Efficacy of artesunate plus pyrimethamine-sulphadoxine for uncomplicated malaria in Gambian children: a double-blind, randomised, controlled trial.

BACKGROUND: Resistance to cheap effective antimalarial drugs, especially to pyrimethaminesulphadoxine (Fansidar), is likely to have a striking impact on childhood mortality in sub-Sharan Africa. The use of artesunate (artesunic acid) [corrected] in combination with pyrimethamine-sulphadoxine may delay or prevent resistance. We investigated the efficacy, safety, and tolerability of this combined treatment. METHODS: We did a double-blind, randomised, placebo-controlled trial in The Gambia. 600 children with acute uncomplicated Plasmodium falciparum malaria, aged 6 months to 10 years, at five health centres were randomly assigned pyrimethaminesulphadoxine (25 mg/500 mg) with placebo; pyrimethamine-sulphadoxine plus one dose of artesunate (4mg/kg bodyweight); or pyrimethamine-sulphadoxine plus one dose 4 mg/kg bodyweight artesunate daily for 3 days. Children were visited at home each day after the start of treatment until parasitaemia had cleared. FINDINGS: The combined treatment was well tolerated. No adverse reactions attributable to treatment were recorded. By day 1, only 178 (47%) of 381 children treated with artesunate were still parasitaemic, compared with 157 (81%) of 195 children in the pyrimethamine-sulphadoxine alone group (relative risk 1.7 [95% CI 1.5-2.0], p<0.001). Treatment-failure rates at day 14 were 3.1% in the pyrimethamine sulphadoxine alone group, and 3.7% in the one-dose artesunate group (risk difference -0.6% [-4.2 to 3.0]) and 1.6% in the three-dose group (1.5 [1.5-4.5], p=0.048). Symptoms resolved faster in children who received artesunate, but there was no additional benefit for three doses of artesunate over one dose. Children given artesunate were less likely to be gametocytaemic after treatment. INTERPRETATION: The combined treatment was safe, well tolerated, and effective. The addition of artesunate to malaria treatment regimens in Africa results in lower gametocyte rates and may lower transmission rates.

PIP: This double-blind, randomized, controlled study investigated the efficacy, safety and tolerability of artesunate plus pyrimethamine-sulphadoxine for uncomplicated malaria among Gambian children. Combined use of artesunate and pyrimethamine-sulphadoxine was hypothesized to delay or prevent resistance, which proved to be effective in reducing childhood mortality in sub-Saharan Africa. A total of 600 children with acute uncomplicated Plasmodium falciparum malaria, 6 months to 10 years old, were randomly administered pyrimethamine-sulphadoxine (25 mg/500 mg) with placebo, 4 mg/kg body weight pyrimethamine-sulphadoxine plus 1 dose of artesunate, or pyrimethamine-sulphadoxine plus 4 mg/kg body weight artesunate for 3 days. Results indicate that combined treatment was well tolerated. On day 1, 178 of 381 children treated with artesunate were still parasitemic compared with 157 of 195 children in the pyrimethamine-sulphadoxine group. On the other hand, failure rates on day 14 were 3.1% in the pyrimethamine-sulphadoxine group and 3.7% in the 1-dose artesunate group and 1.6% in the 3-dose group. Insignificant differences were found among children administered 1-dose and 3-dose artesunate, and were found less likely to be gametocytemic after treatment. In conclusion, this study confirms the safety and efficacy of a combined treatment, which eventually results in lower gametocyte rates and lower transmission rates.

FMRFamide-like and allatostatin-like immunoreactivity in the lateral heart nerve of Periplaneta americana: colocalization at the electron-microscopic level.

Both allatostatin immunoreactivity (AS-IR) and FMRFamide immunoreactivity (FMRFa-IR) have been demonstrated light-microscopically in the lateral heart nerve of Periplaneta americana. The identical labeling of some fibers suggests the coexistence of the two antigens. Electron-microscopically, six granule types in the peripheral part of the lateral heart nerve can be distinguished according to their size and density (types 1-6). These granule types can be subdivided immunocytochemically by means of a new mirror-section technique. Granules of types 4 and 5 always exclusively show FMRFa-IR. In the populations of fibers containing granules of types 1 and 6, axon profiles can be found that contain granules colocalizing FMRFa-IR and AS-IR. Other axon profiles of these populations only contain immunonegative granules of the same ultrastructure. Granules of type 2 can be differentiated immunocytochemically in three forms in the same section: In some fibers, they are nonreactive; in other fibers of the same section, they show FMRFa - IR, whereas in a third fiber type, granules show AS - IR. Finally, granules of type 3 can be observed with FMRFa-IR. In other fibers, they occur with the same ultrastructure but exhibit no immunoreactivity. Two soma types occur in the lateral heart nerve. Soma type I is characterized by the production of electron-dense granules that show FMRFa-IR. Type II is in close contact with various fibers, forming different types of axosomatic synapses, hitherto unknown in Insecta.

