Densitometric Analysis Research Articles

Densitometric analysis of Radiation Absorbed Dose to Patients Undergoing Chest X-ray Examination in a Government Owned Hospital, Kano-Nigeria

Chest X-ray is one the most frequently required diagnostic procedures used in clinical practice to produce an image of organs, tissues and bones of the body with a significant impact on collective dose. However, the radiation dose received by patients from this procedure is likely to show large variations within and among radiodiagnostic centres. This could be traced to be lack of imaging protocols and a lack of local or national diagnostic reference levels. Although the radiation doses resulting from the X-ray procedures are usually measured in terms of effective dose entrance surface dose using electronic personal dosimeter (EPD), pocket dosimeter, thermoluminescent dosimeter (TLD), film badges, and optically stimulated luminescent dosimeter (OSLD). For the sake of radiation protection, densitometric analysis was used to evaluate medical exposure for chest radiographs to determine the radiation doses using a point optical densitometer; PTW-DensiX with a range of 0 to 4.0 optical density. The hospital where this study was carried out is in Kano State. A total of sixty (60) chest radiographic films were collected from the hospital and the optical density of each radiographic film was measured five times at different spots. The mean optical density was obtained and converted by a mathematical relation to absorbed radiation dose in cGy. The results obtained in this study show that the mean absorbed X-ray dose was 0.78 ± 0.13 cGy and is low compared to the previous study held for hospitals in Osun State. Also, when compared with the excess dose of 0.10 Gy recommended by the International Commission on Radiological Protection, it was evident that radiological practice especially as it relates to chest radiographs at the hospital in Kano is within the recommended practice.
 Keywords: Absorbed dose, Densitometer, Optical Density, Radiation, X-ray

Expression Pattern and Immunoregulatory Roles of Galectin-1 and Galectin-3 in Atopic Dermatitis and Psoriasis.

The pathogenesis of atopic dermatitis (AD) and psoriasis (Ps) overlaps, particularly the activation of the immune response and tissue damage. Here, we evaluated galectin (Gal)-1 and Gal-3 levels, which are beta-galactoside-binding proteins with immunomodulatory functions and examined their effects on human keratinocytes stimulated with either interleukin (IL)-4 or IL-17A. Skin biopsies from AD, Ps, and control patients were evaluated using histological and immunohistochemical analyses. Six studies containing publicly available transcriptome data were individually analyzed using the GEO2R tool to detect Gal-1 and Gal-3 mRNA levels. In vitro, IL-4- or IL-17A-stimulated keratinocytes were treated with or without Gal-1 or Gal-3 to evaluate cytokine release and migration. Our findings showed different patterns of expression for Gal-1 and Gal-3 in AD and Ps skins. Densitometric analysis in skin samples showed a marked increase in the protein Gal-1 levels in Ps epidermis and in both AD and Ps dermis compared to controls. Protein and mRNA Gal-3 levels were downregulated in AD and Ps lesional skin compared with the control samples. In vitro, both galectins addition abrogated the release of IL-8 and RANTES in IL-17-stimulated keratinocytes after 24h, whereas IL-6 release was downregulated by Gal-3 and Gal-1 in IL-4- and IL-17-stimulated cells, respectively. Administration of both galectins also increased the rate of keratinocyte migration under IL-4 or IL-17 stimulation conditions compared with untreated cells. Altogether, the immunoregulatory and migration effects of Gal-1 and Gal-3 on keratinocytes under inflammatory microenvironment make them interesting targets for future therapies in cutaneous diseases.

