Brown Staining Research Articles

Correlation of MMP-9 (Matrix Metalloproteinase 9) and SMA (Smooth Muscle Actin) expression with Basal Cell Carcinoma’s risk of recurrence

Basal Cell Carcinoma (BCC) is the most common cancer in many countries, with its incidence steadily rising. Data from the Indonesian Pathologists Association show that among 1,530 skin cancer cases, BCC was the most frequent (39.93%), followed by Squamous Cell Carcinoma (23%) and Malignant Melanoma (7.9%). According to the 2018 WHO classification, BCC is divided into Low-Risk and High-Risk groups based on the risk of recurrence. BCC invades surrounding tissues through extracellular matrix degradation and enhanced cell motility, driven by the expression of MMP-9 (Matrix Metalloproteinase 9) and SMA (Smooth Muscle Actin). This study aims to evaluate the expression of MMP-9 and SMA in BCC using immunohistochemistry and analyze their correlation with the risk of recurrence. A total of 40 paraffin blocks from BCC patients diagnosed between 2019 and 2021 at Dr. Saiful Anwar General Hospital in Malang, Indonesia, were sampled. The blocks were sectioned, stained with MMP-9 and SMA antibodies, and examined for positive expression indicated by brown staining in the cytoplasm of tumor cell and peritumoral stroma. Spearman correlation showed r = 0.811 for MMP-9 and r = 0.857 for SMA (p < 0.05, CI: 95%), indicating a significant relationship between these markers and the risk of recurrence. Higher MMP-9 and SMA expression corresponded to a higher risk of BCC recurrence, suggesting their potential as prognostic biomarkers. However, further research is needed to assess recurrence risk more accurately and determine the most appropriate therapy.

Spatial distribution of remaining movable and non-movable oil fractions in a depleted Maastrichtian chalk reservoir, Danish North Sea: Implications for CO2 storage

Depleted oil and gas fields constitute potentially important storage sites for CO2 in the subsurface, but large-scale injection of supercritical (sc) CO2 in chalk has not yet been attempted. One of the risks is the adverse effect of the substantial amount of remaining oil in the chalk reservoirs on scCO2 injection. In order to counter an undesired effect on injectivity, a fundamental understanding of the spatial distribution and quantity of the movable, semi-movable, and non-movable oil, and solid bitumen/asphaltenes fractions of the remaining oil is critical. In this study a combination of organic geochemistry (gas chromatography of the saturated fraction and programmed pyrolysis), and reflected light microscopy was applied to evaluate and measure the spatial distribution, volume, and saturation of different oil fractions in a well-defined reservoir interval of a waterflooded Maastrichtian chalk reservoir in the Danish Central Graben, North Sea. A total of 127 samples from a slightly deviated vertical well and two ∼5 km-long horizontal wells from the Halfdan and Dan fields were analyzed. An original uneven distribution of oil saturation and composition or different production efficiency of different levels in the reservoir may account for variations in the total oil and oil fraction saturations. Gas chromatography shows that the solvent extractable oil is quite similar in composition, characterized by a dominance of polar compounds and a high content of asphaltenes. Extended slow heating (ESH) pyrolysis reveals that most of the remaining oil saturation consists of semi-movable oil and total non-movable oil (non-movable oil plus solid bitumen/asphaltenes). Reduced oil gravity values (API) are related to evaporation loss of the lightest hydrocarbon fraction during core storage and increase of the relative proportion of the heavier oil fractions by waterflooding during production. Microscopy disclosed three forms of oil: i) Patchy distributed lighter, movable oil showing a bluish fluorescence, ii) Brownish staining with a dark orange to brownish fluorescence, and iii) Dark brown non-fluorescing oil and black solid bitumen/asphaltenes occurring in microfossils and along deformation bands and stylolites, constituting the heavy non-movable oil fractions. There is a general correlation between bulk rock porosity and the total non-movable oil saturation. It thus appears that the heavy non-movable oil fractions preferentially occur in association with low-permeability heterogeneities within high-permeability stratigraphic intervals. These intervals appear to favor accumulation of non-movable oil and solid bitumen/asphaltenes and may carry a higher risk for impeding scCO2 flow.

