Lymph Node Aspirates Research Articles

Accuracy of the direct agglutination test for diagnosis of visceral leishmaniasis: a systematic review and meta-analysis

BackgroundParasitological investigation of bone marrow, splenic or lymph node aspirations is the gold standard for the diagnosis of visceral leishmaniasis (VL). However, this invasive test requires skilled clinical and laboratory staff and adequate facilities, and sensitivity varies depending on the tissue used. The direct agglutination test (DAT) is a serological test that does not need specialised staff, with just minimal training required. While previous meta-analysis has shown DAT to have high sensitivity and specificity when using parasitology as the reference test for diagnosis, meta-analysis of DAT compared to other diagnostic techniques, such as PCR and ELISA, that are increasingly used in clinical and research settings, has not been done.MethodsWe conducted a systematic review to determine the diagnostic performance of DAT compared to all available tests for the laboratory diagnosis of human VL. We searched electronic databases including Medline, Embase, Global Health, Scopus, WoS Science Citation Index, Wiley Cochrane Central Register of Controlled Trials, Africa-Wide Information, LILACS and WHO Global Index. Three independent reviewers screened reports and extracted data from eligible studies. A meta-analysis estimated the diagnostic sensitivity and specificity of DAT.ResultsOf 987 titles screened, 358 were selected for full data extraction and 78 were included in the analysis, reporting on 32,822 participants from 19 countries. Studies included were conducted between 1987–2020. Meta-analysis of studies using serum and DAT compared to any other test showed pooled sensitivity of 95% (95%CrI 90–98%) and pooled specificity of 95% (95%CrI 88–98%). Results were similar for freeze-dried DAT and liquid DAT when analysed separately. Sensitivity was lower for HIV-positive patients (90%, CrI 59–98%) and specificity was lower for symptomatic patients (70%, CrI 43–89%). When comparing different geographical regions, the lowest median sensitivity (89%, CrI 67–97%) was in Western Asia (five studies).ConclusionsThis systematic review and meta-analysis demonstrates high estimated pooled sensitivity and specificity of DAT for diagnosis of VL, although sensitivity and specificity were lower for different patient groups and geographical locations. This review highlights the lack of standardisation of DAT methods and preparations, and the lack of data from some important geographical locations. Future well-reported studies could provide better evidence to inform test implementation for different patient populations and use cases.PROSPERO registrationCRD42021240830

Discrepancies in tumor mutation burden reporting from sequential endobronchial ultrasound transbronchial needle aspiration samples within single lymph node stations - brief report.

Tumour Mutation Burden (TMB) is a potential biomarker for immune cancer therapies. Here we investigated parameters that might affect TMB using duplicate cytology smears obtained from endobronchial ultrasound transbronchial needle aspiration (EBUS TBNA)-sampled malignant lymph nodes. Individual Diff-Quik cytology smears were prepared for each needle pass. DNA extracted from each smear underwent sequencing using large gene panel (TruSight Oncology 500 (TSO500 - Illumina)). TMB was estimated using the TSO500 Local App v. 2.0 (Illumina). Twenty patients had two or more Diff-Quik smears (total 45 smears) which passed sequencing quality control. Average smear TMB was 8.7 ± 5.0 mutations per megabase (Mb). Sixteen of the 20 patients had paired samples with minimal differences in TMB score (average difference 1.3 ± 0.85). Paired samples from 13 patients had concordant TMB (scores below or above a threshold of 10 mutations/Mb). Markedly discrepant TMB was observed in four cases, with an average difference of 11.3 ± 2.7 mutations/Mb. Factors affecting TMB calling included sample tumour content, the amount of DNA used in sequencing, and bone fide heterogeneity of node tumour between paired samples. TMB assessment is feasible from EBUS-TBNA smears from a single needle pass. Repeated samples of a lymph node station have minimal variation in TMB in most cases. However, this novel data shows how tumour content and minor change in site of node sampling can impact TMB. Further study is needed on whether all node aspirates should be combined in 1 sample, or whether testing independent nodes using smears is needed.

Colloid and pigmented histiocytes in lymph node aspirates as a clue to metastasis in patients with a history of papillary thyroid carcinoma.

