Substance P (free acid) adopts different conformation than native peptide in DMSO, water and DPPC bilayers.

The conformation of substance P (free acid) (SPOH) has been investigated in dimethylsulfoxide (DMSO), water and dipalmitoylphosphotidylcholine (DPPC) bilayers by two-dimensional NMR and restraint molecular dynamics simulations. The observed NOE patterns for SPOH in these media are very much different from each other. Molecular modeling of the conformation of SPOH by incorporating NOEs as distance restraints shows wide differences in its conformation in three media. The main structural features for SPOH in DMSO are y-bends at Pro4 and Phe7 along with a non-specific bend around Lys3-Pro4-Gln5-Gln6, which are stabilized by Lys3CO-->Gln5NH, Gln6CO-->Phe8NH hydrogen bonding. The more flexible conformation of SPOH in water is transformed to an ordered structure after incorporation in DPPC bilayers. The conformation of SPOH in DPPC bilayers is characterized by gamma-bends at Pro4, Gln6 and Phe7, which are stabilized by hydrogen bonding between Lys3CO-->Gln5NH, Gln5CO-->Phe7NH and Gln6CO-->Phe8NH, respectively. The absence of biological activity in SPOH has been attributed to the absence of any helix like structure at the central residues and absence of any interresidue interaction with C-terminal OH group, in DPPC bilayers, a feature shown to be an important prerequisite for SP and SP agonists to bind to the NKI tachykinin receptor.

Hypertrophic cardiomyopathy: an autopsy analysis of 14 cases.

BACKGROUND: Hypertrophic cardiomyopathy (HCM) is one of the less common forms of primary cardiomyopathies. There is little data available on HCM in Indian literature. AIMS: To assess the incidence and analyse the clinicopathological features of HCM. SETTINGS: Analysis of data of 15 years from a tertiary care centre. METHODS AND MATERIAL: The clinical and pathological data in fourteen cases of HCM with respect to their gross and microscopic features and clinical presentation were reviewed. RESULTS: Incidence of HCM amongst the autopsied primary cardiomyopathies (N = 101) was 13.9% (n=14). Males were affected more. Common presenting symptoms were exertional dyspnoea, angina and palpitations. Concentric and asymmetric hypertrophy was equally seen. Obliterative small vessel disease was noted in 50% of the cases. Although significant myofibre disarray (>5%) was seen in all fourteen cases, it could be demonstrated in only 40- 50% of an average of twenty sections studied. Type IA myofibre disarray was the commonest. Six of the fourteen patients died suddenly. Cardiac failure was the commonest cause of death. CONCLUSIONS: Myofibre disarray is a highly sensitive and specific marker for HCM only when considered in a quantitative rather than a qualitative fashion. In this context, the rationale for performing endomyocardial biopsy is to rule out mimics of HCM.

Studies on Some New Ru(III) Complexes Using aryl-azo Pentane- 2,4-dione and 2,6-bis (2'-Benzimidazolyl) Pyridine as Ligands: Synthesis, Spectroscopic, Luminescent, Electrochemical and Biological Activities.

Some ruthenium(III) complexes with aryl-azo 2,4-pentanedione as co-ligands (L(1)H - L(3)H(2)) have been synthesized and characterized spectroscopically IR, (1)H NMR, UV/Vis, ESR, conductimetric) along with elemental analysis and FAB-mass data. Their luminescent and redox properties have been studied. The antibacterial, anti-HIV and antitmnour activities have also been reported.

Biomolecular electronics in the twenty-first century.

A relentless decrease in the size of silicon-based microelectronics devices is posing problems. The most important among these are limitations imposed by quantum-size effects and instabilities introduced by the effects of thermal fluctuations. These inherent envisaged problems of present-day systems have prompted scientists to look for alternative options. Advancement in the understanding of natural systems such as photosynthetic apparatuses and genetic engineering has enabled attention to be focused on the use of biomolecules. Biomolecules have the advantages of functionality and specificity. The invention of scanning tunneling microscopy and atomic force microscopy has opened up the possibility of addressing and manipulating individual atoms and molecules. Realization of the power of self-assembly principles has opened a novel approach for designing and assembling molecular structures with desired intricate architecture. The utility of molecules such as DNA as a three-dimensional, high-density memory element and its capability for molecular computing have been fully recognized but not yet realized. More time and effort are necessary before devices that can transcend existing ones will become easily available. An overview of the current trends that are envisaged to give rich dividends in the next millennium are discussed.

A new membrane probing steroidal spin label: synthesis and applications.