7984 A Challenging Case Of Hyperphosphatemic Familial Tumoral Calcinosis

Abstract Disclosure: T.G. de Souza: None. T.J. Weber: None. Introduction: Hyperphosphatemic Familial Tumoral Calcinosis (hFTC) is a rare, autosomal recessive metabolic disorder characterized by elevated serum phosphate levels and the accumulation of calcium phosphate deposits in soft tissues. There are fewer that 150 genetically confirmed cases in the literature. There are no consensus guidelines; therapeutic approaches are guided by case reports or series. This case serves to heighten awareness of therapeutic strategies including global health considerations for the disabled adult with a rare bone disorder. Case: A 32-year-old African-American woman presented to establish care for hFTC, for which she takes ibuprofen for disabling bilateral hip pain. hFTC was diagnosed in early childhood and managed at a large children’s hospital. Therapeutic modalities relied on operative resection of skin lesions on eruption. She describes cosanguinous parentage, consignment to foster care as a child and homelessness as an adult. She has not had adult endocrine care for her hFTC. Her medical comorbidities include major depressive disorder, PTSD and suicide attempt. She reports one uncle with hFTC noting that his phenotype was not significantly disabling and apparent recession of his lesions with age. Her parents did not have symptoms of hFTC. Radiographically, she had amorphous calcium deposits over both hips (her primary sites of severe pain), upper back (site of painless spontaneous eruptions), and left elbow without pain or skin penetration, as well as hyperostosis of the ilium bilaterally. She noticed waxing and waning of her eruptions with her menstrual cycle, such that lesions worsen in the luteal phase. Megestrol acetate also induced new eruptions. Examination shows brown staining to all teeth with several missing teeth. She had a marked lower extremity limb length discrepancy with severe restriction of hip abduction. Blood chemistries were grossly normal, including renal function, calcium, uric acid and PTH. Serum phosphorous was elevated at 5.2 (2.3 - 4.5 mg/dL). CT showed bilateral nephrocalcinosis. We instituted treatment with a low phosphorous diet, sevelamer to reduce intestinal phosphate absorption and acetazolamide to increase urinary phosphate excretion. For her pain and functional disability, we recommended continued NSAIDs, physical therapy and social work consultation to facilitate continued access to care. Conclusions: This case uniquely describes hFTC symptom fluctuation with the menstrual cycle, worsening with increased progesterone, suggesting a possible role of gonadal hormones on FGF-23 biology. It also highlights the socioeconomic burdens that can accompany rare bone disease, which become more pronounced as the patient achieves adulthood. Successful transition of care from pediatric to adult endocrinologist, in addition to engaging multi-disciplinary support, is essential in optimizing life-long care in hFTC. Presentation: 6/1/2024

Effect of Chlorhexidine Gluconate (0.2%), Combination (0.2% Chlorhexidine and 0.05% Sodium Fluoride), and Green Tea Extract (0.5%) Mouth Rinses on Colony Count Changes of Streptococcus mutans and Lactobacillus in Saliva – A Randomized Controlled Trial

Abstract Introduction Streptococcus mutans and Lactobacillus acidophilus are the well-known etiological factors for dental caries. Chlorhexidine (CHX) gluconate is the “gold-standard” antimicrobial agent but having adverse effects such as brown staining of the teeth and altered taste. Thus, there is a need for an alternative herbal product. Objective This study was done to compare the efficacy of CHX gluconate (0.2%) and combination (0.2% CHX and 0.05% sodium fluoride [NaF]) mouth rinses to that of green tea extract (0.5%) mouth rinse in reducing the salivary count of S. mutans and L. acidophilus in dental students. Materials and Methods The sample size of the study was 75 dental students with ≥4 decayed, missing, and filled teeth (DMFT) index. They were divided into three equal groups to test various mouth rinses against S. mutans and Lactobacillus. Unstimulated salivary samples were collected at baseline and postrinse (14 days) and tested for the colony-forming units of S. mutans and Lactobacillus. The Chi-square test was used to ascertain the demographic characteristics of the study population. One-way ANOVA statistics and paired t-test were applied to calculate the inferential statistics between the variables. The statistical constant was fixed at P < 0.05. Results The results of the study indicated that there was a statistically significant reduction in S. mutans and Lactobacilli count in all the three study groups. Conclusion Green tea mouth rinse can be a promising preventive therapy worldwide for the prevention of dental caries but CHX mouthwashes are superior in reduction of S. mutans and Lactobacillus.

Expression of PD-L1 (DAKO 28-8) in Urothelial Carcinoma of Urinary Bladder

Introduction: Around 90% of bladder cancers are urothelial carcinoma. Recent advancements in treatment have shifted towards targeted therapies, particularly immunotherapies targeting PD-L1 (Programmed Cell Death Ligand 1). PD-L1, a ligand for PD-1 receptors, helps cancer cells evade the immune system. Various factors, including genetic alterations, lead to its upregulation in cancer cells. Immunotherapies aim to block this pathway, making cancer cells more susceptible to the immune response. Objective: The study was designed to assess the frequency of PD-L1 expression by immunohistochemistry in urothelial carcinoma of urinary bladder and its association with histological grades. Methodology: This study was conducted at Department of Pathology, Pakistan Institute of Medical Sciences Islamabad. Fifty-six cases of urothelial carcinoma were included. Immunohistochemistry using PD-L1 Dako 28-8 clone was applied to prepared tissue sections. PD-L1 positivity was determined by brown membrane staining in tumor cells, scored using the Tumor Proportion Score (TPS). Results: The median patient age was 63±7 years, with the highest frequency (58.9%) of PD-L1 expression occurring in the age range of 56 to 65 years. However, there was no significant association between PD-L1 expression and age groups. Out of 56 cases, 64.3% had a TPS (Tumor Proportion Score) of ≥5%. Among the 76.7% high-grade cases, 72% had a TPS ≥5%, whereas 28% had a TPS <5%. Among the 23.2% low-grade cases, 61.5% had a TPS <5%, and 38.5% had a TPS ≥5%. Conclusion: Sixty-four percent urothelial carcinoma cases showed PD-L1 expression. Significant association (p value = 0.02) between PD-L1 expression and high-grade urothelial carcinoma was found using Pearson’s Chi-square test.

