Second nationwide anti-tuberculosis drug resistance survey in Namibia.

SETTING: Namibia ranks among the 30 high TB burden countries worldwide. Here, we report results of the second nationwide anti-TB drug resistance survey.OBJECTIVE: To assess the prevalence and trends of multidrug-resistant TB (MDR-TB) in Namibia.METHODS: From 2014 to 2015, patients with presumptive TB in all regions of Namibia had sputum subjected to mycobacterial culture and phenotypic drug susceptibility testing (DST) for rifampicin, isoniazid, ethambutol and streptomycin if positive on smear microscopy and/or Xpert MTB/RIF.RESULTS: Of the 4124 eligible for culture, 3279 (79.5%) had Mycobacterium tuberculosis isolated. 3126 (95%) had a first-line DST completed (2392 new patients, 699 previously treated patients, 35 with unknown treatment history). MDR-TB was detected in 4.5% (95%CI 3.7-5.4) of new patients, and 7.9% (95%CI 6.0-10.1) of individuals treated previously. MDR-TB was significantly associated with previous treatment (OR 1.8, 95%CI 1.3-2.5) but not with HIV infection, sex, age or other demographic factors. Prior treatment failure demonstrated the strongest association with MDR-TB (OR 17.6, 95%CI 5.3-58.7).CONCLUSION: The prevalence of MDR-TB among new TB patients in Namibia is high and, compared with the first drug resistance survey, has decreased significantly among those treated previously. Namibia should implement routine screening of drug resistance among all TB patients.

Impact of dietary phosphorous in diploid and triploid Atlantic salmon (Salmo salar L.) with reference to early skeletal development in freshwater.

In order to assess the effect of dietary phosphorus (P) in reducing vertebral malformations and improving freshwater (FW) performance in triploid Atlantic salmon (Salmo salar), both triploid and diploid Atlantic salmon were fed three different dietary P inclusion levels (low: 4.9, medium: 7.7, and high: 9.7 g available P kg-1) from first feeding until smolt. Somatic and skeletal response was assessed at fry (~0.5 g), parr (~5 g) and smolt (~45 g) stages. Triploid parr initially grew faster on the high P diet, while groups fed low P resulted in a significantly higher weight at smolt. Image analysis of double stained Alcian blue and Alizarin red S fry revealed that low P fed triploid fish presented less well mineralised vertebrae, and significantly more malformed vertebrae in both parr and smolt stages following x-ray radiographic assessment. Triploid parr fed high and medium P had similar numbers of malformed vertebrae relative to their diploid counterparts but greater numbers than at smolt. Low P fed triploids had the highest prevalence of jaw and vertebral malformations as well as the highest number of deformed vertebrae in the central caudal vertebral region, which was more pronounced at parr than at smolt. Shorter vertebrae dorso-ventral lengths were observed throughout the spinal column (R1-R4) in parr fed low P and only in the caudal region (R3) at smolt. In parr, both ploidies showed reduced phosphate homeostasis protein fgf23 gene expression in vertebrae when fed low P diets, while triploids showed greater down-regulation of osteogenic factors (alp, opn and igf1r) between diets relative to diploids, suggesting possible greater active suppression of mineralisation and reduced osteogenic potential in triploids. No effects of diet or ploidy on gene expression were evident at smolt. Comparisons between development stages suggest early P supplementation in triploids is crucial for skeletal development. Ultimately, reducing vertebral deformities observed at smolt with higher P supplementation in triploids could contribute towards improving skeletal performance and welfare of the stocks in the marine phase.

A nutritionally-enhanced oil from transgenic Camelina sativa effectively replaces fish oil as a source of eicosapentaenoic acid for fish.

For humans a daily intake of up to 500 mg omega-3 (n-3) long-chain polyunsaturated fatty acids (LC-PUFA) is recommended, amounting to an annual requirement of 1.25 million metric tonnes (mt) for a population of 7 billion people. The annual global supply of n-3 LC-PUFA cannot meet this level of requirement and so there is a large gap between supply and demand. The dietary source of n-3 LC-PUFA, fish and seafood, is increasingly provided by aquaculture but using fish oil in feeds to supply n-3 LC-PUFA is unsustainable. Therefore, new sources of n-3 LC-PUFA are required to supply the demand from aquaculture and direct human consumption. One approach is metabolically engineering oilseed crops to synthesize n-3 LC-PUFA in seeds. Transgenic Camelina sativa expressing algal genes was used to produce an oil containing n-3 LC-PUFA to replace fish oil in salmon feeds. The oil had no detrimental effects on fish performance, metabolic responses or the nutritional quality of the fillets of the farmed fish.

