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Comprehensive, rapid and sensitive detection of sequence variants of human mitochondrial tRNA genes.

In the present study, a comprehensive, rapid and sensitive method for screening sequence variation of the human mitochondrial tRNA genes has been developed. For this purpose, the denaturing gradient gel electrophoresis (DGGE) technique has been appropriately modified for simultaneous mutation analysis of a large number of samples and adapted so as to circumvent the problems caused by the anomalous electrophoretic behavior of DNA fragments encoding tRNA genes. Eighteen segments of mitochondrial DNA (mtDNA), each containing a single uniform melting domain, were selected to cover all tRNA-encoding regions using the computer program MELT94. All 18 segments were simultaneously analyzed by electrophoresis through a single broad range denaturing gradient gel under rigorously defined conditions, which prevent band broadening and other migration abnormalities from interfering with detection of sequence variants. All base substitutions tested, which include six natural mutations and 14 artificially introduced ones, have been detected successfully in the present study. Several types of evidence strongly suggest that the anomalous behavior in DGGE of tRNA gene-containing mtDNA fragments reflects their tendency to form temporary or stable alternative secondary structures under semi-denaturing conditions. The high sensitivity of the method, which can detect as low as 10% of mutant mtDNA visually, makes it valuable for the analysis of heteroplasmic mutations.

Cell Line↗

Are self-expanding metal mesh stents useful in the treatment of benign esophageal stenoses and fistulas? An experience of four cases.

OBJECTIVE: The aim of this study was to review the long-term results of treating benign esophageal fistula and stenosis using self-expanding metal stents. METHODS: We treated four patients using covered mesh or coiled stents. We removed the stents electively in two patients (one endoscopically and one during planned partial esophagectomy) and unexpectedly in one patient who developed bleeding. One stent migrated and required laparotomy for removal. RESULTS: Placement of self-expanding metal stents successfully sealed the benign fistula in two patients and reestablished swallowing in two other patients with complicated achalasia. Two patients were swallowing normally on long-term follow-up, one died of the underlying disease, and one required gastrostomy. CONCLUSION: Temporary use of self-expanding metal stents as a feasible option for treating benign esophageal stenosis and fistula in patients who have failed other conventional treatments.

Aged↗

Real-time, in vivo analysis of malaria ookinete locomotion and mosquito midgut invasion.

Invasion of the Anopheles mosquito midgut by the Plasmodium ookinete is a critical step in the malaria transmission cycle. We have generated a fluorescent P. berghei transgenic line that expresses GFP in the ookinete and oocyst stages, and used it to perform the first real-time analysis of midgut invasion in the living mosquito as well as in explanted intact midguts whose basolateral plasma membranes were vitally stained. These studies permitted detailed analysis of parasite motile behaviour in the midgut and cell biological analysis of the invasion process. Throughout its journey, the ookinete displays distinct modes of motility: stationary rotation, translocational spiralling and straight-segment motility. Spiralling is based on rotational motility combined with translocation steps and changes in direction, which are achieved by transient attachments of the ookinete's trailing end. As it moves from the apical to the basal side of the midgut epithelium, the ookinete uses a predominant intracellular route and appears to glide on the membrane in foldings of the basolateral domain. However, it traverses serially the cytoplasm of several midgut cells before entering and migrating through the basolateral intercellular space to access the basal lamina. The invaded cells commit apoptosis, and their expulsion from the epithelium invokes wound repair mechanisms including extensive lamellipodia crawling. A 'hood' of lamellipodial origin, provided by the invaded cell, covers the ookinete during its egress from the epithelium. The flexible ookinete undergoes shape changes and temporary constrictions associated with passage through the plasma membranes. Similar observations were made in both A. gambiae and A. stephensi, demonstrating the conservation of P. berghei interactions with these vectors.

Amino Acid Sequence↗

Electromyographic activity of gastrointestinal tract of patient with short-bowel syndrome.

