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Tumor spheroid models have garnered significant attention in recent years as they can efficiently mimic in vivo models, and in addition, they offer a more controlled and reproducible environment for evaluating the efficacy of cancer drugs. In this study, we present the design and fabrication of a micromold template to form multicellular spheroids in a high‐throughput and controlled‐sized fashion. Briefly, polydimethylsiloxane‐based micromolds at varying sizes and geometry were fabricated via soft lithography using 3D‐printed molds as negative templates. The efficiency of spheroid formation was assessed using GFP‐expressing human embryonic kidney 293 cells (HEK‐293). After 7 days of culturing, circularity and cell viability of spheroids were >0.8 and 90%, respectively. At 1500 cells/microwell of cell seeding concentration, the spheroids were 454 ± 15 μm, 459 ± 7 μm, and 451 ± 18 μm when cultured in microwells with the diameters of 0.4, 0.6, and 0.8 μm, respectively. Moreover, the distance between each microwell and surfactant treatment before cell seeding notably impacted the uniform spheroid formation. The centrifugation was the key step to collect cells on the bottom of the microwells. Our findings were further verified using a commercial microplate. Furthermore, Monte Carlo simulation confirmed the seeding conditions where the spheroids could be formed. This study showed prominent steps in investigating spheroid formation, thereby leveraging the current know‐how on the mechanism of tumor growth. [ABSTRACT FROM AUTHOR]

Reactivation of latent viral reservoirs is on the forefront of HIV-1 eradication research. However, it is unknown if latency reversing agents (LRAs) increase the level of viral transcription from cells producing HIV RNA or harboring transcriptionally-inactive (latent) infection. We therefore developed a microfluidic single-cell-in-droplet (scd)PCR assay to directly measure the number of CD4+ T cells that produce unspliced (us)RNA and multiply spliced (ms)RNA following ex vivo latency reversal with either an histone deacetylase inhibitor (romidepsin) or T cell receptor (TCR) stimulation. Detection of HIV-1 transcriptional activity can also be performed on hundreds of thousands of CD4+ T-cells in a single experiment. The scdPCR method was then applied to CD4+ T cells obtained from HIV-1-infected individuals on antiretroviral therapy. Overall, our results suggest that effects of LRAs on HIV-1 reactivation may be heterogeneous-increasing transcription from active cells in some cases and increasing the number of transcriptionally active cells in others. Genomic DNA and human mRNA isolated from HIV-1 reactivated cells could also be detected and quantified from individual cells. As a result, our assay has the potential to provide needed insight into various reservoir eradication strategies.

HIV-1 is a major global epidemic that requires sophisticated clinical management. There have been remarkable efforts to develop new strategies for detecting and treating HIV-1, as it has been challenging to translate them into resource-limited settings. Significant research efforts have been recently devoted to developing point-of-care (POC) diagnostics that can monitor HIV-1 viral load with high sensitivity by leveraging micro- and nano-scale technologies. These POC devices can be applied to monitoring of antiretroviral therapy, during mother-to-child transmission, and identification of latent HIV-1 reservoirs. In this review, we discuss current challenges in HIV-1 diagnosis and therapy in resource-limited settings and present emerging technologies that aim to address these challenges using innovative solutions.

Recent advances in biosensing technologies present great potential for medical diagnostics, thus improving clinical decisions. However, creating a label-free general sensing platform capable of detecting multiple biotargets in various clinical specimens over a wide dynamic range, without lengthy sample-processing steps, remains a considerable challenge. In practice, these barriers prevent broad applications in clinics and at patients' homes. Here, we demonstrate the nanoplasmonic electrical field-enhanced resonating device (NE(2)RD), which addresses all these impediments on a single platform. The NE(2)RD employs an immunodetection assay to capture biotargets, and precisely measures spectral color changes by their wavelength and extinction intensity shifts in nanoparticles without prior sample labeling or preprocessing. We present through multiple examples, a label-free, quantitative, portable, multitarget platform by rapidly detecting various protein biomarkers, drugs, protein allergens, bacteria, eukaryotic cells, and distinct viruses. The linear dynamic range of NE(2)RD is five orders of magnitude broader than ELISA, with a sensitivity down to 400 fg/mL This range and sensitivity are achieved by self-assembling gold nanoparticles to generate hot spots on a 3D-oriented substrate for ultrasensitive measurements. We demonstrate that this precise platform handles multiple clinical samples such as whole blood, serum, and saliva without sample preprocessing under diverse conditions of temperature, pH, and ionic strength. The NE(2)RD's broad dynamic range, detection limit, and portability integrated with a disposable fluidic chip have broad applications, potentially enabling the transition toward precision medicine at the point-of-care or primary care settings and at patients' homes.

