Soaring over open waters: horizontal winds provide lift to soaring migrants in weak thermal conditions.

BACKGROUND: For soaring birds, the ability to benefit from variable airflow dynamics is crucial, especially while crossing natural barriers such as vast water bodies during migration. Soaring birds also take advantage of warm rising air, so-called thermals, that allow birds to ascend passively to higher altitudes with reduced energy costs. Although it is well known that soaring migrants generally benefit from supportive winds and thermals, the potential of uplifts and other weather factors enabling soaring behavior remains unsolved. METHODS: In this study, we GPS-tracked 19 Red Kites, Milvus milvus, from the Central European population that crossed the Adriatic Sea on their autumn migration. Migratory tracks were annotated with weather data (wind support, side wind, temperature difference between air and surface-proxy for thermal uplift, cloud cover, and precipitation) to assess their effect on Red Kites' decisions and soaring performance along their migration across the Adriatic Sea and land. RESULTS: Wind support affected the timing of crossing over the Adriatic Sea. We found that temperature differences and horizontal winds positively affected soaring sea movement by providing lift support in otherwise weak thermals. Furthermore, we found that the soaring patterns of tracked raptors were affected by the strength and direction of prevailing winds. CONCLUSION: Thanks to modern GPS-GSM telemetry devices and available data from online databases, we explored the effect of different weather variables on the occurrence of soaring behavior and soaring patterns of migratory raptors. We revealed how wind affected the soaring pattern and that tracked birds could soar in weak thermals by utilizing horizontal winds, thus reducing energy costs of active flapping flight over vast water bodies.

Global and grain-specific accumulation of glycoside hydrolase family 10 xylanases in transgenic maize (Zea mays).

In planta expression of cell wall degrading enzymes is a promising approach for developing optimized biomass feedstocks that enable low-cost cellulosic biofuels production. Transgenic plants could serve as either an enzyme source for the hydrolysis of pretreated biomass or as the primary biomass feedstock in an autohydrolysis process. In this study, two xylanase genes, Bacillus sp. NG-27 bsx and Clostridium stercorarium xynB, were expressed in maize (Zea mays) under the control of two different promoters. Severe phenotypic effects were associated with xylanase accumulation in maize, including stunted plants and sterile grains. Global expression of these xylanases from the rice ubiquitin 3 promoter (rubi3) resulted in enzyme accumulation of approximately 0.01 mg enzyme per gram dry weight, or approximately 0.1% of total soluble protein (TSP). Grain-specific expression of these enzymes from the rice glutelin 4 promoter (GluB-4) resulted in higher-level accumulation of active enzyme, with BSX and XynB accumulating up to 4.0% TSP and 16.4% TSP, respectively, in shriveled grains from selected T0 plants. These results demonstrate the potential utility of the GluB-4 promoter for biotechnological applications. The phenotypic effects of xylanase expression in maize presented here demonstrate the difficulties of hemicellulase expression in an important crop for cellulosic biofuels production. Potential alternate approaches to achieve xylanase accumulation in planta without the accompanying negative phenotypes are discussed.

Properties of corn-expressed carbohydrase AC1 in swine diets and its effects on apparent ileal digestibility, performance, hematology, and serum chemistry.

Carbohydrases are often incorporated into livestock feed as digestive aids to improve animal performance. AC1 is a thermostable carbohydrase with ß-1,4-glucanase, endo-cellulase, and cellobiohydrolase activity. AC1 has been expressed in corn, where it accumulates in the grain for easy inclusion in animal diets. Incorporating the enzyme in high-fiber diets (corn-soy supplemented with distiller's dry grains with solubles) that were fed to 5-week-old pigs led to a trend of decreasing viscosity of the digesta as the dose of the enzyme increased (P = 0.092). AC1 also tended to increase the apparent ileal digestibility (AID) of neutral detergent fiber (P = 0.076). When fed diets containing 2126 U/kg AC1, pigs experienced no adverse effects in terms of performance metrics (body weights, average daily gain, average daily feed intake and gain-to-feed ratio), hematology, blood chemistry or general health when compared to pigs fed a control diet that lacked AC1.

Determining the phosphorus release of GraINzyme phytase in diets for nursery pigs.

