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Tamoxifen and toremifene are two selective estrogen receptor modulators (SERMs) commonly used to treat breast cancer in women. Toremifene is well-known as a triphenylethylene derivative. Carboxy toremifene is a common metabolite of toremifene and tamoxifen. Since 2005, the World Anti-Doping Agency (WADA) has banned the SERMs category during in and out of competition. These substances are in the S4 category in the WADA prohibited list as "agents with anti-oestrogenic activity." However, there is no commercially accessible carboxy toremifene reference material in the market. This research highlights the novel synthetic procedure, the development of a carboxy toremifene HPLC method, and validation, along with detailed characterization using advanced analytical techniques using

Ethylmorphine is permitted internationally for therapeutic purposes where morphine is not indicated across the globe. Nor-ethylmorphine a major metabolite of ethylmorphine. To differentiate the intake of morphine from ethylmorphine, nor-ethylmorphine stable reference material is desirable. There is no available commercial source and no data for reference material context for this substance. Therefore, nor-ethylmorphine HCl was synthesized and characterized, and purity and potency were assessed using nuclear magnetic resonance (NMR), high-resolution mass spectrometry (HRMS), Fourier transform infrared spectroscopy (FT-IR), thermogravimetry (TGA), and high-performance liquid chromatography (HPLC). Purity and potency were found to be 98.29% and 96.40%, respectively, providing a fit for purpose reference material for doping control analysis in sports.

Prenylamine was initially used for the treatment of angina pectoris and later on withdrawn from the market in 1988 due to cardiac arrhythmias concern. The major phase I metabolite of prenylamine is p-hydroxy prenylamine that has a chiral center in the structure. Even though p-hydroxy prenylamine was synthesized earlier, it lacked complete analytical developments for chiral high-performance liquid chromatography (HPLC) separation. However, p-hydroxy prenylamine reference material is not commercially available. The innovation of this manuscript is the development and validation of a chiral HPLC separation method and more extensive characterization of the reference material than previously reported method. Therefore, it was hypothesized to develop and validate normal phase HPLC method for p-hydroxy prenylamine reference material. p-Hydroxy prenylamine was synthesized in two batches and characterized successfully using 13 C NMR, 1 H NMR, high-resolution mass spectrometry (HRMS), Fourier transform infrared spectroscopy (FT-IR), and thermogravimetric analysis (TGA). A normal phase chiral HPLC method was developed to analyze the p-hydroxy prenylamine purity. Separation of the p-hydroxy prenylamine enantiomers were achieved using ultra-high-performance liquid chromatography (UHPLC) on a ChiralCel ODH column at wavelength of 220 nm. The developed method was validated in terms of its linearity, accuracy, precision, and robustness for purification, purity assessment, and stability studies. Proton and carbon peaks were confirmed by nuclear magnetic resonance (NMR) analysis. Functional groups were confirmed by FT-IR. Loss on drying was 0.3% and 0.6% for Batches 1 and 2, respectively. The purity of the developed reference material for Batches 1 and 2 was found to be 99.59% and 100%, respectively. Therefore, the synthesized batches of p-hydroxy prenylamine can be used in dope testing as reference material.

Insulin-like growth factor 1 (IGF-1) is indicated for growth failure in pediatric patients with primary IGF-1 deficiency and for patients with neutralizing antibodies to growth hormone. IGF-1 was cloned, expressed and purified in-house. Preliminary stability studies prior to the transdermal delivery experiments showed that although stable in contact with stratum corneum, the solution concentration of IGF-1 decreased to 23.63 ± 2.48 and 21.58 ± 2.62% of the initial value upon exposure for 8 h to porcine dermis of 250 and 750 µm thickness. This led to an investigation into how it might be possible to improve the stability of IGF-1 in the presence of porcine/human skin. The stability of IGF-1 in the presence of dermis improved upon heating the skin samples at 60 °C for 2 min suggesting that IGF-1 was subject to enzymatic degradation. Although addition of the protease inhibitor, phenylmethanesulfonyl fluoride (PMSF) alone, did not improve stability, the use of a protease inhibitor cocktail completely blocked proteolytic degradation of IGF-1; the solution concentration after an 8 h exposure to porcine skin was equivalent to the initial level (103.87 ± 9.15%). The results obtained with porcine skin were confirmed with human skin (IGF-1 recovery was 99.31 ± 9.98%). These findings suggest that the inclusion of protease inhibitor cocktails may be useful in limiting the degradation of therapeutic proteins during iontophoresis and transdermal delivery in general – this could be of particular interest for local delivery of peptide/protein therapeutics for dermatological applications.

