Histopathological Tests Research Articles

Transcriptomic Comparison of Non-Hodgkin Lymphomas in Relapsed/Refractory Versus Newly Diagnosed Patients Using Single FFPE Slides

Background Patients newly diagnosed with non-Hodgkin Lymphomas (NHLs) have been historically treated with B-cell depleting antibodies and chemotherapy. Standard of care (SoC) for patients with refractory or relapsed (R/R) disease has rapidly evolved over the past decade to incorporate chimeric antigen receptor T-cell therapy (CAR T), lenalidomide, antibody-directed chemotherapies and stem cell transplantation. This diverse treatment landscape can transform the tumor microenvironment and drive heterogenous tumor escape mechanisms. Understanding this varied tumor contexture will help inform efficacy results from NHL therapeutic studies. Core needle biopsies are used to sample tumor from R/R NHL patients; the bulk of tissue may be consumed for histopathology testing, leaving insufficient sample for transcriptomic profiling. We have shown that HTG EdgeSeq is useful for single-slide analysis of formalin fixed, paraffin embedded (FFPE) tissues (Loya et. al., JCOASCO Suppl. 2022). Here, we use this technique to compare tumors of R/R and newly diagnosed patients with Diffuse Large B-cell Lymphoma (DLBCL) and Follicular Lymphoma (FL). MethodsSamples and Transcriptomics: NHL tumor samples from newly diagnosed, treatment naïve patients (DLBCL n = 99, FL n = 19) and lymph node controls (n = 18) were sourced commercially. Tumor samples from a Phase 1/2a trial of epcoritamab in R/R patients (NCT03625037) were collected prior to treatment (DLBCL n = 38, FL n = 5). Single FFPE slides were processed using the EdgeSeq platform (19,000 gene HT panel, HTG Molecular). Tumors originated from lymphoid organs, gastrointestinal tract, testes, and the pleural cavity. Immunohistochemistry imaging for CD3, PAX5, CD20 was also performed (CellCarta, Belgium). Bioinformatics: Differential expression analysis was performed using linear models in R. Gene set enrichment analysis (GSEA) was performed with GSVA in R (Hänzelmann et. al., BMC Bioinformatics. 2013) using published signature sets (mSigDB: Liberzon et al., Cell Syst. 2015, XCell: Aran, et. al., Genome Biol. 2017). Results Single 5μm slides were analyzed with an average tissue size of 8mm2 for core needle biopsies and 40mm2 for resections. The number of detected genes was comparable to RNAseq, suggesting the HTG platform provides comprehensive data using core needle biopsies obtained during routine histopathology. We compared DLBCL tumor to normal lymph nodes and derived a gene set that was highly correlated to pathologist derived tumor purity. This quantitative proxy allows automated identification of samples that may contain source organ tissue. We applied differential expression analysis to identify gene, pathway and immune cell signatures contrasting newly diagnosed and R/R DLBCL. Top genes upregulated in R/R DLBCL tumors included B-cell markers as expected, T-cell function (CD25/IL2RA), and an intriguing epigenetic mechanism (YTHDC1; a N6-methyladenosine reader with a role in lymphoma proliferation [Liu et al., Front Onco 2022]). MYC pathway signaling was enriched in R/R tumors; consistent with known mutational risk factors for progression (Bisso et. al., Immunol Rev. 2019). Th2 polarized CD4+ T-cell signature was higher in treatment naïve DLBCL compared to R/R and normal lymph nodes. Fibroblast gene signatures were higher in R/R DLBCL, potentially a result of microenvironment remodeling due to chemotherapy or tumor driven adaptation (Dumontet et. al., Front Immunol. 2021). Comparison of DLBCL to FL was also performed in both R/R and untreated patient cohorts. Immune cell signatures for Tregs and CD4+ naïve T-cells were enriched in FL, whereas M1 macrophage signatures were enriched in DLBCL (findings were similar in both R/R and treatment naïve groups). Conclusions The EdgeSeq platform enabled whole-transcriptome profiling from very limited tissue samples in lymphoma clinical trials. This study highlights the importance of characterizing the transcriptional heterogeneity of NHL in relation to genetic alterations and T-cell immunity. Ongoing analysis with large patient cohorts in epcoritamab clinical trials will add statistical power to uncover associations with International Prognostic Score, Cell of Origin, effects of different treatment combinations, and genomic alterations on clinical outcomes.

Concordance of diagnostic modalities in atypical skin and soft tissue infections in hospitalized patients.

