A validated HPLC analytical method for the analysis of solasonine and solamargine in in vitro skin penetration studies

To assess topical delivery studies of glycoalkaloids, an analytical method by HPLC-UV was developed and validated for the determination of solasonine (SN) and solamargine (SM) in different skin layers, as well as in a topical formulation. The method was linear within the ranges 0.86 to 990.00 µg/mL for SN and 1.74 to 1000.00 µg/mL for SM (r = 0.9996). Moreover, the recoveries for both glycoalkaloids were higher than 88.94 and 93.23% from skin samples and topical formulation, respectively. The method developed is reliable and suitable for topical delivery skin studies and for determining the content of SN and SM in topical formulations.

Desenvolvimento de procedimento de extração para análise de sedativos e B-bloqueadores em rim suíno

A procedure was developed for determination of 5 sedatives and 14 β-blockers in swine kidney and subsequent analysis by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Three different procedures for extraction were tested, evaluated through recovery studies. The procedure using acetonitrile for extraction and cleanup with freezing at low temperature and dispersive solid phase extraction using 500 mg celite® 545 before the concentration step presented the better results. The dried samples were redissolved with methanol and analyzed using a LC-MS/MS system with electrospray ionization (ESI) operating in positive MRM mode. The recovery values for this procedure were in the 75-88% range. The robustness of the method was tested against small variations. The method was used to analyze carazolol, azaperone and azaperol in collaborative assay, obtaining results close to designed value.

A simple method for the spectrophotometric determination of cephalosporins in pharmaceuticals using variamine blue

A simple spectrophotometric method for the determination of cefotaxime, ceftriaxone, cefadroxil and cephalexin with variamine blue is presented. The determination is based on the hydrolysis of β-lactam ring of cephalosporins with sodium hydroxide which subsequently reacts with iodate to liberate iodine in acidic medium. The liberated iodine oxidizes variamine blue to violet colored species of maximum absorption at 556 nm. The absorbance is measured within the pH range of 4.0-4.2. Beer's law is obeyed in the range of 0.5-5.8 µg mL-1, 0.2-7.0 µg mL-1, 0.2-5.0 µg mL-1 and 0.5-8.5 µg mL-1 for cefotaxime, ceftriaxone, cefadroxil and cephalexin respectively. The analytical parameters were optimized and the method is successfully applied for the determination of cefotaxime, ceftriaxone, cefadroxil and cephalexin in pharmaceuticals.

Bound state solutions of schrödinger equation for a more general exponential screened coulomb potential via Nikiforov-Uvarov method

The arbitrary angular momentum solutions of the Schrödinger equation for a diatomic molecule with the general exponential screened coulomb potential of the form V(r) = (- a / r){1+ (1+ b )e-2b } has been presented. The energy eigenvalues and the corresponding eigenfunctions are calculated analytically by the use of Nikiforov-Uvarov (NU) method which is related to the solutions in terms of Jacobi polynomials. The bounded state eigenvalues are calculated numerically for the 1s state of N2 CO and NO

A simple spectrophotometric method for the determination of tetracycline and doxycycline in pharmaceutical formulations using chloramine-T

A simple, rapid, accurate and inexpensive spectrophotometric method for the determination of tetracycline and doxycycline has been developed. The method is based on the reaction between these drugs and chloramine-T in alkaline medium producing red color products with absorbance maximum at the Λ = 535 and 525 nm for the tetracycline and doxycycline, respectively. The best conditions for the reactions have been found using multivariate method. Beer´s law is obeyed in a concentration ranges 1.03 x 10-5 to 3.61 x 10-4 mol L-1 and 1.75 x 10-5 to 3.48 x 10-4 mol L-1 for the tetracycline and doxycycline, respectively. The quantification limits were 5.63 x 10-6 mol L-1 and 7.12 x 10-7 mol L-1 for the tetracycline and doxycycline, respectively. The proposed method was successfully applied to the determination of these drugs in pharmaceutical formulations and the results obtained were in good agreement with those obtained by the comparative method at the 95% confidence level.

