Monitoring flap for buried free tissue transfer: its importance and reliability

To improve the success rate of microsurgical flap transfers into a buried area, it is important to monitor the circulation of the flap during the early stage. A monitoring flap includes such advantages as simplicity, reliability, noninvasiveness, and the ability to continuously monitor the vascular status of various buried flaps. This article describes experiences related to the importance and reliability of a monitoring flap. A total of 109 flaps in 99 patients were treated with buried free flaps, including a monitoring flap, between 1990 and 1999. Forty-nine patients received a tubed free radial forearm flap with a skin-monitoring flap, and six received a free jejunal flap with a jejunal segment monitoring flap for the reconstruction of the esophagus. Vascularized fibular grafts with a skin monitoring flap or peroneus longus muscle monitoring flap were used for reconstructing the mandible in six patients and for treating osteonecrosis of the femoral head in 48 flaps in 38 patients. Monitoring flap abnormalities were indicated in 14 flaps; therefore, immediate revisions were performed on the pedicle of the monitoring flap and microanastomosis site. Among these 14 flaps, nine showed true thrombosis and five showed false-positive thrombosis. Among the nine flaps that showed true thrombosis, five were salvaged and four were finally lost. The false-positive thrombosis in the five flaps was attributed to torsion or tension of the perforator of the monitoring flap in three flaps, an unclear determination in one flap because the monitoring flap size was too small, and damage to the perforator in the last flap. The total thrombosis rate was 8.3 percent (nine of 109), and the failure rate of the free tissue transfer was 3.7 percent (four of 109). The overall sensitivity of the monitoring flap was 100 percent, the predictive value of a positive test was 64 percent (nine of 14), and false-positive results occurred in 36 percent (five of 14). The salvage rate was 55.6 percent. To improve the reliability of a monitoring flap, it is recommended that the size of the flap be larger than 1 x 2 cm to assess the arterial status, and that a perforator with the appropriate caliber be selected. When a monitoring flap is fixed to a previous incision line or a newly created wound, any torsion or tension of the perforator should be avoided. In conclusion, the current results suggest that a monitoring flap is a simple, extremely useful, and reliable method for assessing the vascular status of a buried free flap.     

Multiple virus resistance in transgenic plants conferred by the human dsRNA-dependent protein kinase

We have developed a new strategy for engineering resistance to multipleviruses in plants. The strategy exploits the human double stranded (ds)RNA-dependent protein kinase (PKR). PKR is one of theinterferon-induced enzymes. It confers viral resistance in mammals byinhibitingviral replication through the inactivation of the translational initiationfactor, eIF-2, upon activation by dsRNA. The humanPKR gene was fused to the promoter of theArabidopsis blue copper binding protein gene(BCB) that is induced rapidly in response to wounding. Thechimeric gene cassette was introduced into tobacco plants. Expression of thePKR gene in transgenic tobacco plants was demonstrated byRNA gel blot analysis and autophosphorylation assay of anM _r 68,000 protein. The transgenic plantsexpressing the PKR gene showed significantly reduced viralsymptoms or no viral symptoms at all, when challenged by different plant RNAviruses, such as Cucumber mosaic virus, Tobaccoetch virus, or Potato virus Y. Thus, expressionof a single component in the human interferon pathway, thePKR gene, can effectively confer resistance to multipleviruses in transgenic plants.     

High glucose enhances IL-1beta-induced cyclooxygenase-2 expression in rat vascular smooth muscle cells

The changes in vascular prostaglandin production are implicated in the derangement of vascular reactivity in diabetes. However, the mechanism of altered prostaglandin (PG) production in diabetes is largely unknown. In this study, we investigated the effect of high glucose on IL-1beta-induced PG production and the possible underlying mechanism in cultured vascular smooth muscle cell (VSMC). High glucose evoked an augmentation of IL-1beta-induced PG synthesis in a dose dependent manner and enhanced cyclooxygenase (COX) activity, which reached to maximum at 8-12 hours after stimulation. Western blot analysis supported the activity data. Protein kinase C (PKC) inhibitors, H-7 and chelerythrine, significantly inhibited the enhancement of IL-1beta-induced COX-2 expression by high glucose. The activation of PKC by PMA resulted in marked increase of PG production in low glucose group, whilst this was not the case in high glucose group. Furthermore, glucose-enhancing effect was significantly suppressed by zopolrestat, an aldose reductase inhibitor, and sodium pyruvate. These results suggest that the augmenting effect of high glucose on IL-1beta-induced PG production and COX-2 expression is, at least in part, due to increased glucose metabolism via sorbitol pathway following PKC activation.     

