PER2 gene and its association with sleep-related disorders: A review.

The natural circadian rhythm in an individual governs the sleep-wake cycle over 24 h. Disruptions in this internal cycle can lead to major health hazards and sleep disorders. Reports suggest that at least 50 % of people worldwide suffer from sleep-related disorders. An increase in screen time, especially in the wake of the COVID-19 pandemic, is one of the external causative factors for this condition. While many factors govern the circadian clock and its aberrance, the PER2 gene has been strongly linked to chronotypes by many researchers. The current paper provides an extensive examination of key Single Nucleotide Polymorphisms within the PER2 gene and their potential connection to four major types of sleep disorders. This study investigates whether these SNPs play a causative role in sleep disorders or if they are solely associated with these conditions. Additionally, we explore whether these genetic variations exert a lifelong influence on these sleep patterns or if external triggers contribute to the development of sleep disorders. This gene is a crucial regulator of the circadian cycle responsible for the transcription of other clock genes. It regulates a variety of physiological systems such as metabolism, sleep, body temperature, blood pressure, endocrine, immunological, cardiovascular, and renal function. We aim to establish some clarity to the multifaceted nature of this gene, which is often overlooked, and seek to establish the mechanistic role of PER2 gene mutations in sleep disorders. This will improve further understanding, assessment, and treatment of these conditions in future.

Induction of epithelial to mesenchymal transition in HPV16 E6/E7 oncogene transfected C33A cell line.

This study focuses on the induction of EMT by HPV16 in the C33A cell line. Expression of ß-catenin, EMT-transcription factors (EMT-TFs), and c-myc in the nuclei of HPV16 E6/E7 oncogene transfected and non-transfected C33A cells were investigated through immunofluorescence and RT-PCR. Microphotographs of ß-catenin, c-myc, and DAPI-stained nuclei were processed and analyzed by Python and ImageJ respectively. Microphotographs of immunocytochemically stained transfected and control cells were then processed and analyzed with the help of ImageJ and Python programming. The intensity and the integrated density of ß-catenin were computed at the cell membrane area as well as the cytoplasmic area along with the integrated density of c-myc and Renyi entropy of DAPI-stained nuclei was quantified by ImageJ software. Python programming was implemented to determine the total percentage of white pixels depicting the presence of ß-catenin in the cytoplasmic area of cells. The signal of ß-catenin at the cytoplasmic area was found significantly higher in transfected samples which implies the nuclear accumulation of ß-catenin. The expression of the c-myc protein was found significantly higher in transfected cells along with significantly higher nuclear entropy. RT-PCR result shows two folds of up-regulation of EMT-TFs Snail1, Twist1, and Zeb2 and down-regulation of Snail2 and Twist2. The study concludes that HPV16 E6/E7 oncogene can induce EMT.

Simple formulation and characterization of double emulsion variant designed to carry three bioactive agents.

Multiple emulsions are thermodynamically stable systems that mark applications in various fields including drug delivery systems. They allow enhanced availability of drugs, greater absorption, and present reduced toxicity, among other desirable properties. In this work, we aimed to formulate a unique double emulsion (O1/W + W1/O2/W/W) with three bioactive components viz. Ocimum tenuiflorum oil, Cocos nucifera oil and crystalline Cinnamomum camphora. Three surfactants with different HLB values viz. Tween-20, Tween-80 and Triton X-100 were used for the emulsification process. The method followed was simple as compared to current methods employed for formulating multiple emulsions. Formulation was characterized using techniques of bright field microscopy, Dynamic Light Scattering (DLS), High-Resolution Transmission Electron Microscopy (HR-TEM) and Fourier-transform infrared spectroscopy (FTIR). Image processing tools were also used to characterize the formulation, which reliably cross-verified the observations from conventional characterization techniques. The potency of individual components of emulsion was compared with the prepared double emulsion model by testing the activity on two pathologically relevant bacterial strains: Fusobacterium nucleatum (FN) and Porphyromonas gingivalis (PG).

Collagen deposition correlates with loss of E-cadherin and increased p63 expression in dysplastic conditions of oral submucous fibrosis.

