Not so bad after all: retroviruses and long terminal repeat retrotransposons as a source of new genes in vertebrates.

Viruses and transposable elements, once considered as purely junk and selfish sequences, have repeatedly been used as a source of novel protein-coding genes during the evolution of most eukaryotic lineages, a phenomenon called 'molecular domestication'. This is exemplified perfectly in mammals and other vertebrates, where many genes derived from long terminal repeat (LTR) retroelements (retroviruses and LTR retrotransposons) have been identified through comparative genomics and functional analyses. In particular, genes derived from gag structural protein and envelope (env) genes, as well as from the integrase-coding and protease-coding sequences, have been identified in humans and other vertebrates. Retroelement-derived genes are involved in many important biological processes including placenta formation, cognitive functions in the brain and immunity against retroelements, as well as in cell proliferation, apoptosis and cancer. These observations support an important role of retroelement-derived genes in the evolution and diversification of the vertebrate lineage.

The effect of mechanical load on degenerated soft tissue.

OBJECTIVE: To present a form of therapeutic mechanical load, Graston Technique (GT)-an instrument-assisted soft tissue mobilization method) in three case studies including supraspinatus tendinosis, Achilles tendinosis, and plantar fasciosis. METHOD: In each case study, case history and functional testing confirmed the presence of a condition characterized by degenerated soft tissue. Each condition was treated according to the GT protocol. GT is a patented form of treatment using stainless steel instruments designed with a unique curvilinear treatment edge, contoured to fit various shapes of the body. RESULTS: The GT method of load deformation to soft tissue resulted in the elimination of pain and normalization of the positive functional tests that revealed the conditions of supraspinatus tendinosis, Achilles tendinosis, and plantar fasciosis. CONCLUSION: This method of mechanical deformation load on soft tissue lesions is unique for its ability to both detect and treat areas of degenerated tissue. It deserves further consideration for basic research.

Treatment of a case of subacute lumbar compartment syndrome using the Graston technique.

OBJECTIVE: To discuss subacute lumbar compartment syndrome and its treatment using a soft tissue mobilization technique. CLINICAL FEATURES: A patient presented with low back pain related to exercise combined with prolonged flexion posture. The symptoms were relieved with rest and lumbar extension. The patient had restrictive lumbar fascia in flexion and rotation and no neurological deficits. INTERVENTION AND OUTCOME: The restrictive lumbar posterior fascial layers and adjoining restrictive fascia (thoracic, gluteal, hamstring) were treated with a form of instrument-assisted soft tissue mobilization called the Graston technique. Restoration of fascial extensibility and resolution of the complaint occurred after 6 treatment visits. CONCLUSIONS: The posterior spinal fascial compartments may be responsible for intermittent lower back pain. Functional clinical tests can be employed to determine whether the involved fascia is abnormally restrictive. Treatment directed at the restrictive fascia using this soft tissue technique may result in improved fascial functional testing and reduction of symptoms.

Effect of solid and liquid diet on uptake of large particulates across intestinal epithelium in rats.

The effect of diet composition on the uptake of particulates across the gastrointestinal epithelium has been examined in fasted male weanling Sprague-Dawley rats by estimating the systemic uptake of orally administered 2-microns latex polystyrene microspheres. Using a tissue solubilization assay, particle transfer in animals maintained on a fluid diet was determined. A larger number of particles was transferred from the gut lumen to the internal organs, including the mesenteric lymph node, spleen, bone marrow, liver, kidney, and heart of animals fed solid pelleted diet than those maintained on a fluid-diet 4 hr after oral administration of particles. The increase in particle number in rats fed the solid diet was only statistically significant (P < 0.05) for brain tissue in the analysis for trend. However, the number of particles retained in the proximal region of the gut at the end of this period was greater in animals fed the fluid diet. This work demonstrates that diet composition is important in gastrointestinal transepithelial translocation of microspheres.

Supplemental staffing. Nurse manager views of costs, benefits, and quality of care.