Case Study: Polycystic Livers in a Transgenic Mouse Line

Three mice (2 male, 1 female; age, 5 to 16 mo) from a mouse line transgenic for keratin 14 (K14)-driven LacZ expression and on an outbred Crl:CD1(ICR) background, were identified as having distended abdomens and livers that were diffusely enlarged by numerous cysts (diameter, 0.1 to 2.0 cm). Histopathology revealed hepatic cysts lined by biliary type epithelium and mild chronic inflammation, and confirmed the absence of parasites. Among 21 related mice, 5 additional affected mice were identified via laparotomy. Breeding of these 5 mice (after 5 mo of age) did not result in any offspring; the K14 mice with polycystic livers failed to reproduce. Affected male mice had degenerative testicular lesions, and their sperm was immotile. Nonpolycystic K14 control male mice bred well, had no testicular lesions, and had appropriate sperm motility. Genetic analysis did not identify an association of this phenotype with the transgene or insertion site.Abbreviations: K14, keratin 14 promoter; LacZ, bacterial β-galactosidase LacZ reporter; Lsamp, mouse limbic system-associated membrane proteinPolycystic disease is a multiorgan disorder and is the most common genetic life-threatening disease in people, affecting more than 600,000 Americans.¹⁶ Cystic liver disease in people typically is associated with polycystic kidney disease^22,36 but can exist in its absence. Currently, 2 autosomal dominant genes (PRKCSH and SEC63P) that cause a human polycystic liver disease condition without renal involvement have been identified.^4-6,13Numerous rodent models of polycystic kidneys with concurrent polycystic liver exist.^8,34,39,40 However, effective models of polycystic liver without polycystic kidneys would be useful to address clinical and mechanistic issues of polycystic liver not associated with polycystic kidneys.^6,31Here we report multiple cases of a spontaneous polycystic liver phenotype without a kidney phenotype in a transgenic mouse line. We also describe the effect of the transgene on disease expression and our attempts to develop this stock as an animal model.     

Global transcriptional response of pig brain and lung to natural infection by Pseudorabies virus

Background  

Pseudorabies virus (PRV) is an alphaherpesviruses whose native host is pig. PRV infection mainly causes signs of central nervous system disorder in young pigs, and respiratory system diseases in the adult.     

Isoform-specific induction of PKC-epsilon by high glucose protects heart-derived H9c2 cells against hypoxic injury

We investigated which PKC isoforms are involved in high glucose-induced protection against hypoxic injury. Treatment for 48 h with high glucose (22 mM) markedly increased the expression of PKC- epsilon in the particulate fraction (213+/-22.1% of the control) but had no effect on other types of PKC isoforms, suggesting that the high glucose-induced increase in PKC expression is isoform-specific. The mRNA level for PKC- epsilon was also substantially increased, reaching its peak after 4h of high glucose treatment. The high glucose increased PKC-epsilon activity in the particulate fraction up to 183+/-32.2% of the control. During hypoxia, the amount of PKC-epsilon in the particulate fraction was remarkably diminished in the low glucose-treated cells, but remained at a higher level in high glucose-treated cells. The treatment with epsilon V1-2 (10 microM), a specific inhibitor of PKC epsilon, abolished the protective effect of high glucose against hypoxia. These results suggest that isoform-specific induction of PKC-epsilon is involved in high glucose-induced protection against hypoxic injury in heart-derived H9c2 cells.     

Seasonal variations in yields of Hwangchil lacquer and major sesquiterpene compounds from selected superior individuals of<Emphasis Type="Italic">Dendropanax morbifera</Emphasis> Lév.

We studied fluctuations in the production of Hwangchil lacquer and major essential oils byDendropanax morbifera Lev. Considerable seasonal as well as intraspecific (individual-tree) variations were observed. Yields of Hwangchil lacquer as well as ß-elemene, α-selinene, ß-selinene, germacrene D, and δ-cadinene also depended on harvesting time, with levels being generally higher in July and August.     

Dorsalis pedis tendocutaneous delayed arterialized venous flap in hand reconstruction

We report two patients whose acute soft-tissue and tendon defects in the hand were treated with a dorsalis pedis tendocutaneous delayed arterialized venous flap between 1994 and 1997. The surviving surface area was 100 percent in both patients. The flap sizes were 10 x 10 cm and 6 x 6 cm. At 2 weeks postoperatively, active flexion and passive extension commenced, and progressive resistance exercises were performed for an additional 5 weeks. Flaps showed a similar color match and skin texture compared with the normal skin of the hand. Advantages of the tendocutaneous delayed arterialized venous flap are that a larger flap can be obtained than when using a pure venous flap or arterialized venous flap; the survival rate of the arterialized venous flap increases, which permits the use of a composite flap; the main artery of the donor site is preserved; thin, nonbulky tissue is used; and elevation is easy, without deep dissection. The disadvantages are the two-stage operation, donor-site scarring, and weak extension of the toes.     

5S rRNA-assisted DnaK refolding

Although accumulating evidence has revealed that most proteins can fold without the assistance of molecular chaperones, little attention has been paid to other types of chaperoning macromolecules. A variety of proteins interact with diverse RNA molecules in vivo, suggesting a potential role of RNAs for folding of their interacting proteins. Here we show that the in vitro refolding of a representative molecular chaperone, DnaK, an Escherichia coli homolog of Hsp70, could be assisted by its interacting 5S rRNA. The folding enhancement occurred in RNA concentration and its size dependent manner whereas neither the RNA with the reverse sequence of 5S rRNA nor the RNase pretreated 5S rRNA stimulated the folding in vitro. Based on our results, we propose that 5S rRNA could exert the chaperoning activity on DnaK during the folding process. The results suggest an interesting possibility that the folding of RNA-interacting proteins could be assisted by their cognate RNA ligands.