The Expression of Ki-67 and E-cadherin in Triple Negative Breast Cancer: A Hospital Based Study

Introduction: Triple negative breast cancer (TNBC) is a subtype of breast cancer with aggressive nature as it does not respond to targeted chemotherapeutics. Expression of biomarkers is considered as an important factor for selecting treatment strategies and assessing prognosis. Objective: The aim of this study was to assess the expression of Ki-67 and E–cadherin in TNBC tissue. Methods: Immunohistochemical staining was done to assess the expression of proliferative marker Ki-67 and junctional complex marker E-cadherin in the Formalin Fixed Paraffin Embedded tissue (FFPE) of 80 TNBC patients. Ki-67 positive cells were identified on the basis of brown staining nuclei and E-cadherin expressed cells were considered positive on the basis of brown staining cell membrane. Ki-67 positive TNBCs were divided into low and high Ki-67 groups. Results: Ki-67 was expressed in 65% of the TNBC, the median of Ki-67 index was 30.25%. The difference between the medians of Ki-67 expression of low and high groups (7.5% and 53.25% respectively) was highly significant (p = 0.000). E-cadherin was negative in 67.5% of TNBC patients. The mean (±SD) age at diagnosis was 43.79 (±9.10) years. Most of the cancers were invasive ductal cell carcinoma (95%) and of grade II (70%). Lymph node metastasis was present in 73.8% patients and in 51% cases the cancer was in the left breast. Ki-67 expression was significantly associated with lymph node metastasis and E-cadherin expression was more negative in the left sided cancer. Conclusion: The study signifies that the TNBC has an early age at onset and Ki-67 expression is significantly high and associated with lymph node metastasis pointing the importance of this biomarker in TNBC patients. Negative expression of E-cadherin suggests close monitoring of the left sided TNBC.

Pengujian Kualitas Air Sumur Km.10 Kota Sorong Berdasarkan Parameter Fisika dan Kimia

Most residents in Kilo 10, Sorong City, use well water for daily needs. One common issue is the presence of iron in the well water, which causes rust and brown stains on clothes or household items. This study aims to determine the pH, TDS (Total Dissolved Solids), salinity, conductivity, turbidity, TSS (Total Suspended Solids), and temperature in well water. The well water samples were taken from areas with neutral pH (Sumbersari), basic pH (Puger), and acidic pH (Kencong). The results showed that the pH of the well water samples was 6.2, TDS was 185, indicating that the well water met the standards. The salinity was 150 or 1.5 mg/L, and the conductivity of pure water ranged from 0-200 μS/cm (low conductivity), large river water ranged from 200-1000 μS/cm (mid-range conductivity), and saline water was 1000-10000 μS/cm (high conductivity). The conductivity value for potable water is around 42-500 μS/cm. The turbidity result was 19 NTU, indicating that the well water sample was not suitable for drinking. The TDS value was high at 667, and the temperature was 30.5°C.

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.