The first objective was to investigate the transdermal iontophoresis of interferon beta 1b (IFN); the second was to determine whether the addition of 10 Arg residues at the N-terminus, creating a highly charged poly-Arg analogue (Arg10-IFN), increased delivery. Cumulative permeation of IFN and Arg10-IFN after iontophoresis at 0.5 mA/cm2 for 8 h was 6.97 ± 4.82 and 9.55 ± 1.63 ng/cm2, respectively – i.e. >1000-fold less than that of ribonuclease A, cytochrome c and human basic fibroblast growth factor. Co-iontophoresis of acetaminophen showed that, in contrast to lysozyme, neither IFN nor Arg10-IFN interacted with skin to decrease convective solvent flow. Furthermore, there was no statistically significant difference between (i) iontophoretic delivery of IFN across intact or laser porated skin and (ii) passive or iontophoretic delivery of IFN across laser porated skin. Chromatographic characterisation supported the hypothesis that IFN was bound strongly to albumin. The formation of a ~ 86 kDa complex with albumin was probably responsible for the poor cutaneous delivery of IFN/Arg10-IFN despite the use of iontophoresis and/or laser microporation. Biopharmaceuticals might interact with specific proteins during iontophoretic transport and so decrease their (per)cutaneous delivery without affecting electroosmotic solvent flow, which is usually considered as a reliable marker to report on permeant binding during electrotransport across the skin., Graphical abstract Unlabelled Image

The first objective was to investigate the transdermal iontophoresis of interferon beta 1b (IFN); the second was to determine whether the addition of 10 Arg residues at the N-terminus, creating a highly charged poly-Arg analogue (Arg

Most biopharmaceutical formulations use polysorbates: surfactants that are highly efficient but difficult to manage in terms of compositional variability, quality, and stability. Alternatives, such as poloxamers, albumin, and cyclodextrin, are becoming popular and are being explored for their potential to protect biopharmaceuticals against physical and mechanical stresses.

A well-controlled clinical trial is one of the critical steps to evaluate the efficacy and safety of novel medicaments. The use of a placebo is employed in several types of clinical trials in order to set a baseline against which the efficacy of the investigational drug is evaluated. An ideal placebo should match the final formulation as close as possible such that the patient/health care providers are unable to identify any difference. This is difficult for high concentration biologic intended for subcutaneous administration, mainly because of their color and viscosity. Currently, the challenge is overcome by using opaque labels or by unblinded pharmacists, both solutions are expensive. The present study provides an efficient alternative where a protein excipient (recombinant albumin) is used to prepare a placebo for biologicals. We have demonstrated that the use of recombinant albumin can match the color of the active when used in the right quantity. The evaluated solution is highly flexible and has the potential to match the color of different biopharmaceuticals at different concentrations/formulations.

Psoriasis is a chronic autoimmune skin disorder that affects millions of people across the planet. It causes many serious complications to afflicted patients and is indicated by uncontrolled growth and differentiation of keratinocytes. Conventional dosage forms have been used for the topical delivery of antipsoriatic drugs but they possess their own limitations. These include poor patient compliance, inadequate penetration and targeting to diseased skin, and local toxicity. In recent decades nanocarriers like liposomes, micelles, and solid lipid nanoparticles have shown potential in delivering antipsoriatic active compounds to the targeted areas of skin. Such novel formulations improve therapeutic efficacy, increase localization of drug in the skin, and decrease side effects. This chapter sheds light on the current advancements in the field of nanocarriers in treating the psoriatic skin condition.

