clinic at Mbeya Zonal Referral Hospital, Southern Highland Regions of Tanzania

Fungi are a global cause of vaginal infections, with vaginal candidiasis largely afflicting tropical regions including most parts of sub-Saharan Africa. In Tanzania, baseline studies have established the prevalence of Candida species among symptomatic and asymptomatic pregnant women in Dar es Salaam (Namkinga et al., 2005; Namkinga 2012) and in Mwanza (Mushi et al., 2019). However, no study that has been done to determine the above-mentioned enzymes form Candida species in Southern Highland Regions of Tanzania. Several factors have been reported to contribute to the virulence and pathogenicity of Candida, among others the production of extracellular hydrolytic enzymes, particularly phospholipase and proteinase. Therefore, this study was designed to investigate the in vitro production of phospholipases and proteinases enzymes from isolated pathogenic Candida spp from pregnant women.Of the 280 samples collected from symptomatic and asymptomatic pregnant women attending antenatal clinics at Mbeya Zonal Referral Hospital were tested for several microbiological methods such as the wet mount microscopic preparations (hanging drop) and the Gram stained smears for microscopy, cultures on Sabouraud’s dextrose agar and on Conidia enhancing medium (Corn meal agar), germ tube test, biochemical tests, sugar fermentation-assimilation tests and molecular test to characterize Candida species.Phospholipase production was performed in egg yolk medium while the production of proteinase was done in a medium containing bovine serum albumin. All analyses were performed in triplicates. The results showed that; out of the 280 tested, 155 (74.3%) isolates were phospholipase positive while 201 (96.6%) were positive for proteinase activity. C. albicans was the species with the highest number of positive isolates for proteinase and phospholipase 110 (95.6%), and most strains of C. albicans produce both enzymes (phospholipases and proteinases). The non-albicans Candida isolates were also producers of hydrolytic enzymes that, consequently, might be able to cause infections as favorable conditions arises.

Prediction of lymph node status in patients with surgically treated head and neck squamous cell carcinoma via neck lavage cytology: A pilot study.

Neck dissection is a standardized surgical procedure for patients with head and neck squamous cell carcinoma (HNSCC) and plays a critical role in the choice of adjuvant treatment based on histopathological findings. Saline irrigation is routinely performed at the end of surgery. However, this irrigant is not used for diagnostic purposes. Intraoperative irrigation of the neck dissection wound was performed in 56 patients with HNSCC (N=93 neck dissections), and the cytological suspension obtained was processed via the liquid-based cytology (LBC) technique, Papanicolaou staining, and immunocytochemical staining. Microscopic preparations were screened for the presence of tumor cells and classified as positive, borderline, or negative. These results were correlated with the histopathological and clinical data. Neck lavage LBC demonstrated high diagnostic value in detecting lymph node metastases (N+) with extracapsular spread (ECS), with a specificity, sensitivity, negative predictive value, and positive predictive value of 93.1%, 100%, 100%, and 80%, respectively. Tumor cells were detected in 4.8% of N- cases, 20% of N+ cases without ECS, and 100% of N+ cases with ECS. Receiver operating characteristic curve analysis showed an area under the curve of 0.8429 for the prediction of N+ (p<.0001) and 0.9658 for the prediction of N+ with ECS (p<.0001). Differential lavage cytology can provide valid and rapid information on the lymph node status in patients with HNSCC and showed an excellent correlation with histopathology. Thus, neck lavage LBC may facilitate faster and more reasonable planning of adjuvant treatment and help improve the therapeutic management of patients with HNSCC.

Histomorphological Comparison of Tissues Fixed in Conventional Formalin and Eco-friendly Jaggery Solution: A Cross-sectional Study