Mitochondrial сomplexome of etiolated pea shoots

Studies into mitochondrial сomplexomes in various organisms provide an insight into the native organization of proteins and metabolic pathways in the organelles of the subject under study. “Complexome” is a relatively recent concept describing the proteome of protein complexes, supercomplexes, and oligomeric proteins. Complexome analysis is performed using current electrophoretic and mass spectrometric techniques, in particular, by two-dimensional electrophoresis (2D BN/SDS-PAGE) in combination with mass spectrometry (MS). Unlike 2D IEF/SDS-PAGE, this method enables analysis of not only hydrophilic proteins of the mitochondrial matrix, but also membrane proteins and their associations, thus expanding the possibilities of studying the organelle proteome. In the present work, the complexome of etiolated pea shoots was studied for the first time using 2D BN/SDS-PAGE followed by MALDI-TOF MS. To this end, 145 protein spots excised from the gel were analyzed; 110 polypeptides were identified and assigned to different functional groups. A densitometric analysis revealed that the major protein group comprised the enzymes of the mitochondrial energy system (1), accounting for an average of 43% of the total polypeptide content. The remaining 57% was primarily distributed among the following functional categories: pyruvate dehydrogenase complex and citric acid cycle (2); amino acid metabolism (3); nucleic acid processing (4); protein folding (5); antioxidant protection (6); carrier proteins (7); other proteins (8); proteins having unknown functions (9). The obtained data indicate the complex organization of the pea proteome. In addition to the enzymes of the OXPHOS system, the proteins of other functional categories are found to form supramolecular structures. It is suggested that the presence of proteins from other cellular compartments may indicate the interaction of mitochondria with the enzymes or structures of corresponding organelles. In general, the obtained data on the pea complexome represent a kind of a mitochondrial “passport” that reflects the native state of the proteome of organelles corresponding to their physiological status.

Effect of Nesfatin-1 on Rat Humerus Mechanical Properties under Quasi-Static and Impact Loading Conditions.

The investigations on the response of bone tissue under different loading conditions are important from clinical and engineering points of view. In this paper, the influence of nesfatin-1 administration on rat humerus mechanical properties was analyzed. The classical three-point bending and impact tests were carried out for three rat bone groups: control (SHO), the humerus of animals under the conditions of established osteopenia (OVX), and bones of rats receiving nesfatin-1 after ovariectomy (NES). The experiments proved that the bone strength parameters measured under various mechanical loading conditions increased after the nesfatin-1 administration. The OVX bones were most susceptible to deformation and had the smallest fracture toughness. The SEM images of humerus fracture surface in this group showed that ovariectomized rats had a much looser bone structure compared to the SHO and NES females. Loosening of the bone structure was also confirmed by the densitometric and qualitative EDS analysis, showing a decrease in the OVX bones’ mineral content. The samples of the NES group were characterized by the largest values of maximum force obtained under both quasi-static and impact conditions. The energies absorbed during the impact and the critical energy for fracture (from the three-point bending test) were similar for the SHO and NES groups. Statistically significant differences were observed between the mean Fi max values of all analyzed sample groups. The obtained results suggest that the impact test was more sensitive than the classical quasi-static three-point bending one. Hence, Fi max could be used as a parameter to predict bone fracture toughness.

Chromatographic Fingerprinting of an Ayurvedic Formulation by High-Performance Thin Layer Chromatography Method

In the present study, an attempt has been made to develop a high-performance thin layer chromatography fingerprinting of gallic acid, piperine and curcumin from an Ayurvedic formulation, Dekofcyn tablet. Chromatographic separation was achieved on the plate pre-coated with silica gel 60F254 as the stationary phase. The solvent system consisted of toluene:ethyl acetate:formic acid:glacial acetic acid in the ratio (5:3.5:1:1 v/v/v/v) respectively. The retardation factor value was calculated at 0.35 (±0.02), 0.66 (±0.02) and 0.89 (±0.02) for gallic acid, piperine and curcumin respectively. Densitometric analysis was carried out in the isoabsorptive mode at 295 nm. The regression analysis was carried out in the range of 100-700 ng/spot for gallic acid and piperine, 400-1600 ng/spot for curcumin and the regression coefficient for all three markers was found to be 0.99 respectively. Validation of the method was performed according to international conference on hominization guidelines Q2 (R1). This method can also be used to evaluate other formulations containing these markers as one of the constituents, thus conforming to the need for ensuring the quality of an Ayurvedic formulation.

Fractal dimension analysis on CBCT scans for detecting low bone mineral density in postmenopausal women.

