Gastric dilation and air sacculitis in Chinook salmon, Oncorhynchus tshawytscha (Walbaum); correlation of macroscopic and microscopic lesions, and relationship of the syndrome to glomerulonephritis and serum biochemistry.

Macroscopic and microscopic assessment procedures were developed to evaluate the severity and enable diagnosis from histological samples, of gastric dilation and air sacculitis (GDAS) in Chinook salmon, Oncorhynchus tshawytscha. Stomachs and swim bladders were examined from young fish with experimentally induced GDAS and from larger fish with the syndrome held in commercial saltwater net-pens. Fish fed a diet previously known to induce GDAS had significantly wider stomachs with decreased prominence of longitudinal stomach folds that contained larger amounts of fluid (P < 0.001), and thinner stomach walls with greater inter-nerve distances (P < 0.001), than fish fed an alternative commercial diet not associated with the syndrome. These fish also had swim bladders that were more likely to be opaque and contain more fluid (P < 0.001). These observations correlated well with selected criteria for stomach tissue (P < 0.002) and swim bladder (P < 0.04) that could be evaluated microscopically. Four stomach measurements, primarily measurements of wall or partial wall thickness and inter-nerve distances, were suitable for discriminating between affected and non-affected fish. A stomach width ratio, that was independent of fish weight and highly correlated with macroscopic stomach measurements (P < 0.0001), was particularly useful; this ratio was derived from the distances between both the outer border of the muscularis mucosa and mesothelium of the serosal surface to the stratum compactum. Serum biochemistry parameters (osmolality, calcium and magnesium) did not differ between fish fed different diets, but serum creatinine concentration was correlated with the microscopic thickness of the muscularis externa of the stomach wall and the total stomach thickness (P < 0.001 and P < 0.003, respectively). A glomerulonephritis was also noted in these fish. The severity of the lesion was not significantly related to GDAS nor to any serum biochemistry parameter assayed however.

Diagnosis of tuberculosis in ghana: the role of laboratory training.

OBJECTIVES: The laboratory is considered the cornerstone of tuberculosis (TB) control programme. International review of Ghana's programme in the late nineties identified the laboratory services as the weakest component. Sputum smear microscopy (SSM) being the main method of diagnosing pulmonary TB in Ghana, the training objectives were to: (i) strengthen the knowledge and skills of laboratory personnel on SSM (ii) impart necessary techniques in biosafety and (iii) introduce a Quality Assurance (QA) system in order to strengthen SSM services. METHODS: Personnel were selected for training during a nationwide situation analysis of SSM centres in 2000/2001. Four training sessions on SSM/QA were held between 2001/2004. RESULTS: A total of 80 personnel were trained: 10 regional TB coordinators and 70 laboratory personnel. The participants upon return to their respective regions also organized training within their districts. This approach resulted in another 100 district TB coordinators and 200 laboratory personnel being trained. Improvement in smear preparation, staining and reading ability of the participants were observed during the post-test and subsequent visit to their respective laboratories. The training has led to strengthening of TB laboratory services in the country and has contributed to increase in case detection from 10,745 in 2000 to 11,827 in 2004 and 14,022 in 2008. It was observed during the post-training follow-up and quarterly supervision visits that morale of the personnel was high. CONCLUSION: Continuous training and re-training of laboratory personnel on SSM and QA at regular intervals do play an important role for effective and efficient TB control programme.

An immune-complex glomerulonephritis of Chinook salmon, Oncorhynchus tshawytscha (Walbaum).