The objective of this study was to determine the available P (aP) release curve for a new phytase source, GraINzyme Phytase (Agrivida Inc., Woburn, MA), which is expressed in corn containing an engineered Escherichia coli phytase called Phy02. Plant-expressed phytases are created by inserting phytase-encoding genes into plants resulting in their ability to produce seeds with increased concentrations of phytase. A total of 360 pigs (Line 200 × 400, DNA, Columbus, NE, initially 9.9 ± 0.19 kg) were used in a 21-d growth study. Pigs were weaned at approximately 21 d of age, randomly allotted to pens based on initial body weight (BW) and fed common starter diets. From days 18 to 21 postweaning, all pigs were fed a diet containing 0.11% aP. On day 21 postweaning, considered day 0 of the study, pens were blocked by BW and randomly allotted to one of eight dietary treatments with five pigs per pen and nine pens per treatment. Dietary treatments were formulated to include increasing aP derived from either an inorganic P source (0.11%, 0.19%, or 0.27% from monocalcium P) or increasing phytase (150, 250, 500, 1,000, or 1,500 FTU/kg). Diets were corn-soybean meal-based and contained 1.24% standardized ileal digestible Lys. On day 21 of the trial, one pig per pen (weighing closest to the mean pen BW) was euthanized and the right fibula was collected to determine bone ash using the nondefatted processing method. Overall (days 0 to 21), pigs fed increasing aP from inorganic P or phytase had increased (linear, P < 0.002) average daily gain (ADG), average daily feed intake (ADFI), and gain-to-feed (G:F; quadratic, P < 0.05). Bone ash weight (g) and percentage bone ash increased (linear, P < 0.001), with increasing inorganic P or added phytase. Based on the composition of the diets used in this study, the release equations developed for GraINzyme for ADG, G:F, bone ash weight, and percentage bone ash are as follows: aP = (0.255 × FTU)/(1299.969 + FTU), aP = (0.233 × FTU)/(1236.428 + FTU), aP = (45999.949 × FTU)/(462529200 + FTU), and aP = (0.272 × FTU)/(2576.581 + FTU), respectively.

Improved performance of Eimeria-infected chickens fed corn expressing a single-domain antibody against interleukin-10.

Antimicrobial resistance is a significant challenge for human and animal health, and developing effective antibiotic-free treatments is a strategy to help mitigate microbial resistance. The global poultry industry faces growing challenges from Eimeria-induced coccidiosis, a serious enteric disease of chickens that currently requires treatment using ionophore antibiotics. Eimeria stimulates interleukin-10 (IL-10) expression in the small intestine and caecum of infected chickens, suppressing their immune response and facilitating disease progression. Single-domain antibodies raised from llamas immunized with chicken IL-10 (cIL-10) were developed that bind cIL-10 in vitro, block cIL-10 receptor binding and induce interferon gamma (IFN-γ) secretion from cIL-10-repressed primary chicken splenocytes. Single-domain antibodies expressed in transgenic corn demonstrated significant accumulation in phenotypically normal plants. When fed to Eimeria-challenged chickens, the transgenic corn significantly improved body weight gain (equal to that of salinomycin-treated animals), normalized the feed conversion ratio (to the same level as uninfected control animals), lowered E. tenella lesion scores to those of salinomycin-treated control animals, and reduced oocyst counts below those of infected untreated control animals. Here, we propose that transgenic corn may have a role in reducing the use of antibiotics in poultry production and maintaining animal health and productivity, and may contribute to efforts against global antimicrobial resistance.

Effects of feeding corn-expressed phytase on the live performance, bone characteristics, and phosphorus digestibility of nursery pigs.

