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Background: endovascular aneurysm repair (EVAR) reduces morbidity and hospital stay compaired with open surgical repair. Endoleak is a common complication of the procedure. The resulting increase of pressure within the sac may expand the aneurysm with the following risk of rupture. The aim of this study was to recognize the incidence of endoleak in tomographic controls of the patients who underwent endovascular repair at our hospital as well as identify the risk factors associated with this complication., Methods: all consecutive patients who underwent endovascular aneurysm repair at our hospital between 2008, February until 2012, February were restrospectively enrolled in the study, excluding those who were lost at follow-up. 43 patients were included, aged 70.5 ± 6 (men: 88%). The endpoint was endoleak incidence at 1, 6, 12th months after the intervention in the control tomography, and its association with underlying risk factors: hypertension, Smoking, chronic obstructive pulmonary desease and the diameter of the aneurysm., Results: eleven (11%) patients developed endoleak (type I 9%, type III 2%). All were infrarrenal aortic aneurysms. The anteroposterior diameter of the aneurysm (more than 60 mm) showed a trend toward statistical significance as a risk factor (30% vs. 6%; p:0.073). No relationship was found with gender, age, COPD, smoking or hypertension., Conclusion: endoleak after endovascular aneurysm repair is a common complication. The size of the aneurysm might be a risk factor of the event.

Corn offers advantages as a transgenic host for producing recombinant proteins required at large volumes (1,000's of tons per year) and low cost (less than US$50/kg) by generating them as co-products of biorefining. We describe the purification and characterization of a corn grain-derived mammalian structural protein having such market characteristics: a full length recombinant collagen type I alpha 1 (rCI alpha 1) chain. Material properties of interest are gelation behavior, which would depend on as yet unverified ability of corn to carry out post-translational prolyl hydroxylation and formation of triple helical conformation. The starting material was grain where the expression of rCI alpha 1 had been directed by an embryo-specific promoter. Purification consisted of extraction at low pH followed by membrane and chromatographic steps to isolate rCI alpha 1 for characterization. The amino acid composition and immunoreactivity of CI alpha 1 was similar to that of an analogous native human CI alpha 1 and to rCI alpha 1 produced by the yeast Pichia pastoris. Tandem mass spectrometry confirmed the primary sequence of the corn-derived rCI alpha 1 with 46% coverage. Fragments of the rCI alpha 1 chains were also observed, possibly caused by endogenous plant proteases. The corn-derived rCI alpha 1 had a low level of prolyl hydroxylation (approximately 1% versus 11%) relative to animal-derived CI alpha 1 and folded into its characteristic triple-helical structure as indicated by its resistance to pepsin digestion below its melting temperature of 26(o)C. The 29 amino acid foldon fused to the C-terminus to initiate triple helix formation was not cleaved from the rCI alpha 1 chains, but could be removed by pepsin treatment., ((c) 2009 American Institute of Chemical Engineers Biotechnol. Prog. 2009.)

Recombinant DNA technology can be used to design and express collagen and gelatin-related proteins with predetermined composition and structure. Barley seed was chosen as a production host for a recombinant full-length collagen type I alpha1 (rCIa1) and a related 45-kDa rCIa1 fragment. The transgenic barley seeds were shown to accumulate both the rCIa1 and the 45-kDa rCIa1 fragment. Even when the amount of the rCIa1 was just above the detection threshold, this work using rCIa1 as a model demonstrated for the first time that barley seed can be used as a production system for collagen-related structural proteins. The 45-kDa rCI1a fragment expression, targeted to the endoplasmic reticulum, was controlled by three different promoters (a constitutive maize ubiquitin, seed endosperm-specific rice glutelin and germination-specific barley alpha-amylase fusion) to compare their effects on rCIa1 accumulation. Highest accumulation of the 45-kDa rCIa1 was obtained with the glutelin promoter (140 mg/kg seed), whereas the lowest accumulation was obtained with the alpha-amylase promoter. To induce homozygosity for stable 45-kDa rCIa1 production in the transgenic lines, doubled haploid (DH) progeny was generated through microspore culture. The 45-kDa rCIa1 expression levels achieved from the best DH lines were 13 mg/kg dry seeds under the ubiquitin promoter and 45 mg/kg dry seeds under the glutelin promoter. Mass spectroscopy and amino acid composition analysis of the purified 45-kDa rCIa1 fragment revealed that a small percent of prolines were hydroxylated with no additional detectable post-translational modifications.