First Report of Agroathelia rolfsii Causing Southern Blight on Cowpea in China

Cowpea (Vigna unguiculata L.) is a legume consumed as a high-quality plant protein source in many parts of the world. In August 2023, it was observed that a plant disease affected cowpea growth in Yiyang (28.34°N, 112.55°E), China. The average disease incidence was 10%, resulting in 8.5% economic losses in approximately 3,000 m2. The symptoms initially appeared as brown lesions near the stem-soil interface and the lesions were colonized by white mycelia. As the disease progressed, the disease symptoms included constriction and brown staining at the base of the stem, covered with a small amount of white mycelia. Eventually, the entire plants withered and collapsed and many sclerotia were scattered on the ground around the diseased stem. Twenty samples (10 sclerotia and 10 diseased tissue fragments) were collected from symptomatic plants for causal agent isolation. Samples were disinfected with 70% ethanol for 30 s, 5% NaClO for 1 min, rinsed three times with sterile water, dried and placed on potato dextrose agar (PDA) plates at 28℃ in the dark. In total, 20 isolates were obtained by the hyphal tip method (Terrones et al. 2022) and showed a consistent phenotype of white cottony mycelia on PDA with an growth rate of 12.9 to 21.3 mm/day (n = 20). Sclerotia formed at five to eight days post inoculation, were initially whitish, turning beige and eventually dark brown. The diameter of mature sclerotia ranged from 0.89 to 2.13 mm (mean = 1.64±0.29 mm; n =50). For pathogen identification, ITS1/ITS4 (White et al. 1990) and EF1-983F/EF1-2218R (Rehner and Buckley 2005) primers were used to amplify the internal transcribed spacer regions (ITS) and translation elongation factor-1 alpha gene (TEF-1α), respectively. The sequences of all 20 isolates showed 99% to 100% similarity with Agroathelia rolfsii sequences from GenBank by BLAST analysis. The sequences of two representative strains, ID1 and ID4, were deposited in GenBank. The ITS sequences of ID1 (OR689482) and ID4 (OR689481) were ＞99% similar to A. rolfsii strain QJ7 (593/596 bp; MZ750983) and A. rolfsii strain Kale078 (565/568 bp; MN872304), respectively. Also, TEF-1α sequences of ID1 (OR713735) and ID4 (OR713736) were ＞99% similar to the sequences of A. rolfsii strain HS-Sr (1073/1073 bp; OL416131) and A. rolfsii strain MSB1-2 (1070/1075 bp; MN702790), respectively. Phylogenetic analysis based on ITS and TEF1-α sequences indicated that ID1 and ID4 clustered into the A. rolfsii clade. Based on morphology and sequence analyses, the isolates ID1 and ID4 were identified as A. rolfsii (anamorph Sclerotium rolfsii). Pathogenicity tests were conducted three times on healthy 30-day-old cowpea seedlings. Five plants were inoculated with 6-day-old mycelial discs (6 mm) of ID1 or ID4 at the base of the seedlings (n = 30) while four plants were inoculated with a sterile PDA disc as a control (n = 12). All seedlings were cultivated in a greenhouse with a temperature of 26°C to 28°C and relative humidity 60% to 80% with a 14/10 h light/dark photoperiod. Eight days later, all the fungal inoculated seedlings showed symptoms including brown necrosis and collapse of the stems, and eventual withering of the seedlings. Control plants remained asymptomatic. The causal pathogens were reisolated in PDA plates and identified by ITS sequence analysis, completing Koch's postulates. To our knowledge, this is the first report of A. rolfsii causing southern blight on cowpea in China. Early accurate diagnosis will help farmers to adopt suitable practices to control disease outbreaks and reduce losses.

Direct immunofluorescence (DIF) versus immunohistochemical (IHC) staining of complements and immunoglobulins (Ig) in pemphigus group.

Pemphigus is a group of bullous disorders of the skin characterized by the formation of autoantibodies present in the intercellular junction of the epidermis. Diagnosis is made by clinical, histopathological examination, and DIF. As DIF needs frozen sections, fluorescent tagged antibodies, UV light microscope for examination, and trained personnel, its non-availability makes a definitive diagnosis challenging. To evaluate the utility of IHC staining of complements and Ig in cases of Pemphigus. Twenty-six diagnosed cases of Pemphigus were stained by Peroxidase immunohistochemical method using monoclonal antibody to IgG, IgA, IgM, IgG4, C3, C4 d with DAB as chromogen. Pemphigus cases include twenty of pemphigus vulgaris (PV), four cases of pemphigus foliaceous (PF), and two of pemphigus vegetans (Pveg). Positivity was defined as the deposition of Ig and complements as distinct, continuous brown staining of keratinocytes at intercellular junctions. On IHC total of 20 PV 17 showed positivity (85%) for IgG, 11 (55%) C4d, 19 (95%) C3d, and 16 (80%) IgG4 deposits at the intercellular junction of the epidermis. All cases of PF showed a deposit of IgG, with three (75%) cases for IgG4, C3d, and C4d. Both cases of Pveg showed positivity for IgG and C4d while one case was negative for IgG4 and C3d. The overall IgG, C3, IgG4, and C4d expression for pemphigus was seen in 88%, 88%, 76.9%, and 61.5% of cases. The relation between these markers, combination of IgG and C3, was best related to each other ( P value = 0.80). The sensitivities for IgG, IgG4, and C3 were 77.8%%, 73%, and 73% resp. We conclude that IHC is a useful tool in the diagnosis of PV with the highest sensitivity of IgG and C3d. The combination of IgG and C3d could replace the DIF in almost all of our cases, so IHC on FFPE sections be used as an alternative method to DIF.

A Robust Method for the Unsupervised Scoring of Immunohistochemical Staining.