Cystic changes, calcification, colloid material, and multinucleated giant cells are frequently associated with primary and metastatic papillary thyroid carcinoma (PTC). These features are sometimes present in negative lymph node fine-needle aspiration cytology (FNAC). This study aims to review nodal aspirates of PTC to elucidate the significance of these cytological features in aspirates without tumor cells. FNAC specimens from patients with PTC confirmed on thyroidectomy were reviewed for cystic changes, tumor-associated features, and lymphoid components. Histologic follow-up of the lymph nodes were retrieved for correlation. A total of 113 aspirates were retrieved, of which 79 showed tumor cells on the FNAC specimen, and 95 were matched to a positive lymph node histology. At univariable analysis, calcification (n = 18/113, p = .044), colloid material (n = 40/113, p = .001), multinucleated giant cells (n = 29/113, p = .028), tumor cells (n = 79/133, p < .001), foamy histiocytes (n = 36/113, p = .002) and pigmented histiocytes (n = 62/113, p < .001) were associated with a positive histology. Presence of lymphoid fragments (n = 11/113, p < .001) and abundant background lymphocytes correlated with a negative histologic follow-up (n = 45/113, p = .005). In aspirates without tumor cells, multivariable analysis demonstrated colloid material (p < .001) and pigmented histiocytes (p = .003) to be independently predictive of metastatic PTC, whereas lymphoid fragments (p < .001) were independently associated with a negative histologic follow up. Colloid material and pigmented histiocytes, and to lesser degree calcification and multinucleated giant cells, when seen in an aspirate without tumor cells, raises suspicion metastatic disease. On the contrary, the lymphoid fragments can be regarded as supportive evidence of adequate sampling and a true negative result.

Xpert MTB/RIF assay among the extra-pulmonary and non-sputum respiratory samples: One year experience at National Tuberculosis Reference Laboratory, Bangladesh

Background: Extra-pulmonary tuberculosis (EPTB) is a diagnostic and therapeutic challenge due to the paucity of suitable rapid diagnostic tools. Objectives: The aim was to assess the frequency of detection of Mycobacterium tuberculosis using Xpert MTB /RIF assay among the extra-pulmonary and non-sputum respiratory samples (NSRS) along with demographic profile, life style variables and proportion of Rifampicin resistant TB among those specimens. Methodology: This cross-sectional retrospective study was conducted at National Tuberculosis Reference Laboratory (NTRL) from January 2018 to December 2018 for period of 1 year. Total 3918 extra-pulmonary and non-sputum respiratory samples from suspected TB patients were enrolled. Detection of Mycobacterium tuberculosis and Rifampicin resistance was performed by Xpert MTB /RIF assay and culture in both liquid and solid media. Results: Among 3918 samples, 3152 were new cases and 766 were previously treated TB cases. A total of 853 (21.77%) MTB were detected by Xpert MTB /RIF assay. Among them, 713 (83.59%) were extra-pulmonary and 140 (16.41%) were non-sputum respiratory samples. Lymph node (LN) aspirates and tissues were the most frequent (37.9 and 9.9%) samples of EPTB. The proportion of RR cases among EPTB was 2.8% and 6.3% among new and previously treated cases. And 2.6% and 11.1% among NSRS new and previously treated cases respectively. Conclusion: This study will give an insight of EPTB and NSRS profile with a view to diagnostic and therapeutic approach in Bangladesh. Bangladesh Journal of Medical Microbiology, January 2023;17(1):21-28

SAT536 A Case of Papillary Thyroid Cancer in Multiple Cervical Lymph Nodes, But Not Found in Thyroid Tissue on Pathology