The authors identified nurse managers' perceptions of supplemental staffing costs, benefits, and quality of care related to patient care delivery and knowledge of the setting. Qualitative responses related to perceptions of costs and benefits were analyzed. Perceptions of quality of care were more positive when related to actual patient care delivery, but relatively negative for items relating to knowledge of policies, procedures, and care delivery setting.

Decreased incorporation of D-glucosamine into glycosphingolipids of intact Familial Dysautonomia lymphoblasts.

Familial Dysautonomia (FD) is an autosomal recessive Ashkenazi Jewish genetic disease, of unknown etiology, involving deficits in both autonomic and sensory functions. Previously, we found statistically significant increases in globotriaosylceramide (Gb3) in FD fibroblasts and lymphoblasts, and a decrease in ganglioside levels. FD fibroblasts exhibited pleiomorphic changes at the light microscopy level, suggestive of changes in the plasma membrane. We described an increase in Gb3 on the surface of synchronized cells at the G1/S boundary of the cell cycle, based on Gb3-verotoxin (derived from E. coli) interactions. Using D-glucosamine-1-14C as an in vitro precursor, we herein report a marked decrease in the rate of incorporation of D-glucosamine into the sialic acid and the N-acetylgalacto/glucosamine moieties of gangliosides and neutral glycosphingolipids in intact FD compared to control lymphoblasts. The total ganglioside content of FD cells (primarily GM3, measured as incorporation of 3H from NaB3H4) was also decreased. These data indicate differences in the turnover of sialic acid and N-acetylated sugar constituents in FD vs normal cells.

Homozygous presence of the crossover (fusion gene) mutation identified in a type II Gaucher disease fetus: is this analogous to the Gaucher knock-out mouse model?

Gaucher disease (GD) is an inherited deficiency of beta-glucocerebrosidase (EC 3.1.2.45, gene symbol GBA). In type I GD, the CNS is not involved (nonneuronopathic), whereas in type II GD (acute neuronopathic) CNS involvement is early and rapidly progressive, while in type III GD (subacute neuronopathic) CNS involvement occurs later and is slowly progressive. The T6433C (L444P) substitution is prevalent in type GD II. It may occur alone as a single base-pair mutation but often is found as part of a complex allele containing additional GBA nucleotide substitutions, G6468C (A456P) and G6482C (V460V), without (recNciI) or with (recTL) G5957C (D409H). This complex allele is presumed to have formed by recombination (crossover, fusion) of the structural gene with the pseudogene, which contains the mutated sequences. Two complex alleles have never been demonstrated to coexist in any individual. We devised a selective PCR method for the specific amplification of the normal and/or fusion gene. Using this procedure we demonstrated the fusion gene in homozygous form for the first time, in a Macedonian/Ashkenazi Jewish GD type II fetus. Both parents were carriers of the recombination. This was confirmed by direct sequence analysis. A previous conceptus in this family was stillborn at 36 weeks, with features of severe type II GD. Neonates showing a severe clinical phenotype, analogous to the early neonatal lethal disease occurring in mice homozygous for a null allele produced by targeted disruption of GBA, have been described elsewhere, but the specific mutations in these cases have not yet been characterized.(ABSTRACT TRUNCATED AT 250 WORDS)

Genotype-phenotype pitfalls in Gaucher disease.

Gaucher disease (GD), caused by inherited deficiency of beta-glucocerebrosidase (beta-Glc, EC 3.1.2.45), is classified type I if the CNS is not involved (non-neuronopathic), type II if CNS involvement is early and rapidly progressive (acute neuronopathic), and type III if CNS involvement occurs later and is slowly progressive (subacute neuronopathic). The clinical course is not predictable by measurement of residual beta-Glc activity. Patient classification by identification of specific mutations is more promising: homozygosity for the common A5841->G (N370S) mutation invariably predicts type I; homozygosity for the T6433->C (L444P) mutation usually indicates type III (Norbottnian). Type II disease patients often carry the T6433->C allele together with a complex allele derived in part from the downstream pseudogene by crossover or gene conversion, producing a T6433->C substitution, plus 2 or 3 additional single base substitutions (fusion gene). Employing selective PCR amplification of the structural gene, we detected homozygous T6433C (L444P) point mutations in a Caucasian boy, initially classified as having GD type I, who succumbed to severe visceral GD before age 3 years. A second novel PCR procedure for discriminating between the normal gene and the fusion gene confirmed the homozygous point mutation results. Post mortem neuropathological findings showed neuronal complex lipid accumulation consistent with late-onset type III disease. Although in Norbottnian patients it is generally accepted that onset of neurological findings is delayed, patients with the L444P/L444P genotype can only be initially classified as type III with this ancestry. Other patients described sporadically elsewhere are invariably considered type I until neurological findings arise.(ABSTRACT TRUNCATED AT 250 WORDS)