Skin and soft tissue infections (SSTIs) have high rates of morbidity and mortality worldwide but lack reliable standards for diagnostic workup. As a result, atypical infections, more prevalent among immunocompromised patients, can be missed due to deviance from classic features only to be revealed later through inconsistently performed ancillary studies. Our objectives included to evaluate the sensitivities of clinical impression, histopathology, tissue culture, and molecular and non-molecular ancillary tests in diagnosing inpatient SSTIs, as well as to qualitatively discuss the unusual features making a subset of infections "atypical." To do so, we retrospectively reviewed the histopathologic reports and charts of inpatient dermatologic consults at a single tertiary care institution over a 3-year period. We identified a total of 111 cases of SSTIs evaluated by the inpatient dermatology consultation service with concurrent skin or soft tissue biopsy, with 32.4% representing atypical infections. Among these, clinical impression suggested infection in 9(25.0%), routine histopathology in 21(58.3%), specialized stains for microorganisms in 22(68.8%), and tissue culture in 15(68.2%). Due to incomplete picture that each modality by itself creates, we conclude that clinicians and pathologists should carry a low threshold for including SSTIs in their differential diagnoses and should evaluate with skin biopsy, special stains for microorganisms, and ancillary studies, particularly in critically ill individuals who necessitate timely diagnoses.

Ginsenoside Rg1 alleviates learning and memory impairments and Aβ disposition through inhibiting NLRP1 inflammasome and autophagy dysfunction in APP/PS1 mice.

Alzheimer's disease (AD) is a common neurodegenerative disorder. Amyloid β (Aβ) deposition is considered an important pathological feature of AD. Growing evidence has linked neuroinflammation and autophagy to Aβ deposition in the progression of AD. However, there are few drug options for inhibiting neuroinflammation and autophagy to prevent AD. Ginsenoside Rg1 (Rg1), a steroidal saponin extracted from ginseng, has been reported to possess multiple neuroprotective effects. The present study aimed to evaluate whether Rg1 treatment could attenuate cognitive disorders and neuronal injuries by inhibiting NLRP1 inflammasome and autophagy dysfunction in an AD model of APP/PS1 mice. The results of behavioral tests indicated that Rg1 treatment for 12 weeks could significantly improve olfactory dysfunction as well as learning and memory impairments. The results of histopathological tests indicated that Rg1 treatment could reduce Aβ deposition and neuronal damages in APP/PS1‑9M mice. Additionally, the results of immunoblot, reverse transcription‑quantitative PCR or immunohistochemistry demonstrated that Rg1 treatment significantly downregulated the expression levels of inflammation‑related proteins of NLRP1, caspase1, IL‑1β and TNF‑α, as well as autophagy‑related proteins of p‑AMPK/AMPK, Beclin1 and LC3 II/LC3 I, and increased the expression levels of p‑mTOR/mTOR and P62 in APP/PS1‑9M mice. In addition, the molecular docking analysis showed that there was favorable binding result between Rg1 and NLRP1. The present study suggested that Rg1 may alleviate learning and memory impairments and Aβ disposition by inhibiting NLRP1 inflammasome and improving autophagy dysfunction, suggesting that Rg1 may be a potential therapeutic agent for delaying AD.

Preparation and characterization of benzydamine hydrochloride-loaded lyophilized mucoadhesive wafers for the treatment of oral mucositis

Chemo/radiotherapy-induced oral mucositis (OM) has painful ulcers that can impair patients' lives. This study aimed to develop and characterize (in vitro and in vivo) benzydamine (BNZ) freeze-dried mucoadhesive wafers from chitosan (CS) in combination with different hydrophilic polymers as a locally controlled delivery system for OM treatment. The porous (69–83%) optimum wafers (0.02–0.03 g and 3.2–4.3 mm) released 90–95% of their drug content over 8 h and demonstrated 40–53 g mucoadhesion strength. Mechanical evaluations of F1 (CS) and F14 (CS/HPMC (3:1)) selected formulations showed modulus of elasticity, tensile strength, and % elongation at break ranging from 0.2 to 0.3 MPa, 0.01–0.02 MPa, and 15–18%, respectively. The antimicrobial efficacy of BNZ-loaded wafers was significantly higher than drug-free ones. The extract solutions of the drug-free and BNZ-loaded wafers revealed 100 and 20% viability in human gingival fibroblasts (HGFs) exposure. In vivo analysis demonstrated that BNZ-loaded wafers represented a significant reduction in macroscopic evaluation of OM and histopathological tests compared to positive and negative controls and drug-free wafers. Conclusively, lyophilized F1 and F14 mucoadhesive wafers had acceptable structural, mechanical, and biological properties and might be promising drug delivery systems for OM.

Nanotechnology in Cancer Diagnostics and Therapeutics: A Review.