A 41-d feeding trial was conducted to determine the efficacy of a corn-expressed phytase (GZ; GraINzyme, Agrivida Inc., Woburn, MA) on the live performance, bone characteristics, and P digestibility of nursery pigs fed a reduced P diet. Weaned piglets (21 ± 3 d; n = 324) were acclimated on a common diet for 7 d (phase 1) before randomization into 54 single-sex pens (5 gilt and 4 barrow pens per treatment) containing 6 pigs (6.6 ± 1.2 kg) per pen. Six treatments were fed: positive control (PC; 0.4% or 0.32% aP for phase 2 or 3 and 4, respectively), negative control (NC; 0.15% reduction in aP), and 500, 1,000, 2,000, or 4,000 FTU per kg phytase from GZ added to NC in a 3-phase feeding program. Pigs were weighed on day 1, 14, 28, and 41, and feed disappearance recorded per phase. Apparent total tract digestibility (ATTD) of P was determined by feeding chromic oxide marker (day 28 to 35) and collecting fecal samples on day 35. On day 41, 4 pigs per pen were euthanized and metacarpal bones were collected to evaluate bone breaking strength (BBS) and ash. Data were analyzed using PROC GLM of SAS (block, sex, and treatment). Treatment least squares means were separated and linear and quadratic treatment effects evaluated. Other than feed efficiency (G:F) and day 15 to 28 ADFI, the pigs fed PC were superior (P < 0.05) to NC-fed pigs in all other variables. Pigs fed ≥500 FTU per kg phytase had increased (P < 0.05) ADG and ADFI compared to NC pigs and equivalent (P > 0.05) ADG and ADFI as PC pigs from day 0 to 41. Feeding ≥500 FTU per kg phytase resulted in higher (P < 0.05) ATTD of P than both NC and PC pigs and higher (P < 0.05) BBS and bone ash weight than NC. Pigs fed 1,000 or 2,000 FTU per kg phytase had equivalent (P > 0.05) BBS and bone ash weight compared to pigs fed PC diets. Feeding 4,000 FTU per kg phytase resulted in higher (P < 0.05) day 1 to 41 ADG, ATTD of P, and bone ash weight compared to feeding ≤1,000 FTU per kg phytase or PC diets. There were linear (P < 0.05) increases in ADG, ADFI, ATTD of P, BBS, and bone ash characteristics as GZ inclusion increased. In conclusion, ≥500 FTU per kg phytase from GZ improved growth, ATTD of P, BBS, and bone ash when added to a reduced P diet and 4,000 FTU per kg phytase increased growth greater than the PC treatment.

Prenatal nicotine increases pulmonary alpha7 nicotinic receptor expression and alters fetal lung development in monkeys.

It is well established that maternal smoking during pregnancy is a leading preventable cause of low birth weight and prematurity. Less appreciated is that maternal smoking during pregnancy is also associated with alterations in pulmonary function at birth and greater incidence of respiratory illnesses after birth. To determine if this is the direct result of nicotine interacting with nicotinic cholinergic receptors (nAChRs) during lung development, rhesus monkeys were treated with 1 mg/kg/day of nicotine from days 26 to 134 of pregnancy. Nicotine administration caused lung hypoplasia and reduced surface complexity of developing alveoli. Immunohistochemistry and in situ alpha-bungarotoxin (alphaBGT) binding showed that alpha7 nAChRs are present in the developing lung in airway epithelial cells, cells surrounding large airways and blood vessels, alveolar type II cells, free alveolar macrophages, and pulmonary neuroendocrine cells (PNEC). As detected both by immunohistochemistry and by alphaBGT binding, nicotine administration markedly increased alpha7 receptor subunit expression and binding in the fetal lung. Correlating with areas of increased alpha7 expression, collagen expression surrounding large airways and vessels was significantly increased. Nicotine also significantly increased numbers of type II cells and neuroendocrine cells in neuroepithelial bodies. These findings demonstrate that nicotine can alter fetal monkey lung development by crossing the placenta to interact directly with nicotinic receptors on non-neuronal cells in the developing lung, and that similar effects likely occur in human infants whose mothers smoke during pregnancy.

In vitro selection of peptides acting at a new site of NMDA glutamate receptors.