Maternal and perinatal mortality in ward A, Ikosi-Isheri LCDA in Lagos State, Nigeria: from "guestimates" to near actuals.

BACKGROUND: The problem of authentic statistics on maternal and peri-natal mortality has persisted in Nigeria probably because of inadequate involvement of community members in data management. OBJECTIVE: The objective of this study was to improve the accuracy of information on pregnancy outcome in a ward in Lagos state. METHODS: Residents of Ward A of Ikosi-Isheri Local Council Development Area (LCDA) of Lagos State were educated on the importance of accurate health data. Youths selected based on criteria and community-based health care providers, following refresher training on research methods, assisted with the collection, analysis and presentation of data. RESULTS: The records of churches, mosques, Traditional Birth Attendants, the government owned Primary Health Care (PHC) centre and 7 private health facilities in the ward reported no (0) maternal mortality, but 14 out of 633 live births in the latter died, giving a peri-natal mortality of 22.1 per thousand in one year. However, neither the CDAs, nor LCDA nor the state received regular reports on pregnancy outcome. CONCLUSION: Under-reporting or over-estimation of pregnancy outcome can be reduced by involvement of the leaders of Residents Associations and other relevant community members in data management. Data should be collected by smaller units into which a ward is divided and collated at the ward level for transmission to the LGA/LCDA. Feedback to community members will improve awareness about the importance of appropriate antenatal, natal and peri-natal care. Health care providers are also more likely to take prompt action to prevent maternal and peri-natal mortality.

A randomized trial comparing the efficacy of four treatment regimens for uncomplicated falciparum malaria in Assam state, India.

A four-arm drug sensitivity study compared chloroquine, sulfadoxine-pyrimethamine (SP), mefloquine and mefloquine-artesunate in Sonitpur and Karbi Anglong districts in Assam state, India. Two criteria were used to ascertain outcome: success of clinical treatment and parasitologic cure. In Sonitpur, at 14 days, there were 36/56 early and late treatment failures plus late parasitologic failures to chloroquine and 16/56 for SP. In Karbi Anglong, combined treatment failure at 14 days was 16/56 to chloroquine and 8/60 to SP. Mefloquine and mefloquine-artesunate demonstrated 93.9% and 93.6% sustained responses respectively at 42 days. High failure rates to both chloroquine and SP preclude the use of these drugs as first-line treatment for uncomplicated falciparum malaria in this region. A mefloquine-artesunate combination presents an effective alternative utilizing the currently recommended higher dose of mefloquine.

Histidine nutrition and genotype affect cataract development in Atlantic salmon, Salmo salar L.

The aim of this study was to investigate effects of dietary levels of histidine (His) and iron (Fe) on cataract development in two strains of Atlantic salmon monitored through parr-smolt transformation. Three experimental diets were fed: (i) a control diet (CD) with 110 mg kg(-1) Fe and 11.7 g kg(-1) His; (ii) CD supplemented with crystalline His to a level of 18 g kg(-1) (HD); and (iii) HD with added iron up to 220 mg kg(-1) (HID). A cross-over design, with two feeding periods was used. A 6-week freshwater (FW) period was followed by a 20-week period, of which the first three were in FW and the following 17 weeks in sea water (SW). Fish were sampled for weighing, cataract assessment and tissue analysis at five time points. Cataracts developed in all groups in SW, but scores were lower in those fed high His diets (P < 0.05). This effect was most pronounced when HD or HID was given in SW, but was also observed when these diets were given in FW only. Histidine supplementation had a positive effect on growth performance and feed conversion ratio (P < 0.05), whereas this did not occur when iron was added. Groups fed HD or HID had higher lens levels of His and N-acetyl histidine (NAH), the latter showing a marked increase post-smoltification (P < 0.05). The HD or HID groups also showed higher muscle concentrations of the His dipeptide anserine (P < 0.05). There was a strong genetic influence on cataract development in the CD groups (P < 0.001), not associated with tissue levels of His or NAH. The role of His and His-related compounds in cataractogenesis is discussed in relation to tissue buffering, osmoregulation and antioxidation.

