Efficacy and Safety of Skin Radiance Collagen on Skin and Hair Matrix: A Placebo-Controlled Clinical Trial in Healthy Human Subjects.

Purpose: Collagen supplements are rising in the market as collagen has been demonstrated to be an important protein in the human aging process. Also, it is safe and easily absorbed in the body. Hence the aim of this study was to examine the effectiveness and safety of a collagen and antioxidant-rich treatment compared to a placebo in relation to various skin and hair indicators in healthy adult human subjects. Patients and Methods: Forty healthy adult non-pregnant/non-lactating women (aged 38-50 years) provided their informed consent in writing before their participation. Skin Radiance Collagen (SRC) treatment and a placebo were assessed for efficacy before application on Day 1, and post-application on Days 28 and 56, to measure changes in skin elasticity, hydration, brightness, pigmentation; texture, wrinkles, dryness, smoothness, fine lines, changes in the crow's feet region; as well as hair strength and hair fall. Results: It was observed after 56 days that therapy with SRC, compared to placebo, produced a substantial effect on reduction of wrinkle depth and fine lines by 48.11% and 39%, respectively, with p-value <0.01 in the test group. There was a 15.69% improvement in skin hydration observed and 28% reduction in hair fall with p-value <0.01. Conclusion: SRC, a combination of collagen with hyaluronic acid (HA), biotin, and vitamins C and E, showed a significant improvement in skin and hair health, including improvements in skin elasticity, skin hydration, reduction in crow's feet area wrinkles and fine lines, hair fall, and decrease in roughness, leading to improved skin texture. Vitamin C in the formulation also acts as a collagen builder for the body and helps in preventing oxidative stress in the body. The test treatment SRC was found to be efficacious and safe in healthy human adult subjects.

BODIPY-Hg2+ Complex: A Fluorescence "Turn-ON" Sensor for Cysteine Detection.

A BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) based pioneering sensing material (HLPy) having 2-amino pyridine as receptor was synthesized and used for the selective detection of Hg2+ ions. The synthesized HLPy features a high affinity towards Hg2+ (ka = 2.04 × 105 M-1), accompanied by effective quenching of fluorescence in DMF:H2O (1:9 v/v, 10 mM HEPES buffer, pH 7.4) with 54 nM limit of detection (LOD). The emission titration experiments (Job's plot) in the presence of varying mole-fraction of Hg2+ ions reveals the formation of non-fluorescent 2:1 coordination complex [Hg(LPy)2]. The resulting non-fluorescent [Hg(LPy)2] was thoroughly characterized using various spectroscopic techniques and analyses. Interestingly, the non-fluorescent complex [Hg(LPy)2] is able to specifically respond towards Cys over other biothiols and amino acids through a reversible de-complexation mechanism. As a result, the remarkable recovery of the fluorescence can be observed. The limit of detection (LOD) for Cys detection is estimated to be 29 nM in DMF:H2O (1:9 v/v, 10 mM HEPES buffer, pH 8.0). The reversibility and reusability of [Hg(LPy)2] were achieved by the sequential addition of Cys and Hg2+ ions up to five cycles. Moreover, the removal of Hg2+ ions up to 89% from aqueous samples using HLPy was successfully demonstrated.

Understanding the effect of high gas entrance velocity on Chinese hamster ovary (CHO) cell culture performance and its implications on bioreactor scale-up and sparger design.

There are three main potential sources for cell shear damage existing in stirred tank bioreactors. One is the potential high energy dissipation in the immediate impeller zones; another from small gas bubble burst; and third is from high gas entrance velocity (GEV) emitting from the sparger. While the first two have been thoroughly addressed for the scale-up of Chinese hamster ovary (CHO) cell culture knowing that a wide tolerable agitation range with non-damaging energy dissipation exists and the use of shear protectants like Pluronic F68 guard against cell damage caused by bubble burst, GEV remains a potential scale-up problem across scales for the drilled hole or open pipe sparger designs. GEV as high as 170 m/s due to high gas flow rates and relatively small sparger hole diameters was observed to be significantly detrimental to cell culture performance in a 12,000 L bioreactor when compared to a satellite 2 L bioreactor run with GEV of <1 m/s. Small scale study of GEV as high as 265 m/s confirmed this. Based on the results of this study, a critical GEV of >60 m/s for CHO cells is proposed, whereas previously 30 m/s has been reported for NS0 cells by Zhu, Cuenca, Zhou, and Varma (2008. Biotechnol. Bioeng., 101, 751-760). Implementation of new large scale spargers with larger diameter and more holes lowered GEV and helped improve the cell culture performance, closing the scale-up gap. Design of such new spargers was even more critical when hole plugging was discovered during large scale cultivation hence exacerbating the GEV impact. Furthermore, development of a scale down model based on mimicry of the large scale GEV profile as a function of time was proven to be beneficial for reproducing large scale results.

