Effect of verapamil on the non-adrenergic response of the field stimulated rat vas deferens

A comparison has been made, in the present study, between the effects of verapamil (reported to have smooth muscle depolarizing action) and K+ depolarization on the responses of noradrenaline, ATP and those of field stimulation on the vas deferens obtained from reserpinized rats. Field stimulation of the vas using single pulse (1 ms pulse width; supramaximal voltage) resulted in a fast twitch response reaching a maximum at 300 +/- 20 ms. Verapamil (6 X 10(-6) M) significantly potentiated this response. Verapamil potentiated the twitch component of the biphasic response resulting from field stimulation of the intrinsic nerves with repetitive pulses, while the tonic component was markedly inhibited. Verapamil enhanced the ATP (7 X 10(-5) M) response, while the phasic and tonic components of KCl (5.36 X 10(-2) M)-induced biphasic responses were nearly abolished. While the phasic component of the noradrenaline (7 X 10(-6) M) response remained unaltered in the presence of verapamil, the tonic component was markedly inhibited and rhythmicity following phasic component was markedly enhanced. Partial depolarization, achieved by increasing K+ concentration in the normal Krebs by two-fold i.e., to 11.8 mM, enhanced the responses of ATP, noradrenaline and the twitch resulted from the single pulse stimulation. The finding that verapamil potentiates the contractile response to exogenously applied ATP, which is believed to be the "noradrenergic" neurotransmitter in the vas deferens, suggests that this is the mechanism through which verapamil potentiates the twitch responses to field stimulation of the nerve supply.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of K(+)-free solution of contractile responses of rat vas deferens

The effect of K(+)-free solution as well as its modulatory effect on neurogenic and agonist-induced contractile responses in rat vas deferens were investigated in the present study. Isolated rat vas deferens, normally a quiescent smooth muscle, developed spontaneous contractions on exposure to K(+)-free solution which was not affected by tetrodotoxin (3 x 10(-7) mol/l) or prazosin (10(-6) mol/l) but was abolished in Ca(2+)-free medium. Transmural field stimulus (3-30 Hz; 0.5 ms duration; supramaximal voltage; 10 s) induced biphasic responses, consisting of fast and slow components, were markedly inhibited in K(+)-free medium and were restored on readdition of K+ to the medium. Similarly, the twitch responses at 0.1 Hz were also inhibited in K(+)-free solution. In the absence of K+, the sensitivity of the tissues to noradrenaline (10(-6) to 10(-4) mol/l) and ATP (10(-4) mol/l) was not significantly altered. The observations made in the present study suggest that spontaneous contractions induced by K(+)-free solution are myogenic in origin and are dependent on extracellular Ca2+. Selective inhibition of neurogenic responses in K(+)-free medium without any significant effect on exogenous noradrenaline and ATP is suggestive of the involvement of prejunctional inhibition of neurotransmitter release.     

Two complex, adenovirus-based vaccines that together induce immune responses to all four dengue virus serotypes

Dengue virus infections can cause hemorrhagic fever, shock, encephalitis, and even death. Worldwide, approximately 2.5 billion people live in dengue-infested regions with about 100 million new cases each year, although many of these infections are believed to be silent. There are four antigenically distinct serotypes of dengue virus; thus, immunity from one serotype will not cross-protect from infection with the other three. The difficulties that hamper vaccine development include requirements of the natural conformation of the envelope glycoprotein to induce neutralizing immune responses and the necessity of presenting antigens of all four serotypes. Currently, the only way to meet these requirements is to use a mixture of four serotypes of live attenuated dengue viruses, but safety remains a major problem. In this study, we have developed the basis for a tetravalent dengue vaccine using a novel complex adenovirus platform that is capable of expressing multiple antigens de novo. This dengue vaccine is constructed as a pair of vectors that each expresses the premembrane and envelope genes of two different dengue virus serotypes. Upon vaccination, the vaccine expressed high levels of the dengue virus antigens in cells to mimic a natural infection and induced both humoral and cellular immune responses against multiple serotypes of dengue virus in an animal model. Further analyses show the humoral responses were indeed neutralizing against all four serotypes. Our studies demonstrate the concept of mimicking infections to induce immune responses by synthesizing dengue virus membrane antigens de novo and the feasibility of developing an effective tetravalent dengue vaccine by vector-mediated expression of glycoproteins of the four serotypes.     

Inhibition of contractions and 45Ca influx by adenosine in rabbit aorta.

