Scoring tools to identify TB patients facing catastrophic costs in the Philippines.

BACKGROUND: This study was to meet a practical need to design a simple tool to identify TB patients who may potentially be facing catastrophic costs while seeking TB care in the public sector. Such a tool may help prevent and address catastrophic costs among individual patients. METHODS: We used data from the national TB patient cost survey in the Philippines. We randomly allocated TB patients to either the derivation or validation sample. Using adjusted odds ratios (ORs) and ß coefficients of logistic regression, we developed four scoring systems to identify TB patients who may be facing catastrophic costs from the derivation sample. We validated each scoring system in the validation sample. RESULTS: We identified a total of 12 factors as predictive indicators associated with catastrophic costs. Using all 12 factors, the ß coefficients-based scoring system (area under the curve [AUC] 0.783, 95% CI 0.754-0.812) had a high validity. Even with seven selected factors with OR > 2.0, the validity remained in the acceptable range (ß coefficients-based: AUC 0.767, 95% CI 0.737-0.798). CONCLUSION: The ß coefficients-based scoring systems in this analysis can be used to identify those at high risk of facing catastrophic costs due to TB in the Philippines. Operational feasibility needs to be investigated further to implement this in routine TB surveillance.

CONTEXTE: Cette étude visait à répondre à un besoin pratique de concevoir un outil simple pour identifier les patients atteints de TB qui pourraient potentiellement être confrontés à des coûts catastrophiques lorsqu'ils recherchent des soins de TB dans le secteur public. Un tel outil pourrait aider à prévenir et à traiter les coûts catastrophiques chez les patients individuels. MÉTHODES: Nous avons utilisé des données de l'enquête nationale sur les coûts des patients atteints de TB aux Philippines. Nous avons réparti aléatoirement les patients atteints de TB dans l'échantillon de dérivation ou de validation. À l'aide des odds ratio (OR) ajustés et des coefficients ß de la régression logistique, nous avons développé quatre systèmes de notation pour identifier les patients atteints de TB qui pourraient être confrontés à des coûts catastrophiques à partir de l'échantillon de dérivation. Nous avons validé chaque système de notation dans l'échantillon de validation. RÉSULTATS: Nous avons identifié un total de 12 facteurs en tant qu'indicateurs prédictifs associés à des coûts catastrophiques. En utilisant les 12 facteurs, le système de notation basé sur les coefficients ß (aire sous la courbe [AUC] 0,783 ; IC 95% 0,754­0,812) avait une validité élevée. Même avec sept facteurs sélectionnés avec OR > 2,0, la validité est restée dans la plage acceptable (basée sur les coefficients ß : AUC 0,767 ; IC 95% 0,737­0,798). CONCLUSION: Les systèmes de notation basés sur les coefficients ß dans cette analyse peuvent être utilisés pour identifier les personnes à haut risque de faire face à des coûts catastrophiques liés à la TB aux Philippines. La faisabilité opérationnelle doit être étudiée plus avant pour mettre en œuvre cela dans la surveillance de routine de la TB.

Novel excitatory neuropeptides isolated from a prosobranch gastropod, Thais clavigera: The molluscan counterpart of the annelidan GGNG peptides.

The GGNG peptides are excitatory neuropeptides identified from earthworms, leeches and polychaeta. Two structurally related peptides were purified and characterized from a mollusk, Thais clavigera (prosobranch gastropod). The peptides designated as Thais excitatory peptide-1 (TEP-1) (KCSGKWAIHACWGGN-NH2) and TEP-2 (KCYGKWAMHACWGGN-NH2) are pentadecapeptides having one disulfide bond and C-terminal GGN-NH2 structures, which are shared by most GGNG peptides. TEP augmented the motilities of Thais esophagus and penial complex. TEP-like immunoreactivity is distributed in both the neurons of the central nervous system and nerve endings in the penial complex. Thus, the involvement of TEP in the contraction of the digestive and reproductive systems is suggested. Substitution of amino acids in TEP revealed that two tryptophan residues in TEP are important for maintaining bioactivity.

The Aplysia mytilus inhibitory peptide-related peptides: identification, cloning, processing, distribution, and action.