Cancer is the uncontrolled proliferation of cells that involves accumulation of genetic mutations by different types of mutagens including physical, chemical, and biological. Consequently, normal cell cycles get interrupted. Immunological assays, histopathological tests, polymerase chain reaction, computed tomography, magnetic resonance, and radiation therapy are some conventional techniques for cancer diagnostics. However, these techniques are not only expensive, time-consuming, tedious but also toxic to healthy cells. Therefore, these limitations are overcome by nanodevices that show high sensitivity, selectivity, rapidity, and cost-effectiveness in the detection of cancer biomarkers. Electrochemical biosensors are more efficient in the early diagnosis of cancers that help in patients' effective and timely treatment. Distinct types of nanotools viz. inorganic, organic, and polymeric nanomaterials are used in cancer therapeutics. Nano approaches have shown many advantages: they are site-specific, require meager amounts of drugs, limited toxicity, avoid drug resistance, and are more efficient, sensitive, and reliable. Therefore, future research should focus on developing highly inventive nanotools for the diagnosis and therapeutics of cancers.

PSAT023 Not Every Pain Is a Gain! A Rare Case of Adrenal Leiomyosarcoma.

Abstract Introduction Adrenal leiomyosarcoma (AL) is a rare malignant tumor, representing only 0.1% to 0.2% of all retroperitoneal soft tissue sarcomas in adults (1). A young male's acute abdominal and back pain led to an unusual diagnosis of AL. Case Presentation A 38-year-old male patient with no past medical history presented with a history of right sided abdominal pain of several months duration radiating to the back and significant weight loss. His vital signs were stable; however, he appeared mildly distressed and had tenderness on deep palpation in the right mid-upper quadrant. An abdominal sonogram showed a hypoechoic mass between the liver and the right kidney with no evidence of acute cholecystitis. A CT scan of the abdomen with IV contrast showed a lobulated mass invading the right adrenal gland, measuring 5.4×4.3×5.1 cm, with a post-contrast density measurement of 30 Hounsfield units. Laboratory tests showed plasma renin of 1.8 (upper limit of normal ULN 4.3 ng/ml/h), aldosterone of 8.6 (ULN 21 ng/dl), plasma metanephrine of <0.2 (ULN 0.50 nmol/l), and plasma normetanephrine of 0.49 (ULN 0.90 nmol/l). Additionally, his cortisol level after dexamethasone suppression was 0.9 (ULN 22.4 ug/dl). He underwent a right-sided laparoscopic adrenalectomy. The histopathological testing returned as a high-grade undifferentiated malignant neoplasm consistent with leiomyosarcoma. The IHC stains showed positive staining for SMA, CD10, and GATA3. He underwent whole-body PET/CT that showed multiple hypermetabolic abnormalities, including the right adrenalectomy bed, with a possible tumor invading the right upper renal pole. Systemic chemotherapy with doxorubicin and dacarbazine was initiated. Foundation panel testing showed NF1 loss on exons 2-3, CDKN2A/B CDKN2A rearrangement on exon 2, and TP53 loss on exons 1-4. He has received two rounds of chemotherapy, with no evidence of disease progression. Discussion Adrenal leiomyosarcoma usually originates from the smooth muscle cells of the adrenal or adjacent vascular structures. The tumor is asymptomatic until it enlarges and compresses the surrounding structures. Currently, there are about 50 reported valid cases (2) with no reports of NF-1 exon loss to our knowledge. The primary treatment is radical surgery with or without adjuvant chemotherapy and radiotherapy. Foundation panel testing showing NF-1 and TP53 exon loss may guide future targeted therapy. References 1-Fatema Jabarkhel, Henri Puttonen, Lina Hansson, Andreas Muth, Oskar Ragnarsson, Primary Adrenal Leiomyosarcoma: Clinical, Radiological, and Histopathological Characteristics, Journal of the Endocrine Society, Volume 4, Issue 6, June 2020. 2-Bahador Oshidari, Amir Zamani, Hooman Bahrami-Motlagh, Elena Jamali, Setareh Mahmoodi, Manoochehr Ebrahimian, Primary leiomyosarcoma of the adrenal; a case report, International Journal of Surgery Case Reports, Volume 90, 2022, 106707. Presentation: Saturday, June 11, 2022 1:00 p.m. - 3:00 p.m.

The association between antigenemia, histology with immunohistochemistry, and mucosal PCR in the diagnosis of ulcerative colitis with concomitant human cytomegalovirus infection