Immunohistochemistry is a powerful technique that is widely used in biomedical research and clinics; it allows one to determine the expression levels of some proteins of interest in tissue samples using color intensity due to the expression of biomarkers with specific antibodies. As such, immunohistochemical images are complex and their features are difficult to quantify. Recently, we proposed a novel method, including a first separation stage based on non-negative matrix factorization (NMF), that achieved good results. However, this method was highly dependent on the parameters that control sparseness and non-negativity, as well as on algorithm initialization. Furthermore, the previously proposed method required a reference image as a starting point for the NMF algorithm. In the present work, we propose a new, simpler and more robust method for the automated, unsupervised scoring of immunohistochemical images based on bright field. Our work is focused on images from tumor tissues marked with blue (nuclei) and brown (protein of interest) stains. The new proposed method represents a simpler approach that, on the one hand, avoids the use of NMF in the separation stage and, on the other hand, circumvents the need for a control image. This new approach determines the subspace spanned by the two colors of interest using principal component analysis (PCA) with dimension reduction. This subspace is a two-dimensional space, allowing for color vector determination by considering the point density peaks. A new scoring stage is also developed in our method that, again, avoids reference images, making the procedure more robust and less dependent on parameters. Semi-quantitative image scoring experiments using five categories exhibit promising and consistent results when compared to manual scoring carried out by experts.

Analyses of the brown stain on the Parthenon Centaur head in Denmark

In 1688 two sculptural fragments, a head of bearded man and a head of an unbearded youth, arrived in Copenhagen, sent from Athens as a gift to King Christian 5. They were placed in the Royal Kunstkammer, their provenance given as the Temple of Artemis in Ephesos, one of the Seven Wonders of the World. Almost a hundred and fifty years later, in the early 1820’s they were noticed and studied by two scholars independently visiting the Kunstkammer. However, both concluded that the two heads belonged to one of the metopes decorating the south side of the Parthenon temple on the Acropolis in Athens, showing fighting between Greeks and the mythical Centaurs, part man and part horse. In the 1830’s another sculptural fragment, a horse’s hoof, obtained through the German archaeologist and state antiquary of Greece, Ludwig Ross, reached Copenhagen. It was forwarded by the Danish consul to Athens, C.T. Falbe, as a gift to King Christian 8. The inventory reads: ‘… was found on the Acropolis near the Parthenon temple and is supposed to belong to one the Centaurs on the metopes.’ The present paper focuses solely on the head of the Centaur.A brown stain was noticed on the Parthenon marbles as early as 1830 by the British Museum and has ever since eluded a deeper understanding of its genesis despite many investigations and attempts of analyses. A quite similar brown stain can be observed on the Centaur’s head in Copenhagen as well.The present study reports analyses by LA-ICP-MS, SEM–EDX, µXRD, GC–MS, and LC–MS-MS, as well as optical microscopy of five small samples sequestered in 1999 from the Centaur head curated by the National Museum of Denmark. Our analyses show that the brown stain consists of two consecutively added surficial layers of the calcium oxalate minerals whewellite and weddellite. Despite a thorough search using proteomics, we have found no viable organic precursor material for the oxalates. Our results do not solve the mystery of the formation of the brown stain, but they do further qualify the structure and characterization of the brown stain.

Immunohistochemical evaluation of Glut1 in dentigerous cysts, odontogenic keratocysts, and ameloblastoma.

Glucose uptake may be considered the rate-limiting step for the growth and metabolism of the cancer cell. Studies on GLUT1 have shown that GLUT1 is involved in cell survival and proliferation in both healthy and pathological circumstances. GLUT1 expression is regarded as one of the crucial elements in the development of local aggressiveness, tumour invasiveness, and metastasis, particularly in malignant tumours. The role of glut1 in odontogenic cysts and tumours has remained uncertain. The aim of the study is to assess the expression of Glut1 in dentigerous cysts, odontogenic keratocysts, and ameloblastoma. The study was conducted in GSL Dental College. The study design was a resprospective immunohistochemical study. Formalin-fixed, paraffin-embedded blocks of histologically confirmed cases (n = 50), 10 cases of odontogenic keratocysts, dentigerous cysts, ameloblastomas solid, ameloblastomas unicystic, and dental follicles each. Brown colour staining was considered as positive staining for GLUT1. Quantitative analysis was performed by counting the number of labelled cells, and semi-quantitative analysis was conducted by assigning immunostaining intensity scores. Chi-square test was used to compare differences between the groups. A P value of ≤0.05 was considered as statistically significant. Odontogenic keratocysts and unicystic ameloblastoma showed ≥50% of label cells with strong intensity of staining. Odontogenic keratocysts and solid ameloblastoma showed sub-cellular localisation of staining in the cytoplasm and membrane. Dentigerous cysts exhibited combined nucleus, cytoplasm, and membrane sub-cellular localisation of staining. The development of ameloblastomas, odontogenic keratocysts, and dentigerous cysts appears to be influenced by GLUT-1. Variation in its expression may aid in explanation of some of the differences in biological activity of these lesions.