This paper focuses on the status of epithelial markers, E-cadherin, and p63 in the backdrop of an abnormal amount of collagen in the sub-mucosa of dysplastic and non-dysplastic grades of OSF. Histologically confirmed OSF and normal oral mucosa samples were procured. Samples were stained by Van Gieson's stain (VG) and immunohistochemistry. The captured images were analyzed by ImageJ software to quantify their grayscale intensities. There was a gradual increase in the intensity of VG stain from normal to non-dysplastic and dysplastic OSF and the differences in their mean grayscale values were found to be significant (p < 0.00001). The intensity of E-cadherin was found to be the highest in non-dysplastic conditions and lowest in dysplastic conditions. The intensity difference of E-cadherin between normal and non-dysplastic OSF was found to be significant (p < 0.00001). The grayscale scale intensity values for p63 in whole epithelium depicted significant differences between normal and diseased conditions but for its intensity, in basal cells, significant differences were found between non-dysplastic and other classes of tissues. There was a positive correlation observed between VG and p63 staining intensity. The diseased oral epithelium demonstrated greater deposition of sub-epithelial collagen fibers along with subsequent loss of E-cadherin and an increased p63 expression.

Hydrogels and Their Role in Bone Tissue Engineering: An Overview.

An increasing incidence of the bone damage either due to trauma or a wide range of diseases related to bone necessitates the advent of new technologies or modification of the existing pattern of treatment to deliver utmost care to an individual thereby helping them to lead a normal and healthy life. Revolutionary changes in the field of tissue engineering (TE) pave a way from repair to regeneration of human tissues and restoring the health of an individual. Among the numerous biomaterials available, hydrogel emerges as a promising source of scaffold material in the field of bone TE (BTE). This article presents an overview on hydrogels and their role in BTE.

Characterization of alginate-capped nanosilver by fractal and entropy analysis on its transmission electron microphotographs.

The study employs conventional techniques and quantitative image analysis tools to characterize alginate-capped nanosilver synthesized by green methods. Sodium Alginate (0.5 %, 1 % and 2 %) was used as a reducing and stabilizing agent. Presence of particles was confirmed by UV-vis Spectroscopy, with absorbance maxima of 412-413 nm for 0.5 %, 1 % and 2 % of polymer. Hydrodynamic sizes of particles recorded for 0.5 %, 1 % and 2 % polymer were 128.4 ± 1.5, 129.9 ± 3.6 and 148.6 ± 1.0 nm by DLS. TEM revealed roughly spherical to cuboidal particles ranging from 15-20 nm and clusters of 100 nm and Energy Dispersive X-ray Spectroscopy confirmed the presence of silver in the particles. Analysis of the TEM images was done in MATLAB R2016b using histogram equalisation for image enhancement and entropy filtering for image segmentation. These techniques revealed the surface pores and polymer distribution around the particle. Statistical analysis (ANOVA) was performed for the measured fractal dimensions of nanoparticles with polymer coating, width of particle together with polymer coating, and thickness of only polymer coating around the particle for various study groups. Significant differences (p < 0.05) were found both between and within the study groups for fractal dimensions of nanoparticles with polymer coating, width of nanoparticles and thickness of polymer coating alone. The analysis was successful in confirming presence and thickness of polymer layer on particles.

Epithelial Distribution of E-Cadherin, p63, and Mitotic Figures in ApoTome Images to Determine the Oncogenic Potentiality of Oral Submucous Fibrosis.