BackgroundHuman cytomegalovirus (HCMV) colitis can be involved in active ulcerative colitis (UC) in patients refractory to steroid and immunosuppressive drugs. Histological examination with colonic biopsy specimens and antigenemia assays are the standard tests for diagnosing HCMV enterocolitis, and we have previously reported the usefulness of mucosal polymerase chain reaction (PCR) methods. However, the associations among histopathological tests, antigenemia assays, and mucosal PCR are unknown.MethodsWe retrospectively analyzed 82 UC patients who underwent mucosal biopsy from inflamed colonic tissues for histological evaluation and mucosal PCR to detect HCMV. We analyzed the relationships between the HCMV-DNA copy number in colonic mucosa and other HCMV tests.ResultsIn total, 131 HCMV mucosal PCR tests from 82 UC patients were positive. The HCMV-DNA copy number was significantly higher in patients with positive immunohistochemistry (IHC) (p < 0.01) and was correlated with the number of positive cells for the antigenemia (C7-HRP, p < 0.01; C10/11, p < 0.01). Receiver operating characteristic curve analysis confirmed 1300 copies/μg of HCMV-DNA as the best diagnostic cut-off value to predict positive results of antigenemia (area under the curve = 0.80, 95% CI 0.68–0.93). HCMV-DNA copy number also correlated with the total UCEIS score (p = 0.013) and the bleeding score (p = 0.014). For each individual patient, a positive correlation between the change in total UCEIS score and HCMV-DNA copy number was observed (p = 0.040).ConclusionThe antigenemia assay and histopathological test with IHC were significantly associated with the HCMV-DNA copy number in colonic tissues. Moreover, endoscopic examination with the UCEIS can help diagnose the HCMV colitis in UC patients.

Effect of &lt;I&gt;Andrographis paniculata&lt;/I&gt; Leaf Crude Extract Against &lt;I&gt;Edwardsiella tarda&lt;/I&gt; Bacteria and Histopathological Profile of the Liver of &lt;I&gt;Osphronemus gouramy&lt;/I&gt; Juvenil

Highlight Research Paniculata leaf crude extract has antibacterial compounds Paniculata leaf crude extract can inhibit the growth of E. tarda bacteria Gouramy liver infected by tarda bacteria undergoes degeneration, congestion, and necrosis Treatment using paniculata leaf extract can improve Gouramy liver damage Abstract Edwardsiellosis caused by bacterium E. tarda is a serious challenge faced by Gourami farmers. This study aimed to determine the antibacterial activity of A. paniculata leaf crude extract on the growth of E. tarda bacteria and its effect on the histopathology of gourami liver which was carried out in two stages. In the first stage, the active compound was identified in the leaves of A. paniculata followed by the Minimum Inhibitory Concentration (MIC) test at five different doses and two controls. In-silico test was conducted to determine the antibacterial activity of crude extract of A. paniculata leaf. In the second stage, the Lethal Dosage (LD50) and Toxicity (LC50) tests were carried out. Histopathological test of gouramy liver was carried out by taking the liver of fish that had been exposed to E. tarda bacteria. The active compounds contained in the crude extract of A. paniculata leaf are flavonoids, alkaloids, tannins, triterpenoids, and saponins. The Lethal Dosage (LD50) was 106 CFU/ml, and a toxicity test showed a dose of 250 mg/L caused the most death compared to other treatments. Histopathological test of gouramy liver showed that crude extract of A. paniculata leaf could improve liver function optimally at a dose of 300 (mg/L). The results of this study indicate that crude extract of A. paniculata leaf can affect the histology profile of gouramy liver and can cause toxicity if used inappropriately.

Gross necropsy, histopathology, and ancillary test results from neonatal beef calves submitted to a veterinary diagnostic laboratory.

The objective of this study was to evaluate the prevalence of abnormal findings in gross necropsy, histopathology, and ancillary test results from neonatal beef calves submitted to a veterinary diagnostic laboratory. This retrospective clinical case study was conducted by reviewing necropsy reports submitted between 2015 to 2020. Case inclusion criteria were animals had to be a bovine, 2 to 21 days of age, and a nondairy breed. Gross necropsy, histopathology, and laboratory test results were recorded. Identified lesions and abnormal test results were categorized based on body systems and infectious agent type. Age and system affected were analyzed using a 1-way ANOVA and Bonferonni pairwise comparisons. Overall, 1,060 reports were reviewed and 95 met the inclusion criteria. Median age of enrolled calves was 9 days (range, 2 to 21). A total of 252 lesions were identified with a median of 3 lesions/calf (range, 0 to 7) and 2 different body systems involved/calf (range, 0 to 5). The most common disorders were classified as digestive (42.1% [106/252]), respiratory (12.7% [32/252]), and multisystemic (11.1% [28/252]). With respect to age and system affected, calves with neurologic lesions were significantly younger (mean age, 5.1 days) than calves with digestive lesions (mean age 9.6 days). These data suggest a high prevalence of infectious diseases, mainly digestive, respiratory and multisystemic in origin. These findings could help guide producers and veterinarians when assessing factors contributing to neonatal beef calf loss.