Electromyographic recordings were obtained from a patient with short-bowel syndrome. Eight recording sessions were performed in almost 3 mo, during both fasting and fed states. Slow waves were always present, and their frequencies were similar during both states. Phase III of the migrating motor complex (MMC) was not identified in any of the four recordings performed in the 1st mo after the operation. Afterward, phase III was observed in three of four recordings. Ingestion of milk caused substitution of the MMC by the fed pattern in all eight recordings. We conclude that extensive small-bowel resection causes temporary disappearance of phase III of the MMC. A change from a fasting to a fed pattern after food ingestion is normal.

Adult↗

Mechanisms and modulation of intestinal epithelial repair.

The mucosal epithelium of the alimentary tract represents a crucial barrier to a broad spectrum of noxious and immunogenic substances within the intestinal lumen. An impairment of the integrity of the mucosal epithelial barrier is observed in the course of various intestinal disorders including inflammatory bowel diseases (IBD), celiac disease, intestinal infections, and various other diseases. Furthermore, even under physiologic conditions temporary damage of the epithelial surface mucosa may be caused by proteases, residential flora, dietary compounds, or other factors. Generally, the integrity of the intestinal mucosal surface barrier is rapidly reestablished even after extensive destruction because of an enormous regenerative capability of the mucosal surface epithelium. Rapid resealing of the surface epithelium is accomplished by epithelial cell migration, also termed epithelial restitution, epithelial cell proliferation, and differentiation. Healing of the intestinal surface epithelium is regulated by a complex network of highly divergent factors, among them a broad spectrum of structurally distinct regulatory peptides that have been identified within the mucosa of the intestinal tract. These regulatory peptides, conventionally designated as growth factors and cytokines, play an essential role in regulating differential epithelial cell functions to preserve normal homeostasis and integrity of the intestinal mucosa. In addition, a number of other peptide molecules such as extracellular matrix factors and blood clotting factors, and also nonpeptide molecules including phospholipids, shortchain fatty acids, adenine nucleotides, trace elements, and pharmacological agents, have been demonstrated to modulate intestinal epithelial repair mechanisms. Some of these molecules may be released by platelets, adjacent stromal cells, inflammatory cells, or injured epithelial and nonepithelial cells and may play an important role in the modulation of intestinal injury. Repeated damage and injury of the intestinal surface are key features of various intestinal disorders including IBD and require constant repair of the epithelium. Enhancement of intestinal repair mechanisms by regulatory peptides or other modulatory factors may provide future approaches for the treatment of diseases that are characterized by injuries of the epithelial surface.

Cell Division↗

The fallacy of placental migration: effect of sonographic techniques.

A study of 233 consecutive second-trimester sonographic examinations performed between 15 and 20 weeks showed no cases of apparent placenta previa and only one case (0.4%) that appeared to be a marginal placenta previa. This differs from previously reported incidences of false-positive placenta previa of 5.6%-45%. The difference is thought to result from the technique of studying patients with completely empty bladders, taking special views of the internal os, and recognizing the temporary changes produced by myometrial contractions, which can give the false-positive diagnosis of placenta previa.

Female↗

Prostatic stents for the treatment of benign prostatic hyperplasia.

PURPOSE OF REVIEW: The insertion of prostatic stents in the treatment of lower urinary tract symptoms in men secondary to benign prostatic obstruction from benign prostatic hyperplasia has proven to be an effective modality in properly selected patients. We review the current literature on the role of prostatic stents in the treatment of bladder outlet obstruction secondary to benign prostatic hyperplasia. RECENT FINDINGS: Permanent stents such as the Urolume and the Memokath offer efficacious and safe treatment alternatives for men with bladder outlet obstruction from benign prostatic hyperplasia who are deemed high surgical risks for traditional transurethral resection of the prostate. When removal of permanent prostatic stent is necessary, the design of the Memokath allows for an easier removal than that of the Urolume. Temporary stents are being investigated as a means to treat the transient bladder outlet obstruction frequently observed after minimally invasive thermotherapy procedures aimed at treating benign prostatic obstruction, with both biodegradable and retrievable stents shown as successful modalities for this indication. SUMMARY: Prostatic stents remain an option to treat men with benign prostatic obstruction/bladder outlet obstruction; however, stent migration remains an obstacle to their widespread use. The key to obtaining optimal outcomes in men with benign prostatic hyperplasia is careful patient selection and experience with proper stent deployment.

