Strengthening Biorisk Management in Research Laboratories with Security-Sensitive Biological Agents Like SARS-CoV-2.

In this chapter, we discuss potential incidents associated with SARS-CoV-2 experimental work in high containment research laboratories. The risk landscape in high containment laboratories is changing due to the strong innovation drive of the life sciences research. Thus, the WHO has recommended life sciences organizations to incorporate good research practices and ethical principles into a risk-based approach of the biorisk management (BRM). Currently, BRM systems in high containment laboratories are predominantly steered by operational personnel and laboratory professional. It is well known that without having a systematic approach and leadership support from the organization, the BRM system in the high containment laboratory will not be sustainable. Even though the roles of organizations and their leadership in establishing the BRM system are spelt out in many international standards, guidance documents and national legislations, operational aspects of these roles are rarely discussed.It is therefore important for everyone to understand about their roles in organizational processes (communication, decision, and performance evaluation) involved in implementation of BRM related operational activities. In this chapter, discussion is based on operational activities of four main organizational behaviors that are considered to have strengthened BRM systems in high containment laboratories: (1) displaying a visible commitment and support to the BRM system from different levels of management, (2) developing a competent and responsible workforce with BRM technical skills and problem identification/solving skills, (3) integrating learning and improvement principles into the BRM system, and (4) enhancing the continuous motivation of laboratory personnel to avoid complacency. The categorization of these organizational behaviors is based on the International Atomic Energy Agency's principles and guidance for strengthening the safety and security culture in nuclear facilities. Furthermore, we encourage the laboratory management to identify gaps in processes and activities related to those organizational behaviors so that one could rapidly address biosafety and biosecurity vulnerabilities in high containment laboratories.

Biorisk Management for SARS-CoV-2 Research in a Biosafety Level-3 Core Facility.

The emerging severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) presents hazards to researchers and other laboratory personnel in research settings where the live virus is stored and handled. The Biosafety Level-3 (BSL-3) Core Facility (CF) at Yong Loo Lin School of Medicine in National University of Singapore (NUS Medicine) has implemented a biorisk management (BRM) system to ensure that biorisk to employees, the public, or the environment are consistently minimized to an acceptable level while working with SARS-CoV-2. This chapter summarizes how a BRM system can be implemented in academic institutions based on international standards in the context of existing local legislations/regulations and institutional policies/guidelines to minimize the risk of laboratory-acquired infections and deliberate misuse of the newly emerged virus, SARS-CoV-2 in BSL-3 laboratories. The BRM programs prioritize performing risk assessments prior to implementation of work processes and reassessing the risk portfolio of the facilities from time to time, determining root causes and prevention of recurrences. Focusing on awareness-raising and educating the laboratory users in biosafety and biosecurity, and identifying opportunities for improvement are the other key factors for a sustainable and successful BRM system in the NUS Medicine BSL-3 CF.

Management System Approach for Addressing Biosafety and Biosecurity of Emerging Pathogens in a Biosafety Level-3 Core Facility.

Introduction: The biosafety level-3 (BSL-3) core facility (CF) at Yong Loo Lin School of Medicine (NUS Medicine) in National University of Singapore (NUS) has adopted international standards and guidelines to establish a biorisk management (BRM) system that helps to improve its BRM system and consistently minimize the risks to employees, the public, and the environment to an acceptable level while working with SARS-CoV-2. Methods: When the NUS Medicine BSL-3 CF started its operations, the Occupational Health and Safety Assessment Series 18001:2007 and the CEN Workshop Agreement 15793:2011 guidelines were used to establish its first BRM framework. The BRM framework provided the roadmap of how to organize, systematically manage, and structure the various biorisk programs that was then modified according to International Organization for Standardization 35001:2019 during the coronavirus disease 2019 pandemic in 2020 to address the specific circumstances. Results: Adopting a management system approach allowed BSL-3 CF to efficiently manage its BRM even during unpredicted emerging pandemic situations. It resulted in integrating a risk management process into daily laboratory operations and ongoing identification of hazards, prioritization of risks, and the establishment of risk mitigation measures specific to SARS-CoV-2. In addition, the implementation of a BRM system in the BSL-3 CF has increased biorisk awareness among BSL-3 CF users and encouraged every stakeholder to take ownership of their activities, and continual improvements in mitigation of biorisks. Discussion: This article summarizes the systematic approaches and major elements of the BRM systems adopted by NUS Medicine BSL-3 CF for the implementation of biosafety and biosecurity precautions, and control measures to minimize the risk of research activities using various RG3 biological agents including SARS-CoV-2.

Diagnostic challenges in cytology of mucoepidermoid carcinoma: Report of 6 cases with histopathological correlation.