Chinook salmon from New Zealand were shown to have a generalized membranous glomerulonephritis that was most severe in large fish. Marked thickening of the glomerular basement membrane was the most consistent lesion, with the presence of an electron-dense deposit beneath the capillary endothelium.Severely affected glomeruli also had expansion of the mesangium and loss of capillaries,synechiae of the visceral and parietal epithelium and mild fibrosis of Bowmans capsule. Chinook salmon from British Columbia, Canada with bacterial kidney disease caused by Renibacterium salmoninarum had similar histological lesions. They also had thickened glomerular basement membranes that were recognized by rabbit antiserum to rainbow trout immunoglobulin. This was true only when frozen sections of kidney were used and not formalin-fixed tissue. An attempt to experimentally produce a glomerulopathy in rainbow trout by repeated immunization with killed R. salmoninarum was not successful. Case records from the Fish Pathology Laboratory at the University of Guelph over a 10-year period revealed that a range of species were diagnosed with glomerulopathies similar to those seen in Chinook salmon. The majority of these cases were determined to have chronic inflammatory disease. This report has identified the presence of immunoglobulin within thickened basement membranes of Chinook salmon with glomerulonephritis and supports the existence of type III hypersensitivity in fish.

Mycobacterial species causing pulmonary tuberculosis at the korle bu teaching hospital, accra, ghana.

SUMMARY OBJECTIVE: Characterize mycobacterial species causing pulmonary tuberculosis (PTB) at the Korle-Bu Teaching Hospital in Ghana. DESIGN: Sputum smear positive samples, two (2) from 70 patients diagnosed as having tuberculosis, after they had consented, were collected from the Korle-Bu Teaching Hospital Chest Clinic between January and July 2003. SETTING: Korle-Bu Teaching Hospital Chest Clinic, Accra. RESULTS: Sixty-four mycobacterial isolates were obtained and confirmed as members of Mycobacterium tuberculosis complex by colonial morphology and conventional biochemical assays. Forty-seven (73%) were M. tuberculosis, the human strain, 2 (3%) M. bovis, the bovine strain, 13 (20%) M. africanum I (West Africa type), and 2 (3%) M. africanum II (East Africa type). CONCLUSION: The results indicate that, there are various strains causing PTB at the Korle-Bu Teaching Hospital and of great concern is M. bovis, which mostly causes extra-PTB in humans but found to cause PTB in this study. This calls for the need to conduct a nationwide survey using both conventional and molecular techniques to characterize various mycobacterial species causing TB in Ghana. This will result in better understanding of the various strains circulating in the country and inform individual TB treatment regimen especially the inclusion or exclusion of pyrazinamide.

Situation analysis of TB microscopy centres in Ghana.

SETTING: Public health laboratories in Ghana performing tuberculosis (TB) microscopy. OBJECTIVE: To assess the situation of the laboratories in terms of staff strength, technical skills, documentation, biosafety practices, equipment, supplies and disposal systems. DESIGN: Methods used for data collection were interviews using a structured questionnaire, informal observation of laboratory registers, disposal systems and safety measures for sputum handling. RESULTS: Of 114 laboratories visited between 2000 and 2001, 102 (89.5%) were performing TB microscopy. Of the staff working in the laboratories, 9% were medical technologists, 24% laboratory technicians, 37% laboratory assistants and 30% orderlies. Average false-negative and -positive rates were respectively 13% and 14%. Although most of the centres (85.3%) were using the recommended TB laboratory register for recording, in most cases they were not filled in accurately or completely. The majority of the available microscopes had mechanical or optical faults. Availability of other materials for smear preparation and staining ranged from 44% to 82%. The main methods employed for disposal of laboratory waste were burning and burying, but conditions were poor in most of the facilities visited. CONCLUSION: Training of laboratory personnel in TB microscopy and establishment of a quality assurance system are needed in Ghana.

Improving the laboratory diagnosis of TB in Ghana: the impact of a quality assurance system.

SETTING: Greater Accra region, Ghana. OBJECTIVE: To establish a pilot quality assurance (QA) system in sputum smear microscopy and to evaluate its impact. DESIGN: Quarterly supporting visits were paid to participating laboratories between 2000 and 2002. Fifteen examined slides were selected randomly from each laboratory during the visits and blindly re-assessed. Feedback was given promptly to the various laboratories. Training and stakeholder workshops were organised whenever necessary. RESULTS: General improvements in smear preparation and staining as well as the reading ability of the laboratory personnel included in the study were observed. The average marks for specimen quality, staining ability, smear cleanness, thickness, size and evenness increased from 64%, 79%, 69%, 46%, 67% and 60% in the last quarter of 2000 to 81%, 90%, 86%, 79%, 80% and 74%, respectively, 24 months after the establishment of the QA system. Within the same period, the rate of false-positives and -negatives decreased from respectively 14.8% and 20.5% to 0%, and agreements in positivity grade increased from 74% to 95%. The performance of the participating laboratories in keeping the laboratory registers up to date also improved. CONCLUSION: The QA system needs to be extended to the rest of the country.