Introduction: Tissue fixation is an essential step in the microscopic preparation of tissues to preserve them by preventing autolysis, bacterial putrefaction, and increasing the tissue's refractive index. The conventional fixative used is 40% formaldehyde. Short-term exposure to formaldehyde can cause irritation to the eyes and respiratory tract, leading to breathlessness and coughing. The International Agency for Research on Cancer (IARC) has classified formaldehyde as a Class 1 human carcinogen capable of potentially causing various neoplasms, including nasopharyngeal carcinoma. Therefore, an innovative approach is being explored to replace formalin with harmless and eco-friendly jaggery. Jaggery possesses cytoprotective, antioxidant, and tissue-preservative properties. At a low pH, the fructose in jaggery breaks down into aldehydes, which cross-link with tissue amino acids, resulting in tissue fixation similar to formaldehyde. Aim: To compare the histological and gross morphological features of tissues fixed in formalin and jaggery. Materials and Methods: This cross-sectional study was conducted in Department of Pathology of a tertiary care centre. Surgical specimens obtained fresh, which were not suspicious of malignancy, were included. A sample size of 23 was selected. Surgical specimens already placed in formalin were excluded. A 30% jaggery solution was prepared by dissolving 300 grams of finely powdered jaggery in 1000 mL of distilled water, which was then filtered using filter paper. A 40% formaldehyde solution was used to prepare a 10% formalin fixative. Tissue bits from each specimen were cut into two halves and placed in formalin and jaggery separately. After 24 hours of fixation, the tissue bits were evaluated for gross morphological features, including tissue shrinkage, consistency, and colour. Tissue shrinkage was classified as mild, moderate, or marked, while consistency ranged from soft to firm to hard. The colour of the specimens varied case by case. Stained slides from jaggeryfixed and formalin-fixed tissues were assessed for histological parameters, such as nuclear details, cytoplasmic details, cellular outline, and overall staining quality. A blinded method was used to compare the stained slides using a microscope. Ratings were assigned to each case on a scale of 1-4. The data were statistically analysed using R software. Results: When comparing the formalin-fixed and jaggery-fixed specimens, no significant differences were observed in gross morphological features. All jaggery-fixed specimens appeared brown grossly. Histological features also showed no significant difference (p-value&gt;0.05) except for cytoplasmic details. Therefore, it was observed that tissue preservation using the jaggery solution was comparable to that of formalin. Conclusion: The tissue-preservative properties of jaggery are on par with formalin. Hence, jaggery solution can be used as an eco-friendly substitute for formalin. Further research with larger sample sizes can pave the way for the effective replacement of hazardous formalin with natural jaggery

Secretory Structures, Histochemistry, and Antibacterial Activity of Macaranga gigantea

Macaranga gigantea is a member of the family Euphorbiaceae. Anak Dalam tribe, an indigenous people that live in Bukit Duabelas National Park Jambi province, used to utilize the bark of M. gigantea to treat diarrhea. This study aimed to identify the secretory structure, histochemical aspects, and antibacterial potency of M. gigantea bark. Microscopic preparations have been made for the observation of secretory structures. Histochemical testing uses Wagner reagent, cupri acetate, ferric trichloride, and aluminum trichloride. Antibacterial activity was tested using the well diffusion method. The type of secretory structure was carried out. M. gigantea bark has idioblast cells as a secretory structure. The idioblast cells are distributed in the cortex. The idioblast cells contain phenolic compounds, alkaloids, and terpenoids. The well diffusion method was used to test different concentrations of bark extracts. According to the findings, M. gigantea bark extracts at 100 mg/mL had the best inhibitory activity against Staphylococcus aureus and Escherichia coli, with a maximal inhibition zone

Forensic medical cytological examination of the trace evidence on the knife’s blade in the case of multiple penetrating stab/cut wounds in the chest with lung damage: a case report

Various trace evidence are often found on the instruments of mechanical injury, including microparticles of humans damaged organs and tissues. The detection of cells and microparticles of organs and tissues on the instrument of injury confirms its contact with the human body, which is of great evidentiary value.&#x0D; The article presents the results of forensic medical cytological examination of biological evidence taken from the knifes blade in the case of multiple penetrating stab/cut wounds in the chest that result in lung damage. Microscopic preparations were made from the traces on knifes blade, which were stained with fluorochrome solutions (0.0005% aqueous solution of atebrine and 0.01% solution of acridine orange). The study was performed using the luminescent microscope Leica DM2500 (20 and 60 lenses, 10 eyepiece). The examination of microparticles of the striated skeletal muscle revealed areolar connective and fat tissues with capillary and arteriole fragments. The isolated mesothelial cells and type II pneumonocytes were also detected. Fibers of the striated skeletal muscle tissue looked like long multinucleated cells with distinguishing cross-striation. The areolar connective tissue was represented by loosely arranged fibers embedded in the ground substance. Fragments of the fat tissue looked like the clusters of round and oval large cells, tightly pressed together, with capillaries, and arterioles in between. Y-chromatin was detected in the nuclei of most cells. In addition, human blood was found on the knifes blade.&#x0D; Thus, the study revealed that tiny particles of tissues and isolated human cells of the male genetic sex were detected, their organtissue affiliation was established, which is a valuable diagnostic finding, because it confirms the contact of the trauma instrument with the human body.