The applicability of a new steroidal spin label, 3-oxo-androstan-17 beta-yl-(2",2",6",6"-tetramethyl-N-oxyl) piperidyl butan-1',4'-dioate, in studying the phase transition properties of model membrane L-alpha-dipalmitoyl phosphatidyl choline (DPPC) in the presence and absence of drugs has been explored. Its synthesis and characterization has been described herein. Besides, the localization of this spin label in lipid liposomes has been studied using electron spin resonance (ESR), differential scanning calorimetry (DSC) and 1H and 31P NMR spectroscopic techniques. The label has also been used to study the permeability of epinephrine into membrane. The results show that the spin label has a good potential as a spin probe in the study of biomembranes.

A novel steroidal spin label for membrane structure studies: synthesis and applications.

2,2,6,6-Tetramethyl piperidine-N-oxyl nitroxyls are known to partition between aqueous and lipid phases, thus serving as probes to study membrane dynamics. The synthesis of a novel steroidal spin label, 3alpha-hydroxycholan-24-yl-(2",2",6",6"-tetramethyl-N-oxyl)p iperidyl butan-1',4'-dioate, containing 2,2,6,6-tetramethylpiperidine-N-oxyl moiety covalently bonded to the side chain in 3,24-caprostan-diol has been described. The localization of this spin label in model biomembranes has been studied by using electron spin resonance, differential scanning calorimetry, and 1H and 31P NMR spectroscopic techniques. Its applicability in studying the phase transition properties of model membrane L-alpha-dipalmitoyl phosphatidyl choline in the presence and absence of drugs has been described by using electron spin resonance. The label has also been used to study the permeability of epinephrine into membrane. The results have shown the applicability of the spin label as a potential spin probe in the study of biomembranes.

Replacement of phe(8) in substance P by tyr (Tyr(8)-SP) alters the conformation of the peptide in DMSO, water, and lipid bilayers.

The conformation of [Tyr(8)]SP (Y8SP) in dimethylsulfoxide (DMSO), water, and dipalmitoyl phosphatidylcholine (DPPC) bilayers has been investigated by two-dimensional nmr and molecular dynamics simulations. Molecular modeling of the conformation of Y8SP by incorporating nuclear Overhauser effects as distance restraints shows wide differences in its conformation in the three media. In DMSO, the main structural features are gamma-bends along with a nonspecific bend around Gln(6)-Phe(7)-Tyr(8). The random coil structure seen in water is transformed into a beta-turn around the segment Gln(5)-Gln(6)-Phe(7)-Tyr(8) when Y8SP is incorporated into DPPC bilayers. The lower biological activity of Y8SP compared to the native peptide (SP) has been attributed to the absence of any helix like structure at the central residues, a feature shown to be an important prerequisite for SP and SP agonists to bind to the neurokinin 1 tachykinin receptor.

Arginine acts as a protective and reversal agent against glycolytic inhibitors in spermatozoa.

It is known that the amino acid arginine stimulates sperm motility and glycolytic activity. We have earlier studied its efficacy as a stimulator of glycolysis in goat spermatozoa under anaerobic conditions. Here, we have assessed the influence of arginine in reversing the impairment caused by glycolytic inhibitors, iodoacetamide and iodoacetic acid. Glycolysis has been monitored by measuring the consumption of 13C labeled glucose and the amount of 13C labeled lactate produced under different experimental conditions, using 13C NMR. It is observed that both L- and D-arginine are able to prevent and reverse the inhibitory action of glycolytic inhibitors. The reversal effect of arginine gives rise to about eight times higher metabolic activity as compared to the inhibited cells while structurally related amino acids such as nitro-arginine, homo-arginine, lysine and ornithine are ineffective. The energetics of spermatozoa as measured by 31P NMR show a reduction in ATP level in cells incubated with iodoacetamide. Treatment of these cells with both L- and D-arginine restores the ATP level. The results may have significance in the treatment of male infertility.

Interaction of 7-hydroxy-8-(phenylazo)1,3-naphthalenedisulfonate with bovine plasma albumin. Spectroscopic studies.

Interaction of Orange G (OG) with bovine plasma albumin (BPA) has been investigated using NMR, UV-visible absorption, CD, and fluorescence techniques. The bound conformation of OG is a compact structure with N9-N10 bond in a non-planar syn conformation. The binding causes a decrease in the 478-nm absorption band of OG. The analysis of the binding isotherm generated from UV-visible absorption measurements gives a dissociation constant of 10 microM and stoichiometry 1:1 for BPA.OG complex. Dissociation constant is invariant in the pH range 5.0-8.0 and is approximately 20 times higher at pH 4.0 than its value at pH 7.0. Near and far UV-CD studies indicate alterations in the helical content and in the tertiary structure of the protein on complexation. The binding induces (-) and (+) CD at 335 nm and 465 nm, respectively. The binding also results into an increase in the steady state fluorescence anisotropy of OG without affecting emission maximum and quantum yield. Fluorescence data indicate that quenching of Trp fluorescence by OG is static in nature and OG selectively binds near Trp-135. Observation of similar rotational correlation time for BPA and BPA.OG complex indicates that the overall globular structure of BPA remains unaltered on binding despite certain internal rearrangement in the protein structure.