Biologics are becoming increasingly important and currently contribute to more than one-third of all the new drugs approved—and have done consistently for the past decade. They are relatively new compared to well-established small chemical molecules; furthermore, new molecular entities are entering clinical trials and eventually clinics every year. Apart from being complex and large in size, they are also often more fragile to external stress; overall stability and storage consideration are extremely important for biologics. Scientific development is driven by challenges and pharmaceutical companies are working toward overcoming these challenges by employing several innovative strategies. The purpose of this chapter is to provide an overview of the current status of biologics and what to expect in future.

Medical practice can be viewed as the practice of decision making in the medical field. Today, computers have evolved as a crucial component in medical decision making, but the general perception of “computers in medicine” is often merely of computer applications that assist physicians in diagnosis of illnesses. Contrary to this perception, health-care organizations are under increased pressure to constantly improve their operational efficiency using computers while concurrently keeping overall operating costs down, with no degradation in the delivered quality of health care. To achieve this target, researchers as well as practitioners are increasingly relying on knowledge-based systems and automation in the field of medicine. Most applications of these expert or knowledge-based systems in health care imply a tangential or direct impact on the quality of healthcare services. This chapter explores the ways and technologies by which knowledge-based systems support health-care professionals, from examinations through the enormous database of probable diseases and symptoms, to high-precision surgeries and more. It also brings together the subject specifics by concentrating on systems developed specifically to assist health workers in making medical decisions. In this chapter, we discuss various details of techniques that support clinical decision making, their features as well as their availability as a diagnostic tool. We further discuss their effects on the practices of medicine and surgery, medical education, and on relationships among the concerned population as patients and physicians.

The effect of skin barrier impairment on the iontophoretic transport of low (acetaminophen (ACM), lidocaine (LD), ketorolac (KT)) and high molecular weight permeants, (cytochrome c (Cyt c) and ribonuclease T1 (RNase T1)), was evaluated using tape-stripping (TS) and fractional laser ablation for “large-scale” and “localized” barrier disruption. Interestingly, removal of the stratum corneum did not invariably lead to an increase in iontophoretic delivery of the permeants. Decrease of electroosmotic (EO) flow and facilitated transport of Cl- ions in the cathode-to-anode direction, which reduced cation electromigration (EM), both impacted cation delivery by anodal iontophoresis but the effects were partly offset by enhanced passive diffusion. Decrease in EO increased cathodal iontophoresis of KT but not that of RNase T1. Permeability coefficients confirmed the superiority of EM over EO for small molecules, LD > KT > ACM. A combination of fractional laser ablation and iontophoresis was advantageous for both positively and negatively charged small molecules as passive penetration was significantly enhanced. In conclusion, results demonstrated that (i) skin ablation prior to anodal iontophoresis decreased EO and EM but could be advantageous for delivery if the ablative technique enhanced passive penetration thereby compensating reduction of electrotransport and (ii) reduced EO favored cathodal electrotransport.