Neuropeptides are a ubiquitous class of signaling molecules. In our attempt to understand the generation of feeding behavior in Aplysia, we have sought to identify and fully characterize the neuropeptides operating in this system. Preliminary evidence indicated that Mytilus inhibitory peptide (MIP)-like peptides are present and operating in the circuitry that generates feeding in Aplysia. MIPs were originally isolated from the bivalve mollusc Mytilus edulis, and related peptides have been identified in other invertebrate species, but no precursor has been identified. In this study, we describe the isolation and characterization of novel Aplysia MIP-related peptides (AMRPs) and their precursor. Several AMRPs appear to have some structural and functional features similar to vertebrate opioid peptides. We use matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to confirm that all 14 AMRPs predicted by the precursor are processed in isolated neurons. Northern analysis, whole-mount in situ hybridization, and immunohistochemistry are used to map the abundant expression of these peptides in the CNS and peripheral tissues such as the digestive tract, vasculature, and the reproductive organs. Physiological studies demonstrate that the rank order of the inhibitory actions of these peptides is different for three target muscles. These results underscore the importance of using a multidisciplinary approach to identifying and characterizing the actions of neuropeptides in an effort to gain understanding of their role in systems of interest. The widespread distribution of the AMRPs indicates that they may be operating in many different systems of Aplysia.

The enterins: a novel family of neuropeptides isolated from the enteric nervous system and CNS of Aplysia.

To identify neuropeptides that have a broad spectrum of actions on the feeding system of Aplysia, we searched for bioactive peptides that are present in both the gut and the CNS. We identified a family of structurally related nonapeptides and decapeptides (enterins) that are present in the gut and CNS of Aplysia, and most of which share the HSFVamide sequence at the C terminus. The structure of the enterin precursor deduced from cDNA cloning predicts 35 copies of 20 different enterins. Northern analysis, in situ hybridization, and immunocytochemistry show that the enterins are abundantly present in the CNS and the gut of Aplysia. Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry we characterized the enterin-precursor processing, demonstrated that all of the precursor-predicted enterins are present, and determined post-translational modifications of various enterins. Enterin-positive neuronal somata and processes were found in the gut, and enterins inhibited contractions of the gut. In the CNS, the cerebral and buccal ganglia, which control feeding, contained the enterins. Enterin was also present in the nerve that connects these two ganglia. Enterins reduced the firing of interneurons B4/5 during feeding motor programs. Such enterin-induced reduction of firing also occurred when excitability of B4/5 was tested directly. Because reduction of B4/5 activity corresponds to a switch from egestive to ingestive behaviors, enterin may contribute to such program switching. Furthermore, because enterins are present throughout the nervous system, they may also play a regulatory role in nonfeeding behaviors of Aplysia.

Deamidase inactivates a D-amino acid-containing Aplysia neuropeptide.

Membrane-catalyzed degradation of the cardioexcitatory peptide, Asn-D-Trp-Phe-NH(2) (N(d)WF-NH(2)), which was previously isolated from Aplysia, was investigated in relation to its inactivation mechanism. The principal degradation was deamidation of the C-terminal amide, producing biologically inert Asn-D-Trp-Phe-OH (N(d)WF-OH). Among membrane fractions prepared from different organs, the fraction from the ganglia showed the highest specific activity of the deamidation reaction. The deamidase activity was inhibited by Ebelactone B and the serine protease inhibitor, phenylmethanesulfonyl fluoride (PMSF), while the degradation of the synthetic stereoisomer, Asn-Trp-Phe-NH(2) (N(l)WF-NH(2)), was sensitive to the divalent cation-chelating agent, o-phenanthroline, and aminopeptidase inhibitors, amastatin and bestatin. The presence of D-Trp residue in the second position of N(d)WF-NH(2) endows this peptide not only with stereospecific bioactivity but also peptidase stability. The deamidation reaction seems to be the major inactivation mechanism for this peptide.

NdWFamide: a novel excitatory peptide involved in cardiovascular regulation of Aplysia.

Although diverse peptides are known to affect invertebrate cardiac activity, the peptidergic regulation of the cardiovascular system of Aplysia is still poorly understood. Asn-D-Trp-Phe-NH(2) (NdWFamide) is a recently purified cardioactive peptide in Aplysia. Pharmacological experiments showed that NdWFamide was one of the most potent cardioexcitatory peptides among the known endogenous cardioactive peptides in Aplysia. NdWFamide-immunopositive neuronal processes were abundant in the cardiovascular region of Aplysia, and many of them originated from neurosecretory cells in the abdominal ganglion (R3-R13 cells). The data suggest that NdWFamide is a cardioexcitatory peptide utilized by R3-R13 cells of Aplysia.

A novel GGNG-related neuropeptide from the polychaete Perinereis vancaurica.