Peripheral connections of FMRFamide-like immunoreactive neurons in the snail, helix pomatia: an immunogold electron microscopic study.

Using a postembedding immunogold electron microscopic method, the ultrastructure and synaptic connections of FMRFamide-like immunoreactive varicosities were investigated in different peripheral organs of the snail Helix pomatia, including the heart (auricle), intestine, hepatopancreas, upper tentacle and salivary gland. The FMRFamide-like immunoreactive varicosities contained granules and vesicles as described in a previous study of the CNS of this species, and additionally, based on their granule content, two novel types of varicosities were found in the auricle. A selective accumulation of gold particles over the granules could be demonstrated. The FMRFamide-like immunoreactive varicosities formed unspecialized contacts with postsynaptic target cells in all peripheral organs investigated, with the exception of the tentacle retractor muscle. Both the neuro-muscular and the neuro-glandular contacts were characterized by either unspecialized close apposition of the 'pre- and postsynaptic' membranes or the immunoreactive elements faced the target cell(s) across a relatively wide extracellular space. In the tentacle retractor muscle some of the neuromuscular contacts showed appositions of electron dense material along the presynaptic membrane, clustering of agranular synaptic vesicles and intersynaptic cleft material. The present observations support previous electrophysiological findings and suggest a versatile modulatory role of FMRFamide and related substances in the Helix PNS.

An immunogold electron microscopic analysis of FMRFamide-like immunoreactive neurons in the CNS of Helix pomatia: ultrastructure and synaptic connections.

The ultrastructure and synaptic connections of FMRFamide-like immunoreactive neurons were investigated in the CNS of the snail Helix pomatia, following the application of a post-embedding immunogold method. For comparison, first, we analyzed the ultrastructure and granule content of the identified FMRFamide-containing C3 neuron in the cerebral ganglion. Three types of unidentified immunoreactive neuronal perikarya, and five types of varicosities could be distinguished on the basis of granule content. The different granule types revealed a highly selective accumulation of gold particles. One granule type contained by one FMRFamide-like immunoreactive neuron type (N1) and by one varicosity type (T2) showed similar ultrastructure to that of the granules seen in the C3 cell. In the neuropil, the majority of FMRFamide-like immunoreactive varicosities (four of the five varicosity types) established specialized synaptic contacts with unidentified postsynaptic profiles. In the connective tissue sheath around the ganglia, three types of FMRFamide-like immunoreactive varicosities were found to establish unspecialized contacts with smooth muscle fibres or to be free in the mass of collagen fibres. On the basis of these observations, we suggest (1) an extensive diversity of the localization of FMRFamide (and related substances) at the ultrastructural level; (2) the involvement of FMRFamide-like immunoreactive varicosities in synaptic, modulatory and neurohormonal regulatory processes in the Helix nervous system.

A comparative study of the immunohistochemical localization of a presumptive proctolin-like peptide, thyrotropin-releasing hormone and 5-hydroxytryptamine in the rat central nervous system.

A proctolin (PROC)-like peptide was studied immunohistochemically in the hypothalamus, lower brainstem and spinal cord of the rat using an antiserum against PROC conjugated to thyroglobulin. Neuronal cell bodies containing PROC-like immunoreactivity (PROC-LI) were observed in the dorsomedial, paraventricular and supraoptic nuclei of the hypothalamus and in the nucleus raphe magnus, nucleus raphe pallidus, nucleus raphe obscurus and nucleus interfascicularis nervi hypoglossi in the medulla oblongata. Fibers containing PROC-LI were seen in the median eminence and in other hypothalamic nuclei, and in the lower brainstem in cranial motor nuclei including the dorsal motor nucleus of the vagus nerve, the motor trigeminal nucleus, the facial nucleus and nucleus ambiguous, and in lower numbers in the nucleus of the solitary tract and locus coeruleus. Fibers containing PROC-LI were also located in the spinal cord, in the intermediolateral cell column at thoracic levels and in the ventral horns at all levels of the spinal cord. After transection of the spinal cord, all PROC-immunoreactive fibers below the lesion disappeared. Following injection of Fast blue into the thoracic spinal cord, retrogradely labeled cells in the nuclei raphe pallidus, obscurus and magnus and nucleus interfasciculari nervi hypoglossi were seen to contain PROC-LI. PROC-LI had a similar distribution as thyrotropin-releasing hormone (TRH)-LI in the above-mentioned areas and coexistence of TRH-LI and PROC-LI was shown in cell bodies in the hypothalamus and medulla oblongata. PROC-LI could also be shown to coexist with 5-hydroxytryptamine (5-HT)-LI in neuronal cell bodies in the lower brainstem. The results demonstrate the occurrence of a PROC-like peptide in the mammalian nervous system, and these neurons seem to be at least largely identical to previously described TRH systems. A possible involvement of the PROC-like peptide in spinal motor control is discussed in relation to the well-established role of PROC in control of motor behavior in insects and invertebrates.