The cyclic enaminone moiety has been identified as a new scaffold for selective inhibition of cyclooxygenase-2 with anti-inflammatory and analgesic activities. The designed cyclic enaminones have been synthesized conveniently through the development of a new catalyst-free methodology and evaluated for cyclooxygenase (COX-1 and COX-2) inhibitory activities. Three compounds 7d, 8, and 9 predominantly inhibited COX-2 with selectivity index of 74.09, 19.45 and 108.68, respectively, and were assessed for in vivo anti-inflammatory activity in carrageenan induced rat paw edema assay. The anti-inflammatory activity of 7d was comparable to that of celecoxib at a dose of 12.5 mg/kg. However, the compounds 8 and 9 were more/equally effective as anti-inflammatory agent compared to celecoxib at the doses of 12.5 mg/kg and 25 mg/kg and also exhibited anti-inflammatory activity comparable to that of diclofenac. The therapeutic potential of the most active compound 9 was further assessed by performing in vivo thermal and mechanical hyperalgesia tests using various models that revealed its analgesic activity. The in vivo non-ulcerogenicity of 9 revealed the gastrointestinal safety as compared to the non-selective COX inhibitor indomethacin. The in vitro antioxidant activity and in vivo experiments on heart rate and blood pressure provided the cardiovascular safety profile of 9. The molecular docking studies rationalize the COX-2 selectivity of the newly found anti-inflammatory compounds 7d, 8, and 9.

INTRODUCTION: The use of glucocorticosteroids (GCs) through oral, intravenous, intramuscular, or rectal routes is prohibited in sports. Its use is permitted through inhalation, topical and intra-articular route of administration. Methylprednisolone (MP) is available for use by different routes for anti-inflammatory and immunosuppressive purposes. To discriminate its intake by permitted & forbidden routes, a reporting level of 30 ng/ml is set by World Anti-Doping Agency. The aim of this study was to compare MP's excretion profile following oral & intra-articular administration & to evaluate its effect on endogenous GCs profile. MATERIALS & METHODS: The MP was administered through oral and intra-articular route to different patients & urine samples were collected up to 100 h. The urine samples were hydrolyzed, extracted, and analyzed on Liquid chromatography-mass spectrometry/MS. RESULTS: MP levels in urine exceeded the reporting limit of 30 ng/ml after oral (8 mg) and intra-articular administration (80 mg) routes. After oral intake (8 mg), MP levels exceeded the reporting level up to 24 h. However, after intra-articular injection (80 mg), the MP could be detected above the reporting level up to 80 h. CONCLUSION: The findings reveal that the MP can exceed the reporting level in urine even after administration by permitted route (i.a.). Further analysis of four endogenous GCs (Cortisol, Cortisone, TH Cortisone, and 11-deoxycortisol) showed a decreased excretion following administration of MP by oral & intra-articular routes.

Dermatological diseases, including most skin cancers and rare genetic conditions frequently originate in the epidermis. Targeted, topical cell-based therapy is a promising therapeutic strategy. Here, we present the first report demonstrating that fractional laser ablation enables local 'needle-free' intraepidermal delivery of living human cells.The cells penetrated porcine ear skin via microchannels created by Er:YAG fractional laser ablation; cell delivery was quantified using a haemocytometer. Cutaneous distribution was confirmed visually by laser scanning confocal microscopy and histological analysis.Total cell delivery (sum of amounts permeated and deposited) after 24 h increased from 5.7 ± 0.1 x10The results demonstrate the feasibility of using controlled laser microporation to achieve local 'needle-free' cutaneous delivery of living human cells to the epidermis and dermis. This raises the possibility of using this technique for targeted new approaches for dermatological therapy in these regions.

A simple two-step process was used to disperse acid functionalized multi-walled carbon nanotubes (CNTs) in poly(vinylidene fluoride) (PVDF). While the neat solvent-cast PVDF showed coexistence of α- and β-phases; the composite films exhibited only β-phase crystals. Further studies on the crystalline behaviour, using differential scanning calorimetry and small-angle X-ray scattering techniques showed an increase in the percentage of crystalline phase with CNT. The network formed by CNTs in the matrix reduced the macroscopic electrical resistivity of composite films. The dielectric constant increased with CNT loading. Further, these composites were investigated for its electromagnetic wave absorbance (EWA) and strain sensing properties. The EWA properties were studied in the X-band (6–12 GHz) region. A maximum of ~37 dB reflectivity loss at ~9.0 GHz was obtained in a ~25 μm thick PVDF film containing only 0.25 wt% of functionalized CNT. Preliminary studies showed a systematic change in electrical resistance by the application of dynamic bending strain in nanocomposite film. The film also showed a significant improvement in mechanical stiffness owing to efficient stress transfer from matrix to filler, the property desirable for a good strain sensor. In view of the unique combination of EWA and electro-mechanical properties, the nanocomposite films are expected to serve as a multifunctional material for strain sensing in health monitoring as well as in radar absorption.

