Assessing returns to research investments in rice varietal development: Evidence from the Philippines and Bangladesh.

•This study estimates economic returns to investments in rice varietal development in the Philippines and Bangladesh.•The net returns to IRRI and national partners' investments remain strongly positive.•However, the returns are decreasing at a faster rate in the Philippines (24%) than in Bangladesh (6%).•IRRI and national partners should continue investing in rice R&D, especially to develop superior rice varieties.

Field survey for strongyloidiasis in eastern Uganda with observations on efficacy of preventive chemotherapy and co-occurrence of soil-transmitted helminthiasis/intestinal schistosomiasis.

Following our previous field surveys for strongyloidiasis in western Uganda, 120 mothers and 232 children from four villages in eastern Uganda were examined, with two subsequent investigative follow-ups. As before, a variety of diagnostic methods were used: Baermann concentration, Koga agar plate and strongyloidid enzyme-linked immunosorbent assay (ELISA), as well as Kato-Katz faecal smears for detection of eggs of other helminths. At baseline, the general prevalence of Strongyloides stercoralis was moderate: 5.4% as estimated by Baermann and Koga agar methods combined. A much higher estimate was found by ELISA (42.3%) which, in this eastern setting, appeared to be confounded by putative cross-reaction(s) with other nematode infections. Preventive chemotherapy using praziquantel and albendazole was offered to all participants at baseline. After 21 days the first follow-up was conducted and 'cure rates' were calculated for all parasites encountered. Eleven months later, the second follow-up assessed longer-term trends. Initial treatments had little, if any, effect on S. stercoralis, and did not alter local prevalence, unlike hookworm infections and intestinal schistosomiasis. We propose that geographical patterns of strongyloidiasis are likely not perturbed by ongoing praziquantel/albendazole campaigns. Antibody titres increased after the first follow-up then regressed towards baseline levels upon second inspection. To better define endemic areas for S. stercoralis, careful interpretation of the ELISA is warranted, especially where diagnosis is likely being confounded by polyparasitism and/or other treatment regimens; new molecular screening tools are clearly needed.

Gait alterations in rats following attachment of a device and application of altered knee loading.

Animal models are widely used to study cartilage degeneration. Experimental interventions to alter contact mechanics in articular joints may also affect the loads borne by the leg during gait and consequently affect the overall loading experienced in the joint. In this study, force plate analyses were utilized to measure parameters of gait in the rear legs of adult rats following application of a varus loading device that altered loading in the knee. Adult rats were assigned to Control, Sham, or Loaded groups (n ≥ 4/each). Varus loading devices were surgically attached to rats in the Sham and Loaded groups. In the Loaded group, this device applied a controlled compressive overload to the medial compartment of the knee during periods of engagement. Peak ground reaction forces during walking were recorded for each rear leg of each group. Analyses of variance were used to compare outcomes across groups (Control, Sham, and Loaded), leg (contralateral, experimental) and device status (disengaged, engaged) to determine the effects of surgically attaching the device and applying a compressive overload to the joint with the device. The mean peak vertical force in the experimental leg was reduced to 30% in the Sham group in comparison to the contralateral leg and the Control group, indicating an effect of attaching the device to the leg (p<0.01). No differences were found in ground reaction forces between the Sham and Loaded groups with application of compressive overloads with the device. The significant reduction in vertical force due to the surgical attachment of the varus loading device must be considered and accounted for in future studies.

Molecular phylogenetic analysis of the Pneumoroidea (Orthoptera, Caelifera): molecular data resolve morphological character conflicts in the basal acridomorpha.

A key transition in the evolution of the insect suborder Caelifera (Orthoptera; Insecta) was from predominantly non-angiosperm-feeding basal lineages to the modern acridomorph fauna (grasshoppers and related insects). However, because of conflicts in the distribution of several complex morphological characters, the relationships of the presumed intermediates, and in particular of the superfamily Pneumoroidea, are presently unclear. We undertook a phylogenetic study of representatives of all of the transitional acridomorph families using mitochondrial and nuclear DNA sequences. No support for pneumoroid monophyly was obtained from nonparametric bootstrap analysis. Furthermore, adopting a maximum-likelihood approach, specific hypotheses of relationships within the Pneumoroidea were firmly rejected using parametric bootstrapping and Kishino-Hasegawa tests. The results indicate that the Pneumoroidea are at best a grade. This distinction implies that the evolution of the proposed pneumoroid synapomorphies, femoro-abdominal stridulation and simple male genital structure, might previously have been misinterpreted as cases of single character gains or losses within lineages. Reconstructions of character states for the femoro-abdominal stridulation indicate that, in fact, multiple losses or gains are equally likely. An important implication of our findings is that, in grasshoppers, auditory tympana may have evolved before stridulation, supporting the argument that the original function of tympana may have been related not to conspecific communication but to predator detection. Overall, the results of this study emphasize the high information content of these minor groups (in this case, the four intermediate families under consideration contain only 0.2% of extant orthopteran species diversity). Our analyses also demonstrate the advantages of model-based methods in analyzing systematic problems and, in particular, of the importance of testing specific phylogenetic hypotheses when a priori support for groupings (e.g., from nonparametric bootstrapping) is marginal.