A post-embedding immunoelectron-microscopic demonstration of apolipoprotein-B-containing lipoprotein particles in hepatocytes of fetal rats.

We used the protein-A gold technique to demonstrate the presence of apolipoprotein-B in ultrathin sections of fetal rat liver tissue. It was possible to show for the first time that the electron-dense, osmiophilic particles with diameters of 20-40 nm located within the RER cisternae and Golgi complexes of fetal rat hepatocytes contain apolipoprotein-B components and therefore are lipoproteins. After specific labelling an accumulation of gold label was observed on the RER cisternae, Golgi cisternae and the Golgi-associated secretory vesicles of hepatocytes. The specificity of this labelling pattern was assessed by comparison with cytochemical controls. Our qualitative findings were confirmed by a quantitative analysis of the mean labelling intensity (mean number of gold particles per square micron of the surface area of a particular cellular compartment) on the RER, Golgi complexes, mitochondria, nuclei and the remaining cytoplasm of hepatocytes. It is concluded that the hepatocytes of fetal rats are capable of forming apolipoprotein-B-containing lipoprotein particles. With respect to the size-distribution pattern of the observed intra-hepatic lipoprotein particles, we suggest that the hepatocytes of fetal rats produce lipoproteins of the low- and very low-density-lipoprotein type.

Synaptic connections of the nervus connectivus in the frontal ganglion of Periplaneta americana L. (Insecta). An electron microscopic and iontophoretic study.

The course of the Nervus connectivus (N.c.), its branches, and synaptic connections within the frontal ganglion (FG) were investigated electron microscopically after cobalt iontophoresis of the N.c. The subsequent treatment of ultrathin sections with Timm's method was found to be very suitable for identifying the smallest branches. In the neuropil, fibers of the N.c. form Gray-I-type synapses, but also dyads are abundant, whereby the N.c. fibers occur exclusively in postsynaptic position with neurosecretory fibers. The possible role of these relationships is discussed.

The frontal ganglion of Periplaneta americana L. (Insecta): an electron microscopic and immunohistochemical study.

The frontal ganglion, part of the stomatogastric nervous system, contains about 60 to 80 neurons, 25 to 30 micron in diameter. A well developed Golgi system, producing dense-core vesicles, lysosomes, multivesicular bodies and dense bodies are abundant. Glia elements are sparsely distributed. Many nerve fibres contain granules of different size and electron density. Five groups of fibres can be distinguished: Fibres with granules of about 200 nm (type A), fibres with granules of about 160 to 170 nm (type B), fibres with granules of about 80 to 100 nm (type C) and those with synaptic vesicles of 50 nm (type D) respectively. A fifth very small type contains neither vesicles nor granules. Special attention was paid to synaptic contacts. The divergent dyad seems to be the main type in the frontal ganglion. Frequently, neurosecretory endings are observed in presynaptic position. Immunocytochemical "staining" of neurosecretory material closely corresponds to the distribution of type A fibres, as observed electron microscopically. Immunoelectrophoresis of extracts from frontal ganglia with polyspecific anti-neurosecretion-serum reveals a single precipitation line, indicating that the immunocytochemical localization of neurosecretory material is due to reaction with a specific as well as a cross-reagent anitbody.

Sodium pumps and galactose transfer in the short-circuited small intestine.

In a short-circuited preparation of rat jejunum, there are two sodium pumps, one electrogenic and the other neutral. When energy sources are limited, the total sodium transfer is limited. In the presence of a non-metabolized actively transferred hexose, the electrogenic pump is preferentially used. The neutral sodium pump is only able to function when additional energy is available.

Neutral sodium pump in the small intestine.

A non-electrogenic mechanism for Na absorption has been demonstrated in the in vitro short-circuited rat mid-jejunum. This mechanism exists in addition to the electrogenic Na absorption associated with active non-electrolyte transfer. It was not possible to demonstrate the existence of a similar mechanism in rat terminal ileum and it is suggested that this is related to differences in the metabolism of the ileum as compared with the jejunum. the neutrality of the non-electrogenic pump has been shown to be largely due to the absorption of Na accompanied by Cl. These appears to be a common link between the two types of Na pump since both are inhibited by ouabain although the non-electrogenic pump shows greater sensitivity to ouabain than the electrogenic pump.

Ultrastructure of the prothoracic gland cells of the last instar of Galleria mellonella in relation to the state of development.

The prothoracic glands of the last instar of Galleria mellonella undergo characteristic alterations of their cellular fine structure closely related to cellular activity. During progressive secretory activity of the gland cells there are extensive plasmalemmal infoldings and formation of a pronounced lacunar system. Mitochondria of the active cell phase are characterized by a specific increase in size and paler colour of the matrix. In contrast to the alterations, nuclei, ER and Golgi cisterns do not undergo any submicroscopic changes during the different phases of cellular activity. The relationship between the substructural phenomena and the specific phases of cellular activity are discussed.