The exact process of the malignant conversion of oral submucous fibrosis (OSF) to oral cancer is not fully understood. This study aimed to detect and analyze E-cadherin expression, p63 expression, and number of mitotic figures, all correlated to cancer development, in ApoTome images of oral tissues to determine the oncogenic potentiality of OSF. ApoTome images of the study groups (6 normal, 16 OSF with dysplasia, and 10 OSF without dysplasia) were recorded. Cytoplasmic and membranous E-cadherin expression, breakages of the cell membrane, and p63 expression were detected in MATLAB 2016b. The number of mitotic figures detected by MATLAB was correlated with the number of chromosomes detected by ImageJ. A Mann­Whitney U test was done to determine a significant difference between the study groups for cytoplasmic and membranous E-cadherin distribution points. Statistical significant differences were found for cytoplasmic E-cadherin distribution between normal and OSF (with dysplasia) (p = 0.0278). There was an increase in mitotic figures, p63 expression, and cytoplasmic E-cadherin expression and a decrease in membranous E-cadherin expression from normal to diseased condition. Hence, automated detection and quantification of E-cadherin, p63, and mitotic figures in ApoTome images of oral biopsies can help in determining the oncogenic potentiality of OSF.

Treatment of glaucoma by prostaglandin agonists and beta-blockers in combination directly reduces pro-fibrotic gene expression in trabecular meshwork.

Prostaglandin analogues (PG), beta-blockers (BB) or their combination (PG+BB) are used primarily to reduce the intraocular pressure (IOP) pathologically associated with glaucoma. Since, fibrosis of the trabecular meshwork (TM) is a major aetiological factor in glaucoma, we studied the effect of these drugs on fibrosis-associated gene expression in TM of primary glaucoma patients. In the present study, TM and iris of primary open-angle (n = 32) and angle-closure (n = 37) glaucoma patients were obtained surgically during trabeculectomy and categorized based on the type of IOP-lowering medications use as PG, BB or PG+BB. mRNA expression of pro-fibrotic and anti-fibrotic genes was quantified using qPCR in these tissues. The gene expression levels of pro-fibrotic genes were significantly lower in PG+BB as compared to other groups. These observations and underlying signalling validated in vitro in human TM cells also showed reduced fibrotic gene and protein expression levels following PG+BB treatment. In conclusion, it is observed that PG+BB combination rather than their lone use renders a reduced fibrotic status in TM. This further suggests that IOP-lowering medications, in combination, would also modulate fibrosis-associated molecular changes in the TM, which may be beneficial for maintaining aqueous out-flow mechanisms over the clinical treatment duration.

Therapeutic potential of mesenchymal stem cells in treating both types of diabetes mellitus and associated diseases.

Diabetes mellitus is a common lifestyle disease which can be classified into type 1 diabetes mellitus and type 2 diabetes mellitus. While both result in hyperglycemia due to lack of insulin action and further associated chronic ailments, there is a marked distinction in the cause for each type due to which both require a different prophylaxis. As observed, type 1 diabetes is caused due to the autoimmune action of the body resulting in the destruction of pancreatic islet cells. On the other hand, type 2 diabetes is caused either due to insulin resistance of target cells or lack of insulin production as per physiological requirements. Attempts to cure the disease have been made by bringing drastic changes in the patients' lifestyle; parenteral administration of insulin; prescription of drugs such as biguanides, meglitinides, and amylin; pancreatic transplantation; and immunotherapy. While these attempts cause a certain degree of relief to the patient, none of these can cure diabetes mellitus. However, a new treatment strategy led by the discovery of mesenchymal stem cells and their unique immunomodulatory and multipotent properties has inspired therapies to treat diabetes by essentially reversing the conditions causing the disease. The current review aims to enumerate the role of various mesenchymal stem cells and the different approaches to treat both types of diabetes and its associated diseases as well.

Synthesis of ZnO/Fe3O4/rGO nanocomposites and evaluation of antibacterial activities towards E. coli and S. aureus.

Antibacterial activity of nanoparticles (NPs) and nanocomposites (NCs) has received wide spread attention in biomedical applications. In this direction, the authors prepared zinc oxide (ZnO), iron oxide (Fe3O4), and their composite including reduced graphene oxide (rGO) by hydrothermal method. The structural and microstructural properties of the synthesised NPs and NCs were investigated by XRD, FT-IR, UV-Vis, TGA, and TEM analysis. PEG-coated ZnO and Fe3O4 form in hexagonal wurtzite and inverse spinel structures, respectively. ZnO forms in rod-shaped (aspect ratio of ∼3) morphology, whereas well-dispersed spherical-shaped morphology of ∼10 nm is observed in Fe3O4 NPs. The ZnO/Fe3O4 composite possesses a homogeneous distribution of above two phases and shows a very good colloidal stability in aqueous solvent. These synthesised particles exhibited varying antibacterial activity against gram-positive strain Staphylococcus aureus (S. aureus) and gram-negative strain Escherichia coli (E. coli). The nanocomposite exhibits a better cidal effect on E. coli when compared to S. aureus when treated with 1 mg/ml concentration. Further, the addition of rGO has intensified the anti-bacterial effect to a much higher extent due to synergistic influence of individual components.