The GGNG peptides are myoactive peptides so far identified from earthworms and leeches, which are the earthworm excitatory peptides (EEP) and the leech excitatory peptide (LEP), respectively. A novel GGNG peptide was isolated and structurally determined from a marine polychaete, Perinereis vancaurica, using a combination of immunological assay and high performance liquid chromatography (HPLC). The peptide was a pentadecapeptide whose amino acid sequence was similar to that of EEP and LEP, and showed myoactivity on isolated esophagus of P. vancaurica with a threshold concentration of 10(-10)M. The peptide was designated as polychaete excitatory peptide (PEP). Amidation of the alpha-carboxyl group of C-terminal residue occurred in PEP. This is the case for LEP, but not for EEP. The cDNA cloning revealed that the structure of the PEP precursor is more similar to the EEP precursor than to the LEP precursor. Immunohistochemical staining showed the presence of PEP in several neurons of central nervous system (CNS) as somata and neuropile structure, epithelial cells of the pharynx and epidermal cells throughout the body wall. Altogether these results support the physiological significance of PEP in regulation of the CNS neural activity and the peripheral myoactivity.

Distribution and function of an Aplysia cardioexcitatory peptide, NdWFamide, in pulmonate snails.

The distribution and function of an Aplysia cardioexcitatory peptide, NdWFamide, were examined in the nervous system of pulmonate snails. We chemically identified the authentic NdWFamide from a land snail (Euhadra congenita) and a freshwater snail (Lymnaea stagnalis). NdWFamide potentiated the heartbeat of those snails. Immunohistochemistry using anti-NdWFamide antibody demonstrated the distribution of NdWFamide-containing neurons and fibers in the central nervous system, as well as peripheral tissues, such as the cardiovascular region and accessory sex organs. These results suggest that NdWFamide is a neuropeptide mediating the neural regulation of the activity of the cardiovascular and reproductive systems of snails.

Identification of a vasopressin-like immunoreactive substance in hydra.

Vasopressin (VP)-like immunoreactivity has long been known in the hydra nervous system, but has not yet been structurally identified. In this study, using HPLC fractionation and an immunological assay, we have purified two peptides, FPQSFLPRGamide and SFLPRGamide, from Hydra magnipapillata. Both the peptides shared the same C-terminal structure, -PRGamide, with Arg-VP. The nonapeptide proved to be Hym-355, a peptide that stimulates neuronal differentiation in hydra. Detailed evaluation by competitive enzyme-linked immunosorbent assay (ELISA) and double immunostaining using anti-VP and anti-Hym-355 antibodies enabled us to conclude that the two peptides account for a major part of the VP-like immunoreactivity in hydra nerve cells.

Carassius RFamide, a novel FMRFa-related peptide, is produced within the retina and involved in retinal information processing in cyprinid fish.

Carassius RFamide (C-RFa) is a novel peptide, isolated originally from the brain of the Japanese crucian carp and sharing homologies with mammalian prolactin-releasing peptide (PrRP). It has been demonstrated previously that C-RFa mRNA is abundant in the proximal half (fundus) of the Japanese crucian carp eye. In the present work, we localized C-RFa by immunohistochemistry mainly to perikarya, in the proximal half of the inner nuclear layer (amacrine cell layer). This distribution is different from that of FMRFamide, which is confined to axon terminals of terminal nerve efferent fibers in the inner plexiform layer. Electrophysiological recording revealed that C-RFa depolarized some amacrine cells and hyperpolarized L-type horizontal cells in the carp. These results suggest that C-RFa is produced within the cyprinid retina and functions as a transmitter or neuromodulator in retinal image processing.

The background adaptation of the flatfish, Paralichthys olivaceus.

The background adaptation of the flatfish, Paralichthys olivaceus, was studied by sliding beneath the fish a long strip of plastic sheet with various patterns to serve as a background. The body shade and pattern differed depending upon the pattern used. In general, the fish showed maximal pallor on a white background and darkened when lying on a black background. With a pattern of large checks, several black patches became conspicuous. The fish showed the same tendency in terms of response even when the left eye was covered. However, the fish was prevented from manifesting these changes when the right eye was covered. The results suggest that the left eye of the fish does not see the immediate background. This conclusion is supported by an anatomical study.

Immunohistochemical Localization of C-RFamide, a FMRF-related Peptide, in the Brain of the Goldfish, Carassius auratus.