This paper demonstrates that a fibrous, repetitive amino acid sequence collagen-related protein, a 44-kDa fragment of human collagen I alpha 1 (CIalpha1), was expressed in corn grain molecularly equivalent to that produced in recombinant yeast. The recombinant CIalpha1 was extracted and purified from early generation plants having low levels of recombinant protein accumulation. It was selectively extracted at low pH and purified by ion exchange and gel filtration chromatography, resulting in a 44-kDa CIalpha1 with >70% purity and 60% recovery. The N-terminal sequence, amino acid composition, and immunoreactivity closely matched those of an analogous 44-kDa CIalpha1 fragment produced by the yeast Pichia . The corn-derived 44-kDa CIalpha1 had an intact protein mass of 44088 Da, which is within 0.2% of the mass calculated from the expected sequence. Tandem mass spectrometry confirmed the primary sequence with 78% coverage. The amino acid composition analysis indicated a low level of prolyl hydroxylation. Glycoprotein staining revealed no glycosylation.

Forty-eight children (M age = 4.8 years) infected with HIV/AIDS and living in the Dominican Republic were randomly assigned to a massage therapy or a play session control group. The children in the massage therapy group received two weekly 20-min massages for 12 weeks; the children in the control group participated in a play session (coloring, playing with blocks) for the same duration and length as the massage therapy group. Overall, the children in the massage therapy group improved in self-help abilities and communication, suggesting that massage therapy may enhance daily functioning for children with HIV/AIDS. Moreover, the HIV infected children who were six or older also showed a decrease in internalizing behaviors; specifically depressive/anxious behaviors and negative thoughts were reduced. Additionally, baseline assessments revealed IQ equivalence below normal functioning for 70% of the HIV infected children and very high incidences of mood problems (depression, withdrawn) for 40% of the children and anxiety problems for 20% of the children, suggesting the need for better monitoring and alternative interventions in countries with limited resources to improve cognition and the mental health status of children infected with HIV/AIDS.

Background: Canine splenic hemangiosarcoma (HSA) is a fatal malignancy, and most affected dogs die within a few months of diagnosis. Most dogs present with signs from tumor rupture, resulting in hemoabdomen and intra-abdominal dissemination. The abdomen is also the main site of disease recurrence., Hypothesis: Intraperitoneal (IP) administration of doxorubicin will delay or prevent intra-abdominal tumor recurrence and prolong survival in dogs with HSA., Animals: Fourteen dogs with splenic HSA., Methods: A prospective, unmasked, uncontrolled clinical trial. After staging of disease status and splenectomy, pegylated liposomal encapsulated doxorubicin was administered intraperitoneally (1 mg/kg body weight) every 3 weeks for 4 cycles. All dogs were monitored for recurrence of HSA. Samples of plasma and abdominal fluid were collected for measurement of doxorubicin concentration and pharmacokinetic analysis. Nonlinear mixed-effect modeling was used to describe the pharmacokinetics of liposomal doxorubicin administered IP., Results: All 14 dogs died, 12 because of HSA and 2 from other causes. Postmortem examination was performed on 12 dogs. All 12 dogs died because of HSA-related causes and had hepatic metastases and hemoabdomen. The IP-treated dogs had fewer serosal, mesenteric, and omental metastases than historical controls treated with systemic doxorubicin. Results of the postmortem examination and pharmacokinetic analysis confirmed that IP delivery of doxorubicin resulted in an effective drug concentration with a clearance comparable with that after i.v. delivery., Conclusions and Clinical Importance: IP pegylated liposomal encapsulated doxorubicin administration did not prevent intraabdominal recurrence of HSA in dogs.