Auditory event-related potentials in the assessment of auditory processing disorders: a pilot study.

The aim of this pilot study was to investigate whether children with a suspected auditory processing disorder (sAPD) in the presence of normal hearing, differ significantly from normal age-matched controls on particular parameters of auditory event-related potentials. We assessed nine children (mean age 9.5 years) in whom the clinical profile and the results in a screening test for auditory processing disorder (SCAN/SCAN-A) suggested the presence of an auditory processing disorder, and nine age-matched normal control subjects, using auditory event-related potentials (ERP) to phonemes/ba/(standard) and/da/(deviant). Analysis of the auditory ERP recordings revealed an enlarged P85 - 120 and attenuated N1 and P2 in all sAPD children compared to controls. We also found significantly increased N1 peak latency, and a larger peak to peak amplitude of the P85 - 120-N1 and P2-N2 and smaller peak to peak amplitude of the N1-P2 in the sAPD children. Subtraction of the standard auditory ERP from the deviant revealed a mismatch negativity with no significant differences in duration, peak or onset latency between the control subjects and sAPD. Our results indicate that neurophysiological measures may identify a group of children with specific problems suggestive of an auditory processing disorder in the absence of an obvious structural or functional lesion who warrant further study in order to assess whether these findings reflect delayed CNS myelination.

Replacement of fish oil with rapeseed oil in diets of Atlantic salmon (Salmo salar) affects tissue lipid compositions and hepatocyte fatty acid metabolism.

Duplicate groups of Atlantic salmon post-smolts were fed five practical-type diets in which the added lipid was 100% fish oil [FO; 0% rapeseed oil (0% RO)], 90% FO + 10% RO (10% RO), 75% FO + 25% RO (25% RO), 50% FO + 50% RO (50% RO) or 100% RO, for a period of 17 wk. There were no effects of diet on growth rate or feed conversion nor were any histopathological lesions found in liver, heart, muscle or kidney. The greatest accumulation of muscle lipid was in fish fed 0% RO, which corresponded to significantly lower muscle protein in this group. The highest lipid levels in liver were found in fish fed 100% RO. Fatty acid compositions of muscle lipid correlated with RO inclusion in that the proportions of 18:1(n-9), 18:2(n-6) and 18:3(n-3) all increased with increasing dietary RO (r = 0.98-1.00, P < 0.013). The concentrations of eicosapentaenoic acid [20:5(n-3)] and docosahexaenoic acid [22:6(n-3)] in muscle lipid were significantly reduced (P < 0.05), along with total saturated fatty acids, with increasing dietary RO. Diet-induced changes in liver fatty acid compositions were broadly similar to those in muscle. Hepatic fatty acid desaturation and elongation activities, measured using [1-(14)C] 18:3(n-3), were increased with increasing dietary RO. Limited supplies of marine fish oils require that substitutes be found if growth in aquaculture is to be maintained such that fish health and product quality are not compromised. Thus, RO can be used successfully as a substitute for fish oil in the culture of Atlantic salmon in sea water although at levels of RO >50% of dietary lipid, substantial reductions occur in muscle 20:5(n-3), 22:6(n-3) and the (n-3)/(n-6) polyunsaturated fatty acid (PUFA) ratio, which will result in reduced availability of the (n-3) highly unsaturated fatty acids that are beneficial for human health.

Extensive metallosis and necrosis in failed prostheses with cemented titanium-alloy stems and ceramic heads.

We describe three prostheses with cemented titanium-alloy stems and Al2O3 ceramic femoral heads which had to be revised after a mean period of implantation of 78 months. In each case, the neck of the prosthesis had been so severely worn that the profile was elliptical rather than circular. There was severe metallosis of the periprosthetic tissues. Metal particles isolated from the tissues were approximately one nanometre in size and the ratios of titanium, aluminium and vanadium in the particles were the same as in the original alloy. Histologically, the high concentration of metal particles masked the presence of high-density polyethylene (HDP) debris, but again particles about one nanometre in size were isolated from the tissues. The severe necrobiosis and necrosis noted were consistent with other reports of the presence of extensive wear particles in periprosthetic tissues. Wear is presumed to have occurred as a result of mismatch between the shape or size of the taper cone and the femoral head, or to changes in the geometry of loading due to migration of the cup. To facilitate early intervention, patients with this design of prosthesis should be monitored radiologically.