Humans↗

[Cure of recurrent ascites in alcoholic hepatitis following transient placement of a LeVeen shunt].

Three cases of severe alcoholic hepatitis associated with refractory ascites are reported. A LeVeen shunt was inserted which was effective and well tolerated. However, the shunt was removed in all 3 patients 8 months, 12 months and 9 years after insertion because of spontaneous migration (n = 2) or superior vena cava thrombosis (n = 1). At time of shunt removal all 3 patients had micronodular cirrhosis, but none had recurrent ascites during the 1-3 years of observation and despite the absence of diuretics. The eventual transient aspect of refractory ascites associated with alcoholic hepatitis is discussed, together with the potential beneficial and temporary role of LeVeen shunts in this condition.

Adult↗

[The temporary immunodepression observed in trichinosis: is it related to acceptance of this parasite by its host?].

Swiss mice given 200 Trichinella larvae followed by a further 200 larvae 21 days later were found to be immune and did not show any immunodepression to sheep red blood cells (SRBC). When the challenge dose was given 56 days after the first infection, the pattern of infection was unaffected and an immunodepression to SRBC was observed. Therefore, the immunodepression in trichinellosis might be related to the migrating phase only.

Animals↗

Histopathology of rabbit eyes with intravitreous silicone-fluorosilicone copolymer oil.

Silicone-fluorosilicone copolymer oil is characterized by being heavier than water (density, 1.16 g cm-3) and low viscosity (175-185 centistokes) compared with currently used intraocular silicone oils (density, 0.97 g cm-3 and 1000-5000 centistokes). This oil is potentially useful as an operative tool and a tamponade on the inferior retina in complicated retinal detachment. We evaluate the ocular response clinically and histopathologically within 8 weeks in rabbit phakic eyes to the purified silicone-fluorosilicone copolymer oil after vitreous cavity injection, and compared the oil tolerance with purified silicone oil (0.97 g cm-3, 5000 centistokes) and perfluorotetradecahydrophenanthrene for ophthalmic use (Vitreon, 2.03 g cm-3, 8.03 centistokes) which are currently used as operative tools and as internal retinal tamponade agents in retinal detachment surgery. Because of their low viscosity, silicone-fluorosilicone copolymer oil and perfluorotetradecahydrophenanthrene were easier to inject into the eye than silicone oil. Silicone-fluorosilicone copolymer oil and perfluorotetradecahydrophenanthrene occupied the inferior portion in the eye, and silicone oil occupied the superior portion. Fewer discrete oil droplets and weaker vessel attenuation of medullary rays than in the perfluorotetradecahydrophenanthrene-injected eyes were seen in silicone-fluorosilicone-copolymer-oil-injected eyes. Histopathologically, all retinas injected with silicone-fluorosilicone copolymer oil were normal within 4 weeks. The silicone-fluorosilicone copolymer oil dispersion did not induce histopathological changes within 8 weeks. However, thinning or disappearance of the outer plexiform layer was seen in the inferior retina in some silicone-fluorosilicone-copolymer-oil-injected eyes at 6-8 weeks. A similar effect was found in the superior retina of a silicone-oil-injected eye at 8 weeks. More severe changes such as thinning or disappearance of the outerplexiform layer, thinning and disorganization of the photoreceptor layer, and migration of the receptor cell nuclei to the photoreceptor layer were found in the inferior retina of perfluorotetradecahydrophenanthrene-injected eyes after 2 weeks. Intraocular silicone-fluorosilicone copolymer oil tolerance until about 2 months post-injection is similar to silicone oil and better than perfluorotetradecahydrophenanthrene. Silicone-fluorosilicone copolymer oil may be useful intraoperatively and as a temporary vitreous substitute in cases of inferior retinal detachment.

Animals↗

The role of off-farm employment in tropical forest conservation: labor, migration, and smallholder attitudes toward land in western Uganda.