Synthesis and evaluation of some hydroxyproline-derived peptidomimetics as isoprenyltransferase inhibitors.

CA1A2X peptidomimetics containing a modified proline at position A2 were prepared and evaluated for their ability to inhibit farnesyltransferase (FTase) and geranylgeranyltransferase I (GGTase I) in enzymatic and cell-based assays. These compounds inhibited farnesylation of H-ras in vitro in the high nanomolar to low micromolar IC50 range.

Synthesis and evaluation of hydroxyproline-derived isoprenyltransferase inhibitors.

A series of peptidomimetics based on a hydroxyproline scaffold was prepared and evaluated for inhibition of farnesyltransferase and geranylgeranyltransferase I in both enzymatic and cell-based assays. A number of analogs were potent and selective inhibitors of FTase, while one compound (22) was nonselective in the enzymatic assays but eight-fold selective for inhibition of GGTase in the cellular assay (IC50 = 0.39 microM).

Combined molecular phylogenetic analysis of the Orthoptera (Arthropoda, Insecta) and implications for their higher systematics.

A phylogenetic analysis of mitochondrial and nuclear rDNA sequences from species of all the superfamilies of the insect order Orthoptera (grasshoppers, crickets, and relatives) confirmed that although mitochondrial sequences provided good resolution of the youngest superfamilies, nuclear rDNA sequences were necessary to separate the basal groups. To try to reconcile these data sets into a single, fully resolved orthopteran phylogeny, we adopted consensus and combined data strategies. The consensus analysis produced a partially resolved tree that lacked several well-supported features of the individual analyses. However, this lack of resolution was explained by an examination of resampled data sets, which identified the likely source of error as the relatively short length of the individual mitochondrial data partitions. In a subsequent comparison in which the mitochondrial sequences were initially combined, we observed less conflict. We then used two approaches to examine the validity of combining all of the data in a single analysis: comparative analysis of trees recovered from resampled data sets, and the application of a randomization test. Because the results did not point to significant levels of heterogeneity in phylogenetic signal between the mitochondrial and nuclear data sets, we therefore proceeded with a combined analysis. Reconstructing phylogenies under the minimum evolution and maximum likelihood optimality criteria, we examined monophyly of the major orthopteran groups, using nonparametric and parametric bootstrap analysis and Kishino-Hasegawa tests. Our analysis suggests that phylogeny reconstruction under the maximum likelihood criteria is the most discriminating approach for the combined sequences. The results indicate, moreover, that the caeliferan Pneumoroidea and Pamphagoidea, as previously suggested, are polyphyletic. The Acridoidea is redefined to include all pamphagoid families other than the Pyrgomorphidae, which we propose should be accorded superfamily status.

Dapsone at onset of diabetes lowers glycated hemoglobin and delays death in NOD mice.

Dapsone (4,4'-diaminodiphenyl sulfone) is a compound that has a large clinical experience due to its antimicrobial effects against mycobacterium leprae, the causative agent of leprosy. It is increasingly used in a number of clinical situations where inflammation may play an ancillary role. An inhibitory effect of the drug or lack thereof in the cumulative incidence or propagation of diabetes mellitus in the NOD mouse has mechanistic as well as therapeutic implications. We previously showed that dapsone administered continuously as a percentage of food to NOD mice inhibits the cumulative incidence of diabetes in a dose dependent fashion. In the present experiment, female NOD litter mates were randomized to receive dapsone (0.001% w/w as a percentage of food) at onset of diabetes. There were no differences in weight, blood glucose, or glycated hemoglobin at 10 weeks of age among the animals that were ultimately to receive dapsone (n = 10), mouse chow alone (n = 9), or those who did not develop diabetes (n = 3). The mean time to onset of diabetes, mean blood glucose at onset, and mean glycated hemoglobin at onset did not differ between animals who did or did not receive dapsone. Animals receiving dapsone had significantly (p < or = 0.03) lower glycated hemoglobin at weeks 2, 3, and 4 following the onset of diabetes and lived significantly longer following diagnosis of diabetes (7 vs. 4 weeks, p < 0.05). In conclusion, dapsone modulates the progression of autoimmune diabetes in the NOD mouse even when administered after the initiation of hyperglycemia.