Combining Near-Field and Far-Field Microscopy: A new method for nanoscale super-resolution imaging

<b>Lähikenttä- ja kaukokenttämikroskopian yhdistäminen: Uusi korkearesoluutioinen menetelmä nanokuvantamiseen.b> Osteoporoosi on sairaus, jossa luun uudistumisprosessi ei ole enää tasapainossa. Uuden luun muodostuminen on hitaampaa johtuen osteoblastien laskeneesta aktiivisuudesta. Yksi keino estää osteoporoosin syntyä on estää osteoklastien sitoutuminen luun pinnalle, jolloin ne eivät aloita luun syömisprosessia. Tämän Pro gradu -tutkielman tarkoituksena on luoda uusi työkalu osteoklastien sitoutumisen tutkimiseen samanaikaisesti fluoresenssi- ja atomivoimamikroskoopilla. Tätä tarkoitusta varten yhdistettiin atomivoimamikroskooppi sekä STED mikroskooppi. Kirjallisuuskatsauksessa käydään läpi yksityiskohtaisesti molempien mikroskooppitekniikoiden teoriat. Kokeellisessa osiossa esitetään käytetyt metodit ja alustavat tulokset uudella systeemillä. Lisäksi keskustellaan lyhyesti kuvan analysoinnista ImageJohjelmalla. Konfokaalisen fluoresenssimikroskoopin ja atomivoimamikroskoopin yhdistelmä on keksitty jo aikaisemmin, mutta tavallisen konfokaalimikroskoopin erottelukyvyn raja on noin 200 nanometriä johtuen valon diffraktioluonteesta. Yksityiskohdat eivät erotu, jos ne ovat pienempiä kuin puolet käytettävästä aallonpituudesta. STED mikroskooppi mahdollistaa fluoresenssikuvien taltioimisen solunsisäisistä prosesseista 50 nanometrin lateraalisella erotuskyvyllä ja atomivoimamikroskooppi antaa topografista tietoa näytteestä nanometrien erotuskyvyllä. Biologisia näytteitä kuvannettaessa atomivoimamikroskoopin erotuskyky kuitenkin huononee ja yleensä saavutetaan 30-50 nanometrin erotuskyky. Kuvien kerrostaminen vaatii vertauspisteitä ja tätä varten käytettiin atomivoimamikroskoopin kärjen tunnistamista ja referenssipartikkeleita. Kuva-analysointi suoritettiin ImageJ-kuvankäsittelyohjelmalla. Tuloksista nähdään, että referenssipartikkelit ovat hyviä, mutta niiden sijoittaminen tarkasti tietylle kohdealueelle on hankalaa nanoskaalassa. Tästä johtuen kärjen havaitseminen fluoresenssikuvassa on parempi metodi. Atomivoimamikroskoopin kärki voidaan päällystää fluoresoivalla aineella, mutta tämä lisää kärjen aiheuttamaa konvoluutiota mittausdataan. Myös valon takaisinsirontaa kärjestä voidaan tutkia, jolloin konvoluutio ei lisäänny. Ensimmäisten kuvien kerrostamisessa käytettiin hyväksi fluoresoivalla aineella päällystettyä kärkeä ja lopputuloksessa oli vain 50 nanometrin yhteensopimattomuus fluoresenssi- ja topografiakuvan kanssa. STED mikroskoopin avulla nähdään leimattujen proteiinien tarkat sijainnit tiettynä ajankohtana elävän solun sisällä. Samaan aikaan pystytään kuvantamaan solun fyysisiä muotoja tai mitata adheesiovoimia atomivoimamikroskoopilla. Lisäksi voidaan käyttää funktinalisoitua kärkeä, jolla voidaan laukaista signalointitapahtumia solun ja soluväliaineen välillä. Sitoutuminen soluväliaineeseen voidaan rekisteröidä samoin kuin adheesiomediaattorien sijainnit sitoutumisalueella. Nämä dynaamiset havainnot tuottavat uutta informaatiota solun signaloinnista, kun osteoklasti kiinnittyy luun pintaan. Tämä teknologia tarjoaa uuden näkökulman monimutkaisiin signalointiprosesseihin nanoskaalassa ja tulee ratkaisemaan lukemattoman määrän biologisia ongelmia.

Estimation of the potential evapotranspiration by a simplified penman method

The numerous methods for calculating the potential or reference evapotranspiration (ETo or ETP) almost always do for a 24-hour period, including values of climatic parameters throughout the nocturnal period (daily averages). These results have a nil effect on transpiration, constituting the main evaporative demand process in cases of localized irrigation. The aim of the current manuscript was to come up with a model rather simplified for the calculation of diurnal daily ETo. It deals with an alternative approach based on the theoretical background of the Penman method without having to consider values of aerodynamic conductance of latent and sensible heat fluxes, as well as data of wind speed and relative humidity of the air. The comparison between the diurnal values of ETo measured in weighing lysimeters with elevated precision and estimated by either the Penman-Monteith method or the Simplified-Penman approach in study also points out a fairly consistent agreement among the potential demand calculation criteria. The Simplified-Penman approach was a feasible alternative to estimate ETo under the local meteorological conditions of two field trials. With the availability of the input data required, such a method could be employed in other climatic regions for scheduling irrigation.