Carassius RFamide (C-RFa) is a novel peptide found in the brain of the Japanese crucian carp. It has been demonstrated that mRNA of C-RFa is present in the telencephalon, optic tectum, medulla oblongata, and proximal half of the eyeball in abundance. Immunohistochemical methods were employed to elucidate the distribution of the peptide in the brain of the goldfish (Carassius auratus) in detail. C-RFaimmunoreactive perikarya were observed in the olfactory bulb, the area ventralis telencephali pars dorsalis and lateralis, nucleus preopticus, nucleus preopticus periventricularis, nucleus lateralis tuberis pars posterioris, nucleus posterioris periventricularis, nucleus ventromedialis thalami, nucleus posterioris thalami, nucleus anterior tuberis, the oculomotor nucleus, nucleus reticularis superior and inferior, facial lobe, and vagal lobe. C-RFa immunoreactive fibers and nerve endings were present in the olfactory bulb, olfactory tract, area dorsalis telencephali pars centralis and medialis, area ventralis telencephali, midbrain tegmentum, diencephalon, medulla oblongata and pituitary. However, in the optic tectum the immunopositive perikarya and fibers were less abundant. Based on these results, some possible functions of C-RFa in the nervous system were discussed.

A myomodulin-CARP-related peptide isolated from a polychaete annelid, Perinereis vancaurica.

Myomodulin-CARP-family peptides have been isolated only from molluscs. In the present study, a heptapeptide, Ala-Met-Gly-Met-Leu-Arg-Met-NH2, termed Pev-myomodulin, was isolated from a polychaete annelid, Perinereis vancaurica using the esophagus of the animal as the bioassay system. The sequence of the annelid peptide is highly homologous with those of the myomodulin-CARP-family peptides found in molluscs. The annelid peptide is regarded as a member of the myomodulin-CARP family, though all the molluscan peptides have a Leu-NH2 at their C-termini. The annelid peptide showed a potnet contractile action on the esophagus of the annelid. The peptide may be an excitatory neuromediator involved in the regulation of the esophagus. Among various myomodulin-CARP-family peptides and their analogues, the annelid peptide showed the most potent contractile action on the esophagus. Replacement of the C-terminal Met-NH2 of the annelid peptide with a Leu-NH2 decreased its contractile potency, while replacement of the C-terminal Leu-NH2 of myomodulin and CARP with a Met-NH2 increased their potency. The C-terminal Met-NH2 of the annelid peptide seems to be important, but not essential, for exhibiting its contractile activity on the esophagus. On the anterior byssus retractor muscle of the bivalve mollusc Mytilus edulis, the annelid peptide showed catch-relaxing and contraction-modulating effects qualitatively similar to those of the authentic peptide CARP, though the annelid peptide was less potent than CARP.

Immunohistochemical Localization of Annetocin, an Earthworm Oxytocin-Related Peptide, and Identification and Ultrastructural Characteristics of the Annetocin-Secretory Cells in the Oligochaete Earthworm Eisenia foetida.

Annetocin is an egg-laying-inducing oxytocin-related peptide which we have previously isolated from the earthworm, Eisenia foetida. Here we report the results of immunohistochemical and ultrastructural studies on annetocin-secretory cells in the earthworm. Annetocin-immunoreactive (IR) cell-somata were located mainly at the ventro-lateral side of the subesophageal ganglion. Only four annetocin-IR cells were seen in the cerebral ganglion. Some annetocin-IR cells displayed unipolar-like structure with a process directing to the core region (the neuropile) of the ganglion. Annetocin-IR fibers were also observed in the neuropile of the ventral ganglia and the ventral nerve cord between the 4th and the 30th segments including the clitellum, but not in the posterior segments (31-55th). The number of annetocin-IR fibers decreased from the 4th to the 30th segment. The annetocin-secretory cells were identified by the immunogold staining, and filled with gold-labeled vesicles, 200-250 nm in diameter, which included moderately electron dense material. The annetocin-secretory cells possessed a euchromatic nucleus, well-developed rough endoplasmic reticulum and Golgi apparatus. Some of the annetocin-secretory cells were found to form a neurohemal-like structure, where somata or fibers with loose glial investment came in contact with the coelomic space at the ventral side of the subesophageal ganglion. The results suggest that annetocin is a neuropeptide produced and secreted by the neuron in the cerebral and subesophageal ganglia of the earthworm.

Separation of drugs by packed-column supercritical fluid chromatography.

Packed-column supercritical fluid chromatography (pSFC) has been expected to analyze various kinds of compounds. Many researchers have expected a new chromatographic technique that overcomes the limitations of other techniques, HPLC and GC. In pharmaceutical development, chromatography plays an important role in the evaluation of safety and efficacy of a new compound. This article provides an overview of the separation of drugs by pSFC. The effects of the chromatographic parameters were studied for the separation of steroids. In chiral separation, the successful results were shown and compared with HPLC.

Some mechanical properties of skinned fibres of pregnant human myometrium.

