The effects of conformation and intermolecular hydrogen bonding on the structure and IR spectra of flutamide; a study based on the matrix isolation technique, ab initio and DFT calculations.

In this study, stable conformers of flutamide referred to as an anticancer drug were searched through a relaxed potential energy surface scan carried out at the B3LYP/6-31G(d) level of theory. This was followed by geometry optimization and thermochemistry calculations performed with the HF-SCF, MP2, B3LYP methods and the 6-31G(d), 6-311++G(d,p), aug-cc-pvTZ basis sets for each of the determined minimum energy conformers. The results revealed that flutamide has at least five stable conformers and two of them provide the major contribution to the observed matrix isolation infrared (IR) spectra of the molecule. The effects of conformational variety and intermolecular hydrogen bonding interactions on the observed IR spectra of flutamide were interpreted in the light of the vibrational spectral data obtained for the most stable monomer and dimer forms of the molecule at the same levels of theory. Pulay's "Scaled Quantum Mechanical-Force Field (SQM-FF)" method was used in the refinement of the calculated harmonic wavenumbers, IR intensities and potential energy distributions. This scaling method which proved its superiority to both anharmonic frequency calculations and other scaling methods helped us to correctly interpret the remarkable differences between the matrix IR spectra of flutamide in argon and the condensed phase IR spectra of the molecule in solvents such as KBr, H2O, D2O, ethanol and methanol.

An investigation on the structure and group vibrations of balenine molecule by matrix isolation IR spectroscopy, DFT and MP2 based calculations.

Stable conformers of neutral balenine were scanned through molecular dynamics simulations and energy minimizations using Allinger's MM2 force field. For each of the found minimum-energy conformers, geometry optimization and thermochemistry calculations were performed by using B3LYP, MP2, G3MP2B3 methods, 6-31G(d), 6-311++G(d,p) and aug-cc-pvTZ basis sets. The calculation results have indicated that balenine has about twenty stable conformers whose relative energies are in the range of 0-9.5 kcal/mol. Three of these are thought to provide the major contribution to matrix isolation IR spectra of the molecule. Our solvent calculations using the polarized continuum model revealed the stable zwitterion structures which are predicted to dominate IR spectra of balenine in water and heavy water (D2O) solvents. Pulay's SQM-FF method was used in scaling of the harmonic force constants and vibrational spectral data calculated for the neutral and zwitterion structures. These refined calculation data together with those obtained from anharmonic frequency calculations enabled us to correctly interpret the matrix isolation IR spectrum of balenine and the tautomerism-based changes observed in its KBr IR and solution (D2O) IR spectra. The results revealed the crucial role of conformation and zwitterionic tautomerism on the structure and vibrational spectral data of the molecule.

Identifying predictors of ventral hernia recurrence: systematic review and meta-analysis.

BACKGROUND: Ventra hernias are increasing in prevalence and many recur despite attempted repair. To date, much of the literature is underpowered and divergent. As a result there is limited high quality evidence to inform surgeons succinctly which perioperative variables influence postoperative recurrence. This systematic review aimed to identify predictors of ventral hernia recurrence. METHODS: PubMed was searched for studies reporting prognostic data of ventral hernia recurrence between 1 January 1995 and 1 January 2018. Extracted data described hernia type (primary/incisional), definitions of recurrence, methods used to detect recurrence, duration of follow-up, and co-morbidity. Data were extracted for all potential predictors, estimates and thresholds described. Random-effects meta-analysis was used. Bias was assessed with a modified PROBAST (Prediction model Risk Of Bias ASsessment Tool). RESULTS: Screening of 18 214 abstracts yielded 274 individual studies for inclusion. Hernia recurrence was defined in 66 studies (24.1 per cent), using 41 different unstandardized definitions. Three patient variables (female sex, age 65 years or less, and BMI greater than 25, 30, 35 or 40 kg/m2), five patient co-morbidities (smoking, diabetes, chronic obstructive pulmonary disease, ASA grade III-IV, steroid use), two hernia-related variables (incisional/primary, recurrent/primary), six intraoperative variables (biological mesh, bridged repair, open versus laparoscopic surgery, suture versus mesh repair, onlay/retrorectus, intraperitoneal/retrorectus), and six postoperative variables (any complication, surgical-site occurrence, wound infection, seroma, haematoma, wound dehiscence) were identified as significant prognostic factors for hernia recurrence. CONCLUSION: This study summarized the current evidence base for predicting ventral hernia recurrence. Results should inform best practice and future research.