ANATOMY OF THE ENDEMIC ALYSSUM PATERI SUBSP. PATERI

Objective: Alyssum pateri Nyár. subsp. pateri is an endemic plant known as "kanatlı kevke" in Türkiye. The plant is perennial and in semi-shrub form. The plant is traditionally used externally in the treatment of rheumatism in Bingöl (Türkiye). Microscopic analysis of the anatomical structures of plants can provide useful information for taxonomic classification. Light microscopy analysis is a widely used and effective method for the identification of medicinal plants. In this study, the anatomical features of the A. pateri subsp. pateri were examined. Material and Method: The plant material was collected from Ankara (Türkiye). The samples were protected in 70% alcohol. The cross and surface sections were cut by hand with a razor blade into microscopic preparation form. A Leica DM 4000B microscope was used for anatomical analysis and micro photographs. Result and Discussion: The anatomical characters of the leaf, petiole and stem of the Alyssum pateri subsp. pateri were revealed. The leaf is dorsiventral and contains 1-2 rows of palisade parenchyma. Cruciferous stomata and stellate hairs are located on the lower epidermis. The petiole cross-section is sulcate and stellate hairs are observed in the epidermal layer. The main vein is arc-shaped and is accompanied by lateral veins. The stem is disc-shaped. Stem epidermis contains stellate hairs. The interfascicular tissue between the xylem strands of the vascular bundles is composed lignified cells. Vascular bundles are surrounded by a pericyclic sclerenchymatous cap.

Walther Flemming: life path and scientific legacy

The main stages of Walther Flemming life and activity are considered in the article. Walther Flemming (21.04.1843–4.08.1905) is German biologist, anatomist, one of the most famous histologists of the 19th century and the founder of cytogenetics. He studied medicine at four famous universities of the German Empire – in Göttingen, Tübingen, Berlin and Rostock. After defending his thesis, in 1868 he started teaching anatomy and histology at Würzburg, Amsterdam and Rostock universities, Germany and Charles Ferdinand University, the Czech Republic. During the last 25 years of his life, he was a professor of anatomy at the Christian Albrecht University in Kiel, Germany and headed one of its departments – the anatomical institute.&#x0D; At the beginning of W. Flemming scientific activity, his research mostly concerned histology and anatomy, as well as the development of histological methods and fixation of microscopic specimens. However, the main works of the scientist are devoted to study cell division process and the chromosomes distribution in daughter nuclei. He firstly introduced the concept of “mitosis”, which he studied on stained microscopic preparations, using salamander epithelial cells as a source of biological material. W. Flemming described the processes in the nucleus, distinguished “progressive” and “regressive” phases of cell division. He demonstrated that the cells contain areas that absorb basophilic dyes. These areas were later called chromatin; other structures remained unstained under such conditions and were therefore called achromatin. W. Flemming studied the structure and functions of the spindle during cell division, and he also studied the polar structure, which he called centriole. The scientist found that indirect division usually occurs not only in vertebrates, but also in invertebrates and protists. He established that two successive, morphologically distinct divisions of nuclear occur during spermatogenesis, resulting in the division of chromatin into two daughter cells. Thus, W. Flemming’s research contributed to the development of cell biology and its branches.

CELLULAR COMPOSITION OF THE LYMPHOID STRUCTURES OF THE HUMAN INTRAHEPATIC BILE DUCTS

he aim of our study was to investigate the cellular composition of the lymphoid structures of the human intrahepatic bile ducts. Materials and Methods: The object of the study was the lymphoid structures of the walls of the intrahepatic bile ducts obtained from 45 cadavers of different ages. Lymphoid nodules in the walls of the intrahepatic bile ducts were investigated by T. Hellman’s method. The microanatomy of lymphoid structures was studied on microscopic preparations. Microscopic preparations were stained according to Van Gieson, with hematoxylin-eosin, methylene blue. The digital data obtained during the study was subjected to statistical processing. Results: A macromicroscopic study of lymphoid formations in the intrahepatic bile ducts showed that lymphoid formations in the walls of these organs are represented by lymphoid nodules and diffuse lymphoid tissue. The peripheral contours of the lymphoid nodules are clearly defined. Histologically, the cellular composition of lymphoid nodules and diffuse lymphoid tissue was studied. In the mucous membrane and in the submucosa, small, medium sized lymphocytes, and large lymphocytes, macrophages, plasma, reticular, mast cells, cells in a stage of mitosis, degeneratively altered (destructive), and other forms of lymphoid cells are determined. Conclusions: According to our data, newborns have not only lymphoid nodules, but also a typical cellular composition in the lymphoid tissue, which has a completely “mature character”. The results of morphometric studies showed that in early childhood there is a high percentage of small, medium, large lymphocytes, macrophages, cells with signs of mitosis.