PurposeThe aim of this study was to compare the fractal dimension (FD) measured at 2 bone sites (second cervical vertebra and mandible) on cone-beam computed tomography (CBCT). The research question was whether FD could serve as an accessory tool to refer postmenopausal women for densitometric analysis. Therefore, the reliability and accuracy of FD were evaluated.Materials and MethodsIn total, 103 postmenopausal women were evaluated, of whom 52 had normal bone mineral density and 51 had osteoporosis, according to dual X-ray absorptiometry of the lumbar spine and hip. On the CBCT scans, 2 regions of interest were selected for FD analysis: 1 at the second cervical vertebra and 1 located at the mandible. The correlations between both measurements, intra- and inter-observer agreement, and the accuracy of the measurements were calculated. A P value less than 0.05 was considered to indicate statistical significance for all tests.ResultsThe mean FD values were significantly lower at the mandibular region of interest in osteoporotic patients than in individuals with normal bone mineral density. The areas under the curve were 0.644 (P=0.008) and 0.531 (P=0.720) for the mandibular and vertebral sites, respectively.ConclusionFD at the vertebral site could not be used as an adjuvant tool to refer women for osteoporosis investigation. Although FD differed between women with normal BMD and osteoporosis at the mandibular site, it demonstrated low accuracy and reliability.

Temporal Modulation of Differential Alternative Splicing in HaCaT Human Keratinocyte Cell Line Chronically Exposed to Arsenic for up to 28 Wk.

Background:Chronic arsenic exposure via drinking water is associated with an increased risk of developing cancer and noncancer chronic diseases. Pre-mRNAs are often subject to alternative splicing, generating mRNA isoforms encoding functionally distinct protein isoforms. The resulting imbalance in isoform species can result in pathogenic changes in critical signaling pathways. Alternative splicing as a mechanism of arsenic-induced toxicity and carcinogenicity is understudied.Objective:This study aimed to accurately profile differential alternative splicing events in human keratinocytes induced by chronic arsenic exposure that might play a role in carcinogenesis.Methods:Independent quadruplicate cultures of immortalized human keratinocytes (HaCaT) were maintained continuously for 28 wk with 0 or sodium arsenite. RNA-sequencing (RNA-Seq) was performed with poly(A) RNA isolated from cells harvested at 7, 19, and 28 wk with subsequent replicate multivariate analysis of transcript splicing (rMATS) analysis to detect and quantify differential alternative splicing events. Reverse transcriptase-polymerase chain reaction (RT-PCR) for selected alternative splicing events was performed to validate RNA-Seq predictions. Functional enrichment was performed by gene ontology (GO) analysis of the differential alternative splicing event data set at each time point.Results:At least 600 differential alternative splicing events were detected at each time point tested, comprising all the five main types of alternative splicing and occurring in both open reading frames (ORFs) and untranslated regions (UTRs). Based on functional relevance ELK4, SHC1, and XRRA1 were selected for validation of predicted alternative splicing events at 7 wk by RT-PCR. Densitometric analysis of RT-PCR data corroborated the rMATS predicted alternative splicing for all three events. Protein expression validation of the selected alternative splicing events was challenging given that very few isoform-specific antibodies are available. GO analysis demonstrated that the enriched terms in differential alternatively spliced mRNAs changed dynamically with the time of exposure. Notably, RNA metabolism and splicing regulation pathways were enriched at the 7-wk time point, when the greatest number of differentially alternatively spliced mRNAs are detected. Our preliminary proteomic analysis demonstrated that the expression of the canonical isoforms of the splice regulators DDX42, RMB25, and SRRM2 were induced upon chronic arsenic exposure, corroborating the splicing predictions.Discussion:These results using cultures of HaCaT cells suggest that arsenic exposure disrupted an alternative splice factor network and induced time-dependent genome-wide differential alternative splicing that likely contributed to the changing proteomic landscape in arsenic-induced carcinogenesis. However, significant challenges remain in corroborating alternative splicing data at the proteomic level. https://doi.org/10.1289/EHP9676