The use of prednisolone and prednisone is prohibited by the World Anti-Doping Agency (WADA) due to their performance-enhancing effect. The purpose of the present work was to explore the possibility of identification and detection of various metabolites of prednisolone by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in excretion study samples. Ten metabolites of prednisolone could be identified namely prednisone (11-oxo metabolite) [M-1], 6-β-OH-prednisolone [M-2], 20-β-OH-prednisolone [M-3], 20-α-OH-prednisolone [M-4], 20-α-OH-prednisone [M-5], 20-β-OH-prednisone [M-6], 2 tetrahydro epimers of 20-β-OH-prednisolone [M-7], 2 tetrahydro epimers of 20-α-OH-prednisolone [M-8], 2 tetrahydro epimers of 20-β-OH-prednisone [M-9], and 2 tetrahydro epimers of 20-α–OH-prednisone [M-10]. Prednisolone was administered in 10-, 20-, and 40-mg dosage to healthy volunteers to study detection of various metabolites. The parent, M-1, M-2, and M-3 could be detected up to 72 h while rest of the metabolites were detectable up to 24 h after drug administration. The detection of newer metabolites of the drug can further be used for confirmatory purposes. Copyright © 2012 John Wiley & Sons, Ltd.

Growth and nitrilase production by recombinant Escherichia coli cells harbouring pET 21 (b) plasmid, for the expression of Pseudomonas putida nitrilase were improved using response surface methodology. Central composite design was used for obtaining ideal concentration of critical medium components which include fructose, tryptone, yeast extract and lactose. The optimal values for the concentration of fructose, tryptone, yeast extract and lactose were found to be 1.13, 2.26, 3.25 and 0.9 % (w/v), respectively. Here, fructose served as carbon source for the growth while lactose was preferably used as inducer for the expression of foreign protein. Yeast extract in the medium was used as a growth promoter while tryptone was added as a major nitrogen source. Using this optimized medium, an experimental growth of 6.67 (OD at 600 nm) and nitrilase activity of 27.13 U/ml was achieved. This approach for medium development led to an enhancement of the growth and enzyme activity by 1.4 and 2.2 times, respectively, as compared to the un-optimized medium.

Cathodal iontophoresis of anionic macromolecules has been considered a major challenge owing to (i) the presence of a negative charge on the skin under physiological conditions and (ii) the electroosmotic solvent flow in the (opposite) anode-to-cathode direction. Moreover, electroosmosis, and not electromigration, was considered as the likely electrotransport mechanism for high molecular weight cations. However, it was recently shown that electromigration governed anodal iontophoretic transport of Cytochrome c (12.4 kDa) and Ribonuclease A (RNAse A; 13.6 kDa). Thus, the objective of this study was to investigate the feasibility of iontophoresing a negatively charged protein, the enzyme Ribonuclease T1 (RNAse T1, 11.1 kDa), from the cathode across intact skin. Cumulative permeation and skin deposition of RNAse T1 were investigated as a function of current density (0.15, 0.3 and 0.5 mA/cm(2) applied for 8h) using porcine ear skin and quantified by an enzymatic activity assay. Although RNAse T1 permeation was dependent upon current density (22.41 ± 8.10, 76.41 ± 56.98 and 142.19 ± 62.23μg/cm(2), respectively), no such relationship was observed with respect to skin deposition (9.78 ± 2.39, 7.76 ± 4.34 and 8.70 ± 2.94 μg/cm(2), respectively). MALDI-TOF spectra and the activity assay confirmed that RNAse T1 retained structural integrity and enzymatic function post-iontophoresis. Acetaminophen iontophoresis demonstrated the anode-to-cathode directionality of electroosmotic solvent flow confirming that RNAse T1 electrotransport was due entirely to electromigration. Interestingly, despite its lower net charge and higher molecular weight, electromigration of cationic Ribonuclease A was superior to that of RNAse T1 after iontophoresis at 0.5 mA/cm(2) for 8h. These results provide further evidence that charge to mass ratio and hence electric mobility might not alone be sufficient to predict protein electrotransport across the skin; three dimensional structures and the spatial distribution of physicochemical properties must also be considered. The skin extraction data suggest that negatively charged molecules may have fewer potential binding sites in the skin than their cationic counterparts. This was supported by confocal laser scanning microscopy images which showed that whereas fluorescence from RNAse A was distributed throughout the epidermis and dermis, RNAse T1 appeared to be bound to the epidermis alone. In conclusion, this is the first report demonstrating successful non-invasive cathodal iontophoresis of a negatively charged functional protein (RNAse T1) across intact skin.