BACKGROUND: Mucoepidermoid carcinoma (MEC) is a malignant salivary gland neoplasm with extreme morphologic heterogeneity and hence rendering a definitive fine needle aspiration cytology (FNAC) diagnosis of this neoplasm is really challenging. The present study was undertaken to elucidate the cytological features of MEC and explore the diagnostic accuracy and pitfalls by comparing with subsequent histopathology. MATERIALS AND METHODS: The present study was conducted over a period of 2 years wherein we obtained six histopathologically confirmed cases of MEC. These patients were initially subjected to FNAC. The cytologic features studied included presence of mucous cells, intermediate cells, and squamous cells. Presence of background mucinous material was also noted. The cytological features were compared with the subsequent histopathology. RESULTS: Of the 6 cases of MEC, a definite cytological diagnosis was possible only in 2 cases. Of the remaining 4 cases, 2 cases were broadly diagnosed in cytology as neoplasm with cystic degeneration and 2 cases were underdiagnosed as pleomorphic adenoma. CONCLUSIONS: A satisfactory aspirate with all three types of cells; mucous, intermediate and squamous cells may not be obtained in all cases of MEC for providing a definite diagnosis. Hence, a good clinicoradiological correlation, a high index of suspicion and repeated aspirations especially in cystic lesions may be particularly helpful in difficult cases. In addition, while dealing with mucinous cystic lesions with low cellularity, the importance of early excision should be communicated to the clinician since the possibility of low-grade MEC cannot be excluded.

An Unusual Case of Early Onset Persistent Escherichia coli Septicemia Associated with Endocarditis.

Escherichia coli infection is very common cause of early onset septicemia especially in very low-birth-weight newborns, but E. coli endocarditis has not been described in newborns. E. coli endocarditis, even in the adult population, is a rare and not well-characterized disease and is associated with high mortality. We report a very unusual presentation of persistent E. coli infection associated with endocarditis.

Juvenile granulosa cell tumor of testis: case report and review of literature.

Juvenile granulosa cell tumor (JGCT) of testis is extremely rare in childhood. It is considered a benign entity because metastasis has never been reported. Testicular-sparing surgery is the recommended treatment. We reported this case in a newborn who presented with unilateral scrotal swelling. Histopathology and immunohistochemistry confirmed JGCT. Follow-up at 6 months after surgery did not show any recurrence. Even though JGCT is very rare in childhood, it is one of the important differentials of newborn scrotal mass.

Levels of neuropeptides nocistatin, nociceptin/orphanin FQ and their precursor protein in a rat neuropathic pain model.

Neuropeptides nociceptin/orphanin FQ (N/OFQ) and nocistatin (NST) are related to pain modulation. The amounts of these peptides and their precursor protein, prepronociceptin (ppN/OFQ) in the brain, spinal cord and serum samples of rats with partial sciatic nerve ligation (PSNL) were compared with those in naïve rats using radioimmunoassay (RIA). There was a significant rise in the levels of ppN/OFQ, N/OFQ and NST in the brains of PSNL rats. Their spinal cords showed significantly increased ppN/OFQ and NST levels but no change in N/OFQ levels. The PSNL rats also had increased serum NST (statistically significant) and N/OFQ (statistically insignificant) with decreased ppN/OFQ suggesting important roles of these peptides in neuropathic pain mechanism.

Identification of mature nocistatin and nociceptin in human brain and cerebrospinal fluid by mass spectrometry combined with affinity chromatography and HPLC.

Nocistatin (NST) and nociceptin/orphanin FQ (NCP) are two important bio-peptides derived from the precursor protein prepronociceptin (ppNCP), involved in several central nervous system (CNS) functions including pain transmission. Since the actual form of human NST in CNS is not fully characterized, we studied the structure of NST from human brain tissue and cerebrospinal fluid (CSF) samples. NST and NCP were isolated from human brain and CSF samples by affinity chromatography combined with HPLC. Mass spectrometry was used for the identification and characterization of the peptides. The total NST immunoreactivity was detected as 11.5+/-2.3 pmol/g tissue for the brain and 0.44 pmol/ml for the pooled CSF sample after the HPLC purification by radioimmunoassay. The presence of two different forms of mature nocistatin (NST-17 and NST-30) and a possible N-terminal methionine cleaved NST-29 were confirmed by both radioimmunoassay and mass spectrometry. Affinity chromatography, HPLC and mass spectrometry methods used in this study were highly sensitive and suitable for identification of actual chemical structures and quantification of very small amounts of peptides in biological samples. The present findings may help further for search for new treatment of neuropathic pain, which is often poorly managed by current therapies.