Current advances in the development of meniscus tissue engineering: narrative review.

INTRODUCTION: The meniscus plays an important role in maintaining homeostasis to facilitate the normal function of the knee joint. It is one of the most commonly injured areas of the knee joint. Meniscal-related injuries can lead to significantly decreased athletic ability, and their incidence has increased yearly. It has been found that most meniscal injuries are irreparable, and meniscectomy can increase the predisposition to knee osteoarthritis. Tissue engineering technology on meniscus repairing and transplantation has received widespread attention recently. This review aimed to analyse the scientific literature regarding the potential applications of tissue engineering on meniscus repairing and transplantation procedures. METHOD AND MATERIALS: The electronic search was carried out using PubMed/MEDLINEⓇdatabases with the keywords "tissue engineering AND meniscus" spanning the period of publications from Jan 1980 until Dec 2022. RESULTS: The literature search identified 405 references in PubMed/MEDLINE, and 179 were selected following the eligibility requirements. The research analysis showed that the existing meniscal tissue engineering studies used a wide variety of seed cells, cytokines, bioactive materials and 3D structures. Each showed distinct advantages and disadvantages in terms of biocompatibility, degradability, mechanical strength, porosity, and etc. It was noted that 3D printing technology is promising for tissue engineering meniscus research. In addition, the optimal use of compression and hydrostatic pressure to markedly improve the functional properties of tissue-engineering meniscal can serve as an useful strategy. CONCLUSION: This review analysed the different approaches employed for meniscus tissue engineering and regeneration. Meniscal tissue engineering still faces several major challenges in terms of seed cells, choice of materials and 3D printing strategies, which should be effectively overcome to harness the full potential of this technology.

Development of a rapid viability RT-PCR (RV-RT-PCR) method to detect infectious SARS-CoV-2 from swabs.

Since the rapid onset of the COVID-19 pandemic, its causative virus, Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2), continues to spread and increase the number of fatalities. To expedite studies on understanding potential surface transmission of the virus and to aid environmental epidemiological investigations, we developed a rapid viability reverse transcriptase PCR (RV-RT-PCR) method that detects viable (infectious) SARS-CoV-2 from swab samples in <1 day compared to several days required by current gold-standard cell-culture-based methods. The method integrates cell-culture-based viral enrichment in a 96-well plate format with gene-specific RT-PCR-based analysis before and after sample incubation to determine the cycle threshold (CT) difference (ΔCT). An algorithm based on ΔCT ≥ 6 representing ∼ 2-log or more increase in SARS-CoV-2 RNA following enrichment determines the presence of infectious virus. The RV-RT-PCR method with 2-hr viral infection and 9-hr post-infection incubation periods includes ultrafiltration to concentrate virions, resulting in detection of <50 SARS-CoV-2 virions in swab samples in 17 h (for a batch of 12 swabs), compared to days typically required by the cell-culture-based method. The SARS-CoV-2 RV-RT-PCR method may also be useful in clinical sample analysis and antiviral drug testing, and could serve as a model for developing rapid methods for other viruses of concern.

Rapid viability polymerase chain reaction method for detection of Francisella tularensis.

Francisella tularensis, which causes potentially fatal tularemia, has been considered an attractive agent of bioterrorism and biological warfare due to its low infectious dose, reported environmental persistence, and ability to be transmitted to humans via multiple exposure routes. Due to slow growth on even selective culture media, detection of viable F. tularensis from environmental and drinking water samples, usually takes >3 days. Therefore, a rapid viability polymerase chain reaction (RV-PCR) method was developed to detect and identify viable F. tularensis cells in environmental samples. The method uses a change in PCR response during high throughput (48-well) sample incubation in Brain Heart Infusion/Vitox/Fildes/Histidine growth medium to detect viable F. tularensis presence, which is at least two times faster than the current plate culture-based method. Using the method, 101 to 102 live F. tularensis cells were detected in simulated complex sample matrices containing chemical and biological interferences.

Development of a rapid viability polymerase chain reaction method for detection of Yersinia pestis.