The purpose of the study was to demonstrate the feasibility of using transdermal iontophoresis to deliver a functional protein, ribonuclease A (RNAse; 13.6 kDa), non-invasively across the skin. Iontophoretic transport experiments were conducted using porcine skin in vitro and established the effect of current density and protein concentration on delivery kinetics. A methylene blue-based assay was used to quantify RNAse transport and to simultaneously demonstrate that protein functionality was retained post-iontophoresis. The results confirmed that intact functional RNAse was indeed delivered across the skin; cumulative permeation and steady state flux after 8h iontophoresis at 0.3 mA/cm(2) were 224.37+/-72.34 microg/cm(2) and 68.28+/-23.87 microg/cm(2)h, respectively. Significant amounts of protein were also deposited within the membrane (e.g., 1425.13+/-312.09 microg/cm(2) at 0.3 mA/cm(2)). In addition to the evidence provided by the enzymatic assay with regards to RNAse integrity and functionality, SDS-PAGE gels and MALDI-TOF spectra were also used to characterize RNAse present in the receiver phase (MALDI-TOF spectra: RNAse control, 13.690 kDa cf. RNAse from permeation samples, 13.692 kDa). Co-iontophoresis of acetaminophen showed that, despite its molecular weight, electromigration was the predominant electrotransport mechanism, accounting for >80% of RNAse total flux. Increasing RNAse concentration from 0.35 to 0.7 mM in the formulation did not result in a statistically significant increase in delivery. Iontophoretic transport of RNAse across human skin was statistically equivalent to that seen with porcine skin under the same conditions; cumulative permeation across human and porcine skin was 241.48+/-60.01 and 170.71+/-92.13 microg/cm(2), respectively. Laser scanning confocal microscopy was used to visualize the distribution of rhodamine B-labelled RNAse in the epidermis and dermis as a function of depth following 8h iontophoresis (results were compared to control experiments involving passive administration of the same formulation for 8h). Although fluorescence was localized at the skin surface following passive administration, it was visible throughout the membrane after current application. In conclusion, the results demonstrate that non-invasive transdermal iontophoresis can be used to deliver significant amounts of a structurally intact, functional protein across skin.