Benefit of STR-based chimerism analysis to identify TA-GVHD as a cause of death: Utility of various biological specimens.

Transfusion-associated graft-versus-host disease (TA-GVHD) is a rare condition. It can occur after blood transfusion in immune-compromised and occasionally even in immune-competent patients, and is associated with a mortality rate of >90%. The diagnosis of TA-GVHD is often delayed because of its non-specific clinical features. A case of an immune-competent child who developed TA-GVHD is reported here. DNA profiling (short tandem repeat analysis), a technique that has a wide application in forensic medicine, was performed to detect the presence of donor cells in this patient. The findings suggest that more studies are needed with this tool, and the diagnostic potential of using other multiple biological specimens for DNA profiling such as the hair follicle and buccal swab should be evaluated. This is the first case report where the donor's DNA fingerprinting pattern was substantiated from a patient's hair follicle sample. Chimerism was also present in the blood and buccal swab specimens.

Evaluation of blood, buccal swabs, and hair follicles for DNA profiling technique using STR markers.

AIM: To study the short tandem repeat (STR) pattern of DNA from the blood, buccal swabs, and hair follicles of the recipients of allogenic hematopoietic stem cell transplantation to examine whether these tissues contain donor derived cells. METHODS: The study enrolled 25 patients who sustained engraftment. Peripheral blood, buccal swabs, and hair follicles were collected on days 21-30, 90, and 180 after transplantation and the chimeric status of the recipients was evaluated. RESULTS: Donor derived cells existed in the blood and buccal swabs, but not in hair follicles, which can be used to obtain the pre-transplant sample of the recipient after transplant. CONCLUSION: Peripheral blood and buccal swab do not serve as a reliable source of recipient's origin for DNA analysis of individuals who underwent allogenic hematopoietic stem cell transplantation at least within 6 months after transplant.

Detection of Ralstonia solanacearum from asymptomatic tomato plants, irrigation water, and soil through non-selective enrichment medium with hrp gene-based bio-PCR.

Bacterial wilt of tomato caused by Ralstonia solanacearum (Smith) Yabuuchi et al. (Microbiol Immunol 39:897-904, 1995) is a serious disease, which causes losses up to 60 % depending on environmental conditions, soil property, and cultivars. In present investigation, nucleotide sequences of virulence, hypersensitive response and pathogenicity (hrp) gene were used to design a pair of primer (Hrp_rs 2F: 5'-AGAGGTCGACGCGATACAGT-3' and Hrp_rs 2R: 5'-CATGAGCAAGGACGAAGTCA-3') for amplification of bacterial genome. The genomic DNA of 27 isolates of R. solanacearum race 1 biovar 3 & 4 was amplified at 323 bp. The specificity of primer was tested on 13 strains of R. solanacearum with other group of bacteria such as Xanthomonas oryzae pv. oryzae, X. campestris pv. campestris, and X. citri subsp. citri. Primer amplified DNA fragment of R. solanacearum at 323 bp. The sensitivity of the primer was 200 cfu/ml and improved further detection level by using non-specific enrichment medium casamino acids-pepton-glucose broth followed by PCR (BIO-PCR). Out of 130 samples of asymptomatic tomato plants, irrigation water, and soil collected from bacterial wilt infested field in different agro-climatic regions of India, R. solanacearum was detected from 86.9, 88.5, and 90.9 per cents samples using BIO-PCR, respectively. The primer was found specific for detecting viable and virulent strains of R. solanacearum and useful for the diagnosis of R. solanacearum in tomato seedlings and monitoring of pathogen in irrigation water and soil.

ELP-z and ELP-zz capturing scaffolds for the purification of immunoglobulins by affinity precipitation.

The increasing demand of monoclonal antibodies for therapeutic applications along with the high manufacturing cost have made it necessary to evaluate better process options and technologies for their purification. Affinity precipitation is an attractive alternative to traditional chromatographic methods by affording effective purification using a simple environmental trigger. The feature of elastin-like-protein (ELP) fused with antibody binding domains has already been explored for the purification of antibodies. However, ELP when fused with the bulkier domains such as Protein A, resulted in lower protein production. In this study, ELP was fused to smaller synthetic IgG binding domains such as the z or zz domain, resulting in up to 10-fold higher level of production. Both ELP-z and ELP-zz bind tightly to human immunoglobulin (HIgG) with a dissociation constant of 768±142 nM and 68±23 nM, respectively. Owing to the higher binding affinity, the use of ELP-zz resulted in more than 99% recovery of HIgG in four repeated binding and elution cycles with no observable decrease in the purification performance. The same binding and elution cycle was successfully implemented for the purification of monoclonal antibodies from hybridoma culture supernatant with close to 100% recovery.