Validation study of an overall radiation balance estimation method under Ponta Grossa (PR) Brazil weather conditions

Radiation balance is the fraction of incident solar radiation upon earth surface which is available to be used in several natural processes, such as biological metabolism, water loss by vegetated surfaces, variation of temperature in farming systems and organic decomposition. The present study aimed to assess and validate the performance of two estimation models for Rn in Ponta Grossa city, Paraná State, Brazil. To this end, during the period of 04/01/2008 to 04/30/2011, from radiometric data collected by an automatic weather station set at the Experimental Station, of the State University of Ponta Grossa. We performed a linear regression study by confrontation between measurements made through radiometric balance and Rn estimates obtained from Brunt classical method, and the proposed method. Both models showed excellent performance and were confirmed by the statistical parameters applied. However, the alternative method has the advantage of requiring only global solar radiation values, temperature, and relative humidity.

Method adjustment and equation set of maximum rainfall intensity, duration and frequency in the Mato Grosso do Sul State

Due to the lack of information concerning maximum rainfall equations for most locations in Mato Grosso do Sul State, the alternative for carrying out hydraulic work projects has been information from meteorological stations closest to the location in which the project is carried out. Alternative methods, such as 24 hours rain disaggregation method from rainfall data due to greater availability of stations and longer observations can work. Based on this approach, the objective of this study was to estimate maximum rainfall equations for Mato Grosso do Sul State by adjusting the 24 hours rain disaggregation method, depending on data obtained from rain gauge stations from Dourado and Campo Grande. For this purpose, data consisting of 105 rainfall stations were used, which are available in the ANA (Water Resources Management National Agency) database. Based on the results we concluded: the intense rainfall equations obtained by pluviogram analysis showed determination coefficient above 99%; and the performance of 24 hours rain disaggregation method was classified as excellent, based on relative average error WILMOTT concordance index (1982).

Standardization of metachromatic staining method of myofibrillar ATPase activity of myosin to skeletal striated muscle of mules and donkeys

This study aims at standardizing the pre-incubation and incubation pH and temperature used in the metachromatic staining method of myofibrillar ATPase activity of myosin (mATPase) used for asses and mules. Twenty four donkeys and 10 mules, seven females and three males, were used in the study. From each animal, fragments from the Gluteus medius muscle were collected and percutaneous muscle biopsy was performed using a 6.0-mm Bergström-type needle. In addition to the metachromatic staining method of mATPase, the technique of nicotinamide adenine dinucleotide tetrazolium reductase (NADH-TR) was also performed to confirm the histochemical data. The histochemical result of mATPase for acidic pre-incubation (pH=4.50) and alkaline incubation (pH=10.50), at a temperature of 37ºC, yielded the best differentiation of fibers stained with toluidine blue. Muscle fibers were identified according to the following colors: type I (oxidative, light blue), type IIA (oxidative-glycolytic, intermediate blue) and type IIX (glycolytic, dark blue). There are no reports in the literature regarding the characterization and distribution of different types of muscle fibers used by donkeys and mules when performing traction work, cargo transportation, endurance sports (horseback riding) and marching competitions. Therefore, this study is the first report on the standardization of the mATPase technique for donkeys and mules.

Evaluation of bursal lymphoid depletion: comparison between the conventional histology method and digital lymphocyte depletion evaluation system

Abstract: Fifty-five bursa of Fabricius (BF) were evaluated by optical microscopy for three different avian histopathologists (H1, H3 and H4) to determine the degree of lymphoid depletion. One histologist evaluated the same slides at two different times (H1 and H2) with four-months interval between the observations. The same BFs were evaluated using the system of Digital Lymphocyte Depletion Evaluation (ADDL), being performed by three differents operators of the system, not histopathologists. The results showed was a significant difference between the histopathologists and between the scores established by the same expert (H1 and H2). However, there were not significant differences between the scores with the ADDL system, obtained using ADDL. The results make clear the fragility of the subjective lymphocyte depletion score classification by the traditional histologic method, while the ADDL system proves to be more appropriated for the assessment of the lymphoid loss in the BF.