Oligomeric N-methyl D-aspartate receptor (NMDAR) in brain is a ligand-gated ion channel that becomes selectively permeable to ions upon binding to ligands. For NMDAR channel, the binding of glutamate and glycine results in opening of the calcium permeable channel. Because the calcium influx mediated by NMDAR is important for synaptic plasticity and excitotoxicity, the function of NMDA receptors has been implicated in both health and disease. Native NMDA receptors are thought to be heteromeric pentamers with a central ion conduction pathway. There are five genes (NR1, 2A, 2B, 2C, and 2D) encoding various subunits that have been cloned, and NR1 is thought to be the essential subunit since it forms a functional channel by itself. To study NMDAR structure and function, we have searched for peptide modulators of NR1 using random peptide bacteriophage libraries. The peptides were identified based on their specific association with a purified receptor fusion protein that contains the putative ligand binding domain. We report the identification of one group of cyclic peptides (Mag-1) with a consensus sequence of CDGLRHMWFC. Using biochemical binding analysis and patch clamp electrophysiological recording, we show that the synthetic Mag-1 peptides cause noncompetitive inhibition of the receptor channel activity.

PDZ domain of neuronal nitric oxide synthase recognizes novel C-terminal peptide sequences.

PDZ domains are multifunctional protein-interaction motifs that often bind to the C-terminus of protein targets. Nitric oxide (NO), an endogenous signaling molecule, plays critical roles in nervous, immune, and cardiovascular function. Although there are numerous physiological functions for neuron-derived NO, produced primarily by the neuronal NO synthase (nNOS), excess nNOS activity mediates brain injury in cerebral ischemia and in animal models of Parkinson's disease. Subcellular localization of nNOS activity must therefore be tightly regulated. To determine ligands for the PDZ domain of nNOS, we screened 13 billion distinct peptides and found that the nNOS-PDZ domain binds tightly to peptides ending Asp-X-Val. This differs from the only known (Thr/Ser)-X-Val consensus that interacts with PDZ domains from PSD-95. Preference for Asp at the -2 peptide position is mediated by Tyr-77 of nNOS. A Y77D78 to H77E78 substitution changes the binding specificity from Asp-X-Val to Thr-X-Val. Guided by the Asp-X-Val consensus, candidate nNOS interacting proteins have been identified including glutamate and melatonin receptors. Our results demonstrate that PDZ domains have distinct peptide binding specificity.

Monoclonal antibody therapy for resected Dukes' C colorectal cancer: seven-year outcome of a multicenter randomized trial.

PURPOSE: As previously shown, antibody treatment increased survival of patients with resected colorectal cancer of stage Dukes' C. Since the 5-year analysis was criticized because of the wide range (2.7 to 7.5 years) of follow-up time, we performed a 7-year analysis with only four of 189 patients monitored for less than 5 years. PATIENTS AND METHODS: A total of 189 patients with resected Dukes' C colorectal cancer were randomly allocated to infusions of a total of 900 mg 17-1A antibody, 500 mg postoperatively followed by 4 monthly doses of 100 mg (n=99), or to observation only (n=90). Primary end points were overall survival and disease-free interval. Patients were stratified by a dynamic randomization according to center, sex, location of tumor, number of affected lymph nodes, and preoperative carcinoembryonic antigen concentration. RESULTS: Randomization produced balanced distribution of risk factors. After 7 years of follow-up evaluation, treatment had reduced overall mortality by 32% (Cox's proportional hazard, P < .01; log-rank, P=.01) and decreased the recurrence rate by 23% (Cox's proportional hazard, P < .04; log-rank, P=.07). The intention-to-treat analysis gave a significant effect for overall survival (Cox's proportional hazard, P < .01; log-rank, P=.02) and disease-free survival (Cox's proportional hazard, P=.02; log-rank, P=.11 ). While distant metastases were significantly reduced (Cox's proportional hazard, P=.004; log-rank, P=.004), local relapses were not (Cox's proportional hazard, P=.65; log-rank, P=.52). This differential effect of 17-1A antibody on disseminated isolated tumor cells versus occult local satellites may explain the increased significance seen in the overall survival. CONCLUSION: The now-matured study shows that 17-1A antibody administered after surgery prevents the development of distant metastasis in approximately one third of patients. The therapeutic effect is maintained after 7 years of follow-up evaluation.

Recurrence patterns of hepatocellular and fibrolamellar carcinoma after liver transplantation.

