Autologous fibrin glue as a sealant of the common bile duct

Autologous fibrin glue was evaluated as a sealant of the common bile duct (CBD) with the use of experimental animal models. It proved to be highly effective. Since it is a safe product that can be easily obtained from the patient's own blood, it may find useful application in most biliary tract procedures, liver transplants, and hepatectomies.     

The GABAA/benzodiazepine receptor complex in rat brain neuronal cultures. Characterization by immunoprecipitation

The expression of the GABAA/benzodiazepine receptor (GABAR/BZDR) complex in primary neuronal cultures from rat brain embryos has been investigated. The GABAR/BZDR complex was photoaffinity labeled with [3H]flunitrazepam [3H]FNZ and immunoprecipitated with subunit specific antibodies. These were the mAb 62-3G1 which is specific for the 57-kDa GABA binding subunit, and the rabbit antiserum A which recognizes the 51-kDa [3H]FNZ binding subunit. The results indicate that the cultured neurons express 5 different peptides of 51, 53, 54, 57 and 59 kDa that can be photoaffinity labeled with [3H]FNZ and that all of them are physically coupled to the GABAA receptor. Most of the [3H]FNZ photolabeled peptides have similar mobilities to those found in the brain of the newborn rat. Nevertheless, some of the quantitative changes in the photolabeled peptides observed during the normal development of the rat brain were not observed or occurred at much slower pace in the cultured neurons.     

Sialic acid in a complex oligosaccharide chain of the Tc-85 surface glycoprotein from the trypomastigote stage of Trypanosoma cruzi

The carbohydrate moiety of the Tc-85 surface glycoprotein from the infective trypomastigote form of Trypanosoma cruzi was analysed. Tc-85 could be metabolically labeled by incubation of the cells with D-[14C]mannose or D-[14C]glucose. Degradation techniques were performed directly on the polyacrylamide gel band containing labeled Tc-85. A mannobiose was cleaved by beta-elimination and further treatment of the remaining material under conditions which liberate N-asparaginyl linkages, released a complex oligosaccharide. The presence of sialic acid was demonstrated by: mild acid hydrolysis, neuraminidase treatment and periodate oxidation under mild conditions followed by NaB3H4 reduction, hydrolysis, and detection of NANA7 by paper electrophoresis. In addition, the chromatographic behavior of the asialooligosaccharide was significantly different from that of the original sample. Galactose, mannose and glucosamine are the other monosaccharide components of the sialooligosaccharide.     

Inadvertent germline transmission of AAV2 vector: findings in a rabbit model correlate with those in a human clinical trial.

The risk of germline transmission of vector sequences in humans is a major safety concern, because the enrollment of subjects of reproductive age in early-phase clinical trials of gene transfer continues to increase. In a study of adult men with hemophilia B, adeno-associated virus serotype 2 (AAV2) delivered to the liver via the hepatic artery resulted in unexpected transient vector dissemination to the semen. Here we report that intravenous AAV2 injection in rabbits proved a useful model to assess biologic parameters of vector dissemination to the semen. Detectable vector sequences in semen disappeared in a dose-dependent and time-dependent fashion. AAV infectious particles were present only as long as day 4 after injection and were undetectable thereafter. The kinetics of vector clearance was faster in the semen fractions enriched for motile sperm than in the total semen. In addition, increased frequency of semen sampling accelerated the clearance of vector sequences from semen. Long-term follow-up, spanning hundreds of spermatogenesis cycles, showed that there was no recurrence of detectable vector sequences in semen, thus reducing the probability of inadvertent transduction of early spermatogonia not committed to differentiation at the time of vector injection. We conclude that AAV2 presents minimal germline transmission risk for humans.     

Evaluation of ELISA with ABTS, 2-2'-azino-di-(3-ethylbenzthiazoline sulfonic acid), as the substrate of peroxidase and its application to the diagnosis of schistosomiasis