Cervical Cytology-Histopathology Concordance and Role of Dual Immunomarkers in Biopsy Samples of Uterine Cervix: A Cross-sectional Study from a Tertiary Care Hospital of West Bengal, India

Introduction: Cervical cancer is one of the leading causes of cancer deaths. Liquid-based cervical cytology enables the detection and diagnosis of the disease at an early stage. p40 is a specific immunomarker that distinguishes squamous cell carcinomas from other cervical carcinomas with glandular and neuroendocrine differentiation. Aim: To determine the role of Liquid-based Cytology (LBC) in the early detection of premalignant and malignant lesions of the uterine cervix and its concordance with histopathological findings. Materials and Methods: This cross-sectional study was conducted at the Institute of Post Graduate Medical Education and Research (IPGME&R), Kolkata, West Bengal, India, over a period of one year and six months (from October 2020 to March 2022). Cervical samples were collected from a total of 200 females, and the materials were processed using the BD SurePath™ LBC method. The cytologically confirmed cases were biopsied, and histopathological concordance was established. Immunohistochemical (IHC) staining for p40 and p63 was performed using the peroxidase-antiperoxidase method to differentiate between premalignant, malignant squamous, and glandular lesions of the uterine cervix. For quantitative analysis, all cells were counted in 10 random fields at 400x magnification, and p63 and p40 were expressed as a percentage of positive cells per the total number of counted cells. Cases were considered positive if 5% or more of the tumour cell nuclei showed brown nuclear staining. The mean percentage positivity of p63 and p40 nuclear staining of all the cases in each category was calculated for non neoplastic and precursor/neoplastic lesions of the cervix. Results: Among the 200 cases evaluated by LBC, 169 (84.5%) were non neoplastic, and the remaining 31 (15.5%) were precursor/neoplastic lesions. The vast majority (126 cases, 63%) were inflammatory smears, followed by 20 cases (10%) that were Negative for Intraepithelial Lesion or Malignancy (NILM). Total of 40 cases were followed-up with biopsy and histopathology, and concordance with the cytological diagnosis was evaluated. Among 40 cases, 23 were precursor/ neoplastic on histopathology, and seven cases that were non neoplastic on Histopathological Examination (HPE) had been correctly diagnosed on LBC, giving a concordance rate of 75%. The overall sensitivity, specificity, Positive Predictive Value (PPV), and Negative Predictive Value (NPV) of LBC were 92%, 46.6%, 74.1%, and 77.7%, respectively, considering histological diagnosis as the gold standard. Regarding IHC expression on histopathology, the mean positivity of p40 and p63 for non neoplastic lesions was 9.40% and 10.06%, and for precursor/neoplastic lesions, 47.44% and 46.4%, respectively. Adenocarcinoma-in-situ and adenocarcinoma were negative for both p40 and p63. Conclusion: Cervical cytology is a less invasive, cost-effective, and simple procedure to diagnose cervical epithelial cell abnormalities. Cytological-histopathological concordance revealed that LBC is a sensitive diagnostic method. The comparison between non neoplastic and neoplastic lesions of the cervix revealed a statistically significant difference with respect to the mean percent positivity of p40 and p63 IHC staining. Both of these markers can be used to differentiate squamous cell carcinoma from adenocarcinoma of the cervix.

Cervical Cytology-Histopathology Concordance and Role of Dual Immunomarkers in Biopsy Samples of Uterine Cervix: A Cross-sectional Study from a Tertiary Care Hospital of West Bengal, India

Introduction: Cervical cancer is one of the leading causes of cancer deaths. Liquid-based cervical cytology enables the detection and diagnosis of the disease at an early stage. p40 is a specific immunomarker that distinguishes squamous cell carcinomas from other cervical carcinomas with glandular and neuroendocrine differentiation. Aim: To determine the role of Liquid-based Cytology (LBC) in the early detection of premalignant and malignant lesions of the uterine cervix and its concordance with histopathological findings. Materials and Methods: This cross-sectional study was conducted at the Institute of Post Graduate Medical Education and Research (IPGME&R), Kolkata, West Bengal, India, over a period of one year and six months (from October 2020 to March 2022). Cervical samples were collected from a total of 200 females, and the materials were processed using the BD SurePath™ LBC method. The cytologically confirmed cases were biopsied, and histopathological concordance was established. Immunohistochemical (IHC) staining for p40 and p63 was performed using the peroxidase-antiperoxidase method to differentiate between premalignant, malignant squamous, and glandular lesions of the uterine cervix. For quantitative analysis, all cells were counted in 10 random fields at 400x magnification, and p63 and p40 were expressed as a percentage of positive cells per the total number of counted cells. Cases were considered positive if 5% or more of the tumour cell nuclei showed brown nuclear staining. The mean percentage positivity of p63 and p40 nuclear staining of all the cases in each category was calculated for non neoplastic and precursor/neoplastic lesions of the cervix. Results: Among the 200 cases evaluated by LBC, 169 (84.5%) were non neoplastic, and the remaining 31 (15.5%) were precursor/neoplastic lesions. The vast majority (126 cases, 63%) were inflammatory smears, followed by 20 cases (10%) that were Negative for Intraepithelial Lesion or Malignancy (NILM). Total of 40 cases were followed-up with biopsy and histopathology, and concordance with the cytological diagnosis was evaluated. Among 40 cases, 23 were precursor/ neoplastic on histopathology, and seven cases that were non neoplastic on Histopathological Examination (HPE) had been correctly diagnosed on LBC, giving a concordance rate of 75%. The overall sensitivity, specificity, Positive Predictive Value (PPV), and Negative Predictive Value (NPV) of LBC were 92%, 46.6%, 74.1%, and 77.7%, respectively, considering histological diagnosis as the gold standard. Regarding IHC expression on histopathology, the mean positivity of p40 and p63 for non neoplastic lesions was 9.40% and 10.06%, and for precursor/neoplastic lesions, 47.44% and 46.4%, respectively. Adenocarcinoma-in-situ and adenocarcinoma were negative for both p40 and p63. Conclusion: Cervical cytology is a less invasive, cost-effective, and simple procedure to diagnose cervical epithelial cell abnormalities. Cytological-histopathological concordance revealed that LBC is a sensitive diagnostic method. The comparison between non neoplastic and neoplastic lesions of the cervix revealed a statistically significant difference with respect to the mean percent positivity of p40 and p63 IHC staining. Both of these markers can be used to differentiate squamous cell carcinoma from adenocarcinoma of the cervix.