The properties of contractile elements and intracellular Ca2+ storage sites of pregnant human myometrium were studied by recording the mechanical responses in skinned (saponin-treated and membrane-permeable) fibres. Calmodulin increased the amplitude of contractions induced by Ca2+ and the Ca2+ sensitivity for contractile elements in small myometrium strips, but PGF2 alpha, PGE2, oxytocin, or cyclic AMP failed to produce similar effects. After accumulation of Ca2+ in intracellular Ca2+ storage sites, 10 mumol/l PGF2 alpha, 10 mumol/l PGE2, 30 mmol/l caffeine, and 20 mumol/l InsP3 (inositol-trisphosphate) produced contractions by releasing Ca2+ from storage sites. However, 20 nmol/l oxytocin had no effects under the same conditions. The InsP3 sensitive Ca2+ store was much larger than those of PGs or caffeine. These results suggest that pregnant human myometrium contracts with low Ca2+ by a calmodulin sensitive system. The data also indicate that direct application of PGF2 alpha, or PGE2 into the cells discharges Ca2+ from Ca2+ storage sites and that oxytocin extricates Ca2+ via a pathway involving InsP3 by activation of phosphoinositide turnover. We suggest that these agents induce added contractile responses due to a Ca2+ release mechanism from store sites in addition to the influx of Ca2+ from the extracellular space.

[Clinical experience with S-6436 in patients with urinary tract infections (author's transl)].

Forty eight patients with urinary tract infections including a case of acute prostatitis were treated with S-6436 and the following results were obtained: 1. Thirty eight patients with acute cystitis resulted in 19 excellent, 15 good and 4 poor with effectiveness rate of 89.5%. 2. Ten patients with complicated urinary tract infections resulted in 1 excellent, 5 good and 4 poor with effectiveness rate of 60%. 3. Infecting organisms from 38 patients with acute cystitis were occupied by 28 strains of E. coli. The sensitivity rate of the infecting organisms to cephalexin was 96.4%. 4. In a few cases (6.25%) side effects of S-6436 were observed. 5. S-6436 will be one of the good first choice antibiotics for acute cystitis.

Comparative aspects of invertebrate neuropeptides.

1. We searched for bioactive peptides, most of which were considered to be neuropeptides, in various animals of several phyla. These peptides were compared with each other and with peptides identified by many other investigators. Consequently, we found that structures of neuropeptides are generally conserved in each phylum. 2. We also found some exceptional interesting aspects. First, there are a number of peptide groups whose members are distributed among several phyla. Second, there are many structural similarities between molluscan and annelidan peptides as if molluscs and annelids were the animals in a phylum. Third, certain toxic peptides of invertebrates are closely related to vertebrate neuropeptides. 3. In addition to the above phylogenetic aspects, we found some other interesting aspects. A wide structural variety of members of a peptide group is generally found in invertebrate species. Invertebrate muscles seem to be generally regulated not only by some or several classical non-peptidic neuromediators but also by various peptidic neuromediators. Peptides containing a D-amino acid residue are not rare.

Bioactive peptides of annelids are closely related to those of molluscs.

Several bioactive peptides were isolated from some annelids. Each of the annelidan peptides seemed to be a member of a previously found molluscan peptide family.

[Adverse effect of iodine contrast media].

We performed a survey on the iodide test and its adverse effects on a series of 829 DIP patients who had taken the test at our clinic between November, 1984 and August, 1985. Adverse effects were seen in four out of nineteen positive cases of the iodide test. Among the negative 810 cases, 79 were found to present with adverse effects though not statistically significant. Twenty-six out of 131 cases of positive past history of allergy showed a positive reaction whereas 55 out of 698 negative cases showed adverse effects, with a statistical significance at p less than 0.01. The adverse reaction occurred immediately after injection in the majority of cases and in 85%, the reaction was seen after injection of half of the drug. Symptoms also appeared thirty minutes after injection in some cases. Immunological evaluation of the allergy was done in fifteen positive cases. Four cases showed an increase in peripheral eosinophils, three cases showed increased IgE(RIST) and three cases manifested with a decreased CH 50. Urografin-induced lymphocyte blastogenesis (DLST) was positive in six cases. Four cases showed abnormality in more than two tested parameters, and three cases showed complete normal values in all tests. The adverse effects due to contrast medium are considered to occur not as one type or allergic reaction, but to involve other factors as delayed-type reaction which was also seen in 40% of the patients. Close observation of patients after examination with contrast medium is thus mandatory.(ABSTRACT TRUNCATED AT 250 WORDS)