Segmentation and analysis of surface characteristics of oral tissues obtained by scanning electron microscopy to differentiate normal and oral precancerous condition.

Abnormal epithelial stratification is a sign of oral dysplasia and hence evaluation of surface characteristics of oral epithelial region can help in detection of cancerous progression. Surface characteristics can be better visualised by Scanning Electron Microscopy (SEM) in comparison to light microscopy. In our study we have developed automated image processing algorithms i.e. Gaussian with median filtering and Gradient filtering, using MATLAB 2016b, to segment the surface characteristics i.e. the ridges and pits in the SEM images of oral tissue of normal (13 samples) and Oral Submucous Fibrosis (OSF) (36 samples) subjects. After segmentation, quantitative measurement of the parameters like area, thickness and textural features like entropy, contrast and range filter of ridges as well as area of pit and the ratio of area of ridge vs. area of pit was done. Statistical significant differences were obtained in between normal and OSF study groups for thickness (p=0.0107), entropy (p<0.00001) and contrast of ridge (p<0.00001) for Gaussian with median filtering and for all the parameters except thickness of the ridge(p=1.386), for Gradient filtering. Thus, computer aided image processing by Gradient filter followed by quantitative measurement of the surface characteristics provided precise differentiation between normal and precancerous oral condition.

Influence of Anterior Biometry on Corneal Biomechanical Stiffness of Glaucomatous Eyes Treated With Chronic Medication or Filtration Surgery.

PRéCIS:: Anterior chamber depth, IOP, and thickness confounded the assessment of corneal biomechanical properties with noncontact applanation in glaucoma eyes. Compared with normal eyes, glaucoma eyes, which underwent long-term treatment or filtration surgery, had similar properties. PURPOSE: The purpose of this study was to evaluate corneal stiffness in primary angle-closure (PACG) and primary open-angle (POAG) glaucoma eyes that were subgrouped on the basis of the type of topical medication and filtration surgery using noncontact applanation. METHODS: All eyes were retrospectively reviewed for intraocular pressure (IOP) with Corvis-ST (OCULUS Optikgerate Gmbh, Germany). Nonglaucoma (n=140), PACG (n=102 under medication), and POAG (n=154 under medication) eyes were included. Corneal stiffness was calculated using deformation amplitude and a biomechanical model. Multivariate analyses were performed, which evaluated the effect of systemic conditions (diabetes and hypertension), the effect of medication (prostaglandins or beta blockers or combined), and the effect of filtration surgery (PACG: n=23; POAG: n=26). Age, IOP, central corneal thickness (CCT), refractive error, and anterior chamber depth (ACD) were covariates. RESULTS: Diabetes and hypertension did not alter corneal stiffness of glaucoma eyes compared with nonglaucoma eyes (P>0.05). Corneal stiffness of POAG and nonglaucoma eyes was similar but significantly different from the stiffness of PACG eyes (P=0.002), irrespective of the type of topical medication. This difference was strongly correlated with ACD (P=0.003) in addition to IOP and CCT. In eyes treated with filtration surgery, ACD (P=0.04) again impacted the trends between nonglaucoma and glaucoma eyes. CONCLUSIONS: Medication or filtration surgery did not affect the corneal biomechanical parameters differentially from nonglaucoma eyes. However, IOP, CCT, and ACD strongly affected corneal biomechanical parameters in the same glaucoma eyes.

Analysis of images for detection of oral epithelial dysplasia: A review.

