Production of polycolonal antibody against domain 2-4 of protective antigen of Bacillus anthracis in laboratory animals

زمینه و هدف: آنتراکس که مسبب آن باکتری باسیلوس آنتراسیس می‌باشد یک بیماری عفونی حاد است و اغلب در گیاه‌خواران و انسان اتفاق می‌افتد. فرم رویشی باسیلوس آنتراسیس یک اگزوتوکسین سه جزیی شامل آنتی ژن حفاظت کننده (Protective Antigen=PA)، فاکتور کشنده (Lethal Factor=LF) و فاکتور ادم (Edema Factor=EF) می‌باشد. آنتی ژن حفاظت کننده به عنوان یک ایمونوژن اولیه برای توسعه ایمنی حمایتی بر علیه آنتراکس بررسی شده است. این مطالعه با هدف تولید آنتی بادی علیه ناحیه 4-2 آنتی ژن حفاظت کننده این باکتری در حیوانات آزمایشگاهی طراحی و اجرا شده است. روش بررسی: در این مطالعه تجربی ناحیه 4-2 ژن PA از پلاسمید pXOI با جایگاه های آنزیمی BamHI و HindIII به روش PCR تکثیر و در وکتورها کلون و ساب کلون شد. وکتور (pET28a(+ در باکتری اشرشیاکلی سویه BL21(DE3) ترانسفورم گردید. بعد از القاء با IPTG، بیان پروتئین ژن PA مشاهده شد. پروتئین تخلیص شده در 4 نوبت به موش و خرگوش تزریق شد؛ سپس آنتی بادی تولید شده از سرم موش و خرگوش جداسازی و توسط آزمون الایزا تایید گردید. یافته ها: ناحیه 4-2 ژن PAکلون شده در وکتور بیانی pET28a(+) به وسیله ی توالی یابی، PCR، آنالیز آنزیمی، الکتروفورز در ژل پلی اکریل آمید و لکه گذاری وسترن مورد تأیید قرار گرفت. افزایش تیتر آنتی بادی در خون موش و خرگوش توسط آزمون الایزا تایید گردید. نتیجه گیری: با توجه به ایمونوژن بودن پروتئین PA، می‌توان از آن در طراحی واکسن و همچنین به عنوان ادجوانت قوی سیستم مخاطی استفاده نمود

Preoperative assessment of meningioma aggressiveness by Thallium-201 brain SPECT

Introduction: Meningioma is usually a benign brain tumor, but sometimes with aggressive course. The aim of this study was to assess the ability of 201Tl Brain SPECT to differentiate the pathologic grade of meningioma preoperatively. Methods: Thirty lesions in 28 patients were evaluated in this study. Early (20 minutes) and late (3 hours) brain SPECT images were performed and early uptake ratio (EUR), late uptake ratio (LUR) and retention index (RI) were calculated. All patients were operated and pathologic grade of tumors were defined according to World Health Organization grading system. Results: SPECT results were compared in different pathologic groups. Data analysis clarified no significant difference of EUR in benign and aggressive meningioma (P=0.2). However LUR and RI were significantly higher in aggressive tumors (P=0.001 and P=0.02, respectively). Conclusion: According to our data Tl-201 Brain SPECT with early and late imaging has 80 sensitivity and specificity to differentiate malignant from benign meningioma

Oncologic outcomes following radical nephroureterectomy for upper tract urothelial carcinoma: a literature review

Background and Objective: Radical nephroureterectomy (RNU) represents the gold standard treatment for non-metastatic upper tract urothelial cancer. We sought to provide a comprehensive review of reported oncologic outcomes of the RNU procedure and of factors that might impact these outcomes. Methods: A non-systematic review of the literature was conducted by performing an electronic literature search using PubMed with "radical nephroureterectomy"and "oncologic outcomes"as free text search terms. Both original articles and systematic reviews were considered. Search was limited to articles in English that were published in the last 20 years. Key Content and Findings: Open and laparoscopic RNU offer comparable oncologic outcomes. In more recent years, the discussion has de facto shifted towards the "oncological safety"of robotic RNU, which also seems to offer comparable oncologic outcomes. Several studies have looked at the impact of different treatment-, patient- and tumor-related factors. Among treatment-related factors, attention has been given to diagnostic ureteroscopy and the risk of intravesical recurrence. Surgical wait time and perioperative blood transfusion have also been studied. Perioperative chemotherapy, specifically adjuvant therapy, was shown to improve survival. Among patient-related factors, baseline chronic kidney disease, diabetes mellitus, body mass index, and systemic inflammation have gained recent attention. Some tumor related factors, such as stage, grade, location, and multifocality may negatively impact survival outcomes. Lymphovascular invasion and histologic variants are clinically significant pathological findings. Conclusions: RNU is a procedure with measured long-term oncologic outcomes. Minimally invasive techniques have gained an established role as they seem to offer comparable oncologic "safety", although special attention is needed in relation to the method of bladder cuff excision. Robotic RNU is gaining popularity, and while evidence remains limited, the current literature supports the oncologic safety of this procedure. Several factors, which can be categorized as treatment-related, patient-related, and tumor-related, might impact the oncologic outcomes of UTUC patients undergoing RNU. These factors can provide crucial information to stratify patients based on their relative risk of disease recurrence and mortality which may guide clinical decision-making

Robot-assisted nephroureterectomy: surgical and mid-term oncological outcomes in over 1100 patients (ROBUUST 2.0 collaborative group)