Isolation and partial characterization of a calcium-dependent lectin (chiletin) from the haemolymph of the flat oyster, Ostrea chilensis.

A calcium-dependent lectin (chiletin) was isolated from oyster haemolymph by mannose elution from Sepharose CL-6B followed by anion exchange chromatography. Chiletin was predominantly composed of 12 and 24 kDa bands when examined with SDS-PAGE under reducing and non-reducing conditions, respectively. Larger molecular weight bands of 36 and 50 kDa were also variably present under reducing conditions. The NH2-terminal sequence of the 24 kDa band was determined and was not homologous to any known protein from the databases searched. Isolated chiletin was composed of multiple isomers approximately 12 kDa in size and ranging in pI from 5.2 to 6.0. Rabbit antiserum was raised to a synthetic peptide coupled to keyhole limpet hemocyanin and the size of the chiletin subunits was confirmed by Western blot. Two and five different conformational aggregates of chiletin were resolved in oyster haemolymph using size exclusion chromatography in 8 M urea and PBS, respectively. The largest aggregate obtained from size exclusion in 8 M urea was estimated to be greater than 640 kDa. The ability of whole haemolymph and isolated chiletin to agglutinate sheep red blood cells was inhibited by galactose and mannose. Chiletin was identified by immunohistochemistry to be most consistently present in the auricle, followed by the digestive gland, however staining was seen sporadically in haemocytes, gastrointestinal epithelium and interstitial connective tissue cells.

Experimental production of gastric dilation and its association with osmoregulatory stress and biogenic amines in chinook salmon, Oncorhynchus tshawytscha (Walbaum).

Chinook salmon smolt in fresh water fed a commercial diet known to produce minimal gastric dilation and air sacculitis (GDAS) were randomly assigned to four experimental tanks with flow-through sea water. All four groups were acclimatized to sea water for 3 weeks and fed a diet of minced fresh seafood. After 3 weeks the groups were fed either; seafood as before, a different commercial pelleted diet associated with the development of GDAS on farms, or either diet supplemented with 500 mg L(-1) putrescine, 300 mg L(-1) cadaverine and 250 mg L(-1) tyramine. Gastric dilation was produced in fish fed the commercial diet for 1 month but not by feeding a diet of minced seafood. The addition of putrescine, cadaverine and tyramine to either diet had no significant effect on the development of gastric dilation. Fish fed the commercial diet had significantly (P < 0.0001) wider weight-adjusted stomach widths, less prominent longitudinal stomach folds (P < 0.0001) and lower (P < 0.0001) stomach-width ratios than fish fed the fresh seafood diet. There was no significant difference in serum osmolality or sodium concentration between fish from groups with or without gastric dilation or fed biogenic amines.

[Development of a questionnaire to measure the effect of urinary incontinence on female sexuality: a reliability and validity study].

OBJECTIVES: This study developed a measure of the degree of the effect of urinary incontinence on female sexuality and tested the reliability and validity of the developed measure. Effect of Urinary Incontinence on Sexuality Questionnaire (EISQ). METHODS: The EISQ was developed from a thorough literature review and interviews of 20 individuals with urinary incontinence who said their incontinence had affected their dyadic relationship. These items were submitted to an analysis of content validity. This work resulted in a 28 items questionnaire that used a 4-point Likert-type response scale. The reliability and validity of the EISQ was tested with 144 women with incontinence who visited out-patient clinic in six hospitals. RESULTS: Cronbach's alpha as the index of internal consistency was high. The EISQ total was 0.90 and alpha ranged from 0.70 to 0.93 for the six subscales. Six factors each with an eigenvalue greater-than-or-equal-to-one were extracted from a principal components factors analysis with varimax rotation. The cumulative percentage of variance explained by this solution was 69.0%. Concurrent validity analyses confirmed our predictions that the EISQ scores were related to Patient Incontinence Stress (ISQ-P) scores. CONCLUSION: The EISQ proved to be reliable and valid as a self-administered measure for assessing the effect of urinary incontinence on female sexuality. The EISQ has demonstrated sufficient psychometric properties to warrant continued study of other samples and to explore the relationship between EISQ and other outcome variables.