XANTHOGRANULOMATOUS CHOLECYSTITIS: DIFFICULTY IN DIFFERENTIATING FROM GALLBLADDER CANCER

Xanthogranulomatous cholecystitis (XGC) is a benign, rare form of chronic cholecystitis characterized by focal or diffuse inammation of the gallbladder. Its incidence varies in different subjects from 1.2% to 8.9%. It is often misdiagnosed as a carcinoma GB in many cases in clinical and radiological diagnoses, and sometimes even in histopathological tests. XGC was rst described by Christensen and Ishak, as Fibroxanthogranulomatous inammation in the 1970s. This condition was later described as ceroid as histiocytic granuloma of the gall bladder (GB) and biliary granulomatous cholecystitis. It is characterized by focal or diffuse destructive inammatory process that mainly involves endothelial cells, brous tissues and lipid-laden macrophages. In particular it shows brownish-yellow spots with unusual irregular and asymmetrical thickening of the bladder wall. Its pathogenesis is unclear. It is thought to begin as biliary obstruction with severe or chronic cholelithiasis and an increase in intragallbladder pressure.

S3637 Not Your Usual Gastritis - A Case of Iron Deficiency Anemia Leading to a Rare Diagnosis of Collagenous Gastritis

Introduction: Collagenous gastritis (CG) is a rare disease characterized by deposition of subepithelial collagen in the gastric mucosa with concomitant inflammatory infiltrate in the lamina propria on histopathology. We report a rare case of CG in a young adult who was treated successfully with topical open capsule budesonide. Case Description/Methods: An 18-year-old male presented to the clinic for evaluation of iron deficiency anemia associated with exertional dyspnea, fatigue and intermittent episodes of melena. Initial studies were significant for hemoglobin of 5 g/dl, serum ferritin 3 ng/mL, serum iron 19 mcg/dL, and iron saturation of 6%. Prior to our evaluation, he had undergone EGD which was reported as having “gastritis” with negative Helicobacter pylori histopathology testing. The patient had required intravenous iron and packed red cell transfusions. We pursued repeat endoscopic evaluation. EGD showed hematin in the stomach with diffuse nodularity and friability in the stomach. No distinct ulcers were seen. Colonoscopy and video capsule endoscopy did not demonstrate any other abnormalities. Biopsies from the stomach revealed thickening of subepithelial collagen plate associated with chronic inflammation consistent with collagenous gastritis. Trichrome stain highlighted the thickened collagen plate and CD117 stain showed scattered mast cells without clustering. Treatment with budesonide 9 mg daily was subsequently started with clinical and hematological response on follow-up. (Figure) Discussion: CG can be further characterized to two distinct phenotypes seen in children and adults. In young patients, the disease often occurs in isolation and presents with symptoms of abdominal pain and anemia. Endoscopic features include nodular patterns, mucosal atrophy and collagen band deposition seen in both age groups. Treatments of this condition remains a challenge due to lack of established effective therapy. Prior reports with conventional treatments such as anti-secretary agents, iron supplementation, hypoallergenic diet, and systemic corticosteroids have produced limited success. Topical budesonide has emerged as effective treatment options in achieving clinical and histopathological improvement. CG is a rare entity that should be kept in mind in young patients who present with iron deficiency and nodularity seen on endoscopic evaluation. Our case adds to the growing body of literature in support of topical budesonide in treatment of CG.Figure 1.: Image A shows diffuse nodular mucosa in the gastric body on EGD. Image B shows thickening of subepithelial collagen plate associated with marked chronic inflammation.

Histopathological Features of the Lung Parenchyma in a 64-Year-Old Male Patient with Post-COVID-19 Infection with Spontaneous Pneumothorax Dextra Due to AlveolarPleural Fistula (APF): A Case Report

Introduction: Histopathological features of the lung parenchyma due to the Alveolar-Pleural Fistula (APF) in a patient with post-COVID-19 infection have not been reported. APF usually occurs after a spontaneous pneumothorax. Spontaneous pneumothorax is an abnormal traumatic accumulation of air within the pleural space. It is classified as primary or secondary pneumothorax based on multiple risk factors.Case Presentation: A sixty-four-year-old patient was referred from one of the remote hospitals in the west of Bali due to spontaneous simple pneumothorax dextra. A week after being diagnosed with COVID-19 infection, the patient underwent a bullectomy procedure and was clinically diagnosed with recurrent spontaneous pneumothorax dextra due to APF. Then, the bullectomy specimen was sent to the anatomical pathology laboratory. Histopathology test revealed extensive areas of necrosis and fibrosis with scattered lymphocytes and emphysematous alveoli found in APF lesions. Conclusions: In conclusion, the evidence of extended fibrosis, which destroys the pulmonary parenchymal septum and dilated alveoli with diffuse fibrosis in the subpleural and intraparenchymal areas, may cause impairment of both perfusion and ventilation. Unfortunately, viral cytopathic like-changes related to COVID-19, such as multinucleated cells with large nuclei, amphophilic cytoplasm, and prominent nucleoli in alveolar spaces with intranuclear inclusions, were not found in this case. In this case, surgery is needed in case of fistula, either related or unrelated to infection of the pleural cavity indicating the patient’s functional recovery.