Introduction and Development: Chemotherapy protocols for treatment of brain tumors use toxic molecules for killing cancer cells in a similar way that protocols for treating other cancers. Therefore, secondary effects and poor response are the major handicaps. Technological developments based on pharmacogenomics and pharmacoproteomics will predict response and toxicity giving rise to a personalized medicine. However, there are only few studies that correlate chemotherapeutical molecules for brain tumor treatment and prediction of response and toxicity., Conclusions: The development of new technologies based on high-density microarrays allows the progressive identification of genes whose presence will predict the efficacy of therapeutic protocols. Once identified, specific equipments based on low-density arrays will detect exclusively in an easy and fast way the presence of genes in order to predict patient's response and avoid toxicity. Other more sophisticated techniques at present still at an experimental step based on proteomics as MALDI (Matrix-Assisted Laser Desorption Ionization) and SELDI (Surface-Enhanced Laser Desorption Ionization) will allow the identification of proteins that could predict response and toxicity.

Purpose: The effectiveness of massage therapy on immune parameters was evaluated in young Dominican HIV+ children without current access to antiretroviral therapies., Methods: Eligible children, who were followed at the Robert Reid Cabral Hospital (San Domingo, Dominican Republic), were randomized to receive either massage treatment or a control/friendly visit twice weekly for 12 weeks. Blood was drawn at baseline and following the 3-month intervention for determinations of CD4, CD8, and CD56 cell counts and percentage, along with activation markers (CD25 and CD69)., Results: Despite similar immune parameters at baseline in the two groups, significantly more of the control group exhibited a decline in CD4 cell count (>30%, p = 0.03), postintervention. The decrease was particularly evident in older (5-8 years) children in the control arm, who demonstrated a significant reduction in both CD4 and CD8 cell counts compared to massage-treated older children who remained stable or showed immune improvement. Additionally, a significant increase in CD4+CD25+ cells was observed over the 12-week trial in the massage-treated older children (p = 0.04) but not in the control group. In younger massage-treated children, (2-4 years old), a significant increase in natural killer cells was shown., Conclusion: Together these findings support the role for massage therapy in immune preservation in HIV+ children.

Gastroesophageal reflux (GER) is a common finding in the newborn period, which is in general well tolerated, and tends to resolve spontaneously at about 18 months of age. However, a small number of children will present important respiratory manifestations (aspiration and apneic episodes), or increase in the manifestations of pre existing conditions (tracheomalacia, laryngomalacia), which can produce life threatning events. Ph monitoring does not completely discard the possibility of GER and the use of prokinetic drugs, in association with other drugs to protect the esophagus do not prevent serious consecuences produced by reflux to the airway. In this report six children with severe airway disease and marked GER, with no response to initial medical treatment required laparoscopic fundoplication. They have all resolved their airway disease, and are currently well. The laparocopic technique for fundoplication of small children seems to be a secure, well tolerated and definitive treatment in the hands of trained surgeons.