Pre-Clinical Evaluation of Proprietary Lutein, Zeaxanthin, and Rosemary Formulation for Its Dermal Protective Activity in Male Swiss Albino Mice

This study aimed to evaluate the efficacy of the proprietary lutein, zeaxanthin, and rosemary formulation for its dermal protection against ultraviolet (UV) irradiated skin dehydration. A total of 48 male Swiss albino mice of 8∼12 weeks of age were divided into eight groups of 6 mice: mice in group 1 (G1) were considered the normal control, without treatment and without skin shaving; mice in G2 had their skins were shaved, but did not receive treatment; mice in G3 were the pathological control; mice in G4 were treated as standard (hyaluronic acid); mice in G5∼G8 were treated with low and high doses of 2 different test substances, respectively. Mice were anaesthetized and then depilatory was applied on the dorsal skin area (2 cm×2 cm) on alternate days, then UV/blue light irradiation was carried out for 15 min for 6 weeks. Collagen type 1 gene expression was determined via densitometric analysis, skin elasticity was assessed, and stratum corneum water contents were measured using a cutometer and corneometer. Skin hydration was assessed through transepidermal water loss, and several serum biochemical parameters (collagenase, hydroxyproline, hyaluronic acid, and ceramide levels) were determined to assess the skin moisturizing activity of the product. Images for assessing photoaging were considered between different groups on day 42. All these subjective parameters reached statistical significance (P<0.05) in groups treated with the proprietary lutein and rosemary formulation compared with the placebo-treated group. In conclusion, the proprietary lutein, zeaxanthin, and rosemary formulation showed better protection of skin subjected to UV irradiated skin dehydration.

Using high performance thin layer chromatography for the detection of pharmacologically active secondary metabolites in Empetrum nigrum L.

Introduction. The article presents the results of the detection of pharmacologically active secondary metabolites in black crowberry Empetrum nigrum L. using the method of high performance thin layer chromatography (HPTLC).Aim. To show the efficiency of HPTLC for conducting preliminary phytochemical analysis to determine the main groups of metabolites in promising medicinal plant species.Materials and methods. HPTLC analysis was carried out on a CAMAG device (Switzerland), using MERCK HPTLC silica gel 60 F154, 20 × 10 cm plates. For the evaporation of the samples, a Heidolph vacuum rotary evaporator (Germany) was used. The aerial parts (shoots) of Empetrum nigrum were harvested next to St. Petersburg State Chemical and Pharmaceutical University (SPCPU) nursery garden of medicinal plants (Leningrad Region, Vsevolozhsky District, Priozerskoe Highway, 38 km) in August 2019.Results and discussion. In the course of the research, four fractions from the aerial parts of Empetrum nigrum were obtained: hexane, dichloromethane, butanol, and water. Then, these fractions were investigated by HPTLC in two solvent systems – n-butanol : acetic acid : water (BAW) (4 : 1 : 2) and hexane : dichloromethane : methanol (HDM) (1 : 2 : 0.5). After scanning densitometric analysis of the plates eluted in the HDM system, it was revealed, that the hexane and dichloromethane fractions have a similar composition and contain the greatest amount of compounds, compared to the butanol and water fractions, and in the BAW system, it was found, that the butanol fraction contains the greatest variety of metabolites. As a result of UV spectroscopy, it was found, that the main groups of compounds contained in the hexane and dichloromethane fractions are derivatives of chalcones, dihydrochalcones, bibenzyls and 9,10-dihydrophenanthrenes. While in the butanol fraction, the main groups of secondary metabolites were derivatives of flavonoids and tanninsConclusion. The data obtained allow us to note the efficiency, speed and simplicity of HPTLC for conducting preliminary phytochemical analysis to determine the main groups of metabolites of promising medicinal plant species.

Stability-Indicating Analytical Method Development Using Quality by Design (QbD) Approach for Simultaneous Estimation of Budesonide and Levosalbutamol.