Abstract Disclosure: M.A. Green: None. G.Y. Kim: None. Introduction:As technology advances, imaging becomes increasingly frequent and within the thyroid gland nodules can often be detected. After detection comes evaluation and lymph nodes within the neck are an integral part of this step. For that reason, fine needle aspiration (FNA) of cervical lymph nodes is known to lend a hand toward diagnosis. Here we present a patient who was diagnosed with Papillary Thyroid Cancer (PTC) from FNA pathology of a lymph node, but PTC was not noted within pathology of the thyroid tissue after total thyroidectomy. Case Presentation: This patient is a 34 year-old female with a history of ovarian cancer who presented to the clinic for a thyroid nodule and Hashimoto’s disease. A bed side ultrasound noted a right upper pole 1.6 x 0.6 x 0.6 cm thyroid nodule and a solid 1.8 x 1.5 x 0.8 cm nodule superior to the isthmus likely representing a delphian lymph node. As well, significant cervical lymphadenopathy was seen. A biopsy of the right upper pole nodule was nondiagnostic on FNA, but the delphian lymph node superior to the isthmus was positive for PTC. The patient then underwent total thyroidectomy and right and central neck lymph node dissections. Interestingly, PTC was noted in 3 of 27 lymph nodes in the central compartment, 5 of 18 lymph nodes excised with the thyroid tissue with extranodal extension, and 5 of 24 lymph nodes in the right neck compartment, but not noted in the thyroid tissue itself. The largest metastatic dimension of PTC within the cervical lymph nodes was noted to be 3 cm. The pathology noted a 1.0 cm tumor in the right lobe “approaching skeletal muscle” and psammoma bodies within the left lobe. After the patient’s surgery her stimulated thyroglobulin reported as 0.21 NG/mL and thyroglobulin antibody 2 IU/mL. She was then able to receive 30 mCi of I 131 sodium iodide. TSH maintained at goal on levothyroxine 200 mcg daily. Discussion: Diagnosis of PTC within multiple cervical lymph nodes, but not noted on thyroid pathology is very uncommon. It is highly unlikely in our patient that a cancer within the thyroid does not exist and there are theories to postulate this unique finding. These include missed pathology on the dissection, tumor regression prior to removal, or possibly ectopic tissue as the source of PTC. Although in our patient, given the significant amount of positive cervical lymph nodes, it is hard to believe that ectopic tissue away from the neck was the source or that tumor regression occurred. The pathological findings of psammoma bodies suggest the presence of PTC that spread to the lymphatic system. As well, the nature of thyroid removal leads to some destruction of tissue prior to evaluation. This case highlights the importance of thorough lymph node evaluation as a complete part of preoperative PTC assessment. Presentation Date: Saturday, June 17, 2023

FRI223 Ectopic ACTH Syndrome

Abstract Disclosure: S. Mallesara Sudhakar: None. E.S. Story: None. Introduction: Ectopic ACTH Syndrome (EAS) is caused by hypercortisolism from extra-hypophysial tumors. EAS accounts for 10-15 % of Cushing’s Syndrome. Case 1: A 36 year old male with recently diagnosed stage IV well differentiated grade 2 neuroendocrine tumor (NET) of the pancreas with liver metastases developed new hypertension, hyperglycemia, hypokalemia, metabolic alkalosis, fatigue and muscle weakness. There were no obvious signs of Cushing’s Syndrome. ACTH was 679 pg/ml, cortisol 120 ug/ml. MRI pituitary was normal. He required 4 antihypertensives, insulin, and metyrapone was initiated for hypercortisolism. Due to difficulty obtaining medications during the COVID pandemic, intolerance to therapies and lack of improvement in clinical symptoms, decision was made to perform laparoscopic bilateral adrenalectomy. He was started on hydrocortisone and fludrocortisone. Pathology revealed diffuse adrenal hyperplasia. Post-surgery he improved dramatically and was able to discontinue antihypertensives and insulin. Chemotherapy for the underlying tumor was discontinued due to cytopenias. His cancer progressed and he succumbed to disease 10 months from initial diagnosis. Case 2: A 60 year old female presented with severe hypokalemia, metabolic alkalosis, hypercalcemia, cytopenias, HTN, weight loss, muscle weakness, nausea and mental status changes. CT abdomen showed bilateral adrenal hyperplasia. ACTH was 489 pg/ml, cortisol 81 ug/ml. MRI pituitary was unremarkable. PET-CT additionally showed hypermetabolic mediastinal lymph nodes and right upper lobe nodular opacity, and widespread osseous lesions. She was started on ketoconazole and antihypertensives. Bilateral adrenalectomy was considered but could not be performed due to rapid clinical deterioration. Etomidate was initiated. Fine needle aspiration of a mediastinal lymph node showed small cell carcinoma. Patient became too frail to receive systemic chemotherapy and succumbed within 2 weeks of hospitalization. Discussion: Most Ectopic ACTH Syndrome cases are caused by NET of the lung, pancreas and thymus. In children, Ewing’s sarcoma has been reported. Small cell carcinoma of lung, malignant pancreatic NETs, and medullary carcinoma of thyroid often present with metastasis and carry an ominous prognosis. Not all patients have clinical features of classic Cushing's syndrome. The outcome of EAS correlates with tumor type. Optimum treatment is surgical resection of the tumor, often not possible with metastatic disease. Hypercortisolism may be controlled with adrenal enzyme inhibitors and receptor blockers. Bilateral surgical adrenalectomy may be needed when symptoms are not controlled by medical management. Early and rapid control of Cushing’s Syndrome is important to facilitate antineoplastic treatment of the underlying tumor. Presentation: Friday, June 16, 2023