Due to the occurrence of natural plague outbreaks and its historical usage as a biological weapon, Yersinia pestis is considered one of the high-priority biological threat agents. It can remain viable in certain environments including water for >100 days. Because of its slow-growth characteristic, it usually takes three or more days to detect and confirm the identity of viable Y. pestis cells by PCR, serological, or biochemical assays when using the traditional microbiological plate-culture-based analysis, and that too, assuming faster growing microbes present in a water sample do not mask the Y. pestis colonies and interfere with analysis. Therefore, a rapid-viability Polymerase Chain Reaction (RV-PCR) method was developed for detection of Y. pestis. The RV-PCR method combines 24 h-incubation broth culture in a 48-well plate, and pre- and post-incubation differential PCR analyses, thereby allowing for rapid and high-throughput sample analysis compared with the current plate culture method. One chromosomal and two plasmid gene target-based real-time PCR assays were down-selected, showing ca. 10 genome equivalent detection; the chromosomal assay was then used for RV-PCR method development. A 101-cell level (10-99 cells) sensitivity of detection was demonstrated even with complex sample backgrounds including known PCR inhibitors (ferrous sulfate and humic acid), as well as metal oxides and microbes present in Arizona Test Dust (ATD). The method sensitivity was maintained in the presence of dead Y. pestis cells up to 104 cells per sample. While affording high-throughput and rapid sample analysis, the 48-well plate format used in this method for sample enrichment significantly reduced labor requirements and generation of BioSafety Level-3 (BSL-3) laboratory waste as compared to the usual microbiological plate-culture-based methods. This method may serve as a model for other vegetative bacterial pathogens.

Research Challenges in Financial Data Modeling and Analysis.

Significant research challenges must be addressed in the cleaning, transformation, integration, modeling, and analytics of Big Data sources for finance. This article surveys the progress made so far in this direction and obstacles yet to be overcome. These are issues that are of interest to data-driven financial institutions in both corporate finance and consumer finance. These challenges are also of interest to the legal profession as well as to regulators. The discussion is relevant to technology firms that support the growing field of FinTech.

Optimization of a sample processing protocol for recovery of Bacillus anthracis spores from soil.

Following a release of Bacillus anthracis spores into the environment, there is a potential for lasting environmental contamination in soils. There is a need for detection protocols for B. anthracis in environmental matrices. However, identification of B. anthracis within a soil is a difficult task. Processing soil samples helps to remove debris, chemical components, and biological impurities that can interfere with microbiological detection. This study aimed to optimize a previously used indirect processing protocol, which included a series of washing and centrifugation steps. Optimization of the protocol included: identifying an ideal extraction diluent, variation in the number of wash steps, variation in the initial centrifugation speed, sonication and shaking mechanisms. The optimized protocol was demonstrated at two laboratories in order to evaluate the recovery of spores from loamy and sandy soils. The new protocol demonstrated an improved limit of detection for loamy and sandy soils over the non-optimized protocol with an approximate matrix limit of detection at 14spores/g of soil. There were no significant differences overall between the two laboratories for either soil type, suggesting that the processing protocol will be robust enough to use at multiple laboratories while achieving comparable recoveries.

Rapid-viability PCR method for detection of live, virulent Bacillus anthracis in environmental samples.

In the event of a biothreat agent release, hundreds of samples would need to be rapidly processed to characterize the extent of contamination and determine the efficacy of remediation activities. Current biological agent identification and viability determination methods are both labor- and time-intensive such that turnaround time for confirmed results is typically several days. In order to alleviate this issue, automated, high-throughput sample processing methods were developed in which real-time PCR analysis is conducted on samples before and after incubation. The method, referred to as rapid-viability (RV)-PCR, uses the change in cycle threshold after incubation to detect the presence of live organisms. In this article, we report a novel RV-PCR method for detection of live, virulent Bacillus anthracis, in which the incubation time was reduced from 14 h to 9 h, bringing the total turnaround time for results below 15 h. The method incorporates a magnetic bead-based DNA extraction and purification step prior to PCR analysis, as well as specific real-time PCR assays for the B. anthracis chromosome and pXO1 and pXO2 plasmids. A single laboratory verification of the optimized method applied to the detection of virulent B. anthracis in environmental samples was conducted and showed a detection level of 10 to 99 CFU/sample with both manual and automated RV-PCR methods in the presence of various challenges. Experiments exploring the relationship between the incubation time and the limit of detection suggest that the method could be further shortened by an additional 2 to 3 h for relatively clean samples.

Prevalence of overweight and obesity in Indian adolescent school going children: its relationship with socioeconomic status and associated lifestyle factors.