The original aim of the study was to investigate the transdermal iontophoretic delivery of lysozyme and to gain further insight into the factors controlling protein electrotransport. Initial experiments were done using porcine skin. Lysozyme transport was quantified by using an activity assay based on the lysis of Micrococcus lysodeikticus and was corrected for the release of endogenous enzyme from the skin during current application. Cumulative iontophoretic permeation of lysozyme during 8h at 0.5mA/cm(2) (0.7mM; pH6) was surprisingly low (5.37±3.46μg/cm(2) in 8h) as compared to electrotransport of cytochrome c (Cyt c) and ribonuclease A (RNase A) under similar conditions (923.0±496.1 and 170.71±92.13μg/cm(2), respectively) - despite its having a higher electrophoretic mobility. The focus of the study then became to understand and explain the causes of its poor iontophoretic transport. Lowering formulation pH to 5 increased histidine protonation in the protein and decreased the ionisation of fixed negative charges in the skin (pI ~4.5) and resulted in a small but statistically significant increase in permeation. Co-iontophoresis of acetaminophen revealed a significant inhibition of electroosmosis; inhibition factors of 12-16 were indicative of strong lysozyme binding to skin. Intriguingly, lidocaine electrotransport, which is due almost exclusively to electromigration, was also decreased (approximately 2.7-fold) following skin pre-treatment by lysozyme iontophoresis (cf. iontophoresis of buffer solution) - suggesting that lysozyme was also able to influence subsequent cation electromigration. In order to elucidate the site of skin binding, different porcine skin models were tested (dermatomed skin with thicknesses of 250 and 750μm, tape-stripped skin and heat-separated dermis). Although no difference was seen between permeation across 250 and 750μm dermatomed skin (13.57±12.20 and 5.37±3.46μg/cm(2), respectively), there was a statistically significant increase across tape-stripped skin and heat-separated dermis (36.86±7.48 and 43.42±13.11μg/cm(2), respectively) - although transport was still much less than that seen across intact skin for Cyt c or RNase A. Furthermore, electroosmotic inhibition factors fell to 2.2 and 1.0 for tape-stripped skin and heat-separated dermis - indicating that lysozyme affected convective solvent flow through interactions with the epidermis and predominantly the stratum corneum. Finally, cation exchange and hydrophobic interaction chromatography confirmed that although lysozyme had greater positive charge than Cyt c or RNase A under the conditions used for iontophoresis, it also possessed the highest surface hydrophobicity, which may have facilitated the interactions with the transport pathways and encouraged aggregation in the skin microenvironment. Thus, high charge and electrophoretic mobility seem to be inadequate descriptors to predict the transdermal iontophoretic transport of proteins whose complex three dimensional structures can facilitate interactions with cutaneous transport pathways.

It is a combinational logic circuit, it is used in application in which data must be switched from multiple source to a destination as unidirectional device. it represent the simulation of different 2∶1 its comperative analysis on different parameter such as power supply voltage, operating A multiplexer is known as mux. It is a device that helps in selection a number of input signals, frequency temperature and area efficiency and its applications in 1bit full adder cell all the simulation have been followed on cadence tool at 180nm technology at virtuoso.

Human basic fibroblast growth factor (hbFGF; 17.4 kDa) has shown promise in the treatment of several dermatological conditions; symptomatic improvement was also observed in patients with peripheral arterial disease after arterial infusion. The objective of this study was to demonstrate the feasibility of using transdermal iontophoresis to deliver biologically active hbFGF noninvasively into and across the skin. The protein was cloned, expressed and purified in-house. Porcine skin was used to investigate transdermal iontophoretic transport of hbFGF as a function of current density (0.15, 0.3, and 0.5 mA/cm(2)); results were subsequently confirmed using human skin. Cumulative hbFGF permeation and skin deposition were quantified by ELISA. The absence of proteolytic degradation during skin transit was confirmed by SDS-PAGE. Biological activity postdelivery was determined using cell proliferation assays in human foreskin fibroblast (HFF) and NIH 3T3 cell lines. Confocal laser scanning microscopy (CLSM) was used to visualize the distribution of rhodamine-tagged hbFGF in the skin. Cumulative iontophoretic permeation at 0.3 mA/cm(2) was statistically superior to that at 0.15 mA/cm(2); however, there was no further improvement at 0.5 mA/cm(2). Significant skin deposition of hbFGF was observed, and this dominated transport; for example, after iontophoresis for 8 h at 0.5 mA/cm(2), skin deposition (77.74 ± 37.36 μg/cm(2)) was 4.4-fold higher than cumulative permeation (17.64 ± 5.18 μg/cm(2)). The superior skin deposition may be advantageous for dermatological applications. The HFF and NIH 3T3 cell proliferation assays confirmed that biological activity of hbFGF was retained postdelivery. Coiontophoresis of acetaminophen showed that the dominant transport mechanism switched from electroosmosis to electromigration upon increasing current density from 0.15 to 0.3 mA/cm(2). Experiments using human skin confirmed that iontophoretic permeation of hbFGF across porcine and human membranes was statistically equivalent. CLSM images of rhodamine-tagged hbFGF postiontophoresis indicated that the protein was evenly distributed throughout the epidermis and dermis. In conclusion, the results confirmed that transdermal iontophoresis was indeed able to deliver structurally intact, functional hbFGF noninvasively into and across the skin. The amounts of protein delivered were similar to those in reports from preclinical and clinical studies.

