Assessing the safety of del Nido cardioplegia solution in adult congenital cases.

PURPOSE: del Nido cardioplegia solution (CPS) has been successfully used for myocardial protection in the pediatric population. We propose this solution can be used safely in adult congenital patients. The proposed benefit of this solution is the avoidance of the need for repetitive interruption of the operation to administer multiple doses of standard cardioplegia. METHODS: As part of a quality improvement initiative, 47 consecutive adult patients (mean age 40.9 years, range 18-71) undergoing congenital heart surgery were given del Nido CPS. Cardiac function was assessed pre- and post-operatively by echocardiography (ECHO). Inotrope use, troponin levels and restoration of cardiac rhythm were also evaluated. RESULTS: The average duration of the longest ischemic time was 52.5 minutes ± 15.57 minutes. In patients receiving a single dose (40%, n=19) of CPS, the average ischemic time was 49.8 minutes ± 18.8 minutes. No patients demonstrated any ventricular electrical activity while the aorta was cross-clamped. Post-operative ECHO showed that 94% (n=44) had no change in ejection fraction from the pre-operative ECHO. Patients requiring inotropic support at the time of leaving the operating room (OR) was 43% (n=20). The percentage of patients requiring inotropic support twenty-four hours post-operatively was 17% (n=8). Spontaneous restoration of cardiac rhythm (without the need for defibrillation) after cross-clamp removal occurred in 91% (n=43) of patients. The average troponin T level post-op was 1.86 ± 2.9 µg/L. CONCLUSIONS: del Nido CPS can be used for myocardial protection during adult congenital cardiac surgery without any apparent adverse effects. In addition, we were able to change our re-dosing protocol to 45 minutes with del Nido CPS compared to 20 minutes with our adult 4:1 blood CPS.

Comet-tail-like inflammatory infiltrate to polymer filaments develops in tension-free conditions.

BACKGROUND: Mesh reinforcement in hiatal hernia repair becomes more frequent but is charged by complications such as erosion or stenosis of the oesophagus. These complications are accompanied by an intense inflammatory infiltrate around the polymer fibres. To characterize this effect, the response to polypropylene fibres in the absence of tension was examined. METHODS: In rats, polypropylene sutures (USP size 1, 3-0 and 7-0) were placed in the subcutis of the abdominal wall without knot or tension. On postoperative days 3, 7 and 21, specimens were excised. The expressions of c-myc, ß-catenin, Notch3, COX-2, CD68 and Ki-67 were measured by immunohistochemistry. RESULTS: In the absence of tension, sutures were surrounded by a foreign body granuloma with an inflammatory infiltrate not encircling the fibre but forming almost symmetric comet-tail-like infiltrates on opposite sides. The expression of c-myc, ß-catenin, Notch3, COX-2, CD68 and Ki-67 was significantly reduced over time in the comet tail, but not in the granuloma. CONCLUSIONS: Even in tension-free conditions, surgical sutures cause a foreign body response with infiltrates of inflammatory cells. This reaction is shaped like a comet tail, and its extension depends on the diameter of the used fibre. Therefore, for reduction of perifilamental infiltrates, not only absence of tension is required, but also a small-sized fibre textile.

Evaluation of tissue components in the peripheral nervous system using Sirius red staining and immunohistochemistry: a comparative study (human, pig, rat).

Little is known about species differences in the peripheral nerve system and quantitative evaluation of main tissue components has rarely been done. Nevertheless, animal models are used for example in pain research without exact knowledge of degree of fibrosis in pathological states which would determine possible treatment options. It would therefore be of crucial interest to describe the degree of fibrosis and the remaining functional nerve tissue as exact as possible. In the present study we evaluated collagen (stroma) and nerve fiber (parenchyma) composition of peripheral nerves in three species (human, rat, pig) and used digital colour-separation and analysis for collagen type differentiation and quantification of immuno-positive-stained area. We found similar ratios of collagen types I and III in epineurium and similar immuno-positive area for staining of neurofilament and S-100beta. In contrast, we measured significantly different ratios of collagen type I to type III in the endoneurium. This combined analysis of the main tissue components of peripheral nerves could be an easy-to-use tool in evaluating changes during damage caused by scaring, systemic disease or compression syndromes. The calculated collagen type I/III ratio may serve as an objective diagnostic value for the description or as prognostic marker for therapeutic approaches in peripheral nerve pathology. However, in particular studies of collagen accumulation in nerves, species dependant differences have to be considered.

[Importance of abdominal compartment syndrome in Germany: a questionnaire]. / Bedeutung des abdominellen Kompartmentsyndroms in Deutschland: Eine Umfrage.