A micro-technique of enzyme-linked immunosorbent assay (ELISA) using ABTS, 2,2'-azino-di-(3-ethylbenzthiazoline sulfonic acid), as a substrate for horseradish peroxidase (HRP) conjugate was studied. In a comparative study among 4 substrates, namely; 5-aminosalicylic acid (5AS), O-phenylenediamine (OPD), O-tolidine (OT) and ABTS, for HRP in terms of sensitivity, it was found that ABTS was the most sensitive, stable and the best in visuality by its bluish-green color. The precise reproducibility of ELISA readings was attained by making correction of observed absorbance with positive reference serum. Sera from 50 patients positive for S. japonicum (Sj) egg and from 50 healthy residents in non-infested areas were diluted at 1:200 and examined by this procedure of ELISA against crude egg antigen. The upper limit of 99% critical range of the absorbance among negative sera was 0.080. The reactions of egg positive cases were beyond this value, being 0.505 at the minimal absorbance (1.259 +/- 0.278), and were easily distinguished from negatives even by naked eyes. Cross-reactivity to Sj egg antigen was studied using sera from 75 cases with 22 kinds of varied parasitic infections. Among them, S. mansoni and S. haematobium infections showed higher absorbance while the others did a little higher than the normal range. The highest absorbance shown by a Trichobilharzia brevis case was 0.177. The results indicated that micro-ELISA technique with ABTS developed in the present study is applicable to the field survey because of its high sensitivity, specificity and reproducibility.     

Longitudinal study on Schistosoma japonicum infections in the Philippines. Incidence and prevalence among school children in Dagami Area, Leyte.

The annual incidence of Schistosoma japonicum infection was calculated from the record of the yearly examination among school children of Dagami Area, Leyte, Philippines. Data were processed mainly by the computer. Children were examined for eggs by the merthiolate formalin concentration technique and by the circumoval precipitin test (COPT) once a year in 3 consecutive school years (SY), SY 1974/75, 1975/76 and 1976/77. About 600 to 700 children were examined yearly and the ratios of cases rechecked after 1 year were about 30%. A method to calculate the overall incidence in children who were examined at different intervals, was newly established. The incidence appeared to rise rapidly in the survey period, being 25.59% and 41.30% by fecal examination and 10.74% and 21.27% by COPT in SY 1975/76 and 1976/77, respectively. From the combined results of fecal examination and COPT, the reliable values of incidence, however, were shown to be stable during 2 years such as 22.06% and 24.21%, respectively. The incidence estimated by the age prevalence data was 20.25% in SY 1976/77 by the combined results and was found to be lower than the directly calculated value.     

Two consecutive limb lengthenings with the same PRECICE nail: a technical note

Purpose

The most significant advance in our time about limb lengthening is the magnetic lengthening nail, as the first reports appeared to show good results with accurate lengthening rates and good regenerate bone formation. The described complication rate is generally low. They avoid external fixation elements, and are activated transcutaneously, so the patient’s pain and discomfort are reduced and the rehabilitation is faster and more effective. The aim of authors is to describe a special technical issue of the PRECICE system: the nail can be extended inside the patient limb (after the osteotomy), but it also can be retracted inside the limb after achieving the bone union.

Methods

The authors present a case in which the limb lengthening has been performed in consecutive lengthening periods using the same nail. The nail was extended and retracted by altering the settings on the external remote control as well as accurately setting the rate of distraction.

Results

After two consecutive femoral lengthening with the same PRECICE nail, the patient no longer has a significant lower limb length discrepancy and patient satisfaction was high. During this clinical case, we were not confronted with any type of complications.

Conclusion

This technique utilizes the principles and advantages of lengthening over an magnetic lengthening nail, avoids the necessity of nail removal and minimizes the complication rates and the overall time for complete recovery.

Level of evidence

Level IV.

     

Intermittent stretching induces fibrosis in denervated rat muscle

Introduction: Stretching (St) has been used for treating denervated muscles. However, its effectiveness and safety claims require further study. Methods: Rats were divided into: (1) those with denervated (D) muscles, evaluated 7 or 15 days after sciatic nerve crush injury; (2) those with D muscles submitted to St during 7 or 15 days; and (3) those with normal muscles. Muscle fiber cross‐sectional area, serial sarcomere number, sarcomere length, and connective tissue density were measured. MMP‐2, MMP‐9, TIMP‐1, TGF‐β1, and myostatin mRNAs were determined by real‐time polymerase chain reaction. MMP‐2 and MMP‐9 activity was evaluated by zymography. Collagen I was localized using immunofluorescence. Results: St did not prevent muscle atrophy due to denervation, but it increased fibrosis and collagen I deposition at day 15. St also upregulated MMP‐9 and TGF‐β1 gene expressions at day 7, and myostatin at day 15. Conclusions: Stretching denervated muscle does not prevent atrophy, but it increases fibrosis via temporal modulation of TGF‐β1/myostatin and MMP‐9 cascades. Muscle Nerve 53 : 118–126, 2016