Budesonide is a corticosteroid; levosalbutamol is bronchodilator. In combination, they are used in the treatment of asthma and chronic obstructive pulmonary disease. The present article portrays the development of a stability-indicating high-performance thin-layer chromatography (HPTLC) method for the simultaneous estimation of budesonide and levosalbutamol using design of experiment (DOE) principles. DOE was applied in development part in which a Box-Behnken design was used to study the effect of factors on the Rf value of the drugs. Twenty-five experimental runs were performed to optimize the chromatographic conditions. Aluminum sheets precoated with silica gel 60 F254 were used as the stationary phase. The optimized mobile phase composition was found to be toluene-ethyl acetate-methanol-ammonia (4:1.8:1.8:0.2) and saturation time 20 min, quantified by densitometric analysis at 231 nm. Moreover, the drugs were subjected to acid and alkali hydrolysis, oxidation, thermal, and photodegradation. The drugs undergo degradation under mainly acidic and basic conditions. Also, the degraded products were well resolved from the pure drug with significantly different Rf values. Linearity was performed in the ranges of 224-1120 ng/band for budesonide and 280-1400 ng/band for levosalbutamol. The method was validated according to the International Conference on Harmonization guidelines. This approach is useful to expedite method development and optimization activities in analytical laboratories. The experimental data revealed that the volume of toluene and methanol in the mobile phase significantly affected the Rf value of both the drugs, and ammonia significantly affected the Rf value of levosalbutamol.

Assessment of Corneal Densitometry in Big-Bubble Dissection Versus Manual Dissection Deep Anterior Lamellar Keratoplasty.

The aim of this study was to assess corneal densitometry and visual outcomes after big-bubble deep anterior lamellar keratoplasty (BB-DALK) and manual dissection deep anterior lamellar keratoplasty in patients with keratoconus. This retrospective comparative observational study included 40 keratoconic patients who underwent DALK surgery: 22 eyes had BB-DALK (group I) and 18 eyes had failed BB technique and DALK was completed by manual dissection (group II). Best -corrected visual acuity (BCVA), corneal topographic parameters, residual stromal tissue thickness, and endothelial cell count were recorded at 1, 3, 6, and 12 months postoperatively. Densitometric analysis of different corneal layers and zones was performed using Scheimpflug tomography at each visit; values were recorded and compared between the 2 groups. At 1 and 6 months postoperatively, BCVA was better in group I than in group II, but with no statistically significant difference. At 12 months, the visual acuities became nearly similar in both groups (0.30 ± 0.13 vs. 0.30 ± 0.14 logarithm of the minimum angle of resolution, P = 0.888). Regarding corneal densitometric analysis, the recorded values for the posterior corneal layer were significantly higher in group II compared with group I at 3, 6, 9, and 12 months postoperatively in the 0- to 2-mm zone (P < 0.001) and the 2- to 6-mm zone (P = 0.029, 0.028, 0.001, and <0.001). Manual dissection DALK after failed BB technique may affect the interface stromal reflectivity up to 12 months postoperatively. However, this does not significantly affect the visual acuity in comparison with successful BB-DALK.

Herbal standardization of formulation containing curcuminoids, piperine and ascorbic acid by dual detection mode densitometric analysis

Herbal standardization of formulation containing curcuminoids, piperine and ascorbic acid by dual detection mode densitometric analysis

Peculiarities of application of the CT densitometric indices in the treatment of perforative pulmonary tuberculosis

Objective. To determine the possibilities of application of the CT densitometry in treatment of perforative pulmonary tuberculosis.&#x0D; Materials and methods.The work is based on results of analysis ofpostoperative investigation concerning densitometric indices in pulmonary parenchyma and foci of specific affection in 75 patients, suffering hydropneumothorax.&#x0D; Results. There was established, that densitometric indices of pleural content, revealed while analyzing of the CT images, permits to differentiate trustworthily transudate, exudate and coagulated blood, and to determine activity of specific tuberculosis inflammation, emphysema and the pulmonary parenchyma density. Significant correlation connection of the infiltration parameters and vascular structures (97% CI) was revealed as well, what indicates on possibility of application of densitometric analysis for estimation of the treatment efficacy.&#x0D; Conclusion. Application of densitometric indices of CT permits to estimate the inflammatory process course in pathologically changed pulmonary parenchyma, to reveal in it the emphysematic processes and degree of atelectasis in the blocked parts.