A vibrational spectroscopy study on anserine and its aqueous solutions.

In this study based on vibrational spectroscopic measurements and Density Functional Theory (DFT), we aimed for a reliable interpretation of the IR and Raman spectra recorded for anserine in the solid phase and water (H2O) and heavy water (D2O) solutions. Initial DFT calculations at the B3LYP/6-31G(d) searched possible conformers of the anserine zwitterion using a systematic conformational search. The corresponding equilibrium geometrical parameters and vibrational spectral data were determined for each of the stable conformers (in water) by the geometry optimization and hessian calculations performed at the same level of theory using the polarized continuum model (PCM). The same calculations were repeated to determine the most energetically preferred dimer structure for the molecule and the associated geometry, force field and vibrational spectral data. The harmonic force constants obtained from these calculations were scaled by the Scaled Quantum Mechanical Force Field (SQM) method and then used in the calculation of the refined wavenumbers, potential energy distributions, IR and Raman intensities. These refined theoretical data, which confirm the zwitterion structure for anserine in the solid phase or aqueous solvents, revealed the remarkable effects of intermolecular hydrogen bonding on the structural properties and observed IR and Raman spectra of this molecule.

Scaled Quantum Mechanical scale factors for vibrational calculations using alternate polarized and augmented basis sets with the B3LYP density functional calculation model.

The Scaled Quantum Mechanical (SQM) method of scaling calculated force constants to predict theoretically calculated vibrational frequencies is expanded to include a broad array of polarized and augmented basis sets based on the split valence 6-31G and 6-311G basis sets with the B3LYP density functional. Pulay's original choice of a single polarized 6-31G(d) basis coupled with a B3LYP functional remains the most computationally economical choice for scaled frequency calculations. But it can be improved upon with additional polarization functions and added diffuse functions for complex molecular systems. The new scale factors for the B3LYP density functional and the 6-31G, 6-31G(d), 6-31G(d,p), 6-31G+(d,p), 6-31G++(d,p), 6-311G, 6-311G(d), 6-311G(d,p), 6-311G+(d,p), 6-311G++(d,p), 6-311G(2d,p), 6-311G++(2d,p), 6-311G++(df,p) basis sets are shown. The double d polarized models did not perform as well and the source of the decreased accuracy was investigated. An alternate system of generating internal coordinates that uses the out-of plane wagging coordinate whenever it is possible; makes vibrational assignments via potential energy distributions more meaningful. Automated software to produce SQM scaled vibrational calculations from different molecular orbital packages is presented.

Convenient and rapid detection of pesticides in extracts of sheep wool.

A relatively simple, rapid extraction technique based on acetonitrile was combined with the use of screen-printed electrodes bearing cholinesterases to detect organo-phosphate pesticides from an otherwise intractable matrix, sheep wool. It proved possible to separate, for convenience, the exposure of the electrodes from measurement of their (inhibited) activity. The electrodes were used once and then discarded. Estimation of the extent of inhibition is dependent on reference to the activity of control electrodes. The presence of pesticides in the extracts could be detected with any of three commonly available cholinesterases but the most sensitive enzyme was butyryl cholinesterase from horse serum.

Musical scales and evaluations of happiness and awkwardness: effects of pitch, direction, and scale mode.

Participants rated the perceived happiness, brightness, awkwardness, pitch velocity, and tempo change of ascending and descending musical scales in four modes (natural, melodic, and harmonic minor modes and the major mode). Only minor differences between ratings of natural, harmonic, or melodic minor scales or between ratings of parallel and relative major scales were found. Ascending scales were rated as happier, brighter, and more accelerating than were descending scales; ascending minor scales were rated as faster and more awkward than were descending minor scales. Musical keys in each mode were compared, and significant differences were found. Musical keys that started on a higher pitch were rated as happier, brighter, and faster and as speeding up more than were keys that started on a lower pitch. The data were consistent with previous findings and suggest that pitch and direction (contour), rather than mode or key, influence listeners' judgments of musical stimuli.

Evidence that two phenotypically distinct mouse PKD mutations, bpk and jcpk, are allelic.