Idiopathic multicentric castleman’s disease mimicking immunoglobulin G4-related disease responding well to Bortezomib: a case report

BackgroundCastleman’s disease (CD) is a rare disease that has clinical and pathological similarities to lymphoma and is characterized by a high frequency of associated immunological dysfunction. ImmunoglobulinG4-related disease (IgG4-RD) is a collection of systemic disorders that affect numerous organs and are also referred to as IgG4-associated sclerosing diseases. CD and IgG4-RD are difficult to separate because they may manifest similar commin clinical features.Case presentationThis case describes a 53-year-old female who, during routine medical check-up, exhibited a progressive increase in serum globulin levels and a simultaneous worsening of anemia symptoms, raising concern for a clonal plasma cell disease such as myeloma. However, bone marrow punctures did not reveal any abnormal plasma cells. Also, serum and urine immunofixation electrophoresis demonstrated no abnormal monoclonal protein bands. In addition, several laboratory findings excluded chronic liver disease, chronic infections caused by bacteria or viruses. Later, we found elevated serum IgG4 levels (10,700 mg/L), and identified multiple enlarged lymph nodes throughout the patient’s body. Axillary lymph node aspiration revealed no abnormal lymphocytes, ruling out the possibility of lymphoma. Pathological morphology of the axillary lymph revealed a large number of plasma cells in the lymphatic follicles. In addition, there was a reduction in lymphatic follicle size and apoptosis of the germinal centres. Immunohistochemistry revealed IgG4+/IgG + in > 40% of cells, and more than 100 IgG4 + cells per high powered field (HPF) of specimen. As of now, finding strongly suggested IgG4-RD. This patient was treated with glucocorticoids and various immunosuppressive drugs, such as prednisone, cyclosporine, methotrexate, cyclophosphamide, mycophenolate mofetil, azathioprine and hydroxychloroquine. Unfortunately, the patient did not recover. Ultimately, idiopathic multicentric Castleman disease (iMCD) was diagnosed in relation to the patient’s clinical presentation and laboratory tests, and after combination chemotherapy (VCD: Bortezomib, Cyclophosphamide and Dexamethasone), durable remission was achieved without serious adverse effects. During the follow-up period of one year and ten months, the patient remained stable.ConclusionThe diagnosis of Castleman must be distinguished from other disorders such as IgG4-RD, malignant lymphoma, reactive hyperplasia of various lymph nodes (mostly caused by viral infections), plasmacytoma, advanced HIV and rheumatic diseases. Besides observing systemic symptoms, laboratory tests such as immunoglobulin levels, complement levels, interleukin levels, and C-reactive protein levels should also be performed in order to determine a diagnosis.

Tissue Parathyroid Hormone Washout: A Reliable Method for the Localization and Exclusion of Parathyroid Adenomas.

Background and objective Accurate identification of parathyroid adenoma (PA) is essential for minimally invasive treatment of primary hyperparathyroidism (PHPT). The aim of this study is to evaluate the results of parathyroid hormone (PTH) assay in aspirates of suspicious neck lesions and to clarify its reliability in determining whether the lesion is of parathyroid origin. Methods A total of 134 lesions (104 imaging-suspected PA and 30 concomitant thyroid nodules as a control group) of 101 patients were retrospectively analyzed. Patients with positive, negative, or never scintigraphy were included in our study. Ultrasound (US) was performed again and US-guided tissue fine needle aspiration with PTH washout (PTH-WO) was performed from suspicious lesions. A PTH-WO level higher than the patient's serum PTH levels is proposed for a positive test, and a PTH-WO level lower than the upper limit of the laboratory PTH level is proposed for a negative test. A definitive diagnosis was made postoperatively histopathologically. Results PTH-WO levels (median (IQR)) were significantly higher in the positive group (n = 93, 5000 (1600) ng/L) compared to the negative group (n = 11, 17 (13.1) ng/L) and thyroid nodule aspirate group (n = 30, 14 (4.3) ng/L) (p < 0.001). Among 93 PTH-WO-positive lesions, 42 lesions (45.1%) were not identified by parathyroid scintigraphy (PS), 20 (21.5%) lesions were suspiciously PS, and 33 lesions (35.4%) were negative or suspicious byfirst US findings. Of the 93 patients localized with PTH-WO, two were cystic PAsablated by aspiration. The final pathology result of 91 operated patients proved PA. The sensitivity, specificity, negative predictive value, and positive predictive value of the PTH-WO were observed as 1.00, 0.82, 0.09, and 0.91, respectively. Conclusion Comprehensive repeated US and PTH-WO from the lesion increase the accurate localization of PAs. PTH-WO is a highly reliable method for differentiating parathyroid lesions from other cervical lesions.