Zero-Valent Iron Nanoparticles Induce Reactive Oxygen Species in the Cyanobacterium, Fremyella diplosiphon.

Nanoscale zero-valent iron nanoparticles (nZVIs) are known to boost biomass production and lipid yield in Fremyella diplosiphon, a model biodiesel-producing cyanobacterium. However, the impact of nZVI-induced reactive oxygen species (ROS) in F. diplosiphon has not been evaluated. In the present study, ROS in F. diplosiphon strains (B481-WT and B481-SD) generated in response to nZVI-induced oxidative stress were quantified and the enzymatic response determined. Lipid peroxidation as a measure of oxidative stress revealed significantly higher malondialdehyde content (p < 0.01) in both strains treated with 3.2, 12.8, and 51.2 mg L–1 nZVIs compared to untreated control. In addition, ROS in all nZVI-treated cultures treated with 1.6–25.6 mg L–1 nZVIs was significantly higher than the untreated control as determined by the 2′,7′-dichlorodihydrofluorescein diacetate fluorometric probe. Immunodetection using densitometric analysis of iron superoxide dismutase (SOD) revealed significantly higher SOD levels in both strains treated with nZVIs at 51.2 mg L–1. In addition, we observed significantly higher (p < 0.001) SOD levels in the B481-SD strain treated with 6.4 mg L−1 nZVIs compared to 3.2 mg L–1 nZVIs. Validation using transmission electron microscopy equipped with energy-dispersive X-ray spectroscopy (EDS) revealed adsorption of nZVIs with a strong iron peak in both B481-WT and B481-SD strains. While the EDS spectra showed strong signals for iron at 4 and 12 days after treatment, a significant decrease in peak intensity was observed at 20 days. Future efforts will be aimed at studying transduction mechanisms that cause metabolic and epigenetic alterations in response to nZVIs in F. diplosiphon.

Advanced ankylosing spondylitis: a multisite, multimodality densitometric analysis for investigation of bone loss in the axial and appendicular skeleton

The aim of this study was to investigate if there is a bias in bone mineral density measurements among major densitometric techniques across multiple skeletal sites. In 25 advanced ankylosing spondylitis patients, bone mineral density measurements were acquired in the lumbar spine, the hip, and the forearm. In total, 60% of patients had a bone mineral density Z-score of -2 or less at one or more skeletal sites. Dedicated loss of cortical bone was identified at the distal forearm (60% of patients). Differences in bone mineral density measurements across all densitometric techniques were highly significant (p≤0.001). Bone loss was more striking in spinal trabecular bone by three-dimensional quantitative computed tomography [Z-score -2.1] versus dual-energy X-ray absorptiometry [Z-score 0]. A trabecular bone loss quantified by quantitative computed tomography was about twice as much as a cortical bone loss by single-energy X-ray absorptiometry (p=0.001). Low bone mineral density is prevalent in advanced ankylosing spondylitis patients, predominating in the spine. Bone mineral density measurements have systematic differences when compared to each other. Knowledge of these offsets is useful for improved diagnosis of regional bone loss that allows for targeted treatment of osteoporosis. Three-dimensional quantitative computed tomography is more suitable for evaluating spinal osteoporosis in advanced ankylosing spondylitis than dual-energy X-ray absorptiometry, which rather underestimates bone loss.