PURPOSE: Tumor recurrence is the major limitation of long-term survival after liver transplantation for hepatocellular carcinoma (HCC) or fibrolamellar carcinoma (FLC). Understanding tumor-biologic characteristics is important for selection of patients and for development of adjuvant therapeutic strategies. PATIENTS AND METHODS: The study included 69 patients who underwent potentially curative liver transplantation for HCC/FLC and survived for more than 150 days; minimum follow-up was 33 months. Frequency, localization, and timing of recurrence were analyzed and compared with primary tumor and patient characteristics. RESULTS: Tumor recurrence was observed in 39 patients at 67 locations. Hematogenous spread was the major route of tumor recurrence (87%), and the most frequent sites were the liver (62%), lung (56%), and bone (18%). Parameters associated with recurrence were absence of cirrhosis, tumor size greater than 5 cm, more than five nodules, vascular infiltration, and International Union Against Cancer (UICC) stage IVA. Selective intrahepatic recurrence was found in nine patients (23%); it was associated with highly differentiated tumors, lack of vascular infiltration, and male sex. Recurrence at multiple sites was found predominantly in young patients (< or = 40 years) and for multicentric (> 5) primary tumors. Recurrences were observed within a wide time range after transplantation (43 to 3,204 days; median, 441 days); late recurrences (> 1,000 days, n = 8) were associated with highly differentiated or fibrolamellar tumors and low UICC stages. Surgical treatment was the only therapeutic option associated with prolonged survival after recurrence. CONCLUSION: In transplant recipients, hepatocellular carcinomas vary considerably in their pattern and kinetics of metastases. Tumor cells may persist in a dormant state for long time periods before giving rise to clinical metastases. Surgical treatment of recurrence should be considered whenever possible.

Dominance in lambda S mutations and evidence for translational control.

Phenotypic analysis of a collection of point mutations in the lysis gene S of bacteriophage lambda indicates that many of the S alleles exhibit at least partially dominant character, suggesting that the S gene product (gpS) must oligomerize to achieve its lethal membrane effect. Moreover, mutations found 5' to the coding sequence also show a dominant character and appear to define a site, designated sdi (structure directed initiation) where mRNA secondary structure controls the choice of initiation codons. We propose that formation of the sdi structure occludes the consensus Shine-Dalgarno sequence and results in initiation at the Met3 codon, generating a lethal 105 residue polypeptide. The model predicts that, in the absence of the sdi stem-and-loop, initiation occurs at the Met1 codon, generating a 107 residue polypeptide, which is a non-lethal inhibitor of lysis. In support of the model, alteration of the first codon was achieved using site-directed mutagenesis, resulting in an S allele that is more lethal and induces lysis significantly sooner than the wild-type.

Traceless multipole moment densities and transformations in macroscopic electromagnetism.

We consider whether use of traceless multipole moment densities in macroscopic electromagnetism can yield physically acceptable results. For harmonic plane wave fields it is shown that a traceless electric quadrupole density yields linear constitutive relations for which the dynamical material constants (permittivity and magnetoelectric coefficients) and response fields are unphysical. We further show that, within multipole theory, these constitutive relations cannot be transformed into physically acceptable relations. Specifically, the transformed response field D is unphysical for all orders beyond the electric dipole. This contrasts with use of primitive (traced) moment densities, for which unphysical constitutive relations have been successfully transformed up to electric octopole-magnetic quadrupole order, thereby providing also the leading contribution to the ac permeability.

Metabolic and transcriptional patterns accompanying glutamine depletion and repletion in mouse hepatoma cells: a model for physiological regulatory networks.

An important objective in postgenomic biology is to link gene expression to function by developing physiological networks that include data from the genomic and functional levels. Here, we develop a model for the analysis of time-dependent changes in metabolites, fluxes, and gene expression in a hepatic model system. The experimental framework chosen was modulation of extracellular glutamine in confluent cultures of mouse Hepa1-6 cells. The importance of glutamine has been demonstrated previously in mammalian cell culture by precipitating metabolic shifts with glutamine depletion and repletion. Our protocol removed glutamine from the medium for 24 h and returned it for a second 24 h. Flux assays of glycolysis, the tricarboxylic acid (TCA) cycle, and lipogenesis were used at specified intervals. All of these fluxes declined in the absence of glutamine and were restored when glutamine was repleted. Isotopomer spectral analysis identified glucose and glutamine as equal sources of lipogenic carbon. Metabolite measurements of organic acids and amino acids indicated that most metabolites changed in parallel with the fluxes. Experiments with actinomycin D indicated that de novo mRNA synthesis was required for observed flux changes during the depletion/repletion of glutamine. Analysis of gene expression data from DNA microarrays revealed that many more genes were anticorrelated with the glycolytic flux and glutamine level than were correlated with these indicators. In conclusion, this model may be useful as a prototype physiological regulatory network where gene expression profiles are analyzed in concert with changes in cell function.