This paper provides a review of various image analysis approaches that have been previously used for recognition of dysplasia in images of the epithelium of the oral cavity. This domain has become especially admissible with the uncovering of the importance of image analysis which can probably be an aid to subjective diagnosis by histopathologists. Oral malignancy is a rampant form of cancer found among people of the Indian subcontinent due to various deleterious habits like consumption of tobacco, areca nut, betel leaf etc. Oral Submucous Fibrosis, a precancer, whose pathological category falls between normal epithelium and epithelial dysplasia, is caused because of these habits and can ultimately lead to oral cancer. Hence early detection of this condition is necessary. Image analysis methods for this purpose have an enormous potential which can also reduce the heavy workload of pathologists and to refine the criterion of interpretation. This paper starts with a critique of statistics of oral carcinoma in India and distribution of cancer in intra-oral sites and moves on to its causes and diagnostic approaches including causative agents, problems in curative approach and importance of image analysis in cancer detection. The various image analysis methods to appraise the cytological and architectural changes accompanied by Oral Epithelial Dysplasia in the images of the oral epithelial region have been described in relation to 2005 WHO Classification System and it was found that in future, analysis of images based on the mentioned methods has the potential in better interpretation and diagnosis of oral carcinoma.

Amelioration of excision wounds by topical application of green synthesized, formulated silver and gold nanoparticles in albino Wistar rats.

Wound healing, a complex biological process, has attained a lot of attention as dermatologists are primarily interested in stimulated wound closure without formation of scar or a faint scar. The recent upsurgence of nanotechnology has provided novel therapeutic materials in the form of silver and gold nanoparticles which accelerate the wound healing process. The effect of formulated nanoparticles using Coleus forskohlii root extract (green synthesized) has been tried out for ameliorating full thickness excision wounds in albino Wistar male rats. The evaluation of in vivo activity of nanoparticles in wound healing was carried out on open wounds made by excision on the dorsal sides of albino Wistar rats under anesthesia, and the healing of the wounds was assessed. Histological aspects of the healing process were studied by a HE (Hematoxylin and Eosin) staining method to assess various degrees of re-epithelialization and the linear alignment of the granulation tissue whereas Van Gieson's histochemical staining was performed to observe collagen fibers. The healing action shown by the formulated nanoparticles was remarkable during the early stages of wound healing, which resulted in the substantial reduction of the whole healing period. Topical application of formulated gold nanoparticles was found to be more effective in suppressing inflammation and stimulating re-epithelialization compared to silver nanoparticles during the healing process. The results throw light on the amelioration of excision wounds using nanoparticles which could be a novel therapeutic way of improving wound healing in clinical practice. The mechanism of advanced healing action of both types of nanoparticles could be due to their antimicrobial, antioxidant and anti-inflammatory properties.

Computer-aided molecular pathology interpretation in exploring prospective markers for oral submucous fibrosis progression.

BACKGROUND: Evaluation of molecular pathology markers using a computer-aided quantitative assessment framework would help to assess the altered states of cellular proliferation, hypoxia, and neoangiogenesis in oral submucous fibrosis and could improve diagnostic interpretation in gauging its malignant potentiality. METHODS: Immunohistochemical (IHC) expression of c-Myc, hypoxia-inducible factor-1-alpha (HIF-1α), vascular endothelial growth factor (VEGF), VEGFRII, and CD105 were evaluated in 58 biopsies of oral submucous fibrosis using computer-aided quantification. After digital stain separation of original chromogenic IHC images, quantification of the diaminobenzidine (DAB) reaction pattern was performed based on intensity and extent of cytoplasmic, nuclear, and stromal expression. RESULTS: Assessment of molecular expression proposed that c-Myc and HIF-1α may be used as strong screening markers, VEGF for risk-stratification and VEGFRII and CD105 for prognosis of precancer into oral cancer. CONCLUSION: Our analysis indicated that the proposed method can help in establishing IHC as an effective quantitative immunoassay for molecular pathology and alleviate diagnostic ambiguities in the clinical decision process.