The potential for off-farm employment (OFE) to contribute significantly to forest conservation in the tropics is a widely held logic among donors, governments, and social scientists. While an aggregate level examination of OFE cases can support this logic, there is disagreement as to the operative aspects of specific linkages and assumptions. This study examines the case of the tea industry in western Uganda, and uses a combination of fieldwork and remote sensing to pursue a more nuanced examination of the role of migration and non-monetary aspects of OFE on forest conservation in both a national park and unprotected forest contexts. Results indicate that the tea industry does serve as an off-farm employer to a limited number of local smallholders but these benefits are offset by the industry's overwhelming dependence on migrant labor which sees OFE as temporary, then seeks to settle locally. There is also evidence that the tea industry is contributing to conservation efforts of Kibale National Park by unintentionally serving as a physical buffer zone, which inhibits both human encroachment on the park and wildlife encroachment on smallholder crops. The latter represents a site-specific phenomenon that holds much potential for future management plans of the area and exemplifies the importance of considering the site-specific circumstances associated with OFE development.

Agriculture↗

Long-term result of Memokath urethral sphincter stent in spinal cord injury patients.

BACKGROUND: Memokath urethral sphincter stents are used to facilitate bladder emptying in patients with spinal cord injury, but long term follow-up has not been reported. METHODS: Case series of ten men with spinal cord injury who underwent insertion of Memokath stents and were followed for up to nine years. RESULTS: Within four years, the stent had to be removed in nine out of ten patients because of: extensive mucosal proliferation causing obstruction to the lumen of the stent; stone around the proximal end of the stent, incomplete bladder emptying, and recurrent urinary infections; migration of the stent into the bladder related to digital evacuation of bowels; large residual urine; concretions within the stent causing obstruction to flow of urine, and partial blockage of the stent causing frequent episodes of autonomic dysreflexia. In one patient the stent continued to function satisfactorily after nine years. CONCLUSIONS: The Memokath stent has a role as a temporary measure for treatment of detrusor-sphincter dyssynergia in selected SCI patients who do not get recurrent urinary infection and do not require manual evacuation of bowels.

Autonomic Dysreflexia↗

Comparison of temporary and permanent stent placement with concurrent radiation therapy in patients with esophageal carcinoma.

PURPOSE: To assess the clinical effectiveness of temporary metallic stent placement with concurrent radiation therapy in patients with esophageal carcinoma by comparing it with permanent stent placement with concurrent radiation therapy. MATERIALS AND METHODS: Covered retrievable expandable nitinol stents were placed in 47 patients with esophageal carcinoma 1 week before starting radiation therapy; the stents were electively removed 4 weeks after placement in 24 patients (group A), while not electively removed in the other 23 patients (group B). In cases of complications, the stents were also removed from patients in groups A and B. The dysphagia score, complications (severe pain, granulation tissue formation, stent migration, esophagorespiratory fistula, and hematemesis), tumor overgrowth/regrowth, reintervention rates, and dysphagia-progression-free and overall survival rates were compared in the two groups. RESULTS: Stent placement or removal was technically successful and well tolerated in all patients. The dysphagia score was significantly improved in both groups after stent placement (P < .01). Each of the stent-related complications was less in group A than in group B but there was no significant difference. However, the total number of patients with one or more than one complications and who needed related reinterventions was significantly less in group A than in group B (P = .042 and .030, respectively). Tumor overgrowth/regrowth and the total number of patients who required related reinterventions was not significantly different (P = 1.00 and .517, respectively). Dysphagia-progression-free and overall survival rates were significantly longer in group A than in group B (P = .005 and .001, respectively). CONCLUSION: Temporary placement of a covered retrievable expandable metallic stent with concurrent radiation therapy for patients with esophageal carcinoma is beneficial for reducing complications and related reinterventions and for increasing resultant survival rates compared with permanent esophageal stent placement.

Adult↗

Comparison of different intervention procedures in benign stricture of gastrointestinal tract.