Peptide binding consensus of the NHE-RF-PDZ1 domain matches the C-terminal sequence of cystic fibrosis transmembrane conductance regulator (CFTR).

The Na+-H+ exchanger regulatory factor (NHE-RF) is a cytoplasmic phosphoprotein that was first found to be involved in protein kinase A mediated regulation of ion transport. NHE-RF contains two distinct protein interaction PDZ domains: NHE-RF-PDZ1 and NHE-RF-PDZ2. However, their binding partners are currently unknown. Because PDZ domains usually bind to specific short linear C-terminal sequences, we have carried out affinity selection of random peptides for specific sequences that interact with the NHE-RF PDZ domains and found that NHE-RF-PDZ1 is capable of binding to the CFTR C-terminus. The specific and tight association suggests a potential regulatory role of NHE-RF in cystic fibrosis transmembrane conductance regulator (CFTR) function.

Attitudes of patients before and after transplantation towards various allografts.

BACKGROUND: The presence of an allogeneic graft inside the body may have psychological impact on transplant patients. It was the aim of this study to evaluate the attitude of patients before and after different types of organ transplantation towards organ allografts. METHODS: A total of 1,049 patients (722 patients after and 327 patients on the waiting list for either kidney, liver, heart, or lung transplantation) under care of a single transplant center were studied using a questionnaire with blinded analysis of the data. Mental condition of the patients, their attitude towards the allograft and its donor, emotional stress caused by a graft, and correlation of the attitudes with clinical and demographic parameters were analyzed. RESULTS: The self-reported mental condition of the patients was markedly and consistently better after organ transplantation; 27% of patients before and 60% after transplantation were in good mental condition. Sixty-two percent of transplant patients considered the graft as their own organ, 37% regarded it as a foreign organ now belonging to their body, and 1% considered it as a foreign body; among waiting list patients, 40%, 55%, and 5% assumed they would perceive their graft accordingly. The graft caused considerable emotional stress for 2% of transplant patients, whereas for 70% it did not cause any stress; the latter was assumed by 47% of patients before transplantation. Eleven percent of transplant patients frequently think about the origin of their graft, and 30% would like to have information about their donor. Knowledge about different religion, opposite sex, homosexuality, suicidal death, and age above 65 years of their donor would be of moderate or major concern for 0%, 3%, 21%, 24%, and 38% of the patients, respectively. CONCLUSIONS: The comprehensive survey shows that transplant patients incorporate their graft well into their body image. Emotional stress caused by the graft is very low and is generally less than assumed before transplantation. Knowledge about certain characteristics of the donor may cause increased concerns in some patients.

Rescue hepatectomy for initial graft non-function after liver transplantation.

BACKGROUND: Early retransplantation is the therapy of choice in patients with initial graft nonfunction (INF). In rare cases the patients' conditions deteriorate dramatically with severe cardiovascular and/or pulmonary insufficiency while on the waiting list for retransplantation. In this life-threatening situation removal of the graft and temporary portocaval shunt before allocation of a new liver proved to be effective. Our experience with this two-stage hepatectomy and subsequent liver transplantation in patients with complicated INF is reported. METHODS: Hepatectomy was performed in 20 patients with INF associated with severe cardiovascular and pulmonary insufficiency while on the waiting list for emergency liver retransplantation. The mean age was 41.75+/-16.64 years. The time period between primary transplantation and hepatectomy was 2.80+/-2.84 days with a range from 1 to 9 days. RESULTS: Hepatectomy reduced the need for vasopressive agents and improved pulmonary function in the majority of patients. Four patients died before a liver was available due to brain death in one patient and multiorgan failure in three patients. In the remaining 16 patients liver transplantation could be performed after 19.82+/-15.34 hr (range 6.58 to 72.50 hr). Two of the 16 transplanted patients died on the first postoperative day due to multiorgan failure and pneumonia. The remaining 14 of 16 patients survived retransplantation, but 7 died between days 13 and 105 mostly due to sepsis. Seven patients were discharged from the hospital in good condition and show long-term survival. CONCLUSION: Hepatectomy was able to stabilize the cardiovascular and pulmonary function. This study confirms the beneficial effects of hepatectomy and subsequent liver transplantation as a life-saving procedure in patients with INF complicated by cardiovascular and/or pulmonary instability.