Numerous mouse models of polycystic kidney disease (PKD) have been described. All of these diseases are transmitted as single recessive traits and in most, the phenotypic severity is influenced by the genetic background. However, based on their genetic map positions, none of these loci appears to be allelic and none are candidate modifier loci for any other mouse PKD mutation. Previously, we have described the mouse bpk mutation, a model that closely resembles human autosomal recessive polycystic kidney disease. We now report that the bpk mutation maps to a 1.6 CM interval on mouse Chromosome 10, and that the renal cystic disease severity in our intersubspecific intercross progeny is influenced by the genetic background. Interestingly, bpk co-localizes with jcpk, a phenotypically-distinct PKD mutation, and complementation testing indicates that the bpk and jcpk mutations are allelic. These data imply that distinct PKD phenotypes can result from different mutations within a single gene. In addition, based on its map position, the bpk locus is a candidate genetic modifier for jck, a third phenotypically-distinct PKD mutation.

Measurement of soluble L-lactate in dairy products using screen-printed sensors in batch mode.

Amperometric sensors based on lactate oxidase and platinized carbon were constructed entirely by screen-printing. They were used to estimate the lactate concentration in diluted samples of yoghurt and buttermilk. Estimates made in untreated (apart from dilution) solutions were subject to bias, relative to values obtained from spectrophotometric assays. Removal of cations improved the accuracy of the estimates. Calcium may have made a small contribution to the bias but the nature of the inhibition is otherwise unknown. With untreated samples, determinations of any accuracy require sufficient dilution to avoid such bias, although the smaller signals resulting from dilution place more demands on calibration and manufacturing standards. The relative standard deviations of estimates, based on variation among sensors, of the most dilute samples were generally too high for practical use. The errors of estimates at lower dilutions approached those of screen-printed sensors made under industrial conditions.

Narrow complex tachycardias.

Narrow complex tachycardias are those cardiac rhythms with a ventricular rate of more than 100 beats per minute and a QRS complex width of less than 0.12 seconds. They originate either from the SA node, from atrial tissue itself, or from in or around the AV node. The term SVT is generally accurate for such tachycardias. By diagnosing the source of an SVT and appreciating its likely cause, therapy can be more precisely, safely, and effectively guided to treat these patients. Atrial and junctional rhythms can be treated with vagal maneuvers, drugs from classes I to IV and other antiarrythmic agents, magnesium, and cardioversion. Some patients may be candidates for surgical or catheter ablation.

Pharmacological characterization and autoradiographic localization of dopamine receptors in the portal vein.

1. Dopamine (DA) and DA receptor agonists exert a variety of effects on the cardiovascular system through interaction with specific DA receptors, including decreases in blood pressure and heart rate. 2. The decrease in blood pressure is due primarily to arterial vasodilation. This phenomenon is due to the stimulation of both postjunctional (D1-like or DA1) and prejunctional (D2-like or DA2) receptors causing respectively relaxation of arterial smooth muscle and decrease of the sympathetic vasoconstriction tone. 3. In view of the lack of detailed information on the existence of DA receptors in venous tissue, we have analysed D1-like and D2-like receptors in the rat portal vein using radioligand binding techniques associated with light microscope autoradiography. 4. No D1-like receptors were demonstrated in sections of the rat portal vein, whereas the D2-like receptor ligand, [3H]-spiroperidol, was bound to sections of the vein in a manner consistent with the labelling of D2-like sites. Anatomically, D2-like sites were located within the tunica adventitia, including the adventitia-media border, and in the endothelium. 5. These findings suggest the existence of D2-like but not D1-like receptor sites in the rat portal vein. D2-like sites of the tunica adventitia are probably prejunctional and involved in the modulation of sympathetic outflow. The functional significance of endothelial D2-like sites, if any, should be clarified in future studies.

Immunogold ultrastructural localization of calpastatin, the calpain inhibitor, in rabbit skeletal muscle.

The ultrastructural localization of calpastatin, the endogenous inhibitor of the neutral calcium-dependent proteases (calpains), was investigated in rabbit skeletal muscle fibers using a polyclonal antibody against the 34 kDa form of the inhibitor isolated from rabbit. Quantitative studies by pre- and postembedding immunogold techniques revealed that the distribution pattern of the specific immunoreactivity included: 1) the sarcolemma with the adjacent cytoplasm (about 1 micron wide); 2) the myofibrils; 3) the mitochondria and 4) the nuclei (condensed as well as extended chromatin). Other cell substructures, such as lysosomes and the intermyofibrillar cytoplasm, were substantially devoid of immunoreactivity. Furthermore, in accordance to previous light microscope immunohistochemical experiments, an extracellular (endomysial) localization of specific immunoreactivity was confirmed. These results favour the view, which is also supported by a series of biochemical evidences, that calpastatin in rabbit skeletal muscle is present in cell structures also containing calpains and/or their putative substrates. The above multiple patterns of distribution also suggest that the muscular calpain-calpastatin system in skeletal muscle fibers may play different physiological roles in the various subcellular compartments.