Mode B ultrasonography and abdominal Doppler in crab-eating-foxes ( Cerdocyon thous )

Abstract: Annually hundreds of crab-eating foxes (Cerdocyon thous) are referred to rehabilitation centers and zoos in Brazil. The ultrasonographic study of wildlife species is an important tool for a non-invasive and accurate anatomical description and provides important information for wildlife veterinary care. The aim of the present study was to determine the characteristics of the main abdominal organs as well as the vascular indexes of the abdominal aorta and renal arteries of crab-eating foxes (Cerdocyon thous) using mode B ultrasonography and Doppler ultrasonography, respectively. Ultrasonographic features of the main abdominal organs were described and slight differences were noticed between ultrasound imaging of abdominal organs of crab-eating foxes and other species. The bladder presented wall thickness of 12±0.01mm, with three defined layers. Both, the right and left kidneys presented corticomedullary ratio of 1:1 and similarly to the adrenals and the liver, they were homogeneous and hypoechoic compared to the spleen. The spleen was homogeneous and hyperechoic compared to the kidneys. The stomach presented 3 to 5 peristaltic movements per minute, wall thickness of 39±0.05mm and lumen and mucosa with hyperechoic and hypoechoic features, respectively. Small and large intestines presented 2 to 3 peristaltic movements per minute, wall thickness of 34±0.03mm and three defined layers with hyperechogenic (submucosa and serosa) and hypoechogenic (muscular) features. Ovaries of the female crab-eating fox were hypoechoic compared to the spleen and with heterogeneous parenchyma due to the presence of 2x2mm ovarian follicles. Prostates of the six males were regular and with a well defined boundary, with a homogeneous and hyperechoic parenchyma compared to the spleen. Vascular indexes of the abdominal aorta (PSV: 25.60±0.32cm/s; EDV: 6.96±1.68cm/s; PI: 1.15±0.07 e RI: 0.73±0.07) and right (PSV: 23.08±3.34cm/s; EDV: 9.33±2.36cm/s; PI: 1.01±0.65 e RI: 0.65±0.16) and left renal arteries (PSV: 23.74±3.94cm/s; EDV: 9.07±3.02cm/s; PI: 1.04±0.31 e RI: 0.64±0.10) were determined. Thus, conventional and Doppler ultrasonographic imaging provides basic information that can be used as reference for the species as well for other wild canids and it is a precise and non-invasive method that can be safely used to evaluate and diagnose abdominal injuries in these patients.

Method to obtain platelet-rich plasma from rabbits (Oryctolagus cuniculus )

Abstract: Platelet-rich plasma (PRP) is a product easy and inxpesnsive, and stands out to for its growth factors in tissue repair. To obtain PRP, centrifugation of whole blood is made with specific time and gravitational forces. Thus, the present work aimed to study a method of double centrifugation to obtain PRP in order to evaluate the effective increase of platelet concentration in the final product, the preparation of PRP gel, and to optimize preparation time of the final sample. Fifteen female White New Zealand rabbits underwent blood sampling for the preparation of PRP. Samples were separated in two sterile tubes containing sodium citrate. Tubes were submitted to the double centrifugation protocol, with lid closed and 1600 revolutions per minute (rpm) for 10 minutes, resulting in the separation of red blood cells, plasma with platelets and leucocytes. After were opened and plasma was pipetted and transferred into another sterile tube. Plasma was centrifuged again at 2000rpm for 10 minutes; as a result it was split into two parts: on the top, consisting of platelet-poor plasma (PPP) and at the bottom of the platelet button. Part of the PPP was discarded so that only 1ml remained in the tube along with the platelet button. This material was gently agitated to promote platelets resuspension and activated when added 0.3ml of calcium gluconate, resulting in PRP gel. Double centrifugation protocol was able to make platelet concentration 3 times higher in relation to the initial blood sample. The volume of calcium gluconate used for platelet activation was 0.3ml, and was sufficient to coagulate the sample. Coagulation time ranged from 8 to 20 minutes, with an average of 17.6 minutes. Therefore, time of blood centrifugation until to obtain PRP gel took only 40 minutes. It was concluded that PRP was successfully obtained by double centrifugation protocol, which is able to increase the platelet concentration in the sample compared with whole blood, allowing its use in surgical procedures. Furthermore, the preparation time is appropriate to obtain PRP in just 40 minutes, and calcium gluconate is able to promote the activation of platelets.