Comparison of CBNAAT and AFB screening using Ziehl Neelsen stain and fluorescent stain on FNAC sample for rapid diagnosis of tubercular lymphadenitis

: Tuberculous lymphadenitis is the most common etiology of cervical lymphadenopathy in endemic countries. Fine needle aspiration of palpable lymph nodes is done for rapid diagnosis of tuberculous lymphadenitis This study presents a comparative evaluation of fine needle aspiration cytology (FNAC) with acid fast bacilli screening using Ziehl Neelsen stain, Fluorescent stain with Cartridge based nucleic acid amplification test (CBNAAT) for the rapid diagnosis of tubercular lymphadenitis.: An observational cross sectional study was done over a period of 15 months from January 2020 to March 2021. All newly diagnosed cases of tubercular lymphadenitis irrespective of age and sex were included. Fine needle aspiration was performed from the palpable lymph node. Smears were prepared using Giemsa, Papanicolaou, Ziehl Neelsen and Auramine O stain.Rest of the sample was used for Mycobacterial growth indicator test (MGIT) and CBNAAT. Statistical Analysis was performed using McNemar test. For gold standard, MGIT as well as a composite reference standard on parameters that included MGIT, radiological findings of tuberculosis, Positive Mantoux test and Positive response to ATT seen in the form of complete resolution of clinical and radiological findings.: The diagnostic value of CBNAAT differed with respect to the chosen gold standard. With MGIT as gold standard, CBNAAT had the highest sensitivity, specificity, positive predictive value and negative predictive value. The diagnostic accuracy of CBNAAT was also the highest. Using CRS (Composite Reference standard) as gold standard, CBNAAT showed the highest specificity and positive predictive value.: With CBNAAT showing statistically significant data of a higher diagnostic value in our study as well as showing rapid result, being automated and not subjected to observer interpretation, we conclude that CBNAAT is more efficient in the diagnosis of tubercular lymphadenitis as compared to AFB screening methods. The only limitation of CBNAAT in our study was its ability to show positive results for three atypical mycobacteria due to possible cross contamination.

Canine Somatic Mutations from Whole-Exome Sequencing of B-Cell Lymphomas in Six Canine Breeds-A Preliminary Study.

Canine lymphoma (CL) is one of the most common malignant tumors in dogs. The cause of CL remains unclear. Genetic mutations that have been suggested as possible causes of CL are not fully understood. Whole-exome sequencing (WES) is a time- and cost-effective method for detecting genetic variants targeting only the protein-coding regions (exons) that are part of the entire genome region. A total of eight patients with B-cell lymphomas were recruited, and WES analysis was performed on whole blood and lymph node aspirate samples from each patient. A total of 17 somatic variants (GOLIM4, ITM2B, STN1, UNC79, PLEKHG4, BRF1, ENSCAFG00845007156, SEMA6B, DSC1, TNFAIP1, MYLK3, WAPL, ADORA2B, LOXHD1, GP6, AZIN1, and NCSTN) with moderate to high impact were identified by WES analysis. Through a Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of 17 genes with somatic mutations, a total of 16 pathways were identified. Overall, the somatic mutations identified in this study suggest novel candidate mutations for CL, and further studies are needed to confirm the role of these mutations.

Canine Leishmania spp. infection in two distinct foci of visceral and cutaneous leishmaniasis in Tunisia.