Effect of denervation on lamellar cells of Meissner-like sensory corpuscles of the rat. An immunohistochemical study.

The denervation-induced changes on S-100 protein, glial fibrillary acidic protein (GFAP) and vimentin immunoreactivity (IR) of the lamellar cells from cutaneous Meissner-like sensory nerve formations (SNF), or corpuscles, of the adult rat hind limb foot-pads were studied, using combined immunohistochemical and image analysis (optic microdensitometry) techniques. Animals were allowed to survive for 1, 3 and 7 days following sciatic and saphenous nerves transection. Lamellar cells of Meissner-like corpuscles displayed S-100 protein- and vimentin-IR, but not GFAP-IR. Denervation caused a marked time-dependent decrease of S-100 protein IR whereas vimentin-IR did not change or weakly increased. No positive GFAP-IR was observed in denervated SNF. These findings suggest that continuity of SNF with nerve fibers supplying them is necessary to maintain some of the immunohistochemical characteristics of the non-neuronal cells of SNF.

Localization of 5-hydroxytryptamine-like immunoreactive cells and nerve fibers in the rat female reproductive system.

The presence of 5-hydroxytryptamine (5-HT)-like immunoreactivity (IR) was studied in the rat female reproductive system using polyclonal antibodies directed against 5-HT. Moreover, 5-HT levels in the ovary, oviduct, uterus, and cervix were measured by high-pressure liquid chromatography with electrochemical detection. The highest 5-HT concentrations were found in the oviduct, followed in descending order by the cervix, the ovary, and the uterus. Most 5-HT-like IR was observed in the cytoplasm of mast cells. These cells were found in the connective tissue around the fimbria, in the oviduct, in the uterus, and in the ovary. Mast cells are clustered in the proximity of the parenchymal blood vessels. Moreover, a few 5-HT-like nerve fibers were found distributed mainly perivascularily in the uterine cervix and in the uterine horns as well as in the oviduct. IR nerve fibers were rarely seen within the ovary. The present data provide direct evidence that 5-HT in the female reproductive system not only is associated with mast cells but is located in nerve fibre-like structures as well. The functional significance of this probable 5-HT-ergic innervation of the female reproductive tract discovered in the present study should be clarified in future investigations.

Changes in glutathione content and localization in rat heart as a function of age.

The influence of aging on glutathione levels and distribution in the heart was studied in male Sprague-Dawley rats of 3 (young), 12 (adult) and 24 (old) months of age using biochemical and histofluorescence techniques, respectively. Biochemical assays of reduced glutathione (GSH) in the right and left ventricles and in the septum showed a significant decrease in GSH levels in adult in comparison with young animals. No further changes were noticeable between adult and old rats. GSH histofluorescence revealed a rather homogeneous distribution of the product of histochemical reaction within both right and left atria in 3-month-old rats. In 12-month-old rats a reduction of GSH histofluorescence in comparison with younger animals was noticeable. The loss is more consistent in the epicardial portion of the right atrium and in the endocardial region of the left atrium. In the atria of 24-month-old rats GSH reactivity was homogeneously distributed throughout the atrial wall and was significantly lower than in young or adult rats. In 3-month-old rats GSH histofluorescence was slightly lower in the epicardial than in the endocardial portions of both ventricles. In adult rats a significant decrease of GSH histofluorescence was noticeable in comparison with 3-month-old rats. The loss is particularly pronounced within the endocardial region of the left ventricle. In 24-month-old rats GSH histofluorescence showed no significant differences between adult rats. However, GSH was more homogeneously distributed throughout the ventricular wall than in adult animals. The significance of these data is discussed in relation to the role that GSH plays in protecting the myocytes against free radical damage.

Endothelial dopamine DA-1 receptor sites in the rabbit pulmonary artery: autoradiographic demonstration.