Application of Central Composite Design for screening and Optimization of HPTLC method for simultaneous quantitation of Aprepitant, Dexamethasone and Ondansetron in their synthetic mixtures

New HPTLC method was developed and optimized for estimation of Ondansetron (OND), Dexamethasone (DEX) and Aprepitant (APT) in laboratory prepared ternary mixtures by using Central composite design (CCD). The independent variables used for the optimization were the acetone content in mobile phase (%mL), distance of developing solvent (cm) and saturation time (min). HPTLC Separation was performed on Precoated silica gel F254 aluminum plate (10X10 cm, 100μm thickness) with a mobile phase consisting of chloroform: methanol: acetone: ethyl acetate: ammonia (9:4:2:5:0.2 % v/v/v/v). Quantification of OND, APT and DEX were achieved based on a Densitometric analysis over the concentration range of 200-1200 ng/band, 500-1000 ng/band and 1000-2000 ng/band, respectively, at 254nm. The method was yielded dense and well-resolved bands at Rf values of 0.54± 0.02, 0.79±0.02 and 0.23±0.01 for OND, APT and DEX, respectively. The linear regression analysis for the calibration plots produced r2= 0.9997, r2= 0.9998 and r2=0.9997 for OND, APT and DEX, respectively. The method was validated according to the ICH guidelines. The robustness test was determined that the selected factors have an insignificant effect on the responses. The results indicated that the method is suitable for the routine quality control testing of OND, APT and DEX in their bulk form.

Development and Validation of HPTLC Method for Simultaneous Estimation of Resveratrol and Piperine in Its Pharmaceutical Dosage Form

Resveratrol is a plant compound that work as antioxidant and it also has anti-aging properties whereas Piperine is a alkaloid and it majorly used in spices. A HPTLC (High performance thin layer chromatography) study is conducted for estimation of Resveratrol and Piperine. The Mobile phase used was Chloroform: Ethyl Acetate (50:50 v/v). Rf Value of Resveratrol 0.59 and Piperine 0.79 was found. Stationary phase of Silica gel 60 F254 was used. Densitometric analysis was performed at 325 nm. The method was found to be linear. Recovery (ranging from 97% to 102.24 %), limit of detection (40.26 ng/spot, 1.64 ng/spot respectively for resveratrol &amp; piperine), limit of quantification (122.02 ng/spot, 4.98 ng/spot respectively for resveratrol &amp; piperine) and precision (≤ 2.00%) were found to be satisfactory. Validation performed with linearity, accuracy, precision, specificity, robustness, limit of detection and limit of quantification. Every parameter found within the range. The developed method allows to confirm that an accurate and reliable potency measurement of a pharmaceutical preparation can be performed.

Activity of the glycosidases β-galactosidase, α-l-fucosidase, β-N-acetyl-hexosaminidase, and sialidase in uterine tissues from female dogs in diestrus with and without pyometra

This study aimed to compare the activity of selected glycosidases (β-galactosidase, α-l-fucosidase, β-N-acetyl-hexosaminidase, and sialidase) in homogenates of uterine tissues obtained from female dogs with and without pyometra. In addition, it examined the availability of substrates for these glycosidases in the homogenates. The study was carried out on female dogs undergoing ovariohysterectomy for pyometra (n = 10) and clinically healthy dogs (n = 10) undergoing elective spaying. The activity of β-galactosidase, α-l-fucosidase, and β-N-acetyl-hexosaminidase was analyzed using a spectrofluorometer and that of sialidase using a colorimetric method. Sodium dodecyl sulphate–polyacrylamide gel electrophoresis with Alcian Blue (AB) and Periodic Acid–Schiff (PAS) staining was performed to determine the presence of substrates for these glycosidases in the homogenates of uterine tissues. The results revealed that the activity of all the examined glycosidases was significantly higher (P < 0.05) in the uterine tissues isolated from dogs with pyometra in comparison to healthy dogs. The electrophoretic patterns of the selected samples showed several proteins, which contained different sugar moieties stained by AB and PAS and the profiles differed significantly between the pyometra group and the healthy group. Densitometric analysis of AB staining showed patterns between 233 and 148, 86 and 55, and 43 and 20 kDa, which differed markedly in sugar content between the examined groups of animals. Similarly, PAS staining analysis revealed patterns of different molecular weights, between 233 and 117 and between 55 and 32 kDa, which also differed in sugar content. These findings suggest that canine pyometra is accompanied by the increase in the activity of selected glycosidases in the uterus. This could potentially modify the glycan structures of uterine glycoproteins and in result their biological functions. Further studies are needed to elucidate the potential role of the increased activity of glycosidases in the pathogenesis of this disease.