Inferences about orthopteroid phylogeny and molecular evolution from small subunit nuclear ribosomal DNA sequences.

We determined DNA sequences of SSU rRNA genes in twenty-nine polyneopteran insect species and aligned these with homologues from eight other insects. In a phylogenetic analysis we recovered the classic divisions of Palaeoptera and Neoptera, with the latter divided into monophyletic Paraneoptera and Polyneoptera. The polyneopterans divided into three lineages: one includes the Grylloblattodea, Dermaptera and Plecoptera, the second contains the Blattodea, and the third (Orthopteroidea sensu Hennig) contains the Embiidina, Phasmida, and Orthoptera, in that order. The monophyly of the Orthoptera is supported by the analyses, as is the separation between taxa from its suborders Caelifera and Ensifera. The Caelifera are not always supported as a monophyletic group; the basal Tridactyloidea are separated from the rest of the Caelifera in some analyses. Inside of Tridactyloidea, the Acridoidea, Pamphagoidea, Pneumoroidea and Trigonopterygoidea are always recovered as a monophyletic group. We also examined the basal orthopteran relationships, with the specific aim of assessing the antiquity of the Ensifera. Character state reconstructions indicated that the ancestral ensiferan sequence is very similar to the ancestral orthopteran sequence. However, likelihood ratio tests rejected the null hypothesis of a molecular clock and we conclude that a change in substitution rate has occurred within the Orthoptera and several of the other polyneopteran orders. Similar observations have been made in holometabolous insects, suggesting that variation in substitution rate is a general feature of insect nuclear rRNA evolution.

The effectiveness of mitochondrial rRNA gene sequences for the reconstruction of the phylogeny of an insect order (Orthoptera).

We investigated the value of mitochondrial rRNA sequences for analyzing pre-Cainozoic divergence events in insects. Using small subunit and large subunit rRNA sequences from 38 orthopteroid species, we examined several aspects of sequence evolution including secondary structure, substitution rate, and base composition. Substitution matrices calculated from the two genes were very similar, though differences were detected in rates of C-T transitions between paired and unpaired positions in secondary structures. By contrast, extreme disparities between substitution frequencies at different phylogenetic levels make character-transition weighting essential in parsimony reconstruction. The analysis of base composition indicated that branch attraction of at least two important lineages was due to shared base composition biases and to long branch attraction. The importance of taxonomic sampling and sequence length for the effectiveness of phylogenetic recovery using the rRNA fragments was also assessed. Significantly, combining the two sequences seemed both justifiable and necessary for this taxonomic sample. However, for reconstructing deep branches of phylogeny, it seems that increasing either or both the number of taxa or nucleotide positions will not necessarily solve all problems. Instead, the resolution of ancient branching events using mtDNA sequences probably depends upon the development and application of better specified reconstruction methods.

The phylogeny of the Caelifera (Insecta, Orthoptera) as deduced from mtrRNA gene sequences.

Fragments of both mitochondrial ribosomal RNA genes of 32 caeliferan taxa (representing six of the seven superfamilies) and six outgroup Orthopteroids were sequenced. The combined alignment length was 630 bp after removal of all ambiguously aligned positions. Separation for the basal taxa was problematic and analysis using the LogDet transformation indicated that shared base composition biases were a confounding factor. The suborder Caelifera and all traditional caeliferan superfamilies except the Pamphagoidea are retrieved as monophyletic groups, though the Eumastacoidea lack significant bootstrap support. Of the traditional pamphagoid taxa, the Pamphagidae is embedded between classically acridid subfamilies, whereas Pyrgomorphidae is placed close to the Pneumoroidea. The morphological similarities of the Pyrgomorphidae and Pamphagidae may thus be homoplasic. A consensus tree based on five different methods of analysis indicated the following order: (Tridactyloidea, Tetrigoidea (Eumastacidae, Proscopiidae (Pneumoridae, Pyrgomorphidae (Acrididae + Pamphagidae)))).

Direct demonstration of geranylgeranylation and farnesylation of Ki-Ras in vivo.

It has recently been reported that Ki-Ras protein can be modified in vitro by farnesylation or geranylgeranylation. However, a previous analysis of Ki-Ras prenylation in vivo found only farnesylated Ki-Ras. In this report it is shown that under normal conditions, Ki-Ras is farnesylated in vivo and when cells are treated with the farnesyl transferase inhibitors B956 or B957, farnesylation is inhibited and Ki-Ras becomes geranylgeranylated in a dose dependent manner. These results have strong implications in the design of anticancer drugs based on inhibition of prenylation.

