Spectrophotometric and atomic absorption spectrometric determination of ramipril and perindopril through ternary complex formation with eosin and Cu(II)

Two sensitive, spectrophotometric and atomic absorption spectrometric procedures are developed for the determination of ramipril and perindopril. Both methods are based on the formation of a ternary complex, extractable with chloroform, between copper(II), eosin and the two cited drugs. Spectrophotometrically under the optimum condition, the ternary complexes showed an absorption maximum at 535 nm, with apparent molar absorptivities of 6.55 and 4.00 x 10(3) mol(-1) x cm(-1) and Sandell's sensitivities of 5.80 x 10(-2) and 1.04 x 10(-1) microg x cm(-2) for perindopril and ramipril, respectively. The solution of ternary complex obeyed Beer's law in concentration ranges 10-60 and 20-100 microg x ml(-1) for perindopril and ramipril, respectively. The proposed method was applied to the determination of the two cited drugs in pharmaceutical tablets. The atomic absorption spectrometric method, directly through the quantitative determination of copper content of the organic extract of the complex, was also investigated for the purpose of enhancing the sensitivity of the determination. The spectrophotometric and atomic absorption spectrometric procedures hold their accuracy and precision well when applied to the determination of ramipril and perindopril dosage forms.     

Peripheral synapses at identified mechanosensory neurons in spiders: three-dimensional reconstruction and GABA immunocytochemistry

The mechanosensory organs of arachnids receive diverse peripheral inputs. Little is known about the origin, distribution, and function of these chemical synapses, which we examined in lyriform slit sense organ VS-3 of the spider Cupiennius salei. The cuticular slits of this organ are each associated with two large bipolar mechanosensory neurons with different adaptation rates. With intracellular recording, we have now been able to correlate directly the staining intensity of a neuron for acetylcholinesterase with its adaptation rate, thus allowing us simply to stain a neuron to identify its functional type. All rapidly adapting neurons stain more heavily than slowly adapting neurons. Immunostaining of whole-mount preparations reveals GABA-like immunoreactive fibers forming numerous varicosities at the surface of all sensory neurons in VS-3; peripheral GABA-like immunoreactive somata are lacking. Sectioning the leg nerve procures rapid degeneration of most fiber profiles, confirming that the fibers are efferent. Punctate synapsin-like immunoreactivity colocalizes to these varicosities, although some synapsin-like immunoreactive puncta are GABA-immunonegative. Fibers with similar immunoreactivities are also associated with trichobothria, tactile hairs, internal joint receptors, i.e. other types of spider mechanosensory organs. In organ VS-3, immunoreactivity is most dense across the initial axon segment. The exact distribution of peripheral synapses was reconstructed from a 10-microm-long electron micrograph series of the dendritic, somatic, and initial axon regions of acetylcholinesterase-stained VS-3 neurons. These reveal a pattern similar to that of the synapsin-like immunoreactivity. Two different types of synapse were distinguished on the basis of their presynaptic vesicle populations. Many peripheral synapses thus appear to derive from efferent GABA-like immunoreactive fibers and probably provide centrifugal inhibitory control of primary mechanosensory activities.     

Compliance in pulmonary tuberculosis patients using directly observed treatment short course

Five hundred newly diagnosed cases of pulmonary tuberculosis patients were studied at Iwo Local Government tuberculosis (TB) control clinic Alaye, Iwo, Osun state of Nigeria. The study was to determine (a) the rate of compliance to antituberculous drugs in newly diagnosed pulmonary tuberculosis (PTB) patients over a 2-year period in Iwo, Nigeria treated with directly observed treatment short-course (DOTS). DOTS is a short course treatment lasting 6-8 months and (b) the effect of DOTS and the use of home visitors on compliance with a view to formulating a model that can be used all over Nigeria to stem the resurgence of PTB. All newly diagnosed cases of PTB at Iwo, Osun state of Nigeria were treated with DOTS over a two-year period. TB home visitor provided with a motorcycle was used throughout the treatment period with free drug provision by Damien Foundations of Belgium based in Ibadan, Nigeria. The rate of compliance was noted in all cases. Total (100 percent) compliance and cure rate were recorded with the use of DOTS and TB home visitor in this study. The home visitor carried out 50 visitations during the study period. The use of DOTS with free drug provision and the use of home visitor as used in this study confirmed its effectiveness in enhancing compliance and hence cure of PTB. The study is a model that can be adopted for the whole of Nigeria and other countries globally if tuberculosis must be controlled.     

Kalirin inhibition of inducible nitric-oxide synthase

Nitric oxide (NO) acts as a neurotransmitter. However, excess NO produced from neuronal NO synthase (nNOS) or inducible NOS (iNOS) during inflammation of the central nervous system can be neurotoxic, disrupting neurotransmitter and hormone production and killing neurons. A screen of a hippocampal cDNA library showed that a unique region of the iNOS protein interacts with Kalirin, previously identified as an interactor with a secretory granule peptide biosynthetic enzyme. Kalirin associates with iNOS in vitro and in vivo and inhibits iNOS activity by preventing the formation of iNOS homodimers. Expression of exogenous Kalirin in pituitary cells dramatically reduces iNOS inhibition of ACTH secretion. Thus Kalirin may play a neuroprotective role during inflammation of the central nervous system by inhibiting iNOS activity.     