The use of prednisolone and prednisone is prohibited by the World Anti-Doping Agency (WADA) due to their performance-enhancing effect. The purpose of the present work was to explore the possibility of identification and detection of various metabolites of prednisolone by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in excretion study samples. Ten metabolites of prednisolone could be identified namely prednisone (11-oxo metabolite) [M-1], 6-β-OH-prednisolone [M-2], 20-β-OH-prednisolone [M-3], 20-α-OH-prednisolone [M-4], 20-α-OH-prednisone [M-5], 20-β-OH-prednisone [M-6], 2 tetrahydro epimers of 20-β-OH-prednisolone [M-7], 2 tetrahydro epimers of 20-α-OH-prednisolone [M-8], 2 tetrahydro epimers of 20-β-OH-prednisone [M-9], and 2 tetrahydro epimers of 20-α-OH-prednisone [M-10]. Prednisolone was administered in 10-, 20-, and 40-mg dosage to healthy volunteers to study detection of various metabolites. The parent, M-1, M-2, and M-3 could be detected up to 72 h while rest of the metabolites were detectable up to 24 h after drug administration. The detection of newer metabolites of the drug can further be used for confirmatory purposes.

Nitrilases have attracted tremendous attention for the preparation of optically pure carboxylic acids. This article aims to address the production and utilization of a highly enantioselective nitrilase from Pseudomonas putida MTCC 5110 for the hydrolysis of racemic mandelonitrile to (R)-mandelic acid. The nitrilase gene from P. putida was cloned in pET 21b(+) and over-expressed as histidine-tagged protein in Escherichia coli. The histidine-tagged enzyme was purified from crude cell extracts of IPTG-induced cells of E. coli BL21 (DE3). Inducer replacement studies led to the identification of lactose as a suitable and cheap alternative to the costly IPTG. Effects of medium components, various physico-chemical, and process parameters (pH, temperature, aeration, and agitation) for the production of nitrilase by engineered E. coli were optimized and scaled up to a laboratory scale bioreactor (6.6 l). Finally, the recombinant E. coli whole-cells were utilized for the production of (R)-(−)-mandelic acid.

Caffeine (1,3,7-trimethylxanthine) is a xanthine alkaloid compound that acts as a stimulant and was banned for use in sports. Caffeine was removed from the World Anti-Doping Agency (WADA) prohibited list from January 2004. As per the requirement of WADA, the WADA-accredited labs are monitoring the presence of caffeine for signs of possible abuse. National Dope Testing Laboratory (NDTL), India, is monitoring caffeine levels in in-competition urine samples of sports persons from different sports disciplines. The aim of the present study is to find the distribution of caffeine levels in urine in different sport disciplines in Indian and other south-east Asian countries. Caffeine was measured using an alkaline extraction with tertiary butyl methyl ether followed by gas chromatographic nitrogen-phosphorus specific detection. A comparison of statistical data from the urine samples done from January 2008 to July 2010 in NDTL, India, for different discipline of sports since January 2008 to July 2010 was performed. The study proves beneficial in monitoring the abuse of caffeine in different disciplines of sports.