EEA1, an early endosome-associated protein. EEA1 is a conserved alpha-helical peripheral membrane protein flanked by cysteine "fingers" and contains a calmodulin-binding IQ motif.

Early endosomes are cellular compartments receiving endocytosed material and sorting them for vesicular transport to late endosomes and lysosomes or for recycling to the plasma membrane. We have cloned a human cDNA encoding an evolutionarily conserved 180-kDa protein on early endosomes named EEA1 (Early Endosome Antigen1). EEA1 is associated with early endosomes since it co-localizes by immunofluorescence with the transferrin receptor and with Rab5 but not with Rab7. Immunoelectron microscopy shows that it is associated with tubulovesicular early endosomes containing internalized bovine serum albumin-gold. EEA1 is a hydrophilic peripheral membrane protein present in cytosol and membrane fractions. It partitions in the aqueous phase after Triton X-114 solubilization and is extracted from membranes by 0.3 M NaCl. It is a predominantly alpha-helical protein sharing 17-20% sequence identity with the myosins and contains a calmodulin-binding IQ motif. It is flanked by metal-binding, cysteine "finger" motifs. The COOH-terminal fingers, Cys-X2-Cys-X12-Cys-X2-Cys and Cys-X2-Cys-X16-Cys-X2-Cys, are present within a region that is strikingly homologous with Saccharomyces cerevisiae FAB1 protein required for endocytosis and with Caenorhabditis elegans ZK632. These fingers also show limited conservation with S. cerevisiae VAC1, Vps11, and Vps18p proteins implicated in vacuolar transport. We propose that EEA1 is required for vesicular transport of proteins through early endosomes and that its finger motifs are required for this activity.

Enamel surfaces after orthodontic bracket debonding.

The enamel surfaces of extracted teeth were studied clinically and with a scanning electron microscope following debonding of orthodontic attachments and subsequent polishing. Excess orthodontic resin was removed with tungsten carbide burs and abrasive discs. Several combinations of polishing agents were evaluated. The no. 30 fluted tungsten carbide bur appeared to be the most efficient method of removing highly filled resin, and it produced the least amount of scarring. A polishing sequence was developed which used resin points and cups followed by a water slurry of fine pumice and brown and green cups. This procedure was tested clinically and appeared to return the enamel to an acceptable condition. This procedure is fast, efficient, and comfortable for the patient.

Insulin-like growth factor-I and its association with binding proteins in bovine milk.

Immunoreactive insulin-like growth factor-1 (IGF-I in bovine milk was quantified. IGF-I was principally assoication with an approximately 45 kDa binding protein. In addition, a small fraction of IGF-I occurred at a molecular weight approximately the same as that of unbound IGF-I. Available binding sites existed on the approximately 45 kDa binding protein. Bound IGF-I was readily dissociated from binding protein by acid treatment. When IGF-I was estimated in milk obtained from primiparous and multiparous cows, mutiparous cows had a higher concentration (40 nmol/1) [corrected] at parturition than primiparous cows (19.2 nmol/1) [corrected]. By day 2 of lactation, IGF-I concentrations were 30 and 50% of initial estimates for multiparous and primiparous cows respectively. the final IGF-I concentration, on day 56 of lactation, was 4.5 nmol/1 [corrected] for combined parity groups. At parturition in multiparous cows, the mass of IGF-I was estimated at 183 and 157 nmol [corrected] for blood and milk pools respectively. Milk, therefore, represents a substantial pool of IGF-I in the cow. The mechanism of the appearance of IGF-I in bovine milk is unknown.

B cell stimulatory factor 1 induces lobster agglutinin 1-separated mouse thymocytes to express cytotoxic T lymphocyte activity.

Mouse thymocytes low in surface sialic acid were prepared by using the lectin lobster agglutinin 1 (LAg1). These LAg1-thymocytes do not become CTL when incubated with Con A or Con A plus mouse rIL-2, whereas unseparated thymocytes and thymocytes with high levels of surface sialic acid develop good levels of polyclonal CTL activity under these conditions. However, LAg1- thymocytes developed high levels of CTL activity when incubated with B cell stimulatory factor-1 (BSF-1), provided as the supernatant of the rBSF-1-secreting T cell hybridoma D9-C1.12.17. Affinity-purified BSF-1 from D9-C1 supernatant and rBSF-1 also stimulated these cells to become CTL, but they were not as active as the D9-C1 supernatant. The ability of D9-C1 supernatant and of affinity-purified BSF-1 to induce CTL activity was inhibited by the anti-BSF-1 mAb 11B11. Moreover, this mAb inhibited the ability of 24-h Con A-stimulated spleen cell supernatant to induce these cells to express CTL activity. 11B11 also inhibited LAg1+ thymocytes from becoming CTL when stimulated with Con A alone. These experiments suggest that BSF-1 is required for LAg1- and LAg1+ thymocytes to become CTL.