Objective: To determine the value of CT-guided bone core biopsy and investigate factors that affect diagnostic yield and biopsy outcome. Materials and methods: The single-centre retrospective analysis included 447 patients who had CT-guided core biopsy with a 13-G needle (Bonopty®) from January 2016 to December 2021. Histological results or ≥ 6 months of clinical and radiological follow-up served as outcome references. A successful biopsy was classified as "diagnostic" when a definitive diagnosis was made and "adequate" when only the malignant or benign nature of the tumour could be determined. Biopsies were "nondiagnostic" when the nature of the lesion could not be determined. The occult lesions were defined as not seen on CT but visible on other modalities. Results: In 275 (62%) females and 172 (38%) males, the overall success rate was 85% (383 biopsies), with 314 (70%) diagnostic biopsies and 69 (15%) adequate biopsies. There was no relationship between biopsy success and the localisation of the lesion, length of biopsy material, or number of biopsy attempts. The lesions' nature had a statistically significant effect on biopsy success with lytic and mixed lesions having the highest success rate. Occult lesions had the lowest success rate. Conclusion: CT-guided bone core biopsy is an effective method in the workup of musculoskeletal diseases with the highest success rate in lytic and mixed lesions. No apparent relationship was found between biopsy success and biopsy length, number of attempts, or localisation of the lesion. [ABSTRACT FROM AUTHOR]

Acetic acid bacteria (AAB) are ubiquitous wine spoilage microorganisms causing significant economic damage to winemakers. Considering difficulties in their isolation through traditional microbiological methods, it would be advantageous to detect them using molecular methods at all stages of winemaking and, thus, prevent wine spoilage. In this research, we analyzed wines, musts and grapes of 13 varieties grown in different regions of the Republic of Moldova. The DNA was extracted and analyzed via PCR using home-designed primers to detect Acetobacter aceti and Acetobacter pasteurianus. Generally, samples with no detectable amounts of AAB in either musts or wine had volatile acidity within the acceptable limits. Only one grape (Rara Neagra) had detectable amounts of AAB (A. pasteurianus) at all analyzed stages (grape, must, wine), and this sample had the highest amount of volatile acidity (2.11 g/L), exceeding the maximum acceptable limit for red wines of 1.2 g/L. A. pasteurianus was more common than A. aceti, both in musts and wines. Samples positive for AAB but containing low amounts of them in wine (Cq value > 35) did not have volatile acidity above the acceptable level. Samples that were wine-negative but must-positive for AAB had volatile acidity close to the acceptable limit. This study shows the utility of PCR diagnostics for predicting the risks of wine spoilage by AAB. [ABSTRACT FROM AUTHOR]

Aim: To evaluate the role of MMP-13 rs2252070 in patients with non-small cell lung cancer (NSCLC) in the Turkish population. Method: A total of 95 NSCLC patients and 94 healthy controls were included in this study. The MMP-13 rs2252070 variant was genotyped by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The results of the analyses were evaluated for statistical significance. Results: There was no G/G homozygous genotype in the patient or control groups. The prevalence of genotypes of A/A and A/G profiles for the MMP-13 rs2252070 variant was 34.7% and 65.3%, respectively, in patients and 46.8% and 53.2%, respectively, in the control group. No significant difference was found between the patient and control groups in terms of MMP-13 rs2252070 genotype distribution and allele frequency (p= 0.091, OR: 0.605, CI 95%:0.337-1.086; p: 0.199, OR: 1.337, CI 95%: 0.858-2.083, respectively). Conclusions: Our results in this study showed no association between MMP-13 rs2252070 and NSCLC. To fully comprehend the mechanisms underlying NSCLC development, more research is required. [ABSTRACT FROM AUTHOR]

Psychiatric findings may be seen in Fibromyalgia (FM) patients. We aimed to evaluate the relationship between anxiety, depression, and pain catastrophizing in FM patients and normal individuals. The study group consisted of FM patients and a healthy control group. Socio-demographic data form, Beck Depression Inventory (BDI), Beck Anxiety Inventory (BAI), Visual Analogue Scale (VAS), Fibromyalgia Impact Questionnaire (FIQ), and Pain Catastrophizing Scale (PCS) were administered to the participants. It was evaluated whether there was a statistical difference between the groups in terms of BDI, BAI, PCS, VAS, and FIQ. Of the 178 patients in our study, 85 were FM patients and 93 were control groups. The BDI, BAI, PCS, PCSH (PCS Helplessness), PCSM (PCS Magnification), and PCSR (PCS Rumination) were found to be higher in FM patients compared to the control group. The FM patients were found to have moderate depression according to the BDI score (18.7±10.6) and moderate anxiety according to the BAI score (16.5±9.9). There was a positive correlation between BDI and BAI, FIQ and VAS, PCS and BDI, BAI and FIQ, and BAI and FIQ. It was found that the FM patients had "moderate" depression and anxiety, and their level of pain catastrophizing increased with the increase in depression and anxiety severity. [ABSTRACT FROM AUTHOR]