Automated and robust extraction of genomic DNA from various leftover blood samples.

With the development of genomic technologies, the isolation of genomic DNA (gDNA) from clinical samples is increasingly required for clinical diagnostics and research studies. In this study, we explored the potential of utilizing various leftover blood samples obtained from routine clinical tests as a viable source of gDNA. Using an automated method with optimized pre-treatments, we obtained gDNA from seven types of clinical leftover blood, with average yields of gDNA ranging from 3.11 ± 0.45 to 22.45 ± 4.83 µg. Additionally, we investigated the impact of storage conditions on gDNA recovery, resulting in yields of 8.62-68.08 µg when extracting gDNA from EDTA leftover blood samples stored at 4 °C for up to 13 weeks or -80 °C for up to 78 weeks. Furthermore, we successfully obtained sequenceable gDNA from both Serum Separator Tube and EDTA Tube using a 96-well format extraction, with yields ranging from 0.61 to 71.29 µg and 3.94-215.98 µg, respectively. Our findings demonstrate the feasibility of using automated high-throughput platforms for gDNA extraction from various clinical leftover blood samples with the proper pre-treatments.

Plasma Rich in Growth Factors as an Adjuvant Treatment for the Management of Frontal Fibrosing Alopecia: A Retrospective Observational Clinical Study.

BACKGROUND: Frontal fibrosing alopecia (FFA) is a scarring alopecia in which the exact etiopathogenesis has not been completely elucidated and the available treatments are not very effective. Plasma rich in growth factors (PRGF) has shown to induce folliculogenesis in hair loss related disorders. However, the scientific evidence when facing FFA is scarce. OBJECTIVES: The aim of this study was to retrospectively analyze the adjuvant use of PRGF compared to the conventional treatment in the management of FFA. METHODS: Participants with clinically diagnosed FFA who had been treated with either conventional therapy (Control Group) or conventional therapy combined with PRGF (PRGF Group) were identified from the center's medical records. The clinical assessment was based on the "Frontal Fibrosing Alopecia Severity Score" (FFASS), which was fulfilled during a period of two and 4 years. RESULTS: This study included 118 patients with clinically diagnosed FFA (Control Group: 57 and PRGF Group: 61). No adverse effects related to the treatments were observed. Both treatments showed to halt the steady progression of hair loss compared to baseline. PRGF treatment also induced significant hair regrowth compared to the Control Group. The scalp inflammation was reduced in response to treatments. The FFASS score indicated that PRGF Group improved the symptoms and severity of FFA in a significant manner. CONCLUSIONS: The adjuvant use of PRGF may exert long-term beneficial effects on hair loss reduction and might reduce the symptoms and severity of FFA.

Protocol for Biospecimen Collection and Analysis Within the BACPAC Research Program.

The Biospecimen Collection and Processing Working Group of the National Institutes of Health (NIH) HEAL Initiative BACPAC Research Program was charged with identifying molecular biomarkers of interest to chronic low back pain (cLBP). Having identified biomarkers of interest, the Working Group worked with the New York University Grossman School of Medicine, Center for Biospecimen Research and Development-funded by the Early Phase Pain Investigation Clinical Network Data Coordinating Center-to harmonize consortium-wide and site-specific efforts for biospecimen collection and analysis. Biospecimen collected are saliva, blood (whole, plasma, serum), urine, stool, and spine tissue (paraspinal muscle, ligamentum flavum, vertebral bone, facet cartilage, disc endplate, annulus fibrosus, or nucleus pulposus). The omics data acquisition and analyses derived from the biospecimen include genomics and epigenetics from DNA, proteomics from protein, transcriptomics from RNA, and microbiomics from 16S rRNA. These analyses contribute to the overarching goal of BACPAC to phenotype cLBP and will guide future efforts for precision medicine treatment.

Longitudinal in Utero Analysis of Engrailed-1 Knockout Mouse Embryonic Phenotypes Using High-Frequency Ultrasound.

Large-scale international efforts to generate and analyze loss-of-function mutations in each of the approximately 20,000 protein-encoding gene mutations are ongoing using the "knockout" mouse as a model organism. Because one-third of gene knockouts are expected to result in embryonic lethality, it is important to develop non-invasive in utero imaging methods to detect and monitor mutant phenotypes in mouse embryos. We describe the utility of 3-D high-frequency (40-MHz) ultrasound (HFU) for longitudinal in utero imaging of mouse embryos between embryonic days (E) 11.5 and E14.5, which represent critical stages of brain and organ development. Engrailed-1 knockout (En1-ko) mouse embryos and their normal control littermates were imaged with HFU in 3-D, enabling visualization of morphological phenotypes in the developing brains, limbs and heads of the En1-ko embryos. Recently developed deep learning approaches were used to automatically segment the embryonic brain ventricles and bodies from the 3-D HFU images, allowing quantitative volumetric analyses of the En1-ko brain phenotypes. Taken together, these results show great promise for the application of longitudinal 3-D HFU to analyze knockout mouse embryos in utero.