The tools of recombinant protein expression are now being used to provide recombinant sources of both collagen and gelatin. The primary focus of this review is to discuss alternatives to bovine collagen for biomedical applications. Several recombinant systems have been developed for production of human sequence collagens. Mammalian and insect cells were initially used, but were thought to be too costly for commercial production. Yeast have been engineered to express high levels of type I homotrimer and heterotrimer and type II and type III collagen. Co-expression of collagen genes and cDNAs encoding the subunits of prolyl hydroxylase has lead to the synthesis of completely hydroxylated, thermostable collagens. Human types I and III collagen homotrimers have been expressed in transgenic tobacco plants, while transgenic mice have been engineered to produce full-length type I procollagen homotrimer as well as a alpha2 (I) homotrimeric mini-collagen. Most recently, a transgenic silkworm system was used to produce a fusion protein containing a collagenous sequence. Each of these transgenic systems holds great promise for the cost-effective large-scale production of recombinant human collagens. As seen in other recombinant expression systems, transgenic silkworms, tobacco, and mice lack sufficient endogenous prolyl hydroxylase activity to produce fully hydroxylated collagen. In mice and tobacco, this was overcome by over-expression of prolyl hydroxylase, analogous to what has been done in yeast and insect cell culture. In addition to recombinant alternatives to bovine collagen, other sources such as fish and sponge collagen are discussed briefly. Recombinant gelatin has been expressed in Pichia pastoris and Hansenula polymorpha in both non-hydroxylated and hydroxylated forms. Pichia was shown to be a highly productive system for gelatin production. The recombinant gelatins produced in yeast are of defined molecular weight and physio-chemical properties and represent a new biomaterial not previously available from animal sources. Genetic engineering has made great progress in the areas of recombinant collagen and gelatin expression, and there are now several alternatives to bovine material that offer an enhanced safety profile, greater reproducibility and quality, and the ability of these materials to be tailored to enhance product performance.

Evaporation of water from a 1/1 mixture of trehalose and sucrose gives rise to optically clear glasses that are transparent in the UV and visible ranges and do not crystallize when they are prepared at ambient temperatures. Two proteins, liver alcohol dehydrogenase and parvalbumin, and the tryptophan derivative N-acetyl-tryptophanamide were incorporated into the glasses. Infrared spectroscopy of the amide I band reveals that the proteins retain secondary structure in the glass over a temperature range of 20-300K. The amide II band of the protein and the HOH bending band of residual water in the glass shift with temperature changes, consistent with increased H-bonding strength as temperature is lowered. Phosphorescence of tryptophan can be seen from the proteins at room temperature, which shows the immobilization of the protein by the glass and the curbing of oxygen diffusion. It is suggested that using mixed sugars to form glasses is a way to immobilize proteins over a wide temperature range without distortions from solvent crystals.

Although clinical studies have shown that posterior cut angle affects tibial component stability, biomechanical studies are lacking. Fifteen Sawbones tibiae were divided into 5 groups and prepared with 0 degrees , 3 degrees, 6 degrees, 9 degrees, and -5 degrees tibial surface cuts, and a tibial component was implanted with cement. Using a standard and then a highly congruent polyethylene insert, the knee was loaded at 0 degrees and 30 degrees of flexion. There were statistically significant increases in anterior micromotion of the standard polyethylene component for each increase in posterior slope cut angle, which increased for the highly congruent polyethylene component. The anterior slope cut (-5 degrees) led to significant posterior micromotion of the tibial polyethylene component. Increased posterior slope cut angle significantly decreased tibial anterior compressive strains and significantly increased tibial posterior compressive strains. The highly congruent insert significantly increased this posterior strain. The results indicate that cutting the articular surface of the tibia at a 0 degrees or 3 degrees posterior slope provides the greatest tibial component stability.

Objective: To characterize imaging findings in cats with confirmed inflammatory bowel disease (IBD) of the upper gastrointestinal tract (i.e., stomach and small intestine) and relate these findings to clinical signs and histologic changes., Design: Retrospective study., Animals: 32 cats with clinical and histopathologic diagnoses of IBD., Procedure: Medical records were reviewed for signalment, clinical signs, clinicopathologic findings, radiographic and ultrasonographic findings, and results of endoscopic examination. Histologic findings were reviewed and characterized by severity and type of inflammatory infiltrate., Results: All cats had 1 or more clinical signs (e.g., vomiting, diarrhea, weight loss, and anorexia) consistent with IBD. Lymphocytic and plasmacytic infiltrates were observed in histologic sections of gastrointestinal tissue. Crypt distortion, villous blunting and fusion, and fibrosis were most commonly seen in cats with moderate or severe IBD. Clinicopathologic findings of some cats included anemia, leukocytosis or leukopenia, hypocholesterolemia, and hyper- or hypoproteinemia. Abnormalities were not found on abdominal radiographic views in 9 of 9 cats. However, contrast studies using barium revealed radiographic abnormalities in 1 of 3 cats. In 13 of 17 cats, abdominal ultrasonography revealed several intestinal abnormalities (e.g., poor intestinal wall layer definition, focal thickening) and large mesenteric lymph nodes with hypoechoic changes consistent with IBD. Endoscopic observation revealed findings (e.g., erythema, plaques, mucosal friability) consistent with inflammation in 9 of 18 cats., Conclusions and Clinical Relevance: Compared with endoscopy of the gastrointestinal tract or abdominal radiography, clinical signs and ultrasonographic findings appear to have the best association with histologic grade of IBD in cats.