Rapid electroblotting of small DNA fragments from polyacrylamide gels.

With the recent advances in PCR technology, the need for a simplified analysis of small double-stranded DNA fragments (less than 1.5 kb) has increased dramatically. An easy and rapid procedure has been developed for the separation, transfer and probe analysis of small double-stranded DNA fragments from polyacrylamide gels.

Invasive differentiated thyroid carcinoma: tracheal resection and reconstruction procedures in the hands of the endocrine surgeon.

BACKGROUND: Although differentiated carcinoma of the thyroid gland is a relatively benign tumor, up to 20% of patients are endangered by potentially fatal complications resulting from infiltrating tumor growth into the upper aerodigestive tract. METHODS: This study included 33 patients who underwent 34 tracheal or laryngotracheal procedures for invasive differentiated thyroid carcinoma under the direction of a single surgeon (G.F.W.S.). From 1990 to 1994, radical tumor extirpation was performed by "window" resection, and from 1995 to 1998, radical surgery consisted of either circumferential sleeve resection or laryngotracheal "step" resection--a novel method of reconstruction in cases of unilateral tumor infiltration into the larynx and trachea. Resection was limited to laminar ablation in 17 cases. The mean follow-up of 16 patients who survived was 42.5 months (range, 2 months to 8.9 years). RESULTS: Procedures resulting in primary end-to-end anastomosis of the upper airways were associated with lower perioperative morbidity and improved recurrence-free survival when compared with "window" resections with muscle flap reconstruction. In cases of superficial tracheal tumor infiltration, laminar ablations were sufficient for local tumor control. CONCLUSIONS: Radical eradication of differentiated thyroid carcinoma infiltrating the upper airways followed by radioiodine application should be considered the treatment of choice. Laryngotracheal "step" resection allows tumor extirpation with preservation of neural and muscular structures of the larynx.

Long-term experience after ex situ liver surgery.

BACKGROUND: Ex situ liver surgery allows liver resection and vascular reconstruction in patients who have liver tumors located at critical sites. Only a small series of studies about ex situ liver surgery is available in the literature. No long-term results have been published. METHODS: Twenty-four patients were considered for ex situ liver surgery because conventional liver surgery was considered impossible or too hazardous. The patient's ages were 51.3 +/- 7.5 years. Indications were various primary and secondary liver malignancies and benign liver tumors in 2 patients. RESULTS: In 22 of 24 patients, the ex situ liver resection and subsequent autotransplantation were performed. The anhepatic periods in these patients lasted for 5.6 +/- 1.1 hours. In the remaining 2 patients, autotransplantation was not possible and allogenic liver transplantation was performed 17 and 19 hours after hepatectomy. In 4 patients, liver failure occurred after autotransplantation and required transplantation. The confluens between hepatic veins and the inferior vena cava was reconstructed in 5 patients. Fifteen patients survived the postoperative period and were discharged after 36.5 +/- 16 days. The median survival time of 6 patients who had metastases of colonic carcinoma was 21 months. The 2 patients with benign liver disease are alive 9 and 5 years after ex situ surgery. CONCLUSIONS: Extended liver resections with difficult reconstructions of the hepatic venous confluens are feasible by ex situ liver surgery and subsequent autotransplantation. However, the early postoperative mortality rate is high, especially in patients with cholestatic livers. Early tumor recurrence remained the problem in these patients with extended local tumor spread. Ex situ liver surgery should only be performed in selected patients.