AIM: To determine the most effective intervention procedure by evaluation of mid and long-term therapeutic efficacy in patients of stricture of the gastrointestinal tract (GIT). METHODS: Different intervention procedures were used to treat benign stricture of GIT in 180 patients including pneumatic dilation (group A, n=80), permanent (group B, n=25) and temporary (group C, n=75) placement of expandable metallic stents. RESULTS: The diameters of the strictured GIT were significantly greater after the treatment of all procedures employed (P<0.01). For the 80 patients in group A, 160 dilations were performed (mean, 2.0 times per patient). Complications in group A included chest pain (n=20), reflux (n=16), and bleeding (n=6). Dysphagia relapse occurred in 24 (30%) and 48 (60%) patients respectively during 6-and-12 month follow-up periods in group A. In group B, 25 uncovered or partially covered or antireflux covered expandable metallic stents were placed permanently, complications included chest pain (n=10), reflux (n=15), bleeding (n=3), and stent migration (n=4), and dysphagia relapse occurred in 5 (20%) and 3 patients (25%) during the 6-and-12 month follow-up periods, respectively. In group C, the partially covered expandable metallic stents were temporarily placed in 75 patients and removed after 3 to 7 days via gastroscope, complications including chest pain (n=30), reflux (n=9), and bleeding (n=12), and dysphagia relapse occurred in 9 (12%) and 8 patients (16%) during the 6-and-12 month follow-up periods, respectively. The placement and withdrawal of stents were all successfully performed. The follow-up of all patients lasted for 6 to 96 months (mean 45.3+/-18.6 months). CONCLUSION: The effective procedures for benign GIT stricture are pneumatic dilation and temporary placement of partially-covered expandable metallic stents. Temporary placement of partially-covered expandable metallic stents is one of the best methods for benign GIT strictures in mid and long-term therapeutic efficacy.

Adolescent↗

Collagen containing neonatal astrocytes stimulates regrowth of injured fibers and promotes modest locomotor recovery after spinal cord injury.

The use of collagen as a vehicle to transplant neonatal astroglial cells into the lesioned spinal cord of the adult rat allows a precise application of these cells into the lesion gap and minimizes the migration of the transplanted cells. This approach might lead to anatomical and functional recovery. In the present study, 20 adult female Wistar rats were subjected to a dorsal hemisection at thoracic spinal cord levels. Cultured cortical neonatal rat astrocytes were transplanted into the lesion with collagen as a vehicle (N = 10). Prior to transplantation, the cultured astroglial cells were labelled with fast blue. Control rats received collagen implants only (N = 10). During 1 month of survival time, functional recovery of all rats was continuously monitored. Histological data showed that the prelabelled astroglial cells survived transplantation and were localized predominantly in the collagen implant. Virtually no fast blue-labelled GFAP-positive astroglial cells migrated out of the implant into the adjacent host spinal cord. The presence of transplanted neonatal astroglial cells resulted in a significant increase in the number of ingrowing neurofilament-positive fibers (including anterogradely labeled corticospinal axons) into the implant. Ingrowing fibers were closely associated with the transplanted astroglial cells. The implantation of neonatal astroglial cells did result in modest temporary improvements of locomotor recovery as observed during open-field locomotion analysis (BBB subscore) or during crossing of a walkway (catwalk).

Absorbable Implants↗

[Bone grafts in hip prosthesis revisions].