[Experimental retinal branch vein occlusion. 4. Pathological changes in the middle and late stage].

Retinal branch vein occlusion in macaca monkeys (Macaca irus) was produced by dye laser photocoagulation, and observed histopathologically from five days to one year after photocoagulation. Ten days later, retinal edema and hemorrhage observed at an early stage gradually decreased. Two weeks later, capillary bed closure areas were observed in fluorescein angiography. The capillary closure was not reversible when disturbance of the retinal circulation continued for more than three or four days after photocoagulation. Three months later, dry retina was observed in the capillary bed closure areas. One year later, the retina was severely degenerated and thinned. In these retinal areas, capillary lumens observed microscopically were occluded by cellular components. Sheathing of large veins was observed in these retinal areas. The walls of these large veins were thick and fell into hyaline degeneration. Their lumens were narrowed or obstructed. During the period of observation, retinal neovascularization was not observed.

[Recanalization of venule following experimental retinal branch vein occlusion].

Retinal branch vein occlusion was experimentally produced in monkey (macaca irus) by dye laser photocoagulation. Venous dilatation, retinal edema and punctate hemorrhages were the ophthalmoscopic changes immediately after the occurrence of venous occlusion by photocoagulation. Fluorescein angiography revealed delay of retinal circulation and dye leakage from these venules. Fluorescein angiography on the 2nd day revealed caliber irregularities in the venules distal to the photocoagulated points. Histopathologically, thrombus formation and partial defect of the endothelial cells were observed in the venules in the areas which showed recovery with immature endothelial cells on the 4th day. Young endothelial cells proliferating along the endothelial basement membrane, which remained around the thrombi, and recanalization were observed on the 7th day. These results reveals a process of recanalization in the thrombogenous retinal venous occlusion.

Linear lichen planus mimicking creeping eruption.

A 42-year-old woman was referred to our hospital with a linear eruption on her right flank of two months duration. Because she had eaten loach-fish a month before she noticed the eruption, a creeping eruption due to Gnathostoma spp. was initially suspected, but the histological findings of the biopsy specimens showed typical features of lichen planus. Linear lichen planus is discussed based on the cases accumulated in the literature regarding the distribution of Blaschko lines.

[Experimental retinal branch vein occlusion].

Experimental retinal branch vein occlusion in monkeys (macaca irus) were produced by yellow wavelength (577nm) dye laser photocoagulation. The clinical sequence was observed in terms of the structural and ultrastructural findings occurring during the natural history of experimental branch vein occlusion, particularly in the early stage. Immediately after the occlusion, venous dilatation and retinal edema and hemorrhage appeared. Extravascular leakage of dye in fluorescein angiography and hemorrhage appeared from small venules at an early stage. Leakage from the larger vessels was observed later. Opening of the intercellular junctional complexes, which signifies breakdown of the blood-retinal barrier was detected by electron microscopy. Subsequently degenerative or necrotic changes in endothelial cells appeared with the formation of intravascular thrombi and extravascular leakage of blood components appeared from these vessels.

Studies on the enhanced effect of acupuncture analgesia and acupuncture anesthesia by D-phenylalanine (2nd report)--schedule of administration and clinical effects in low back pain and tooth extraction.