Objective: To analyse surgical, functional, and mid-term oncological outcomes of robot-assisted nephroureterectomy (RANU) in a contemporary large multi-institutional setting. Patients and Methods: Data were retrieved from the ROBotic surgery for Upper tract Urothelial cancer STtudy (ROBUUST) 2.0 database, an international, multicentre registry encompassing data of patients with upper urinary tract urothelial carcinoma undergoing curative surgery between 2015 and 2022. The analysis included all consecutive patients undergoing RANU except those with missing data in predictors. Detailed surgical, pathological, and postoperative functional data were recorded and analysed. Oncological time-to-event outcomes were: recurrence-free survival (RFS), metastasis-free survival (MFS), cancer-specific survival (CSS), and overall survival (OS). Survival analysis was performed using the Kaplan–Meier method, with a 3-year cut-off. A multivariable Cox proportional hazard model was built to evaluate predictors of each oncological outcome. Results: A total of 1118 patients underwent RANU during the study period. The postoperative complications rate was 14.1%; the positive surgical margin rate was 4.7%. A postoperative median (interquartile range) estimated glomerular filtration rate decrease of −13.1 (−27.5 to 0) mL/min/1.73 m2 from baseline was observed. The 3-year RFS was 59% and the 3-year MFS was 76%, with a 3-year OS and CSS of 76% and 88%, respectively. Significant predictors of worse oncological outcomes were bladder-cuff excision, high-grade tumour, pathological T stage ≥3, and nodal involvement. Conclusions: The present study contributes to the growing body of evidence supporting the increasing adoption of RANU. The procedure consistently offers low surgical morbidity and can provide favourable mid-term oncological outcomes, mirroring those of open NU, even in non-organ-confined disease

Decisional and prognostic impact of diagnostic ureteroscopy in high-risk upper tract urothelial carcinoma: A multi-institutional collaborative analysis (ROBUUST collaborative group)

Background: Diagnostic ureteroscopy (URS) with or without biopsy remains a subject of contention in the management of upper tract urothelial carcinoma (UTUC), with varying recommendations across different guidelines. The study aims to analyse the decision-making and prognostic role of diagnostic ureteroscopy (URS) in high-risk UTUC patients undergoing curative surgery. Materials and methods: In this retrospective multi-institutional analysis of high-risk UTUC patients from the ROBUUST dataset, a comparison between patients who received or not preoperative URS and biopsy before curative surgery was carried out. Logistic regression analysis evaluated differences between patients receiving URS and its impact on treatment strategy. Survival analysis included 5-year recurrence-free survival (RFS), metastasis-free survival (MFS), cancer-specific survival (CSS) and overall survival (OS). After adjusting for high-risk prognostic group features, Cox proportional hazard model estimated significant predictors of time-to-event outcomes. Results: Overall, 1,912 patients were included, 1,035 with preoperative URS and biopsy and 877 without. Median follow-up: 24 months. Robot-assisted radical nephroureterectomy was the most common procedure (55.1%), in both subgroups. The 5-year OS (P = 0.04) and CSS (P < 0.001) were significantly higher for patients undergoing URS. The 5-year RFS (P = 0.6), and MFS (P = 0.3) were comparable between the 2 groups. Preoperative URS and biopsy were neither a significant predictor of worse oncological outcomes nor of a specific treatment modality. Conclusions: The advantage in terms of OS and CSS in patients undergoing preoperative URS could derive from a better selection of candidates for curative treatment. The treatment strategy is likely more influenced by tumor features than by URS findings

Prognostic Significance of Grade Discrepancy Between Primary Tumor and Venous Thrombus in Nonmetastatic Clear-cell Renal Cell Carcinoma: Analysis of the REMEMBER Registry and Implications for Adjuvant Therapy

BACKGROUND: Further stratification of the risk of recurrence of clear-cell renal cell carcinoma (ccRCC) with venous tumor thrombus (VTT) will facilitate selection of candidates for adjuvant therapy. OBJECTIVE: To assess the impact of tumor grade discrepancy (GD) between the primary tumor (PT) and VTT in nonmetastatic ccRCC on disease-free survival (DFS), overall survival (OS), and cancer-specific survival (CSS). DESIGN, SETTING, AND PARTICIPANTS: This was a retrospective analysis of a multi-institutional nationwide data set for patients with pT3N0M0 ccRCC who underwent radical nephrectomy and thrombectomy. OUTCOMES MEASUREMENTS AND STATISTICAL ANALYSIS: Pathology slides were centrally reviewed. GD, a bidirectional variable (upgrading or downgrading), was numerically defined as the VTT grade minus the PT grade. Multivariable models were built to predict DFS, OS, and CSS. RESULTS AND LIMITATIONS: We analyzed data for 604 patients with median follow-up of 42 mo (excluding events). Tumor GD between VTT and PT was observed for 47% (285/604) of the patients and was an independent risk factor with incremental value in predicting the outcomes of interest (all p < 0.05). Incorporation of tumor GD significantly improved the performance of the ECOG-ACRIN 2805 (ASSURE) model. A GD-based model (PT grade, GD, pT stage, PT sarcomatoid features, fat invasion, and VTT consistency) had a c index of 0.72 for DFS. The hazard ratios were 8.0 for GD = +2 (p < 0.001), 1.9 for GD = +1 (p < 0.001), 0.57 for GD = -1 (p = 0.001), and 0.22 for GD = -2 (p = 0.003) versus GD = 0 as the reference. According to model-converted risk scores, DFS, OS, and CSS significantly differed between subgroups with low, intermediate, and high risk (all p < 0.001). CONCLUSIONS: Routine reporting of VTT upgrading or downgrading in relation to the PT and use of our GD-based nomograms can facilitate more informed treatment decisions by tailoring strategies to an individual patient's risk of progression. PATIENT SUMMARY: We developed a tool to improve patient counseling and guide decision-making on other therapies in addition to surgery for patients with the clear-cell type of kidney cancer and tumor invasion of a vein