Visceral leishmaniasis (VL) and zoonotic cutaneous leishmaniasis (ZCL) caused by Leishmania (L.) infantum and L. major, respectively, are endemic in Tunisia. The aim of the study was to assess canine Leishmania spp. infection prevalence as well as to identify the Leishmania species involved in two well-documented and geographically distinct VL and ZCL foci. One hundred seventy-six dogs were randomly recruited in the VL focus of Sbikha-Zaghouan (n = 100) and the ZCL focus of Echrarda-Nasrallah (n = 76). Physical examination and blood collection were systemically performed. Needle aspiration was done in case of lymph node (LN) enlargement. All sera were tested by ELISA. kDNA RT-PCR was performed on DNA extracts from (i) buffy coats of seropositive dogs and (ii) LN aspirates. Leishmania species identification was done by ITS1 PCR-sequencing. Thirty-three dogs (18.8%) were infected by Leishmania; 30 having anti-Leishmania antibodies and 3 were seronegative dogs with Leishmania DNA in LN aspirates. Prevalence of infection was significantly higher in VL foci than in ZCL foci (27% versus 7.9%, p = 0.002). Leishmania species was identified in 11 dogs and corresponded to L. infantum. Combination of serology and qPCR on LN aspirates seems to be the best option for canine leishmaniasis diagnosis. Infection is more frequent in VL foci and L. infantum is the only identified species.

Clinicopathologic Outcomes of Thyroid Nodule Aspirates Cytologically Suspicious for Hurthle Cell Neoplasms: A Retrospective Institutional Review

Clinicopathologic Outcomes of Thyroid Nodule Aspirates Cytologically Suspicious for Hurthle Cell Neoplasms: A Retrospective Institutional Review

Prevalence and incidence of canine visceral leishmaniasis and its clinical-immunological features in an endemic area of the Brazilian Amazon.

A cohort study for 2 years period analysed the prevalence, incidence and clinical-immunological features of canine Leishmania (L.) chagasi-infection in 316 mongrel dogs in a visceral leishmaniasis-endemic area in Pará State, Brazil. Diagnosis of infection was performed by the indirect fluorescent antibody test (IFAT-IgG), the leishmanin skin test (LST) and a parasite search (from the popliteal lymph node aspiration) at the beginning of the study and at 6, 12 and 24 months intervals. IFAT/LST revealed three immune profiles of infection: (I) IFAT(+) /LST(-) (81), (II) IFAT(-) /LST(+) (17) and (III) IFAT(+) /LST(+) (13). Prevalence of profiles I, II and III were 25.6, 5.4 and 4.1%, and an overall prevalence 35.1%. Incidence of profiles I, II and III were 5.4, 0.3 and 0.0%, and an overall incidence 5.7% dogs per month. Incidence at the age ranges <1 year, ≥1 year, <7 years and ≥7 years evidenced a highest rate in the age range <1 year (6.6% dogs per month). Parasitological diagnosis was positive in 19% dogs at the prevalence (85.7% profile I), and in 11% at the incidence (100% profile I). The clinical picture of 179 infected dogs showed 145 (81%) of profile I (82% subclinical); 21 (11.7%) of profile II (100% subclinical); and 13 (7.3%) of profile III (84.6% subclinical). Conversion from subclinical to sick dogs was higher (p<0.05) in profile I (40.2%) than in profiles II (5.8%) and III (9%). Immunological conversion showed that only 3.2% of profile I dogs (prevalence) converted to LST(+) (two at the end of the first 6 months and 1 after 24 months), while 82.3% of profile II dogs converted to IFAT(+) (11 in the first 6 months, whereas three after 12 months). A 100% death rate was observed in dogs from profile I alone. These results reinforce the need of adopting preventive strategies against CVL as early as in the first semester of the dog's life.

Baseline tumor gene expression signatures correlate with chemoimmunotherapy treatment responsiveness in canine B cell lymphoma.

Pet dogs develop spontaneous diffuse large B cell lymphoma (DLBCL), and veterinary clinical trials have been employed to treat canine DLBCL and to inform clinical trials for their human companions. A challenge that remains is selection of treatment to improve outcomes. The dogs in this study were part of a larger clinical trial evaluating the use of combinations of doxorubicin chemotherapy, anti-CD20 monoclonal antibody, and one of three small molecule inhibitors: KPT-9274, TAK-981, or RV1001. We hypothesized that significant differential expression of genes (DEGs) in the tumors at baseline could help predict which dogs would respond better to each treatment based on the molecular pathways targeted by each drug. To this end, we evaluated gene expression in lymph node aspirates from 18 trial dogs using the NanoString nCounter Canine Immuno-oncology (IO) Panel. We defined good responders as those who relapsed after 90 days, and poor responders as those who relapsed prior to 90 days. We analyzed all dogs at baseline and compared poor responders to good responders, and found increased CCND3 correlated with poor prognosis and increased CD36 correlated with good prognosis, as is observed in humans. There was minimal DEG overlap between treatment arms, prompting separate analyses for each treatment cohort. Increased CREBBP and CDKN1A for KPT-9274, increased TLR3 for TAK-981, and increased PI3Kδ, AKT3, and PTEN, and decreased NRAS for RV1001 were associated with better prognoses. Trends for selected candidate biomarker genes were confirmed via qPCR. Our findings emphasize the heterogeneity in DLBCL, similarities and differences between canine and human DLBCL, and ultimately identify biomarkers that may help guide the choice of chemoimmunotherapy treatment in dogs.