Meropenem-loaded Cement Is Effective in Preventing Gram-negative Osteomyelitis in an Animal Model.

Low-dose antibiotic-loaded acrylic cement is routinely used for preventing skeletal infection or reimplantation in patients with periprosthetic joint infections. However, few reports about the selection of antibiotics in acrylic cement for antigram-negative bacteria have been proposed. (1) Does the addition of antibiotics (tobramycin, meropenem, piperacillin, ceftazidime, ciprofloxacin, and aztreonam) to acrylic cement adversely affect compressive strength before and after elution? (2) Which antibiotics have the highest cumulative release within 28 days? (3) Which antibiotics showed antimicrobial activity within 28 days? (4) Does meropenem-loaded cement improve body weight, temperature, and other inflammatory markers compared with control unloaded cement? This is an in vitro study that assessed the mechanical strength, antibiotic elution, and antibacterial properties of antibiotic-loaded cement, combined with an animal study in a rat model that evaluated key endpoints from the animal study. In the in vitro study, we added 2 g of tobramycin (TOB), meropenem (MEM), piperacillin (PIP), ceftazidime (CAZ), ciprofloxacin (CIP), and aztreonam (ATM) to 40 g of acrylic cement. The compressive strength, elution, and in vitro antibacterial properties of the antibiotic-loaded cement were detected. Thirty male rats were randomly divided into two groups: CON (antibiotic-unloaded cement) and MEM (meropenem-loaded cement, which had the most stable antibacterial properties of the six tested antibiotic-loaded cements in vitro within 28 days). The right tibia of all rats underwent arthroplasty and was implanted with the cement, followed by inoculation with Pseudomonas aeruginosa in the knee. General status, serum biomarkers, radiology, microbiological assay, and histopathological tests were assessed over 14 days postoperatively. The compressive strength of all tested antibiotic cement combinations exceeded the 70 MPa threshold (the requirement established in ISO 5833). The cumulative release proportions of the raw antibiotic in cement were 1182.8 ± 37.9 µg (TOB), 355.6 ± 16.2 µg (MEM), 721.2 ± 40.3 µg (PIP), 477.4 ± 37.1 µg (CAZ), 146.5 ± 11.3 µg (CIP), and 372.1 ± 14.5 µg (ATM) within 28 days. Over a 28-day period, meropenem cement demonstrated antimicrobial activities against the four tested gram-negative bacteria ( Escherichia coli , P. aeruginosa , Klebsiella pneumoniae , and Proteus vulgaris ). Ciprofloxacin cement inhibited E. coli growth, ceftazidime and aztreonam cement inhibited K. pneumonia growth, and tobramycin cement inhibited P. aeruginosa . Only meropenem demonstrated antimicrobial activity against all gram-negative bacteria on agar diffusion bioassay. Rats treated with meropenem cement showed improved body weight (control: 280.1 ± 4.2 g, MEM: 288.5 ± 6.6 g, mean difference 8.4 [95% CI 4.3 to 12.6]; p < 0.001), improved knee width (control: 13.5 ± 0.3 mm, MEM: 11.8± 0.4 mm, mean difference 1.7 [95% CI 1.4 to 2.0]; p < 0.001), decreased inflammatory marker (control: 316.7 ± 45.0 mm, MEM: 116.5 ± 21.8 mm, mean difference 200.2 [95% CI 162.3 to 238.2]; p < 0.001), decreased radiographic scores (control: 17.7 ± 2.0 mm, MEM: 10.7± 1.3 mm, mean difference 7.0 [95% CI 5.4 to 8.6]; p < 0.001), improved bone volume/total volume (control: 8.7 ± 3.0 mm, MEM: 28.5 ± 5 .5 mm, mean difference 19.8 [95% CI 13.3 to 26.2]; p < 0.001), decreased Rissing scale scores of the knee gross pathology (control: 3.3 ± 0.5, MEM: 1.1 ± 0.7, mean difference 2.2 [95% CI 1.7 to 2.7]; p < 0.001), decreased Petty scale scores of knee synovium (control: 2.9 ± 0.4 mm, MEM: 0.7 ± 0.7 mm, mean difference 2.1 [95% CI 1.7 to 2.5]; p < 0.001), and decreased bacterial counts of the bone and soft tissues and negative bacterial cultures of cement (p < 0.001, p < 0.001, p < 0.001, p < 0.001, respectively). In this current study, MEM cement had the most stable in vitro antimicrobial activities, effective in vivo activity while having acceptable mechanical and elution characteristics, and it may be an effective prophylaxis against skeletal infection caused by gram-negative bacteria. Meropenem-loaded acrylic cement is a potentially effective prevention measure for skeletal infection caused by gram-negative bacteria; however, more related clinical research is needed to further evaluate the safety and efficacy.