PURPOSE OF THE STUDY: We describe the collection, storage and examination of allogenic bone grafts and their use in revision hip arthroplasty, indicated because of acetabular loosening, that involved either an uncemented cup or an augmentation device with a cemented cup according to the extent and nature of acetabular defects. MATERIAL: Allogenic bone grafts, usually long bones, were collected from deceased humans within 12 hours of death. They were processed according to the standards of US tissue banks. Using ELISA, the blood of each donor was serologically examined for antibodies against selected antigens (HIV I, HIV II, syphilis, HbsAg, HCV, and p24HIV I, HTLV I and II, and HbcAg, if required). Microbiological examination of each bone sample was also included. The samples were kept in a quarantine box until the examination results were known. When these were negative, the bones were cut, under aseptic conditions, to obtain parts (condyle, metaphysis, diaphysis, patella, etc.) weighing from 80 to 150 gr. Subsequently, another quality check, involving microbiological and histological examination, was performed. The allografts were stored at -80 degrees C. METHODS: Between October 1999 and October 2000, 67 revision total hip replacements were performed. Of these, allogenic bone grafting was used for acetabular reconstruction in 30 cases (45%). The average amount of graft tissue per operation was 107 gr (range, 43 to 163 gr). Eleven grafts had been harvested from the tibial condyle, nine from the trochanters, eight from the femoral condyle and two from the patella. The graft bone, ground into coarse chips, was washed with an antibiotic solution. Two techniques were used in graft implantation. 1) Impaction grafting, under pressure, of the acetabulum whose surface was free from any granulation tissue and scratched to make the bone bleed, and implantation of a cemented cup (LOR or Spotorno CLS). 2) Filling of large defects with a bulk of allograft bone chips, using an augmentation ring, mesh or Burch-Schneider cage, and implantation of a cemented polyethylene acetabular component. Uncemented implants were used in 17 and cemented cups with an augmentation device in 13 cases. The hips were checked by X-ray at 3, 6 and 12 months and then at yearly intervals. RESULTS: In the postoperative period of 26 to 38 months, an early recurrent infection led to temporary removal of the implant in one case, one hip showed superficial infection that responded to therapy and there was one case of aseptic acetabular loosening. A recurrent dislocation led to femoral stem exchange for a modular component with a longer neck in one case. The remaining 28 cases showed good incorporation of the graft without any signs of implant migration. DISCUSSION: Segmental defects of the acetabulum can be treated by implantation of specific types of acetabular cups or by massive bone grafting. Cavitary defects can be filled with cancellous allograft bone or bioactive, artificial materials. We prefer morselized allograft bone from the tissue bank and this was used in 45% of repeat surgeries for failure of the acetabular component. An uncemented cup is more convenient, supposing sufficient primary stability can be ensured. When defects are extensive, augmentation devices and filling with graft bone are necessary. Cadaver allografts offer advantages over autologous bone because they are available at any amount and thus enable us to avoid extensive damage to the iliac crest by autologous tissue harvesting. Cadaver grafts are bones of good quality, free from degenerative or dystrophic changes because they are usually taken from young subjects. The size of 7 to 10 mm is optimal because it permits the grafts to maintain both structure and strength. The strict procedure of sample collection and examination minimizes the risk of infection transfer. The progress of graft bone ingrowth was evaluated on radiographs that showed incorporation of allograft into the surrounding bone during 3 to 6 months and no radiolucent areas at host bone-implant contact. CONCLUSIONS: Our patients showed good allogenic bone ingrowth in relation to the acetabular bed and good bone restructuring and incorporation in relation to the implant. Neither mechanical failure of implants nor graft rejection were recorded. The risk of infection transfer was low and comparable with routine blood transfusion. The necessary prerequisite for revision hip arthroplasty is a well-operating bone tissue bank.

Acetabulum↗

Acute dislocation of the acromioclavicular joint. Traumatic anatomy and the importance of deltoid and trapezius.

We report a prospective study of 46 patients with acute complete dislocation of the acromioclavicular joint. They were all treated by suture of the deltoid and trapezius over the clavicle with no repair of the coracoclavicular ligaments, using only temporary fixation with two wires. At operation 43 patients (93.5%) had damage to the trapezius or deltoid or both. The coracoclavicular ligaments were intact in six (13%). Follow-up was from 2 to 7.9 years (mean 5.8), and at the latest review only five patients (10.9%) had redisplacement, due to premature removal of wires for infection in one, to migration of the wires in another and to partial failure of the muscle repair in three. We consider that the deltoid and trapezius attachments are important clinical stabilizers of the clavicle and that their repair, with reinforcement, is a useful addition to any method of surgical treatment.

Acromioclavicular Joint↗

Accord No. 000244, 30 June 1988.

This Accord publishes in the Registro Oficial the text of the Andean Instrument on Labour Migration, adopted by the National Congress of Ecuador on 24 June 1987. Under the Instrument, member countries are not to impede the entry and departure of migrant workers who have been contracted in conformity with the Instrument and the immigration laws of the country of immigration. Migrant workers are not to be discriminated against on the basis of race, sex, religion, or nationality and are to have, in general, the same rights of workers of the country of immigration. Workers and their families are also to have the same rights as nationals to education, housing, health care, and social security. Further provisions of the Instrument set forth specific rules with respect to temporary workers, border workers, and undocumented workers.

Americas↗