Development of a stability indicating HPTLC method for the estimation of olanzapine in pharmaceutical dosage forms

We report herein an accurate, precise, and economical stability indicating high performance thin layer chromatographic (HPTLC) method developed to assess the safety of olanzapine in pharmaceutical formulations. Olanzapine was subjected to forced degradation studies to assess the effect of environmental conditions on its stability. Stress conditions such as hydrolysis under acidic and alkaline environment, degradation and oxidation by heat, light and air were used to study the stability of olanzapine. Mobile phase comprising of toluene: methanol (5:5 v/v) and aluminum plate pre-coated with silica gel 60 F254 as a stationary phase were used for the development of chromatogram by HPTLC technique. Densitometric analysis of olanzapine carried out at 297 ​nm gave sharp symmetrical peak with Rf value of 0.50 and a satisfactory baseline resolution for all components. The drug was found to undergo degradation under acidic, alkaline and oxidative conditions. A single distinct peak in acidic and alkaline media while two peaks obtained as a result of oxidative degradation were well resolved along with the parent drug. The degradation products and parent drug showed significantly different Rf values. The developed HPTLC method gave quick and reproducible results for the olanzapine content in the tablets. The mean recoveries were 100.75% which confirms accuracy of the proposed method. The method was further validated for specificity, ruggedness and robustness. Based on the results, it can be suggested that the developed HPTLC method is quite efficient in separating the olanzapine from its degradation products; hence it can be used by pharmaceutical industries and regulatory bodies for the routine analysis of olanzapine in various pharmaceutical dosage forms.

Phytochemical and Network Pharmacology Based Evaluation of Antiepileptic Potential of Identified Metabolites in Argimone mexicana

Pharmacognosy Research,2021,13,4,208-217.DOI:10.5530/pres.13.4.13Published:October 2021Type:Original ArticleAuthors:Vinod Gahlot, and Dinesh Kumar Yadav Author(s) affiliations:Vinod Gahlot1, Dinesh Kumar Yadav2,* 1Department of Pharmacology, SGT College of Pharmacy, SGT University, Gurugram, Haryana, INDIA. 2Department of Pharmacognosy, SGT College of Pharmacy, SGT University, Gurugram, Haryana, INDIA. Abstract:Background: Quality-based assessment of herbal drugs or products is being more concerning for their quality, safety and regulatory purpose. A. mexicana is a traditional herbal medicine that has a long history in the treatment of arthritis, anti-fungal anti-cancer and brain disorders. Aim: Due to lack of scientific evidence based on phytopharmacology, the study is aimed for phytochemical and antiepileptic evaluation of identified metabolites in A. mexicana. Materials and Methods: Phytochemical identification was done using MS, FTIR and 1H-NMR and quantitated using HPTLC densitometric analysis. Further, ADME and network pharmacology studies were performed to evaluate biological response of identified metabolites. Results: The resulted outcomes of the spectral analysis suggest isolated compounds as ferulic acid, caffeic acid, berberine and angoline. In HPTLC quantitative analysis, the content of ferulic acid, caffeic acid, berberine and angoline was found as 3.475 ± 0.028, 1.036 ± 0.013, 0.714 ± 0.014 and 0.738 ± 0.081 μg/mg of A. mexicana extract. In ADME analysis, berberine and angoline showed good bioavailable response while in network pharmacology analysis, except angoline, all the metabolites significantly interacted with several genes (SOD1, NOS, MAPK3, UGT1As, G6PD, ACOT2, BAAT etc) associated with brain ischemia, oxidative and inflammatory stress or the genes response for elimination of toxins from the body. Conclusion: Hence, the study enlightens that A. mexicana possess several major and minor metabolites and which can be the key parameter for further quality and regulatory based assessment of A. Mexicana and biological role against oxidative and inflammatory stress induced epileptic seizure. Keywords:ADME analysis, Argemone mexicana L., HPTLC, Network pharmacology, PhytoconstituentsView:PDF (1.81 MB)