BACKGROUND: The abdominal compartment syndrome (ACS) is a life-threatening condition and may affect any critically ill patient. Little is known about the recognition and management of the ACS in Germany. METHODS: A postal questionnaire was sent to departments of surgery and anesthesia of German hospitals with more than 450 beds. RESULTS: From the 222 eligible hospitals a total of 113 replies were received. Most respondents (95%) indicated that the ACS plays a role in their clinical practice. Measurement of intra-abdominal pressure (IAP) is not performed by 26% while it is routinely done by 30%. Intra-abdominal pressure is mostly (94%) assessed via the bladder pressure. Of the respondents 41% measure IAP only in those patients thought likely to develop ACS. Risk factors of the ACS would lead to IAP monitoring in 10-23% of cases. The majority (86%) would require signs of organ dysfunction together with exceeding the IAD threshold in order to opt for a surgical decompression. The attitude towards the critical threshold (>20 mmHg or >25 mmhg) divided respondents into two groups of similar size (39% compared to 47%, respectively). CONCLUSIONS: German anesthesiologists and surgeons are familiar with the ACS. However, about one-quarter never measure IAP and there is a considerable variance as to which patients are at risk to develop ACS and how often IAP should be measured in these patients. This could indicate a lack of acceptance or simply a persisting need for more data concerning the avoidance and treatment of the ACS.

The Gist of literature on pediatric GIST: review of clinical presentation.

AIM/BACKGROUND: To provide a review of existing literature on pediatric GIST with focus on clinical presentation. METHODS: A MEDLINE search was conducted in July 2007 to give an overview on literature concerning pediatric gastrointestinal stromal tumors (GISTs) with a focus on clinical presentation, using keywords "gastrointestinal stromal tumor" and one of the following "young/boy/girl/child/children/pediatric." Two of the authors sorted the resulting abstracts by relevance for a review on clinical aspects of pediatric GIST if they were in English language, not explicitly only reporting of adults and describing clinical features of patients. RESULTS: One hundred and six articles were found, 43 of which were excluded because they did not match the criteria mentioned above. We found 97 patients in the articles meeting our criteria, of which 38 cases had to be excluded, because of lacking clinical data, negative staining for CD117 or syndromal occurrence. This left 59 patients for analysis of clinical symptoms in the presentation of nonsyndromal CD117-positive GIST in children. DISCUSSION: Clinical feature most frequent was anemia in 86.4% (n=51) symptomatic either through acute or subacute bleeding. There was no palpable tumor in 88.1% (n=52), no abdominal pain in 84.7% (n=50), and no vomiting in 88.1% (n=52). Girls tend to show more high-grade tumors and existing case reports show a 2.7-fold higher incidence in females. Altogether epithelioid cell tumors are most frequent, although in boys spindle-cell tumors are reported more often. On the basis of National Institute of Health criteria (6) tumors were low grade in 22% (n=13), medium grade in 37.3% (n=22), and high grade in 35.6% (n=21). There were more high-grade tumors in girls than in boys (40.5% vs. 28.6%). Local excision was the operation most often performed, but details of surgery were missing in most cases. CONCLUSIONS: Pediatric GIST is a rare but considerable diagnosis in chronic anemia, which is the most frequent clinical finding with this tumor entity. Recent review articles focus on histopathologic criteria but omit clinical features and course of disease. In nonsyndromal CD117-positive GIST, girls tend to show more high-grade tumors and existing literature on pediatric GIST shows a 2.7-fold higher incidence in females. Altogether epithelioid cell tumors are most frequent, although in boys spindle-cell tumors are reported more often. Together with known differences in molecular changes and local as well as systemic tumor behavior this strongly suggests that pediatric GIST represents a different entity than adult GIST. After establishment of clear-cut pathologic features in the past, reports on preoperative diagnostic findings, long-term follow-up, and therapy have to be emphasized to clarify the relationship of these entities.

A clinical evaluation of the Dideco Kids D100 neonatal oxygenator.

In August 2006, Duke University Perfusion Services had the opportunity to be the first institution in the United States to clinically evaluate the Dideco D100 Neonatal Oxygenator. The device was used on six pediatric patients to facilitate correction or palliation of their cardiac defects, which included two arterial switch operations, two truncus arteriosus repairs, one stage 1 Norwood and one repair of total anomalous pulmonary venous return. The average patient weight was 3.1 kg. The average cardiopulmonary bypass (CPB) time was 135 minutes and the average cross-clamp time was 61 minutes. Arterial and venous blood gasses were drawn and used to calculate oxygen transfer. The average oxygen transfer was 14.8 +/- 10.3 ml/O2/min. The Dideco D100 Oxygenator is the first oxygenation device designed specifically for neonates. The Dideco D100 is a microporous hollow-fiber device. It has a static priming volume of 31 ml and a maximum rated flow of 700 ml/min. The integral hard-shell venous reservoir has a minimum operating level of 10 ml and a reservoir capacity of 500 ml. For this evaluation, the Dideco Kids D100 Neonatal Oxygenator performed adequately on patients weighing up to 5 kg. This device provides an excellent first step towards offering very small children appropriate circuitry without having to sacrifice safety or performance.