Identification of low-molecular-weight compounds in goat epididymis using multinuclear nuclear magnetic resonance.

Multinuclear nuclear magnetic resonance (NMR) (1H, 13C, and 31P) studies have been performed on aqueous solutions of lyophilysates of cell-free extract, epididymal fluid, and intact cells from caput and cauda regions of epididymis of sacrificed goats. Identification of low-molecular-weight compounds present in different maturation phases of spermatozoa has been carried out. Several low-molecular-weight compounds have been identified by assigning 600 MHz 1H NMR spectra with the help of two-dimensional homonuclear and heteronuclear correlation spectroscopy such as double quantum filtered correlation spectroscopy and heteronuclear single quantum correlation spectroscopy. Homonuclear coupling constants have also been used to get unambiguous assignments of resonances. NMR data were compared with those of standard samples measured at same pH and with those reported in the literature. Identification of several amino acids, carbohydrates, and lipids have been made and their presence has been discussed in relation to their relevance to sperm functions. The presence of beta-alanine and hypotaurine has been reported for the first time in goat epididymis.

1-[2-Hydroxy-3-octadecan-1'-oate]propyl-2'',2'',5'',5''-tetramethyl pyrolidine-N-oxyl-3''-carboxylate as a potential spin probe for membrane structure studies.

The synthesis of a new minimum steric perturbing proxyl nitroxide, which is a derivative of glycerol and contains a stearic acid moiety, has been carried out. Its localization in model membrane L-alpha-dipalmitoyl phosphatidyl choline (DPPC) was ascertained with the help of ESR, DSC, 1H and 31P NMR techniques. The nitroxide was used for detecting the changes in the phase transition temperature of the model membranes in the presence and absence of drugs. The permeation of the vasodilating drug epinephrine has also been studied using this spin label. The results prove the potential applicability of the new spin probe in the spin labeling of biomembranes.

Arginine activates glycolysis of goat epididymal spermatozoa: an NMR study.

The present study explores the mechanism underlying the action of L-arginine on the metabolic activity of spermatozoa. Goat epididymal spermatozoa were incubated with different concentrations of L-arginine to determine its effect on the utilization of glucose, fructose, and pyruvate. NMR techniques have been applied to elucidate the effect of L-arginine, L-lysine, and L-ornithine on the glycolysis of epididymal goat spermatozoa. Whereas 31P NMR has been used to estimate the change of pH in the presence of different concentrations of L-arginine, 13C NMR has been used to estimate the substrate consumption and lactate production. At optimal concentration of L-arginine, the forward metabolic rates have been found to increase by two to three times over control experiments. Arginine is not consumed in these reactions, but acts as an activator. Longitudinal relaxation time (T1) measurements indicate that the guanidino group of L-arginine plays an active role in binding to cells. The amino acid L-lysine is less effective, and L-ornithine is ineffective.

Iron coordination by catechol derivative antioxidants.

Iron complexes of nitrocatechols with different substituent groups [1: -CH = CR2; 2: -CH2-CHR2; 3: -CH = CR'(R")] were synthesized and their effects on iron-induced free radical reactions of biological importance investigated. Catechol and nitrocatechol derivatives effectively inhibited iron-induced lipid peroxide-dependent lipid peroxidation. In the Fenton-like reaction, iron-catechol generated hydroxyl radicals more strongly than did iron citrate, and iron-nitrocatechol derivative 2 generated a small amount of hydroxyl radicals. The iron complexes of derivatives 1 and 3 did not generate hydroxyl radicals. Iron-catechol had the highest ratio of reduction to oxidation rate constants and the second was iron-nitrocatechol 2, suggesting that iron chelated by nitrocatechols 1 and 3 may be most difficult to reduce. To elucidate the structure and physical properties of the iron complexes, UV/vis absorption spectroscopic, ESR and 1H NMR studies were performed in aqueous and DMSO solutions. In aqueous solution at pH 7.4, iron complexes of the nitrocatechol derivatives were high-spin tris(nitrocatecholato)ferrate(III) with a characteristic ligand-to-metal charge transfer absorbance (pi -> d pi). The lambda max of iron-nitrocatechol derivative 2 was shorter than those of iron-nitrocatechol derivatives 1 and 3, suggesting that the reduction potential of iron-nitrocatechol 2 is higher than that of iron-nitrocatechols 1 and 3. Nitrocatechol derivatives with a conjugation structure can sequester the chelated iron more effectively than catechol and the derivative without the conjugation against free radical generation by keeping the iron in the ferric state, probably because of the reduction potentials.

Membrane fluidization by animycotic bifonazole.