The effects of adenosine on contractility and 45Ca-uptake in rabbit aorta have been examined and compared with those of nifedipine. Both adenosine (10(-5) M) and nifedipine (10(-8) M) inhibited contractions caused by CaCl2 (0.1-10 mM) in K(+)-depolarized preparations. Nifedipine (10(-7) M) significantly inhibited the contractions of noradrenaline (NA) at all doses (10(-8)-10(-4) M), while adenosine inhibited the contractions induced only by lower doses of NA (10(-8) and 10(-7) M) without affecting those at higher doses (greater than 10(-7) M). Adenosine (10(-8) M) and nifedipine (10(-7) M) inhibited K(+)-stimulated 45Ca-uptake by aortic strips by 75 and 100%, respectively. Similarity, NA (10(-4) M)-stimulated 45Ca influx was inhibited by about 70% by both these agents. The results suggest that adenosine inhibits inward Ca2+ movement involving both voltage-operated and receptor-operated calcium channels in rabbit aortic smooth muscle.     

Nasopalpebral lipoma coloboma syndrome

Nasopalpebral lipoma-coloboma syndrome is characterized by nasopalpebral lipoma and eyelid coloboma. We report a case of a 16-year-old Indian girl who reported to us with this rare syndrome. Computed tomography scan showed a significantly hypodense lesion on the right side of nose which was confirmed to be a lipoma on histopathological examination. This condition should be included in differential diagnosis of conditions with congenital eyelid coloboma.     

Integrated TK–TD modeling for drug-induced concurrent tachycardia and QT changes in beagle dogs

Drug-induced cardiotoxicity, including tachycardia and QT prolongation, remains a major safety concern that needs to be identified and its risk mitigated in early stages of drug development. In the present study, an integrated toxicokinetic–toxicodynamic (TK–TD) modeling approach within a nonlinear mixed-effect modeling framework is applied to investigate concurrent abnormal heart rate and QT changes in three beagle dogs, using a Novartis internal compound (NVS001) as the case example. By accounting for saturable drug absorption, circadian rhythms, drug-effect tolerance, and nonlinear rate-dependency of QT interval, the dynamic TK–TD model captures the experimentally observed drug effects on heart rate and QT interval across a wide dosing range of NVS001 in beagle dogs. Further analyses reveal that the NVS001-induced QT prolongation observed in the low-dose groups is potentially caused by direct drug inhibition on the hERG channel, while the apparent QT shortening in the high-dose groups may be due to strong rate-dependency of QT at high heart rates. This study also suggests that the TK–TD model can be used to identify direct drug effects on the non-rate-dependent QT component by dissociating QT changes from tachycardia and deriving a new QT correction method. The integrated TK–TD model presented here may serve as a novel quantitative framework for evaluating drug-induced concurrent changes in heart rate and QT to potentially facilitate preclinical and clinical safety studies.     

Tetravalent neutralizing antibody response against four dengue serotypes by a single chimeric dengue envelope antigen

We employed DNA shuffling and screening technologies to develop a single recombinant dengue envelope (E) antigen capable of inducing neutralizing antibodies against all four antigenically distinct dengue serotypes. By DNA shuffling of codon-optimized dengue 1-4 E genes, we created a panel of novel chimeric clones expressing C-terminal truncated E antigens that combined epitopes from all four dengue serotypes. DNA vaccines encoding these novel chimeras induced multivalent T cell and neutralizing antibody responses against all four dengue serotypes in mice. By contrast, a mixture of four unshuffled, parental DNA vaccines failed to produce tetravalent neutralizing antibodies in mice. The neutralizing antibody titers for some of these antigens could be further improved by extending the sequences to express full-length pre-membrane and envelope proteins. The chimeric antigens also protected mice against a lethal dengue-2 virus challenge. These data demonstrate that DNA shuffling and associated screening can lead to the selection of multi-epitope antigens against closely related dengue virus serotypes and suggest a broad utility for these technologies in optimizing vaccine antigens.     

Antinociceptive and antipyretic activities of Pongamia pinnata leaves

In the present study, the antinociceptive activity of a 70% ethanol extract of Pongamia pinnata leaves (PLE) was investigated in different models of pain in mice and rats. Further, PLE was also evaluated for its antipyretic activity in Brewer's yeast-induced pyrexia in rats. Per os (p.o.) administration of the PLE (100-1000 mg/kg) produced significant antinociceptive activity in the hotplate and tail flick (central) as well as in acetic acid writhing and Randall-Selitto (peripheral) nociceptive tests suggesting the involvement of both central and peripheral mechanisms in alleviating the pain response. In addition, PLE also exhibited a significant antipyretic response in Brewer's yeast-induced pyrexia in rats. These results demonstrated that PLE possesses marked antinociceptive as well as antipyretic activities and thus scientifically validated its use in the treatment of pain and pyretic disorders.