Combined Therapy with Laser and Autologous Topical Serum for Facial Rejuvenation: A Multiple Case Series Report.

Background: Lasers require several sessions to achieve significant results and may lead to adverse reactions. Platelet-rich plasma (PRP) therapy is a good adjuvant to laser therapies; however, repeated blood extractions and invasive injections are needed. A 100% autologous topical formulation based on the patient's proteins has been recently developed, known as Endoret-Serum (ES). Unlike other PRPs, ES provides a home care, storable, and topical needle-free application. Objective: Preliminarily assess the clinical performance of a single session of the combined therapy with nonablative laser and ES in the management of cutaneous pigmented and vasculature lesions. Materials and Methods: Nine patients with clinical signs of skin aging received a single session of nonablative laser. ES was topically applied twice daily. They were clinically assessed after 1 and 8 weeks. VISIA-CR System was used for high-resolution topographic analysis. Subjects were asked to complete a self-assessment questionnaire and an impression of improvement survey. An investigator's global assessment scale was fulfilled. Results: The combined treatment improved cutaneous spots, wrinkles, and texture after 8 weeks, whereas significant pore reduction was observable at 1 week. Ultraviolet (UV) spots and porphyrins decreased at 1 week, whereas red area improvement was noticeable after 8 weeks. Overall wrinkle amelioration, periorbital hyperpigmentation decrease, softened skin, and tone recovery was observed. Patients referred to be very satisfied and felt that their cutaneous condition was much better. At the end of the study, subjects presented minimal dermatological symptoms like pigmented lesions, redness, or capillaries. No side effects were reported. Conclusion: Results presented herein suggest that one session of laser in combination with ES provides a good clinical outcome.

Combined therapy with Endoret-Gel and plasma rich in growth factors vs Endoret-Gel alone in the management of facial rejuvenation: A comparative study.

BACKGROUND: Skin suffers progressive decrement. An endogenous regenerative technology has been developed that has the versatility to provide an autologous injectable gel (Endoret-Gel) or a liquid plasma rich in growth factors (PRGF) based on the patient´s own platelet-rich plasma. AIMS: To compare the efficacy of the combined therapy with Endoret-Gel and PRGF versus Endoret-Gel alone in the management of facial rejuvenation. METHODS: Twenty clinically diagnosed patients with aged skin received either Endoret-Gel monotherapy or Endoret-Gel + PRGF combined therapy. Patients underwent three sessions at one-month intervals and were clinically assessed for six months. Corneometry, sebumetry, and high-resolution topographic analysis were carried out. Patient self-assessment questionnaires and clinical improvement scores were also performed. RESULTS: The combined therapy showed to promote a higher hydration index. These results were also significant for spot improvement at three months, while conversely, monotherapy with Endoret-Gel demonstrated higher UV spot improvement. A significant decrease of sebum production and wrinkle development was observed for both treatment groups. Red areas also improved in a similar way at the end of the follow-up period. After Endoret-Gel or Endoret-Gel + PRGF therapy, 30% and 70% of patients referred to be very satisfied, respectively. Accordingly, 40% and 80% showed a "very improved" esthetic performance. None of the patients reported a negative change and no adverse events were recorded. CONCLUSION: Both Endoret-Gel monotherapy and the combined treatment with PRGF were shown to promote facial rejuvenation and to palliate the age-related cutaneous atrophy. The combined therapy may exert a synergistic effect that addresses both skin quality improvement and soft tissue restoration in a shorter period.

ImagCell: a computer tool for cell culture image processing applications in bioimpedance measurements.

This paper presents a computer tool for automatic analysis of cell culture images. The program allows the extraction of relevant information from biological images for pre- and postsystem analysis. In particular, this tool is being used for electrical characterization of electrode-solution-cell systems in which bioimpedance is the main parameter to be known. The correct modeling of this kind of systems enables both electronic system characterization for circuit design specifications and data decoding from measurements. The developed program allows cell culture image processing for geographic information extraction and generates cell count and equivalent circuit descriptions useful for system simulations.