AIMS AND OBJECTIVES: Obesity and overweight have become a worldwide epidemic, and there is an urgent need to examine childhood obesity and overweight across countries using a standardized international standard. In the present study we have investigated the prevalence of obesity and overweight and their association with socioeconomic status (SES) and the risk factors like diet, physical activity like exercise, sports, sleeping habit in afternoon, eating habits like junk food, chocolate, eating outside at weekend, family history of diabetes and obesity. MATERIAL AND METHODS: The study was carried out in 5664 school children of 12-18 years of age and having different SES. The obesity and overweight were considered using an updated body mass index reference. SES and life style factors were determined using pre-tested questionnaire. RESULTS: Age-adjusted prevalence of overweight was found to be 14.3% among boys and 9.2% among girls where as the prevalence of obesity was 2.9% in boys and 1.5% in girls. The prevalence of overweight among children was higher in middle SES as compared to high SES group in both boys and girls whereas the prevalence of obesity was higher in high SES group as compared to middle SES group. The prevalence of obesity as well as overweight in low SES group was the lowest as compared to other group. Eating habit like junk food, chocolate, eating outside at weekend and physical activity like exercise, sports, sleeping habit in afternoon having remarkable effect on prevalence on overweight and obesity among middle to high SES group. Family history of diabetes and obesity were also found to be positively associated. CONCLUSION: Our data suggest that the prevalence of overweight and obesity varies remarkably with different socioeconomic development levels.

Study of process induced polymorphic transformations in fluconazole drug.

The polymorphic form-I of the fluconazole drug commonly crystallized from the solution phase could be obtained by the solid state transformation of form-II employing different process parameters. As received fluconazole-II drug melted at 138.4 degrees C. The molten drug undercooled almost to ambient temperature of 30 degrees C and solidified to a glassy mass which, on ageing for 48 h transformed to a white powder which could be identified as fluconazole-I. The same glassy mass on heating at 5 degrees C/min, without ageing, also underwent polymorphic transformation to fluconazole-I above 81 degrees C. The application of uniaxial pressure of 200 kg/cm2 on as received fluconazole-II sample also yielded form-I of the drug. This phase transformation was enhanced by the application of pressure (200 kg/cm2) on the as received sample aged for 36 months. The phase transformation was concluded from the difference in differential scanning calorimetric (DSC) curves of the original sample (form-II) and the products obtained by adopting the different processing routes. The DSC patterns of fluconozole-I obtained by different methods were found to be identical. The phase transformation in the as received drug (form-II) induced by different process parameters, concluded from the DSC data was corroborated by X- ray diffraction (XRD) studies and scanning electron microscope (SEM) photographs of the two polymorphic forms. The intrinsic dissolution rates of polymorphic form-I and -II and the influence of crystal habit on the drug dissolution process have also been studied.

Study of polymorphic transformation of ornidazole drug by differential scanning calorimetry and other complementary techniques.

Differential scanning calorimetric (DSC) curves recorded for ornidazole drug during heating and cooling showed that the drug which melted around 86.1 degrees C undercooled to well below ambient room temperature of 27 degrees C during the cooling cycle. The undercooled melt kept in the freezer at 0 degree C for 10 days duration also remained in the viscous liquid form. This liquid on taking out from the freezer after ten days and ageing at ambient room temperature of 27 degrees C for 12 h transformed into white powder. The DSC pattern recorded for this white powder consisted of two prominent endothermic peaks beginning at 73.2 and 85.9 degrees C, respectively, suggesting that the powder consisted of a mixture of more than one phase. The X-ray diffraction (XRD) pattern recorded for this powder showed it to be a mixture of semi-crystalline phase and the original compound. The semi-crystalline phase melted at 73.2 degrees C prior to the melting of original compound at 85.9 degrees C. This phase on further ageing for 7 days transforms almost completely to its original form. DSC observations were corroborated by XRD and scanning electron microscopy (SEM) techniques.

Synthesis and biological activity of mixed ligand dioxouranium(VI) and thorium(IV) complexes.

Mixed ligand complexes of dioxouranium(VI) and thorium(IV) in the proportion 1:1:1 and 1:2:1 have been synthesized using 8-hydroxyquinoline as a primary ligand and L-proline and 4-hydroxy-L-proline as secondary ligands, respectively. The metal complexes have been characterized on the basis of elemental analysis, molar conductance, magnetic, spectral and thermal studies. The molar conductance studies of the complexes in DMF at 10(-3) M concentrations indicate their non-electrolytic nature. Room temperature magnetic susceptibility measurements revealed diamagnetic nature of the complexes. Electronic absorption spectra of the complexes show intra-ligand and charge transfer transitions, respectively. The thermal analysis data of the complexes indicates the presence of a coordinated water molecule/molecules. The tube dilution method has been used to study the antibacterial activity of the complexes against the pathogenic bacteria Staphylococcus aureus and Escherichia coli. The results have been compared against those of control tetracycline, which was screened simultaneously. The complexes have been screened for in vitro cytotoxicity (IC50) studies against Ehrlich ascites cells and Dalton's lymphoma ascites cells, respectively.