Surface coil phosphorus-31 nuclear magnetic resonance studies of the intact eye.

The feasibility of employing the surface coil probe technique for the non-invasive study of ocular tissue metabolism by phosphorus-31 nuclear magnetic resonance spectroscopy (31P NMR) in enucleated bovine, rabbit, human and rat globes is demonstrated. An assessment of individual phosphorus-metabolite contributions from ocular tissues, including the cornea, lens and iris, to the overall 31P NMR spectrum (NMR spectral acquisition parameters optimized for the lens region of the globe) was accomplished through the combination of surgical ablation and difference spectroscopy. The NMR measurements also provided tissue pH values for the lens and cornea. The strengths and limitations of the surface coil NMR method, which is particularly appropriate for in vivo metabolic studies of ocular tissues such as the lens, are discussed.

Studies on the structure and carbohydrate binding properties of lobster agglutinin 1 (LAg1), a sialic acid-binding lectin.

Lobster agglutinin 1 (LAg 1) was isolated from the hemolymph of the American lobster (Homarus americanus) by a sequential combination of ammonium sulfate precipitation, preparative starch block electrophoresis, gel filtration and affinity chromatography on Sepharose-Fetuin and Sepharose-Colominic acid columns. Two types of protomeric structures with molecular weights of 700 and 500 Kilodaltons respectively were isolated. These molecules are composed of noncovalently held subunits with a molecular weight of 70 Kilodaltons. Analysis of preparations by double immunodiffusion, polyacrylamide gel electrophoresis and isoelectrofocusing indicates that the LAg 1 obtained was a single molecular species. Hemagglutination inhibition experiments indicated that the best inhibitors were bovine mucin, glycophorin, fetuin and human IgM in that order. The desialylated forms of some of these proteins still bound lectin, although to a lesser degree than their intact sialylated counterparts. Affinity chromatography experiments indicated that LAg 1 binds to N-acetylneuraminic acid, N-acetylglucosamine and N-acetylgalactosamine. LAg 1 does not contain sialic acid nor neuraminidase activity: oligosaccharides associated with it appear to be either of the oligomannosyl or biantennary type. The sialic acid binding specificity of this lectin was used to separate immature mouse thymocytes (low sialic acid content) from mature thymocytes (high sialic acid content).

Self-reported dark red bleeding as a marker comparable with occult blood testing in screening for large bowel neoplasms.

The study was designed to determine the prevalence of large bowel symptoms in an adult working population and to assess their value in screening for large bowel neoplasms. A symptom questionnaire was sent to workers in two large organizations and the results were compared with faecal occult blood testing in the same individuals using Haemoccult (Eaton Laboratories). Completed symptom questionnaires and Haemoccult test kits were returned by 916 out of a total of 1805 employees over the age of 40 (compliance 50.7 per cent). Twenty-eight (3.1 per cent) were Haemoccult positive and 114 (12.4 per cent) had one or more symptoms. All positives (129 persons) were examined by flexible sigmoidoscopy and barium enema. No cancer was found but 7 patients with adenomas greater than 10 mm diameter were discovered. Each of the 7 patients reported at least one symptom (dark red bleeding in 4, bright red bleeding in 2 and diarrhoea in 1) and 6 were Haemoccult positive. There was no individual with a Haemoccult positive adenoma without symptoms. Predictive values for adenomas over 10 mm for Haemoccult positive tests (21 per cent), self-reported dark bleeding (16 per cent) and diarrhoea (17 per cent), were significantly higher than for other symptoms. The predictive value rose significantly to 46 per cent for Haemoccult-positive patients who in addition had at least one symptom (P less than 0.05) and to 57 per cent for Haemoccult positive with dark bleeding. These combinations of Haemoccult- and symptom-positive results increased the specificity of 97.6 per cent for Haemoccult alone to values over 99 per cent without reducing sensitivity.(ABSTRACT TRUNCATED AT 250 WORDS)