Construction and characterization of a plant transformation-competent BIBAC library of the black Sigatoka-resistant banana Musa acuminata cv. Tuu Gia (AA).

A plant transformation-competent binary bacterial artificial chromosome (BIBAC) library was constructed from Musa acuminata cv. Tuu Gia (AA), a black Sigatoka-resistant diploid banana. After digestion of high-molecular-weight banana DNA by HindIII, several methods of DNA size selection were tested, followed by ligation, using a vector/insert molar ratio of 4:1. The library consists of 30,700 clones stored in 80 384-well microtiter plates. The mean insert size was estimated to be 100 kb, and the frequency of inserts with internal NotI sites was 61%. The majority of insert sizes fell into the range of 100+/-20 kb, making them suitable for Agrobacterium-mediated transformation. Only 1% and 0.9% of the clones contain chloroplast and mitochondrial DNA, respectively. This is the first BIBAC library for banana, estimated to represent five times its haploid genome (600 Mbp). It was demonstrated by hybridization that the library contains typical members of resistance gene and defense gene families that can be used for transformation of disease susceptible banana cultivars for banana genetic improvement.

Use of ECMO without the oxygenator to provide ventricular support after Norwood Stage I procedures.

Extracorporeal membrane oxygenation (ECMO) has been found effective in supporting infants with severe cardiac dysfunction following open heart surgery. Centers using this mode of support can also, in instances of single ventricle morphology, consider the option of eliminating the oxygenator from the standard ECMO set-up and thereby provide roller pump ventricular assist. In these cases, the infant's own lungs can provide excellent oxygenation simply by leaving the aortopulmonary shunt open. Since ventricular support ensures maintenance of normal cardiac output, manipulation of pulmonary versus systemic flows is not necessary. This configuration retains the safety features of the ECMO system and is easily staffed by the ECMO support personnel. There may be several benefits to employing this type of management.

Techniques of paediatric modified ultrafiltration: 1996 survey results.

In September 1996, perfusionists from 50 paediatric open-heart surgery programmes were contacted to identify centres that are currently using the technique of modified ultrafiltration (MUF). Of the 50 centres contacted, 22 (44%) were utilizing the technique. These centres were surveyed on the following: neonatal circuit description, patient entry criteria, MUF circuit description, conduct of MUF, use of extracorporeal safety devices and/or modifications, and technical complications. All 22 centres used roller pumps and membrane oxygenators. In 19 centres, MUF was utilized exclusively in the arteriovenous mode (86%), while two centres (9%) used the venovenous mode and one centre (5%) used both methods. Most (82%) of the 22 MUF centres used a blood cardioplegia system for myocardial preservation. After cardiopulmonary bypass (CPB), these blood cardioplegia systems were often converted for use as MUF circuits in a variety of ways. Other methods of accessing the CPB circuit for MUF included utilizing either a recirculation line or a dedicated port added to the circuit specifically for MUF. Blood flow rates during MUF, pump strategies, haemoconcentrator vacuum levels and endpoints were variable from centre to centre. Technical complications related to MUF were reported by 82% of the surveyed MUF centres. The most common complication, air cavitating into the circuit, was reported by 15 centres. From these data, we propose recommendations on the integration of MUF into CPB circuits, the conduct of perfusion during MUF, and appropriate safety considerations to minimize technical complications.

Experimental use of an ultra-low prime neonatal cardiopulmonary bypass circuit utilizing vacuum-assisted venous drainage.

In adult cardiopulmonary bypass surgery, vacuum assisted venous drainage has become a popular technique to augment venous return to the bypass circuit. The application of this technique in neonatal cardiopulmonary bypass surgery could be beneficial to the further miniaturization of neonatal circuitry by coupling radical respositioning of the oxygenator and pump console with decreasing line length. This report communicates the use of an investigational, vacuum assisted venous drainage neonatal circuit that is positioned at patient level utilizing a modified pump console with elevated double head twin roller pumps. The circuit, including the oxygenator, arterial line, venous line, raceway tubing, and a functional level in the venous reservoir has a priming volume of 107 ml. Initial bench and animal tests have demonstrated that this technique may be clinically feasible in CPB applications. With vacuum assisted venous drainage, the goal of asanguinous neonatal cardiac surgery could become a reality. Safety issues must be adequately addressed to ensure that this technique does not impose unacceptable risks.