CD56 Expression on Natural Killer Cells and its Correlation with Clinical Outcomes in Oral Squamous Cell Carcinoma Patients

Background: Squamous cell carcinoma (SCC) of the oral cavity can grow quickly and the response to therapy in each patient is different. The immune system plays a major role in the development of cancer cells, especially in SCC of the oral cavity. Natural killer cells (NK cells) play a major role in both innate immunity and adaptive immunity. Objective: To evaluation CD56 expression on NK cells and the correlation with clinical outcomes in oral squamous cell carcinoma. Methods: We collect the literature from Journal, Pubmed, Web of Science, Scopus, Crossref, and Google Scholar. CD56 expression was examined by immunohistochemical staining in cancer tissue. On each slide, the number of positive stained cells was read on 5 high power fields (HPF) 400x, then the number of cells stained on each HPF was added up and section 5. Cells with positive CD56 showed brown staining in the intracytoplasm regardless of the intensity of the staining. Results: High CD56 expression was associated with good overall survival (OS) (HR 0.27; 95%CI 0.12-0.60; p=0.001), good progression-free survival (PFS). (HR 0.35; 95%CI 0.17-0.74; p=0.005), and good distant metastasis-free survival (DMFS) (HR 0.27; 95%CI 0.13-0.55; p=0.0004) in head and neck SCC. Other study in oropharyngeal SCC, that high CD56 expression had good OS (HR 0.32; 95%CI 0.10-0.96; p=0.04). The meta-analysis study showed that the CD56 marker on NK cell was a good prognostic marker for OS in head and neck SCC (HR 0.19, 95%CI 0.11-0.35, p<0.00001). Conclusion: NK cells with CD56 expression will increase cytotoxic function. The presence of CD56 can increase infiltration into tumor tissue. Based on these data, CD56 on NK cells can be used as an indicator of cancer prognosis in patients with SCC of the oral cavity. Apart from that, it can be used as an immunotherapy target which functions to restore the immune response against cancer cells.

Potential of purple eggplant skin fraction (Solanum Melongena Var. Serpentinum L.) as an in vitro sunscreen

Sunscreen is a product that is used as skin protection against sun exposure. The skin of purple eggplant is known to contain secondary metabolites which are used as sunscreens. This study aims to determine the activity of sunscreen fraction of the purple eggplant (Solanum melongena var. serpentinum L.) in vitro. Testing of sunscreen activity was carried out by calculating the value of SPF (Sun Protection Factor), %TE (Percent Transmission of Erythema) and %TP (Percent Transmission of Pigmentation) using the UV-Vis Spectrophotometry method. The purple eggplant skin fractionation method is by using LVC (Liquid Vacuum Chromatography). The results of the fractionation were identified using phytochemical screening and combined TLC. The fraction results were tested for SPF (Sun Protection Factor), %TE and %TP values. The results showed that the extract yield was 26.33%. The fractions used are fraction 9; 10; 11; and 12 which showed the same stain after being evaporated, which was a brownish yellow stain with a yield of 21,24%. The purple eggplant skin fraction (Solanum melongena var. serpentinum L.) has SPF, %TE and %TP values with optimal protection at a concentration of 400 ppm with each value obtained being 19; 1.35 and 9.79. The results of the statistical test of the purple eggplant skin fraction showed that the data were normally distributed but not homogeneous. The results of the homogeneity test for each concentration of results showed that the data were not significantly different.