Combined in vivo radioreceptor binding and autoradiographic techniques were used to characterized the pharmacological profile and to study the anatomical localization of dopamine (DA) DA-1 receptor sites in sections of rabbit pulmonary artery. [3H]R-(+)-8-chloro-2,3,4,5-tetrahydro-5-phenyl-1H-3-benzazepin-7- alhemimaleate (SCH 23390), which was used as a ligand, was bound by sections of rabbit pulmonary artery in a manner consistent with the binding of DA DA-1 sites. The Kd value was 4.75 nM, whereas the Bmax value was 78.3 +/- 5.7 fmol/mg tissue. Light microscope autoradiography demonstrated specific [3H]SCH 23390 binding sites primarily in the endothelium of the rabbit pulmonary artery. Moreover, sparse receptor sites were visualized in the medial layer. Mechanical removal of endothelium caused the disappearance of [3H]SCH 23390 binding sites showing the endothelial localization, but was without effect on the receptor sites of the medial layer. The present findings suggest that differently from systemic arteries, where DA-1 receptor sites are localized in the medial layer, probably within smooth muscle, the majority of DA-1 sites in the rabbit pulmonary artery are endothelial. The possible significance of these sites visualized in the present study for the first time is discussed.

Calpain inhibitor in rabbit skeletal muscle: an immunochemical and histochemical study.

Calpastatin, the endogenous inhibitor of calcium-activated neutral proteases (calpains; EC 3.4.22.17), was studied in the rabbit vastus lateralis muscle by means of immunochemical and immunohistochemical techniques. Immunoaffinity chromatography using an antibody raised against the 34-kDa monomer of the 68-kDa dimeric inhibitor allowed us to isolate three main proteins (130-, 100- and 80-kDa). These proteins strongly inhibited calpain activity in muscle homogenate (I50 at about 50 micrograms/ml). Immunohistochemical experiments showed that calpastatin-related immunoreactivity was present in all fibre types (oxidative, glycolytic, oxidative-glycolytic) at both surface and cytoplasmic level. However, a few (20%) of the slow-twitch, oxidative fibres (5% of the total fibres), did not contain the cytoplasmic inhibitor. Specific immunoreactivity for calpastatin was also associated with the interstitial connective tissue. These results suggest that (i) calpastatin in skeletal muscle, as in other tissues, is present as a mixture of proteins of various molecular weights and (ii) the inhibitor may act not only in the cytoplasm but also at the surface or extracellular level.

Autoradiographic localization of the gamma-aminobutyric acid type A receptor agonist 3H-muscimol in the rat superior cervical ganglion.

The anatomical localization of gamma-aminobutyric acid type A (GABA-A) receptor sites in the rat superior cervical ganglion was studied using combined radioreceptor binding and autoradiographic techniques. 3H-Muscimol was used as a ligand of GABA-A receptor sites. The binding was consistent with the labelling of GABA-A sites. The dissociation constant value was 6.4 nmol/l, and the maximum density of binding sites was 146 +/- 7.8 fmol/mg tissue. Light microscope autoradiography revealed the accumulation of 3H-muscimol mainly in superior portions of the ganglion. Binding sites are located primarily in the neuropil rather than within ganglionic neurons. It is probable that the sites revealed by autoradiography are involved in the inhibition of acetylcholine release from ganglionic neurons.

Neurotransmitters, neuroreceptors and aging.

At first glance, it is satisfying to see the progress which has been made in the study of neurotransmitters. We have learned a great deal in the last number of years. First, we have been able to identify previously unknown compounds which affect the nervous system or associated peripheral organs. We now know a great deal about the metabolism of these molecules including their synthesis and catabolism. We have learned to identify and to classify their receptors. We have learned that alterations in the effects of neurotransmitters may be responsible for certain pathologies or may be a function of normal aging. Yet, we still have far to go in our research. There are neurotransmitters still to be discovered. We need to continue our efforts because there is still a large amount of confusion in the literature, for example, far too many contradictory reports concerning the effects of age confuse rather than clarify. Possibly order may return to the literature if investigators can agree on some basic tenets. For example, we need a basic definition of old. Some research groups consider 12-month-old rats as old while other groups consider them to be young individuals. We need to have standardization of methodology so that the conclusions can have validity. Once again certain investigators use whole brain homogenates while others use only discrete portions. We need to consider whether the effect we see in our experiment is primary or secondary to aging. We can be certain that due to the aging population, the importance of basic research of age-dependent changes in neurotransmitters and neuroreceptors will increase in the future.