Multilocus DNA fingerprinting and genetic relatedness in plants: a case study with banana and tomato.

The technique of DNA fingerprinting is frequently used for studies of genetic diversity and relatedness in a wide range of organisms. In humans and animals, multilocus fingerprints are mainly applied to paternity and identity test cases, behavioral ecology, and the analysis of population structures. In plants and fungi, the frequent occurrence of "low-variability" fingerprint patterns additionally allows to use multilocus fingerprinting for studying taxonomical problems at an intraspecific level. In the present article, we (1) present an overview of such approaches in a series of plant species, (2) summarize our attempts to estimate genetic relationships within two cultivated plant species, banana and tomato, by band sharing data derived from oligonucleotide fingerprints, and (3) discuss the limitations and potentials of multilocus fingerprinting for the determination of genetic relatedness.

DNA fingerprinting in sugar beet (Beta vulgaris) - identification of double-haploid breeding lines.

The distribution and abundance of simple repetitive sequences complementary to the synthetic oligonucleotides (GACA)4, (GATA)4, (GTG)5 and (CA)8 in the genomes of several cultivars of Beta vulgaris and in the wild beet B. vulgaris ssp. maritima were investigated. Hybridization experiments revealed that all four motifs were present, though at different abundances, in the genomes of all of the investigated beet cultivars. Considerable intraspecific variation of the resulting DNA fingerprints was observed. The extent of polymorphism depends on the oligonucleotide probe. The most informative banding patterns were obtained with the (GATA)4 probe hybridized to HinfI-, HaeIII-, or RsaI-restricted DNA, respectively. DNA fingerprinting with (GATA)4 allowed a clear differentiation of double-haploid breeding lines (DH lines). We demonstrated that the application of oligonucleotide probes for DNA fingerprinting is a sensitive tool for genome diagnosis in cultivated beet.

Oligonucleotide and amplification fingerprinting of wild species and cultivars of banana (Musa spp.).

DNA oligonucleotide and amplification fingerprinting have been successfully used to detect genetic polymorphisms in 15 representative species and cultivars of the genus Musa, comprising AA, AAA, AAAA, AAB, ABB, and BB genotypes. In-gel-hybridization of Hinf I-digested genomic banana DNA to the 32P-labeled synthetic oligonucleotides (GATA)4, (GTG)5, and (CA)8 revealed considerable polymorphisms between Musa species and cultivars. The fingerprint patterns proved to be somatically stable and did not show differences between individual plants of 'Grand Nain' (AAA genotype). Dendrograms based on oligonucleotide fingerprint band sharing data proved to be consistent with most of the known features of the history of banana and plantain cultivation and evolution, respectively. DNA samples from the same banana species and cultivars were also amplified by PCR using single or pairwise combinations of short oligonucleotide primers. Amplification products were separated on agarose or polyacrylamide gels and visualized by ethidium bromide or silver staining, respectively. Polymorphic patterns were obtained with some but not all primers. By using the CCCTCTGCGG primer in simplex and/or duplex PCR, the induced mutant 'GN60A' was clearly recognized from its original variety 'Grand Nain'. Both fingerprint techniques allowed the detection of bands characteristic for the A and B genome. This DNA fingerprinting technology has potential application in several areas of Musa improvement.

Contamination of highly purified human serum cholinesterase by dipeptidyl peptidase IV causing hydrolysis of substance P.

The purification and kinetic characterization of cholinesterase from blood plasma (pseudocholinesterase; butyrylcholinesterase: EC 3.1.1.8) is described. The hydrolysis of the artificial peptide substrate Lys-Pro-p-nitroanilide served as a model of the second step in degradation of substance P by dipeptidyl peptidase IV. The substrate is hydrolyzed by a gel-electrophoretic homogeneous cholinesterase preparation with a reaction rate of 5.8 mumol/min X mg and a KM value of 0.12 mmol/l. The proteolytic reaction could not be affected with typical cholinesterase inhibitors NaF and dibucain. On the other hand Lys (pNO2-Z)-Pro and a specific suicide substrate (diacylhydroxylamine derivative) inhibit the activity in a manner analogous to dipeptidyl peptidase IV. Though these active site-directed inhibitors also influenced the benzoylcholine hydrolyzing activity of serum cholinesterase, we conclude from the data that dipeptidyl peptidase IV was the true Lys-Pro-p-nitroanilide cleaving activity. Furthermore, the conclusion can also be drawn that hydrolysis of substance P reported by Lockridge 1982 is caused by the contamination that cannot be completely separated from the esterase during the purification method used.