Calorimetry, nuclear magnetic resonance and X-ray diffraction techniques have been used to obtain thermodynamic and structural information on dipalmitoyl phosphatidylcholine (DPPC) liposomes doped by the antimycotic drug bifonazole in the range 0 < R < 1, where R = moles of bifonazole/moles of DPPC. The technique of spin labeling electron spin resonance (ESR) has also been used to study permeability and fluidity properties. The decrease of the cooperativity at the gel to liquid crystalline phase transition, as shown by ESR an DSC measurements, indicates that bifonazole imparts higher fluidity to the lipid matrix. Increase in permeability of ascorbate ions, after incorporation of bifonazole in the membrane, has been detected by ESR experiments using spin label 5-SASL. 13C NMR spectra indicate that the drug molecule is highly immobilized. X-ray diffraction and freeze fracture TEM results show that the equilibrated phase at room temperature is lamellar and unidimensional together with the presence of small particles and pits of uniform size. A marked hysteresis is evident in the formation of this phase.

Riboflavin: a potential material for molecular electronics applications.

Riboflavin (Rbf), the simplest manifestation of the isoalloxazine ring, undergoes redox reactions during biochemical processes. The redox reaction can be simulated under electrochemical conditions. The two oxidation states of the isoalloxazine molecule have different optical properties. Rbf embedded in a polyurethane matrix has been shown to undergo colour change on application of an external electric field. Rbf has been modified to form TARbf and TPRbf--the amphiphilic analogues for the convenience of forming ordered two-dimensional organization. The modification has been found to leave native characteristics of Rbf undisturbed. TPRbf forms monolayers at the air-water interface with a collapse pressure of 10 mNm-1.

Immobilization of enzymes/coenzymes for molecular electronics applications.

A brief survey of immobilization strategies and methods has been presented. Relative merits and demerits are discussed with reference to their end use.

2'-(3 alpha-Benzyloxy-24-norcholan-23-yl)-2',4',4'-trimethyl-4',5'- dihydrooxazoline-N-oxyl as a potential spin probe for model membranes.

A new steroidal doxyl (4,4-dimethyloxazolidine-N-oxyl) nitroxide (SDN) viz. 2'-(3 alpha-benzyloxy-24-norcholan-2'-yl)-2',4',4'-trimethyl-4',5'- dihydrooxazoline-N-oxyl has been synthesized. This is expected to have higher mobility over other spin labels reported earlier. The localization of this spin probe in lipid bilayers has been determined using 1H NMR and 31P NMR techniques. The alterations induced by drugs in the membrane characteristics such as phase transition and permeability have been investigated using electron paramagnetic resonance (EPR) techniques. The results show the applicability of SDN as a potential spin probe in the study of biomembranes.

Proxyl nitroxide of lithocholic acid: a potential spin probe for model membranes.

A new steroidal proxyl (2,2,5,5-tetramethylpyrrolidine-N-oxyl) nitroxide (SPN), with the proxyl nitroxide moiety in the pendant side chain of the steroid, has been synthesized. Its localization in lipid bilayers was ascertained with the help of 1H NMR and 31P NMR experiments. The effects of the nitroxide group in SPN incorporated into the bilayer on 13C relaxation times are interpreted qualitatively in terms of localization of the nitroxide group within the bilayer structure. The nitroxide SPN was used to monitor changes in membrane fluidity and permeability induced by local anaesthetics, mepivacaine and xylocaine and the antikeratinizing agent, azelaic acid. The results conclusively proved the applicability of the new steroidal proxyl nitroxide (SPN) as a potential spin probe for spin labeling studies.

Maturation, capacitation, and metabolism of goat spermatozoa using magnetic resonance methods.

The ability of intact cells to reduce spin labels has been utilized to characterize the activity of spermatozoa of goat. The kinetics of reduction of TEMPO has been found to be sensitive to the quantity, quality and state of epididymal maturation of the spermatozoa. Presence of alcohol caused activation and Gossypol acetic acid left sperm activity unaltered. Electron spin resonance (ESR) and 31P Nuclear magnetic resonance (NMR) indicate that the period of in vitro capacitation requires optimization. 31P NMR spectra indicate a good correlation with the progressive maturation of the cells.

Maturation of rat spermatozoa: ESR spin labeling studies.

The ability of spermatozoa to reduce nitroxide spin--TEMPO has been used as a parameter to understand maturation, capacitation and calcium uptake of sperm obtained from Holstman strain rats. The rate of spin label reduction by sperm follows the trend--caput greater than cauda greater than corpus. With the increase in age, the electron donating capability shows first a gradual increase, for 60- to 85-day-old rats, peaking at 85 days (corresponding to puberty) and leveling off after 92 days. Calcium uptake takes place in two phases which corresponds to accumulation of and activation by calcium. The presence of polyclonal antibody which is known to cause agglutination, does not adversely affect the sperm activity.