The sequence, organization, and evolution of the Locusta migratoria mitochondrial genome.

The sequencing of the cloned Locusta migratoria mitochondrial genome has been completed. The sequence is 15,722 bp in length and contains 75.3% A+T, the lowest value in any of the five insect mitochondrial sequences so far determined. The protein coding genes have a similar A+T content (74.1%) but are distinguished by a high cytosine content at the third codon position. The gene content and organization are the same as in Drosophila yakuba except for a rearrangement of the two tRNA genes tRNAlys and tRNAasp. The A+T-rich region has a lower A+T nucleotide content than in other insects, and this is largely due to the presence of two G+C-rich 155-bp repetitive sequences at the 5'end of this section and the beginning of the adjacent small rRNA gene. The sizes of the large and small rRNA genes are 1,314 and 827 bp, respectively, and both sequences can be folded to form secondary structures similar to those previously predicted for Drosophila. The tRNA genes have also been modeled and these show a strong resemblance to the dipteran tRNAs, all anticodons apparently being conserved between the two species. A comparison of the protein coding nucleotide sequences of the locust DNA with the homologous sequences of five other arthropods (Drosophila yakuba, Anopheles quadrimaculatus, Anopheles gambiae, Apis mellifera, and Artemia franciscana) was performed. The amino acid composition of the encoded proteins in Locusta is similar to that of Drosophila, with a Dayhoff distance twice that of the distance between the fruit fly and the mosquitoes. A phylogenetic analysis revealed the locust genes to be more similar to those of the Dipterans than to those of the honeybee at both the nucleotide and amino acid levels. A comparative analysis of tRNA orders, using crustacean mtDNAs as outgroups, supported this. This high level of divergence in the Apis genome has been noted elsewhere and is possibly an effect of directional mutation pressure having resulted in an accelerated pattern of sequence evolution. If the general assumption that the Holometabola are monophyletic holds, then these results emphasize the difficulties of reconstructing phylogenies that include lineages with variable substitution rates and base composition biases. The need to exercise caution in using information about tRNA gene orders in phylogenetic analysis is also illustrated. However, if the honeybee sequence is excluded, the correspondence between the other five arthropod sequences supports the findings of previous studies which have endorsed the use of mtDNA sequences for studies of phylogeny at deep levels of taxonomy when mutation rates are equivalent.

Inhibition of human tumor xenograft growth by treatment with the farnesyl transferase inhibitor B956.

ras oncogenes are present in several types of cancers but are most frequently described in colon and pancreatic carcinomas. Consequently, ras is being targeted for drug development as a means to develop therapies for these types of cancer. The ras protein is posttranslationally modified by the addition of a farnesyl group, followed by cleavage of the COOH-terminal 3 amino acids and methylation of the prenylated cysteine. Because the posttranslational addition of farnesyl is obligatory not only for the remaining modifications to take place but also for ras control of cell growth, inhibitors of farnesylation are being developed as potential antitumor agents. In this report, a new peptidomimetic inhibitor of farnesyl transferase is described. This compound, B956, and its methyl ester B1086, inhibit the formation of colonies in soft agar of 14 human tumor cell lines expressing different ras oncogenes at concentrations between 0.2 and 60 microM. Higher concentrations of B956 (10-80 microM) were required to inhibit colony formation by 5 tumor cell lines without ras mutations. B956/B1086 at 100 mg/kg also inhibited tumor growth by EJ-1 human bladder carcinoma, HT1080 human fibrosarcoma, and to a lesser extent by HCT116 human colon carcinoma xenografts in nude mice. Furthermore, inhibition of tumor growth by B956 is shown to be correlated with inhibition of ras posttranslational processing in the tumor. Thus, peptidomimetic inhibitors of ras farnesylation have the potential to be developed as therapy for ras-dependent tumors.

Peptidomimetic inhibitors of Ras farnesylation and function in whole cells.

The ras protooncogene is involved in regulation of cell growth. Mutations that activate the protein result in uncontrolled cell growth. Ras undergoes a series of posttranslational processing events, the first of which, farnesylation, is crucial for the function of the protein. Inhibitors of the farnesyltransferase enzyme are therefore potential candidates for the development of anticancer drugs. Tetrapeptides have been reported to be good inhibitors of this enzyme in vitro. We have synthesized analogs of the tetrapeptide Cys-Val-Phe-Met by replacement of the amino-terminal amide bonds. One inhibitor, B581, is permeable to the cell membrane. In the cell, it inhibits processing of two farnesylated proteins, H-ras and lamin A, but it does not inhibit processing of a geranylgeranylated protein, Rap 1A. Microinjection of B581 into frog oocytes inhibits maturation induced by activated, farnesylated H-ras but not maturation induced by activated, geranylgeranylated H-ras or by progesterone. These results demonstrate that this peptide mimic inhibits farnesylation selectively in the cell. The inhibition of farnesylation results in inhibition of H-ras function.