A case report of hepatic actinomycosis: A rare form of presentation

Hepatic Actinomycosis (HA) is a very rare abdominal actinomycosis that can be confused with hepatic involvement due to a tumor. Liver involvement can occur from an abdominal focus or by blood dissemination from another focus. This disease is much more common in men between 50 - 70 years and in a situation of immunosuppression. Symptoms are nonspecific and diagnosis includes histopathology, cultures, and imaging test. Treatment includes prolonged antibiotic therapy with antibiotics such as penicillin and drainage of abscesses. We present a case of a 54-year-old man patient with a record of three years of chronic pancreatitis of probably alcoholic origin, who developed hepatic actinomycosis, requiring drainage of liver abscesses and directed antibiotic treatment.

Dapsone improves the vincristine-induced neuropathic nociception by modulating neuroinflammation and oxidative stress.

Peripheral neuropathy is a dose-limiting adverse effect of vincristine (VCR) in cancer chemotherapies. Dapsone is commonly used for the prevention of opportunistic infections following cancer therapies. Therefore, a high rate of VCR and dapsone co-administration has occurred in leukemias. Recently neuroprotective effects of dapsone have been reported in various diseases. Regarding the physiopathology of VCR-induced peripheral neuropathy (VIPN) and dapsone neuroprotection, this study evaluated the effect of dapsone on VIPN. VIPN was induced by VCR injection (0.5mg/kg IP, every other day, 1week) in male Wistar rats. In the treatment group, dapsone(12.5mg/kg IP, 1week) was injected 30min before VCR. Hot plate, Von Frey, motor neuron conduction velocity (MNCV), and histopathological tests were applied. The levels of TNF-α and NF-kB in the sciatic nerve and caspase-3 activity in dorsal root ganglion were measured by the ELISA method. The levels of malondialdehyde (MDA) and Glutathione (GSH) in the sciatic nerve were measured by spectrophotometry and colorimetric assays. VIPN was observed as araised thermal and mechanical threshold, reduced MNCV, and sciatic nerve demyelination. However, dapsone reduced the mechanical and thermal threshold and improved the MNCV. Also, dapsone reduced TNF-α, NF-kB, MDA, and Caspase-3 activity, and increased the GSH level in the sciatic nerve. Moreover, dapsone prevented VCR-induced demyelination in the sciatic nerve. This research demonstrated that dapsone could be used as a protective drug against VIPN. It improves the impaired thermal and mechanical sensations by reducing inflammatory, oxidant, and apoptosis factors and preventing demyelination in the sciatic nerve.

The effects of 21 cytological and nucleic acid binding dyes on autosomal and Y-chromosomal STR markers using new generation STR amplification kits

Before DNA fingerprinting analysis, many biochemical and histopathological testing are conducted on a forensic biological sample. The samples are exposed to cytological and nuclear binding dyes for their visualization under a microscope. The present study assessed the usability of these biological samples exposed to 21 routinely used histological and nuclear binding dyes for forensic DNA examination in a simulated experiment. Wright's stain, Hematoxylin, Coomassie Brilliant Blue, and Evan's Blue did not show any effect on the amplification of autosomal STR markers. Y-STRs showed higher robustness in presence of 10 dyes. Wright's stain showed the least effect on the generation of complete autosomal and Y-STR profiles. Mobility issues in capillary electrophoresis were observed in the presence of Eosin yellowish, Coomassie Brilliant Blue, and Hematoxylin for autosomal STRs and Wright's stain, Orcein, Malachite Green, Nigrosin, Bromophenol blue, and Fast green for Y STRs. Peak heights of the respective Y STR markers showed a mixed result in comparison to the positive control. In autosomal STR analysis, the peak heights showed a sharp decrease in comparison to the positive control.

Tyrosinase Inhibitory Ability and In Vitro, In Vivo Acute Oral and In Silico Toxicity Evaluation of Extracts Obtained from Algerian Fir (Abiesnumidica de Lannoy ex CARRIERE) Needles.

This study was designed to evaluate the tyrosinase inhibitory effect, in vitro, in vivo, and in silico toxicity of fractions isolated from A. numidica de Lannoy needles. The cytotoxicity of extracts was examined against Artemia salina larvae, while the toxicity of these extracts was tested by acute oral toxicity in mice; by administration of a dose of 2000 mg/kg b.w A. numidica leaves extracts. The blood samples were collected from the eye orbital sinus for further analysis of biochemical parameters. The absorption, distribution, metabolism, elimination, and toxicity (ADMET) properties were identified by the pkCSM web server. The data stated that ethyl acetate (EA) presented strong anti-tyrosinase apt. The results reported that ethyl acetate extract exhibited a strong inhibitory capacity against A. salina larvae with LD50 of 75.004 µg/mL. The data also showed that no mortality occurred, and no toxicity symptoms were observed in mice. The biochemical parameters revealed that both extracts significantly affected the hepatic profile by increasing ALT, AST, and alkaline phosphatase. Histopathological tests also confirmed that both fractions were toxic at this concentration on hepatic and renal tissues, with necrosis observed. The toxicity of molecules in silico revealed no effect on all examined biomolecules.It can be concluded that this plant was toxic on the liver and renal profiles and tissues at the dose studied.