Connecting cyto-nano-architectural attributes and epithelial molecular expression in oral submucous fibrosis progression to cancer.

OBJECTIVE: Problems in pre-cancer diagnosis complicate cancer theragnosis as well as life expectancy. There is uncertainty regarding malignant transformation of oral submucous fibrosis (OSF), an oral pre-cancer with dysplastic (OSFWD) and non-dysplastic (OSFWT) subtypes. Understanding the structural, molecular and physical aspects of epithelial homeostasis may be useful. MATERIALS AND METHODS: Histopathological grading of biopsy sections was performed using H&E staining. Alterations in epithelial surface architecture in different groups was evaluated using scanning electron microscopy (SEM). The expression of crucial epithelial genes (p63, CK-5/6, CK-10, E-cadherin and ß-catenin) was studied by immunohistochemistry, Western blot and RT-PCR analysis. RESULTS: SEM observations revealed that the surface epithelial ridge pattern became thick and dense, and pit pattern gradually decreased in OSFWD and oral squamous cell carcinoma (OSCC). p63, ΔNp63 and CK-5/6 were up-regulated in OSFWD and OSCC but down-regulated in OSFWT. CK-10 was down-regulated in OSFWD compared to OSFWT. Cytoplasmic expression of E-cadherin and ß-catenin was elevated in dysplastic and cancerous conditions. Moreover, statistical correlation between SEM features (ridges and pits) and molecular attributes demonstrated a significant positive relationship between the ridge-to-pit ratio and p63 population density (r=0.85) and the ridge-to-pit ratio and CK-5/6 intensity (r=0.63). CONCLUSIONS: Molecular changes related to epithelial progressive maturation and cellular proliferation are correlated with concomitant alteration of epithelial surface architecture which helps to predict the malignant potentiality of OSF.

Correlated analysis of semi-quantitative immunohistochemical features of E-cadherin, VEGF and CD105 in assessing malignant potentiality of oral submucous fibrosis.

Oral submucous fibrosis, a potentially premalignant condition for oral squamous cell carcinoma, manifests both non-dysplastic and dysplastic grades. Early and specific identification of its malignant potentiality suffers from diagnostic limitations that may be addressed by correlated molecular pathology attributes having histopathological backdrop. Present study correlates expressional alteration in prime epithelial marker E-cadherin, with neo-angiogenic molecules viz. VEGF and CD105 for elucidation of malignant potentiality in different stages of oral submucous fibrosis. Sixty-eight incision biopsies from normal oral mucosa (n = 10), non-dysplastic (n = 18) and different dysplastic grades (n = 40) of oral submucous fibrosis were semi-quantitatively analyzed for immunohistochemical expressions of E-cadherin (membranous and cytoplasmic), VEGF and CD105 which were further statistically correlated. The loss of membranous E-cadherin with increase in cytoplasmic accumulation in differentiative layers of epithelium through the progression of dysplasia was noted along with up-regulation in VEGF expressions. The number of CD105(+) blood vessels and their major axis also showed significant increase from non-dysplasia toward higher grades of dysplasia. The positive correlation between deregulated expression of epithelial cell-cell adhesion molecule and increase in neo-angiogenic attributes of oral submucous fibrosis with increase in dysplastic grades indicated elucidatory potential of molecular expression features in assessment of malignant potentiality in oral submucous fibrosis.

Epithelio-mesenchymal transitional attributes in oral sub-mucous fibrosis.