CARTRIDGE-BASED NUCLEIC ACID AMPLIFICATION TEST, AN IMPORTANT DIAGNOSTIC TOOL FOR TUBERCULOSIS BOTH PULMONARY AND EXTRA-PULMONARY TUBERCULOSIS

Background and Aims: Tuberculosis (TB) is severe infectious disease caused by Mycobacterium tuberculosis and imposes significant morbidity and mortality. Early diagnosis of active TB forms pulmonary and extrapulmonary is very important and the diagnostic methods such as culture test and Ziehl-Neelsen (ZN) stain displayed low diagnostic accuracy. Cartridge-based nucleic acid amplification test (CBNAAT) is widely used for the diagnosis of TB with good accuracy rate. Hence, the current study was commenced to assess the diagnostic usefulness of CBNAAT for the diagnosis of mycobacterium TB (MTB) and compared with ZN staining. Methods: This was a prospective study conducted on 250 sputum samples of pulmonary TB (new cases) and also patients who were receiving the treatment for TB. 5 mL of sputum sample including CSF, pericardial or ascitic fluid, pus, or lymph nodes aspirates were collected and subjected to ZN staining and CBNAAT. Results: Mean age observed was 48.12±8.76 years. 30 (12%) showed smear positive and 220 (88%) were smear negative for ZN staining. Further, samples have been subjected to CBNAAT showed 60 (24%) MTB positive, and 190 (74%) showed MTB negative. CBNAAT showed sensitivity and specificity of 91.5% and 89%. Out of 60 positives for MTB by CBNAAT, MTB has been identified in 28 patients. Conclusion: CBNAAT is a great tool in the early diagnosis of pulmonary and extrapulmonary TB.

Pattern and characteristics of mutations conferring resistance to second line drugs in Mycobacterium tuberculosis isolates of pulmonary and extrapulmonary TB samples

Background & objectivesThe purpose of present study is to analyse the distribution and pattern of genetic mutations in PRE-XDR-TB and extensive drug resistant Mycobacterium tuberculosis (XDR-TB) using second-line line probe assay and to compare them with different parameters. MethodSputum, Lymph node aspirate and cold accesses from patients with rifampicin resistant Tuberculosis were subjected to first line and second line Probe Assay (Genotype MTBDRsl by Hain Life Science, Germany) to assess additional drug resistance to fluroquinolones (Levofloxacin & Moxifloxacin) and Aminoglycosides (Amikacin, Ofloxacin and Kanamycin). The genetic mutation pattern was analysed and compared with demographic, clinical and other parameters. ResultsThe final study population included 123 fluoroquinolone resistant isolates including 14 isolates with additional second line aminoglycosides drug resistance. The most frequent mutation observed among Gyr A drug resistance mutation was D94G (Gyr A MUT3C, 50/123,40%) corresponding to high level resistance to levofloxacin and moxifloxacin. The most frequent wild type mutant among Gyr A gene locus was WT 3 (85/123,69%). The most common mutation among second line aminoglycoside resistant isolates was at eis WT2 (7/14,50%) followed by rrs MUT 2 (4/14,29%). ConclusionsGyrA MUT3C (Asp94Gly) was the most common mutation in Gyr A gene locus in M. tuberculosis causing high level levofloxacin and moxifloxacin resistance. Patients with Asp94Gly mutation was significantly associated with underweight body mass index (p = 0.026). This study also observed that history of anti-tuberculosis therapy is a risk factor for FQ drug resistance mutations (p < 0.001).

Comparison of Diagnostic Sensitivity and Procedure-Related Pain of Concurrent Ultrasound-guided Fine-needle Aspiration and Core-needle Biopsy of Axillary Lymph Nodes in Patients with Suspected or Known Breast Cancer.