The organization of exteroceptive sensory inputs to interneurons of the flight neuropil in locusts.

1. Thirty-seven pairs of mesothoracic interneurons respond selectively to visual or ocellar stimuli corresponding to deviations from course in flight, expressed as angular rotation around the three spatial axes. 2. Sensitivities to roll and yaw are very strongly associated. All interneurons showing a directional preference for yaw rotations showed the same preference for roll rotations. A few roll-sensitive cells were not directionally sensitive to yaw. Some interneurons respond exclusively to pitch rotations, most to both pitch and roll/yaw. 3. Approximately equal numbers of interneurons prefer pitch up, pitch down, roll/yaw to the ipsilateral side and roll/yaw to the contralateral side. All four possible combinations of pitch (up or down) with roll/yaw (ipsilateral or contralateral) preferences occur with equal probability. 4. No relationship between neuronal structure and directional properties could be discerned. 5. The average latency of the ocellar EPSPs recorded in the interneurons is not significantly different from the average latency of the ocellar spike in the descending neurons (at the same temperature and in the same ganglion). The average ocellar IPSP latency is 8.5 ms longer. The data support the hypothesis that most EPSPs are derived from monosynaptic inputs from the DNs, and most IPSPs from polysynaptic inputs. A few EPSPs are also derived from polysynaptic inputs. 6. Most of these neurons are sensitive to wind, at least some directionally so, in a manner functionally compatible with their visual or ocellar directionality, and most are excited. Two neurons respond to movement of small objects in the visual field, and 5 to high frequency sound.

The nosocomial wound infection report. Its impact in the OR.

1. Reporting surgical wound infection rates to the OR nurse is valued and used as a feed-back mechanism to monitor aseptic technique in the operating room. 2. At the institution surveyed, the chief influences on OR personnel's surgical aseptic practice were the OR nurse, followed by the infection control nurse and then the attending surgeon. 3. If the clean surgical wound infection rate was above 3%, the OR personnel would review preoperative and intraoperative risk factors at the institution surveyed.

Microbial contamination of arterial infusions used for hemodynamic monitoring: a randomized trial of contamination with sampling through conventional stopcocks versus a novel closed system.

Arterial catheters are now commonly used to monitor blood pressure and obtain blood samples for arterial blood gas and other laboratory determinations. Stopcocks inserted into the pressure monitoring circuit have been the primary means of obtaining blood from arterial catheters. However, these stopcock systems have been associated with nosocomial contamination and bacteremias. Because of the problems of bacterial contamination and blood wasting with the stopcock sampling systems, we compared the frequency and extent of contamination of external sampling ports and the monitoring tubing fluid in stopcocks with that of a novel closed needle-sampling system (Lab-Site, Migada Ltd, Rehovot, Israel), incorporated into pressure monitoring tubing (Abbott Laboratories Inc., North Chicago, Illinois). We found that use of the novel sampling system resulted in significantly fewer episodes of internal bacterial contamination of the arterial monitoring line (7%) than did the use of a stopcock system (61%). External contamination of the sampling port was also lower in the novel system (8%) than in the stopcock system (37%). This suggests that the closed system may reduce the risk of nosocomial infections in patients requiring arterial pressure monitoring.

The taxonomy of invertebrate neurons: a plea for a new field.

It is generally accepted that the 'identified cell' concept and its practical application are responsible for the appeal of invertebrate preparations (and perhaps in the near future of certain vertebrate preparations too) for general neurobiology. The considerable number of neurons contained in many such preparations, and the number of investigations to which they are subjected, has led to a minor crisis: it is increasingly hard to determine whether a given neuron has been previously described, and if so, under what name. The taxonomy of identified interneurons requires a far more serious and rigorous approach than has so far been the rule. The main proposals made in this article are for the application of the normal procedural rules of classical organism taxonomy, which are highly applicable to the neural situation, and for a standardized nomenclature. Also recommended is the establishment of networked computer data bases for each of the popular invertebrate preparations (e.g. locusts, leeches) and of international committees for their supervision, and the increased use of confocal fluorescence microscopy to increase the amount of anatomical data which can be gathered from a normal physiological preparation.