Evaluating molecular attributes in association with its epithelial and sub-epithelial changes of oral sub-mucous fibrosis is meaningful in exploring the plausibility of an epithelio-mesenchymal transition (EMT) and malignant potentiality of this pathosis. In this study histopathological and histochemical attributes for basement membrane and connective tissue in biopsies of oral sub-mucous fibrosis (n = 55) and normal oral mucosa (n = 16) were assessed and expressions of p63, E-cadherin, ß-catenin, N-cadherin and TWIST were analyzed immunohistochemically. The p63 and its isoforms (TA and ∆N), PARD3, E-cadherin and ß-catenin were also assessed transcriptomically by q-PCR and EMT players like TWIST1, ZEB1, MMP9 and micro-RNA 205 were searched in gene expression microarrays. Oral epithelium demonstrating impairment in progressive maturation in oral sub-mucous fibrosis concomitantly experienced an increase in basement membrane thickness and collagen deposition along with alteration in target molecular expressions. In comparison to non-dysplastic conditions dysplastic stages exhibited significant increase in p63 and p63∆N expressions whereas, E-cadherin and ß-catenin exhibited loss from the membrane with concurrent increase in cytoplasm. Further the N-cadherin and TWIST were gained remarkably along with the appearance of nuclear accumulation features of ß-catenin. The microarray search had noticed the up-regulation of TWIST1, ZEB1 and MMP9 along with down regulation of micro-RNA 205. The simultaneous increase in basement membrane thickness and sub-epithelial collagen deposition were the plausible indicators for increased matrix stiffness with expected impact on oral epithelial functional homoeostasis. This was corroborated with the increase in expressions of epithelial master regulator p63 and its oncogenic isoform (∆N) along with membranous loss of E-cadherin (EMT hallmark) and its associate ß-catein and gain of mesenchymal markers like N-cadherin and TWIST. These also became indicative for the induction of epithelial to mesenchymal transitional mechanism in oral sub-mucous fibrosis when connoted here with the relevant modulation in expressions of EMT regulators.

Noninvasive characterisation of foot reflexology areas by swept source-optical coherence tomography in patients with low back pain.

Objective. When exploring the scientific basis of reflexology techniques, elucidation of the surface and subsurface features of reflexology areas (RAs) is crucial. In this study, the subcutaneous features of RAs related to the lumbar vertebrae were evaluated by swept source-optical coherence tomography (SS-OCT) in subjects with and without low back pain (LBP). Methods. Volunteers without LBP (n = 6 (male : female = 1 : 1)) and subjects with LBP (n = 15 (male : female = 2 : 3)) were clinically examined in terms of skin colour (visual perception), localised tenderness (visual analogue scale) and structural as well as optical attributes as per SS-OCT. From each subject, 6 optical tomograms were recorded from equidistant transverse planes along the longitudinal axis of the RAs, and from each tomogram, 25 different spatial locations were considered for recording SS-OCT image attributes. The images were analysed with respect to the optical intensity distributions and thicknesses of different skin layers by using AxioVision Rel. 4.8.2 software. The SS-OCT images could be categorised into 4 pathological grades (i.e., 0, 1, 2, and 3) according to distinctness in the visible skin layers. Results. Three specific grades for abnormalities in SS-OCT images were identified considering gradual loss of distinctness and increase in luminosity of skin layers. Almost 90.05% subjects were of mixed type having predominance in certain grades. Conclusion. The skin SS-OCT system demonstrated a definite association of the surface features of healthy/unhealthy RAs with cutaneous features and the clinical status of the lumbar vertebrae.

Assessment of molecular events during in vitro re-epithelialization under honey-alginate matrix ambience.

Re-epithelialization is one of the most important stages of cutaneous regeneration and its success requires supportive micro-ambience which may be provided with suitable bio-matrix. Biocompatibility and efficacy of such bio-matrix in re-epithelialization could be explored by multimodal analysis of structural and functional attributes of in vitro wound healing model including evaluation of prime molecular expressions of the epithelial cells during repair. Present study examines the influence of honey-alginate and alginate matrices on re-epithelialization in keratinocyte (HaCaT) population in a 2-D wound model. Cellular viability, proliferation and cell-cell adhesion status were assessed during wound closure using live/dead cell assay and by evaluating expressions of Ki67, p63 and E-cadherin along-with % change in cellular electrical impedance. Efficacy of honey-alginate matrix in comparison to only alginate one was demonstrated by a quicker reduction in wound gap, improved cellular viability, enhanced expressions of Ki67, p63 and its isoforms (TAp63, ΔNp63) as well as E-cadherin. Faster restoration of electrical attribute (% of impedance change) after wounding also indicated better impact of honey-alginate matrix in re-epithelialization.