Conflicting data exist on the diagnostic sensitivity of core-needle biopsy (CNB) compared to fine-needle aspiration (FNA) in the evaluation of axillary lymph node metastasis from breast cancer. Our purpose was to evaluate the sensitivity of CNB and FNA using subsequent axillary surgery as the gold standard and to compare the patients' subjective pain levels for each biopsy method. This IRB-approved prospective study enrolled 140 patients from February 2014 to May 2019 with known or suspected breast cancer. Patients underwent both US-guided FNA and 14-gauge CNB of the same node with clip placement and rated their pain level using a verbal numerical rating scale of 0 to 10. The diagnostic sensitivities were determined by pathology of the surgically excised lymph node using the McNemar test of correlated proportions. Changes in pain scores for CNB and FNA were determined using the Wilcoxon rank sum test. A total of 94 patients had confirmatory excision of the biopsied node with nodal metastasis detected in 71.3% (67/94). The sensitivity of CNB for detection of nodal metastasis was 95.5% (64/67), while the sensitivity of FNA was 67.2% (45/67) (P < 0.05). Overall pain score ratings for CNB increased by 0.6 from baseline on an 11-point numerical rating scale, while overall pain score rating for FNA decreased by 0.2 from baseline (P < 0.05). Our study demonstrates that 14-gauge CNB has superior sensitivity for detection of axillary nodal metastases and mildly increased pain compared with 25-gauge FNA in patients with breast cancer.

Cervical lymph node metastasis of bladder cancer: a case report and review of literature

Objective To report an extremely rare case of bladder cancer patient with cervical lymph nodes, abdominal lymph nodes, and bone metastases at the same time. Methods and results The case was investigated by follow-up and immunohistochemistry was used in the pathological part. Result The patient was diagnosed with bladder cancer (high-grade urothelial metastatic epithelial cell carcinoma) by pathology and immunohistochemistry after transurethral resection of bladder tumor (TURBT) and metastatic bladder cancer by pathology and immunohistochemistry after cervical lymph node aspiration due to neck lymph node enlargement 1 year later, and a CT of the chest and abdomen suggested that the patient also had abdominal lymph node and bone metastases.At the 2.5-year regular chemotherapy follow-up, the patient showed that the abdominal lymph node metastasis disappeared, the cervical lymph node fusion shrank, and the bone metastasis still existed. Conclusion 1. Regular postoperative review is particularly important; 2.For patients with UCB who undergo TURBT, a effective regular perfusion program should be performed throughout the postoperative period; 3. For patients with postoperative metastatic symptoms of UCB, Complex treatment has a positive effect on patient prognosis; 4.The presence of enlarged head and neck lymph nodes in patients with bladder cancer should also be considered as metastatic of UCB.

Case report on an outbreak of theileriosis in a dairy herd: Could it be a co-infection of a virulent strain of Theileria mutans with Theileria parva?

This case report aimed to alert veterinarians of the possibility of simultaneous infections with more than one Theileria species in cattle herds in Zimbabwe. Cows (N=7) and a postmortem specimen from a dairy farm located in Chivhu, Mashonaland East province, Zimbabwe were presented for clinical and postmortem examination, respectively. On history taking, the herd had a decrease in milk yield and high cow mortalities. The presented live animals had a heavy tick burden and were febrile and lethargic. Post-mortem findings revealed strong evidence suggestive of theileriosis. Theileria schizonts and piroplasms were identified in stained lymph node aspirate, impression, and blood smears. The herd was managed using tetracyclines and buparvaquone together with an intensive dipping protocol until deaths had stopped. Both the clinical and postmortem findings were strongly suggestive of the possibility of a simultaneous infection with more than one Theileria species in this herd. Thus, these finding highlight the need to characterise the Theileria species profile in Zimbabwe.

Whole Genome Sequence of a Treponema pallidum Strain From a Formalin-Fixed, Paraffin-Embedded Fine Needle Aspirate of a Cervical Lymph Node.

A patient with unilateral cervical lymphadenopathy suspicious for malignancy underwent a fine needle aspiration. Histology demonstrated mixed inflammatory infiltrates with abundant spirochetes. Sufficient spirochete DNA was extracted from paraffin-embedded tissue sections to obtain the near-complete genome sequence of a macrolide-resistant strain belonging to the SS14 omega strain of Treponema pallidum .