D-phenylalanine (DPA) is known to block the activity of carboxypeptidase, an enzyme which degrades enkephalins, endogenous morphine-like substances. Therefore, it is considered that DPA administered as an inhibiting drug of this degrading enzyme might prolong analgesia induced by acupuncture. 1) Thirty patients suffering from chronic low back pain were treated with acupuncture 30 minutes after the oral administration of 4.0 grams of DPA. The results were: excellent in 7 cases, good in 11, fair in 6 and poor in 6. Cases graded excellent and good were then compared with a placebo group. The effect was increased 26% in the DPA-acupuncture group, which shows no statistically significant difference (P less than 0.1). 2) In 56 patients, tooth extraction was performed under acupuncture anesthesia: 18 had received 4.0 gram of DPA (P.O.) 30 minutes earlier. The results were excellent in 8, good in 6, fair in 3, and poor in 1. The excellent and good cases were compared with 38 placebo group cases. The effect in the DPA-acupuncture anesthesia group was significantly increased by 35% (P less than 0.01). 3) In order to determine the optimum time for the administration of DPA, two schedules of administration were compared. [1] DPA was given on the previous day in three 0.5 gram doses (26 cases). [2] A single 4 gram dose was administered 30 minutes before treatment (30 cases). The results from the "excellent", "good" and "fair" cases showed a 16% increase in effectiveness when DPA was administered the day before, not a statistically significant difference (P less than 0.1), but a clear tendency to increase was observed. The above findings show that DPA has an enhancing effect on acupuncture analgesia and anesthesia in clinical practice.

[Experimental retinal branch vein occlusion].

Experimental retinal branch vein occlusion was produced in monkeys (macaca irus) by dye laser photocoagulation (yellow wavelength 577nm The 920 ARGON/DYE Laser System, Coherent Radiation). The Clinical process was observed based on ophthalmoscopic and fluorescein angiographic alterations which occur during the natural history of experimental branch vein occlusion. Immediately after occlusion, venous dilatation, retinal edema and punctate hemorrhage, were recognized by ophthalmoscopy, and leakage of dye from postcapillary venules, followed later by additional leakage from large veins were observed frequently by fluorescein angiography. Circulatory disturbances remained over 3-4 days, and the retinal branch vein occlusion were persisted. Ten days later, a non-perfusion area was observed by fluorescein angiography. More than 1 year later, these nonperfusion area remained.

Studies on the enhanced effect of acupuncture analgesia and acupuncture anesthesia by D-phenylalanine (first report)--effect on pain threshold and inhibition by naloxone.

It has been claimed that the mechanism of acupuncture analgesia can be explained in part by endogenous opioids. If so, it might be possible to enhance the analgesic effect of acupuncture by the administration of endorphins. If D-phenylalanine (DPA), an inhibitor of the endorphin degrading enzyme, is administered, the analgesic effect of acupuncture should be prolonged due to the increased level of endorphins. From the changes of the pain threshold (PT), we investigated whether or not the pre-administration of DPA can enhance the analgesic effect of acupuncture in humans. In addition, we examined the inhibitory effect of naloxone. 1) In all five subjects whose PT was raised after acupuncture anesthesia (respondents), the rise in PT was significantly prolonged by DPA. 2) Out of 10 subjects whose PT remained almost unchanged after acupuncture anesthesia (non-respondents), the PT was increased by DPA in 5 cases. 3) The rise in PT was most prominent when DPA was administered 30 minutes before the start of acupuncture anesthesia. 4) In all 4 respondents in whom the rise in PT persisted after DPA and acupuncture anesthesia, their raised PT dropped after the intravenous injection of naloxone (10 mg). 5) These findings show that DPA enhances the analgesic effect of acupuncture by the "endorphin mechanism."

An experimental study on the enhancing effects of phenylalanine on acupuncture analgesia.

In this preliminary study we examined the enhancing effect of D-phenylalanine on acupuncture anesthesia. We made 4 different kinds of experiments with 3 volunteers. The results show that D-phenylalanine extends the analgesic effect of acupuncture analgesia remarkably, with no exception in 3 cases. According to these facts, we believe that these findings have an important meaning for those who are engaged in acupuncture treatment or research.