Comparison of the effect of aqueous extract of Artemisia herba alba and procaine penicillin on healing induced defect of the ear cartilage

This study was undertaken to investigate the effect of aqueous extract of Artemisia herba alba (Asteraceae) and procaine penicillin on healing induced defect of the ear cartilage. A total of (25) female rats which used in this study were distributed randomly into five treatment group(five animals per each). Under routine surgery, all rats had circular holes in the elastic cartilage of their ears. All treatment groups were treated with local application daily for one week. Control group(C) treated with normal saline only. First treatment group(T1) treated with 5% Artemisia aqueous extract, second treatment group(T2) treated with 10% Artemisia aqueous extract, third treatment group(T3) treated with 15% Artemisia aqueous extract, fourth treatment group(T4) treated with procaine penicillin. The results showed that there was no significant differences between all treatment group except between (T3) and other treatment groups (1.857± 0.293). It could be concluded that concentration of aqueous extract at 15% (T3)affected significantly on reduce the diameter of ear defect after three weeks of treatment, the result of histo pathological test reveal that complete healing, absence of scar tissue, both edges of cartilage and fused together with marked fibrosis and formed large blood vessels.

Molecular classification of soft tissue sarcomas for adequate diagnosis: A study on the northeast population of Morocco

BackgroundSoft tissue sarcomas (STS) are a heterogeneous group of tumors. For adequate therapeutic management, an accurate diagnosis is necessary. In Morocco, the diagnosis is essentially based on the morphological and immunohistochemical study. Compared to other techniques, fluorescence in situ hybridization (FISH) is easier to develop and less expensive. This study aims to assess the feasibility and utility of implementing FISH technique to improve diagnostic accuracy and establish a good classification. Material and methodsThis is a retrospective cohort study. 211 cases of mesenchymal tumors were included. Hematoxylin Eosin Safran (HES) staining was performed in all cases followed by immunohistochemistry (IHC). FISH was performed in all cases with suspected STS. The probes used were EWSR1, MDM2 and SS18. The performance of FISH and histopathological test were evaluated by the ROC curve method (receiver operating characteristic). We evaluated the concordance between FISH and real time PCR by Cohen test. ResultsThe real-time PCR technique showed good agreement with the FISH test by a Kappa coefficient of 60% (p = 0.035). FISH was able to confirm that it is more accurate (Youden’s Index = 91%) than histological/immunohistochemical analysis (Youden’s Index = 51%), as well as the positive predictive value was higher (100%) with an ROC curve finding a larger area under the curve of 0.953 (95% CI: 0.918–0.988), p = 0.000 which supports that FISH shows high performance to present an accurate final diagnosis. ConclusionThis is the first and the largest Moroccan series for the molecular diagnosis of STS by FISH. Our study shows that paraffin FISH is a sensitive and specific ancillary tool in the diagnosis of STS when used in the appropriate clinicopathological context.

Formulation and Evaluation of Pulp Devitalization Paste Combination of Jatropha Curcas L. and Piper Crocatum Leaves Extract

Dental caries require operative techniques such as pulp devitalization, but pulp devitalization materials that are often used by dentists have serious adverse effects that need to be considered, such as gingival injury and alveolar bone necrosis. Hence, the aim of the present study was to develop natural ingredients as an alternative to pulp devitalization from a combination of Jatropha curcas (Jatropha curcas L.) and red betel leaf extract (Piper crocatum) in the form of a paste and then conducting physical evaluation tests, Cyclooxygenase-2 (COX-2) tests, and also histopathological picture tests to the sample. The formulation of the pasta sample was carried out by the trituration method. Formulation I (FI) contained 25% of Jatropha resin, 0.25% red betel leaf extract, and 25% Jatropha resin, 0.5% red betel leaf extract for Formulation II (FII). The paste produced was then evaluated for physical properties which consisted of organoleptic, homogeneity, pH, spreadability, adhesion, and anti-inflammatory power using test animals with COX-2 expression number parameters. The FI and FII treatment groups had met the requirements as paste formula. In the COX-2 expression test, FI has a COX-2 expression percentage value of 0.38% and a COX-2 suppression percentage value of 0.62%, while FII has a COX-2 expression percentage value of 0.59% and a COX-2 expression percentage of 0.59% and suppression of COX-2 of 0.41%. The optimal concentration of the paste formulation is the paste with a combination of 25% jatropha latex and 0.25% red betel leaf extract had been shown to have potential as an alternative to pulp devitalization