Application of narrow band imaging and Lugol's iodine staining in screening for nasopharyngeal carcinoma

BackgroundTo investigate the diagnostic value of conventional white light endoscopy (WLE), narrow band imaging (NBI) endoscopy, and Lugol's iodine staining under WLE (endoscopic iodine staining) in the screening and early diagnosis of nasopharyngeal carcinoma.MethodsPatients with nasopharyngeal lesions requiring biopsy attending the Department of Otolaryngology Head and Neck Surgery in our hospital between January 2021 and April 2023 were included in this study. Before biopsy, all subjects underwent conventional WLE, NBI endoscopy, and endoscopic iodine staining. On WLE, according to nasopharyngeal lesion morphology and color, patients were diagnosed with nasopharyngeal carcinoma ( +) or chronic hyperplastic nasopharyngitis (-). On NBI endoscopy, according to nasopharyngeal lesion vascular morphology, patients with type V manifestations (nasopharyngeal carcinoma) were categorized as NBI ( +) and patients with type I-IV manifestations (chronic hyperplastic nasopharyngitis) were categorized as NBI (-). Endoscopic iodine staining (1.6% Lugol's iodine solution) was positive ( +) if the mucosal surface was brown with no white patches, or negative (-) if there was no or light brown staining of the mucosal surface. Patients were divided into 2 groups based on histopathological diagnosis: nasopharyngeal carcinoma or chronic hyperplastic nasopharyngitis. Endoscopic diagnoses were compared with histopathological findings. The diagnostic performance of WLE, NBI endoscopy and endoscopic iodine staining for nasopharyngeal carcinoma were determined.ResultsThis study included 159 patients. On histopathology, 29 patients were diagnosed with nasopharyngeal carcinoma, and 130 patients were diagnosed with chronic hyperplastic nasopharyngitis. There were no significant differences in the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), accuracy, and area under the receiver operating characteristic (ROC) curve (AUC) of conventional WLE, NBI endoscopy or endoscopic iodine staining for differentiating nasopharyngeal carcinoma and chronic hyperplastic nasopharyngitis. The diagnostic performance of the combination of conventional WLE, NBI endoscopy and endoscopic iodine staining was significantly improved compared to any procedure alone.ConclusionsConventional WLE, NBI endoscopy or endoscopic iodine staining had good diagnostic performance for differentiating nasopharyngeal carcinoma and chronic hyperplastic nasopharyngitis. In particular, NBI endoscopy and endoscopic iodine staining alone or combined had clinical utility for identifying patients with nasopharyngeal lesions that are eligible for a watch-and-wait strategy.

Rock - soil transition during weathering of rhyolite

Concentric layers of weathered materials around core-stones show the rhyolite to experience sequential, but gradational, changes in visible features, textures and mineralogy as it transforms from ‘rock’ into ‘soil’. The changes start with the opening-up of grain boundaries and micro-cracks (stage 1) followed by their dark brown staining (stage 2) and the subsequent alteration (to sericite and clay minerals) of plagioclase feldspar groundmass grains and phenocrysts (stage 3). Biotite flakes are then bleached and altered (to chlorite and clay minerals) (stage 4) before there starts alteration (to sericite and clay minerals) of groundmass alkali feldspar grains (stage 5) and finally alteration of alkali feldspar phenocrysts (stage 6). Quartz grains are not altered during these stages of weathering but disaggregate and reduce in size due to continual opening-up of grain boundaries and micro-cracks. Increasing stages of weathering are marked by decreasing dry unit weights, dry densities and uniaxial compressive strengths, but increasing apparent porosities. The transition between ‘rock’ and ‘soil’ occurs during weathering stage 6 when all plagioclase groundmass grains and phenocrysts have been altered as have been all alkali feldspar groundmass grains and most phenocrysts. Stage 6 is marked by large apparent porosities (>14%) but low values of dry unit weight (<21.90 kN/m3) and dry density (<2,232 kg/m3).

Interpretation of Diverse Physico-Chemical Water Quality Parameters of Jaipur (Rajasthan) Sampling Sites for Plantation, Irrigation and Fluorosis Concern

The present study is aimed to evaluate the hydro chemical characteristics and quality of groundwater. Groundwater samples were collected from selected sampling sites like Shivdaspura (S1), Mansarovar (S2), Tonk Phatak (S3) and Chand pole (S4) in Jaipur District, Rajasthan and analyzed for major physico-chemical parameters. Such as Colour vis, Chemical Oxygen Demand (COD), Bio-Chemical Oxygen Demand (BOD), Lead, Cadmium, Chloride, Fluoride, Dissolved Oxygen (DO), Total Hardness (TH), Total Alkalinity, Total Solids, Total Dissolve Solids (TDS), Total Suspended Solids, Taste and Odour, Turbidity, Electrical Conductivity (EC), Magnesium, pH, Nitrate, Calcium, Sulphate Were analysed fora period from 29March 2021 to 09 December 2021. Fluoride content in the water samples of study area varied from 0.57mg/l to 7.97mg/l. Due to the higher fluoride levels in drinking water of Shivdaspura area showing dental fluorosis in which the teeth of the children were damaged and characterized by brown and black stains.