Increased globotriaosylceramide in familial dysautonomia.

Familial Dysautonomia (FD) is an autosomal recessive disease of unknown etiology, occurring primarily in Ashkenazi Jews. Patients are neurologically impaired, with deficits primarily in autonomic and sensory functions. The biochemical and genetic defects have remained elusive, precluding carrier detection and prenatal diagnosis. High-performance liquid chromatography data indicated up to a threefold increase in the neutral glycosphingolipid globotriaosylceramide in Dysautonomic fibroblasts and lymphoblasts. Total ganglioside values, measured by colorimetric, fluorometric or specific sodium borohydride incorporation, were decreased. Affected fibroblasts exhibited a range of pleomorphic phenotypes, such that the usual swirl-like confluent growth pattern of normal fibroblasts was distorted to varying degrees, suggesting abnormalities in the FD plasma membrane, possibly affecting cell-cell contacts. The glycosphingolipid increase could not be accounted for on the basis of markedly decreased alpha-galactosidase activity, as in Fabry's disease, where patients also display decreased autonomic function.

The Hymenoptera venom study. III: Safety of venom immunotherapy.

One thousand four hundred ten (44%) of the 3236 subjects in the Hymenoptera venom study accepted venom immunotherapy (VIT). Time to maintenance averaged 95 days, and the largest number achieved maintenance (147 subjects, 10.4%) at day 56. Ninety-two percent of the treated subjects achieved maintenance, and 84% continued therapy, most subjects (91%) until the study was terminated. One hundred seventy-one subjects (12%) experienced 327 treatment systemic reactions (Srs). The incidence of pruritus and angioedema/urticaria was similar with mild, moderate, or severe SRs. The SR severity did not correlate with the severity of the most recent sting before entry into the Hymenoptera-venom study, the most severe historical sting SR, the most severe SR during venom skin tests, the total dose of venom, the degree of skin test reactivity, or the lowest concentration yielding a positive skin test. Most SRs occurred between 1 and 50 micrograms and at maintenance; honeybee or wasp venoms were most likely to produce SR. This study, the largest of its kind with the use of standardized extracts, demonstrates (1) that there was good compliance, (2) that various historical and diagnostic criteria did not predict SRs to VIT, (3) that SRs to VIT were most likely to occur between 1 and 50 micrograms and at maintenance, (4) that honeybee or wasp venoms were most likely to produce an SR, and (5) that VIT is relatively safe.

Steroid sulfatase activity in the skin of the external ear canal.

Steroid sulfatase is an enzyme which has an important role in the complex process of cell desquamation. The specific activity of this enzyme was found to be higher in the epithelium of the deep external auditory canal than in the epithelium of the superficial external canal. We speculate that this steroid sulfatase activity gradient is involved in the outward migration of the keratinocytes of the canal epithelium.

The Hymenoptera venom study. II: Skin test results and safety of venom skin testing.