Human immunodeficiency virus type 1 (HIV-1) protease was expressed in Escherichia coli as a fusion protein with the N-terminal sequence of IGF-2. The protein accumulated in inclusion bodies as a 40:60 mixture of unprocessed fusion protein and processed protein. A simple purification procedure was developed that yielded 30-40 mg of active protease per liter of fermentation broth with a recovery of 30-40%. The purification process involved the selective extraction of HIV-1 protease from E. coli inclusion bodies with 50% acetic acid and fractional diafiltration to remove impurities and low-molecular-weight protease-related fragments. No chromatographic steps were employed, yet the HIV-1 protease produced by this procedure was greater than 95% pure by SDS-PAGE, reverse-phase HPLC, and N-terminal sequence analysis.

The endobronchial anatomy of 12 lung specimens from horses and 12 healthy, standing, sedated horses was evaluated, using a 200-cm-long, 9.5-mm-diameter videoendoscope. On the basis of these findings, the nomenclature system of Amis and McKiernan was modified for identification of airways of horses during bronchoscopy. Lobar bronchi are identified on the basis of the side of the bronchial tree on which they were found and the order in which they originated from the primary bronchus. Thus, RB1, RB2, and RB3 referred to right cranial lobar bronchus, respectively. On the left side, the designation of LB1 and LB2 refer to the left cranial lobar bronchus and the left caudal lobar bronchus, respectively. Segmental bronchi are identified by consecutive numbers in the order of origination from the lobar bronchus. The direction of the segmental bronchus was denoted by the capital letter D (dorsal), V (ventral), L (lateral), M (medial), R (rostral), and C (caudal). Subsegmental bronchi were identified in the order of origination from the segmental bronchi, using lower case letters (eg, RB2, 1V, a or RB2, 1V, aV). For subsequent branching of the subsegmental bronchi, the branches were numbered consecutively by their order of origination (eg, RB2, 1V, aV, 1D).

T4 RNA ligase catalyzes the ATP-dependent addition of a single 2'-deoxyribonucleoside 3',5'-bisphosphate to the 3'-hydroxyl of an oligodeoxyribonucleotide. The bisphosphate is joined to the deoxyoligomer by a 3' leads to 5' phosphodiester bond and the product, which is terminated by a 3'-phosphate, is one nucleotide longer than the substrate. Bisphosphates of dAdo, dCyd, dGuo, dThd, and dUrd are donors and oligodeoxyribonucleotides with dA, dC, dG, dT, or dU 3' termini act as acceptors. The preferred residue for both donor and acceptor is dCyd. Deoxyoligomers from 3 to 12 residues in length are active as acceptors. To obtain good yields, high concentration of enzyme, long incubation time at low temperature, and manganous rather than Mg(II) ion are required. Under optimal conditions, yields calculated with respect to deoxyoligomer converted to product vary from 40 to greater than 95%. The turnover number of the enzyme for DNA joining is extremely low but, because the preparation is nearly free of DNases, there is less than 3% degradation of substrate or product after 6 days of reaction. We anticipate that this reaction will serve as the basis for a method for the stepwise enzymatic synthesis of DNA of defined sequence.