Segmental antigen challenge increases fibronectin in bronchoalveolar lavage fluid

Fibronectin may contribute to asthma pathogenesis by recruitment and activation of inflammatory cells, and by promotion of subepithelial fibrosis. Fibronectin is produced by several types of airway cells, including epithelial cells, fibroblasts, and alveolar macrophages. To test the hypothesis that antigen-induced airway inflammation is associated with increased local generation of fibronectin, segmental bronchoprovocation (SBP) with antigen and saline was performed in 17 atopic patients. Bronchoalveolar lavage (BAL) was performed at 5 min and 48 h after segmental challenge with saline or antigen. Fibronectin concentrations in BAL fluid, measured by enzyme-linked immunosorbent assay (ELISA), increased more than 5-fold 48 h after antigen challenge (65 [47 to 110] versus 407 [240 to 697] ng/ml, median and 25 to 75% interquartiles, p < 0.05). Fibronectin concentrations 48 h after antigen challenge correlated with histamine concentrations 5 min after antigen challenge and numbers of eosinophils, neutrophils, macrophages, and total cells in BAL fluid 48 h after antigen challenge. BAL was more enriched in fibronectin 48 h after challenge than would be predicted solely from increased permeability of plasma proteins. Western blot analysis showed that fibronectin in BAL fluid was largely intact and contained the extra domain-A (ED-A) splice variant of cellular fibronectin, indicative of local production. We conclude that antigen challenge in atopic subjects causes increased production of fibronectin by airway cells and speculate that this response may contribute to airway remodeling in allergic inflammation.     

Potential difference across subcellular particle membranes. II. Compensation method of measurement

"Zero-loop" of the molecular potential transformer of submitochondrial particles (SMP) is separated from the remaining electron transfer chain by rotenone, and its e.m.f. ET=0,003+RT/2F in [NADP X H] [NAD+]/[NADP+] [NAD X H] volts is used in the compensative method of measurement of the potential difference across the SMP membrane (delta USMP). The phospholipid membrane, measuring the concentration of the penetrating anions in the solution contained SMP, is used as "zero-indicators". This concentration drops monotonically with increase in delta USMP. Delta USMP is equal to ET when the addition of substrates of transhydrogenase reaction with definite ET does not change the potential across phospholipid membrane.     

The clinical significance of granulocyte colonies in bone marrow cultures

The culture system for in vitro evaluation of "colony forming units - culture (CFU-c)" is briefly outlined. This method offers a new approach to studies of proliferation and differentiation of hemopoietic progenitor cells, especially in disorders of granulopoiesis. From available published data it is evident that quantitation of CFU-c is also an indicator of diagnostic and prognostic value for assessment of various types of leukemia. The CFU-c assay has furthermore been introduced to test the viability and proliferating capacity of cryopreserved bone marrow, especially with a view to possible transfusion of stored autologous bone marrow as an adjuvant to cytostatic therapy.     

Fusion and protein-mediated phospholipid exchange studied with single bilayer phosphatidylcholine vesicles of different density

A novel method has been developed for the study of phospholipid exchange and fusion of phospholipid vesicles. Two homogeneous populations of single bilayer phosphatidylcholine vesicles of similar size but markedly different density have been prepared. "Dense" vesicles were made from brominated dioleoyl phosphatidylcholine. "Light" vesicles were prepared from dioleoyl phosphatidylcholine. The two populations were easily separated by density gradient centrifugation. Phosphatidylcholine exchange protein from beef liver was used to promote lecithin exchange between the vesicle populations. Only the lecithin of the external monolayers of the vesicles was available for exchange by exchange protein, implying that flip-flop of vesicle phosphatidylcholine did not take place at a detectable frequency. No spontaneous intervesicle phosphatidylcholine exchange was observed. However, the dense and light vesicles did spontaneously fuse, over several hours, to produce particles of hybrid density.     

The changing presentation of pyloric stenosis

A triplet pregnancy in a 23-year-old woman was terminated at 15 weeks of gestation because of her severe hypertension, lung edema, and secondary hyperthyroidism. The pregnancy consisted of a hydatidiform mole with a 46,XY karyotype and two fetuses each with 46,XX and a 46,XY karyotype. To determine the zygosity and genetic origin of the mole and fetuses, PCR- and computer-assisted genotyping were performed at 27 CA-repeat marker loci that were distributed evenly over the genome. As a result, genotypes of the three pregnancy products were distinct from each other, indicating that the triplets were trizygotic. The mole lacked any maternal alleles but inherited both of the paternal alleles and/or one paternal allele in duplicate. This, along with the XY sex chromosome constitution, indicated that the mole resulted from dispermic androgenesis. The mother developed a persistent trophoblastic tumor thereafter.     

Proinflammatory macrophage-activating properties of the novel phospholipid diacylglycerol pyrophosphate

We have found that the novel phospholipid diacylglycerol pyrophosphate (DGPP), identified in bacteria, yeast, and plants, but not in mammalian cells, is able to potently activate macrophages for enhanced secretion of arachidonate metabolites, a key event in the immunoinflammatory response of leukocytes. Macrophage responses to DGPP are specific and are not mediated by its conversion into other putative lipid mediators such as phosphatidic acid, lysophosphatidic acid, or diacylglycerol. The responses to DGPP are compatible with a receptor-recognition event because they are blocked by suramin. Intracellular signaling initiated by DGPP includes phosphorylation and activation of the Group IV cytosolic phospholipase A2 and of the extracellular-signal regulated p42 mitogen-activated protein kinase (MAPK) and p44 MAPK, and membrane translocation of the protein kinase C isoenzymes alpha, epsilon, delta. These results establish DGPP as a novel macrophage-activating factor and suggest a potential role for this compound in triggering homeostatic cellular responses.