Data are summarized in this Hymenoptera venom study (HVS) article on the safety of skin testing with venom extracts. Of the 3236 subjects studied, 89% had experienced an historical sting systemic reaction (SSR). Seventy-four percent of all subjects and 76% of subjects who had experienced an historical SSR had a positive skin test to at least one venom. More subjects tested positive to yellow jacket venom (51.8%) than to any other venom. There were no significant differences of the wheal and erythema sizes associated with different venoms or different historical sting reactions. Forty-five percent of subjects with positive venom skin tests (VST) were positive to wasp, and 89% of these subjects were also positive to at least one of the following venoms: yellow jacket, yellow hornet, or white-faced hornet. Sixty-four of the 3236 subjects studied (2%) had a systemic reaction (SR) during VST; 13 of the SRs (0.4%) were severe. Thirteen of 64 adverse reactions (20%) were possibly vasovagal, and six other subjects (9%) demonstrated no symptoms of immediate-type hypersensitivity. Thus, 45 (1.4%) of the 3236 subjects tested had an SR that was considered to be a reaction of hypersensitivity, of which eight reactions (0.25%) were severe. Allergic SRs are associated with VST but are unusual and are rarely severe.

Simultaneous fractionation of four placental neutral glycosphingolipids with a continuous gradient.

We describe a method for isolating milligram quantities of the four neutral glycosphingolipids, glucocerebroside, lactosylceramide, traiosylceramide, and globoside, from human placental tissue. This procedure is carried out on a silicic acid column eluted with a continuous chloroform-methanol gradient (19:1 to 4:1); the four glycosphingolipids elute as separate fractions with no need for further separation. The method is simple, rapid, and yields sufficient material to use as analytical standards for several hundred runs. The lipids have been identified by NMR spectroscopy. Placental tissue is freely available in most centers and is an excellent untapped source for these compounds. Given that lactosylceramide is not commercially available and that triaosylceramide (ceramide trihexoside) cannot be obtained in a reliable state, this technique represents an effective solution to this dilemma.

The Hymenoptera venom study I, 1979-1982: demographics and history-sting data.

The Hymenoptera venom study, a study based on case histories, skin test results and adverse reactions, immunotherapy and adverse reactions, and treatment efficacy, for 3236 Hymenoptera-allergic subjects, was begun in 1979 after the Food and Drug Administration approval of Hymenoptera venoms. Eighty-four Fellows and Members of the American Academy of Allergy and Immunology participated in the study. All subjects had a history of an adverse reaction to one or more Hymenoptera insects. The mean age was 30 1/2 years (range 1 to 83 years). Male subjects accounted for 61.5% and female subjects, 38.5%; 3.1% were beekeepers and 32.3% were atopic. Demographic data were similar for subgroups. There was an average of 2.7 history stings per subject. At least one systemic reaction (SR) was reported by 2866 subjects (89%); 2219 (69%) experienced an SR after their most recent sting before entry into the study, and 70% had experienced only a single SR. Moderate to severe SRs were equally likely after stings of yellow jacket, white-faced hornet, and yellow hornet (65%), honeybee (67%), or wasp (70%), although historical SRs were reported more often after stings of yellow jacket, white-faced hornet, or yellow hornet (30%) than after honeybee (19%) or wasp (14%) stings. No association was noted between the number of stings per episode and severity of the SR. Fifty-one percent of SRs were reported as occurring within 10 minutes of sting; however, the onset of a moderate to severe SR sometimes occurred at 301 or more minutes after a sting episode. Of 2219 subjects with an SR after their most recent sting before entry into the study, 68% received epinephrine for treatment.

Regulation of renal sulfoglycolipid biosynthesis.

The in vitro activity of the renal galactolipid sulfotransferase and the level of sulfated glycolipids in the rat kidney have been correlated as a function of age. The galactolipid sulfotransferase was found to be greatly reduced in the young as compared with the adult animal. The relatively minor changes in the sulfated glycolipid content of the kidney with age suggests that an increase in sulfoglycolipid turnover occurs during growth. An inhibitory activity was detected in the homogenate supernate of the young animal capable of reducing the in vitro sulfotransferase activity of the adult. Assay of the human renal galactolipid sulfotransferase showed that this enzyme activity is deleted in samples of the blastematous form of Wilm's renal tumor. The results suggest that the rate of synthesis of renal sulfoglycolipids may prove a marker of renal development, perhaps by post translational regulation.