Validation of a KHV antibody enzyme-linked immunosorbent assay (ELISA).

Koi herpesvirus (KHV) causes KHV disease (KHVD). The virus is highly contagious in carp or koi and can induce a high mortality. Latency and, in some cases, a lack of signs presents a challenge for virus detection. Appropriate immunological detection methods for anti-KHV antibodies have not yet been fully validated for KHV. Therefore, it was developed and validated an enzyme-linked immunosorbent assay (ELISA) to detect KHV antibodies. The assay was optimized with respect to plates, buffers, antigens and assay conditions. It demonstrated high diagnostic and analytical sensitivity and specificity and was particularly useful at the pond or farm levels. Considering the scale of the carp and koi industry worldwide, this assay represents an important practical tool for the indirect detection of KHV, also in the absence of clinical signs.

Microbiological stability of solutions containing local anesthetics and opioids in closed infusion systems used for epidural analgesia.

Nine solutions containing opiod analgesics and local anesthetics as typically use in epidural catheters were tested for antimicrobial stability. Administration via a pefusor syringe requires several refill processes. It was shown that repetitive refilling of the syringes did not result in any microbiological contamination.

Carbonic anhydrase in the gills of seawater- and freshwater-acclimated flounders Platichthys flesus: purification, characterization, and immunohistochemical localization.

Flounders Platichthys flesus were investigated with respect to isolation, purification, and cellular localization of carbonic anhydrase (CA) in the respiratory system. CA was purified from gills and erythrocytes and was shown to exclusively represent a soluble enzyme with an apparent molecular weight of 30 kD. Inhibition constants (KI) towards acetazolamide (ACTZ) were 8.4.10(-9) M for erythrocyte CA and 7.6.10(-9) M for gill CA, indicating a high sensitivity to sulfonamides, as exhibited by human CA II. Specific CA activity did not differ significantly in seawater- and freshwater-acclimated fish. Antibodies were raised against purified gill and erythrocyte CA. Both antisera crossreacted and were used to localize CA in the gills of seawater and freshwater flounders at the light microscopic level. Independent of the salinity, a positive reaction of variable intensity was found in the following cell types: pavement cells (PVCs), forming the gill epithelial surface layer; mucous cells (MCs); pillar cells (PCs), bordering the vascular channels of the secondary lamellae; and chloride cells (CCs), mitochondria-rich cells located in the primary epithelium, the interlamellar regions, and at the bases of the secondary lamellae.(J Histochem Cytochem 47:43-50, 1999)

[Intestinal pouches: gastric reconstruction]. / Intestinale Pouches: Magenersatz.

Reconstruction of the intestinal passage after a total gastrectomy is usually based on a direct esophagojejunostomy with end-to-side implantation of the afferent loop. The second principle of reconstruction is based on preservation of the duodenal passage. Long-term problems such as weight loss and malnutrition are further considerations that lead to the concept that gastric reconstruction should have the form of a reservoir. In addition to the construction of the reservoir itself, the clinical concern of avoiding gastroesophageal reflux is a further requirement for the choice of reconstruction type. Diversion of the duodenal content via a Roux-en-Y end-to-side anastomosis is considered to be the standard procedure. Interposition of a sufficiently long duodenal loop with maintenance of the duodenal passage also has the effect of preventing duodenal reflux. A theoretical advantage of this procedure is the linking of the motility of the duodenum with that of the interposed segment with improved synchronization of the aboral nutrient passage. When one considers complicated reconstructive procedures, the present literature suggests construction of a pouch is definitely functionally superior to the simple esophagojejunostomy. Whether the duodenal passage should be maintained or whether a Roux-Y technique should be used is a question that is still open for discussion.

[Lymphadenectomy in tumors of the upper gastrointestinal tract]. / Lymphadenektomie bei Tumoren des oberen Gastrointestinaltrakts.

Similar to other tumor entities, complete tumor removal with an adequate safety margin in all three dimensions (the oral margin, the aboral margins and the tumor bed) must be the primary aim of any surgical approach to carcinoma of the upper gastrointestinal tract. The same goal has to be achieved in the area of the lymphatic drainage. All positive nodes and nodes with a so-called 'microinvolvement' have to be removed together with the primary tumor. The safety margin of lymphadenectomy can be estimated by the lymph node ratio, i.e., the ratio between the number of removed and positive nodes. Several studies have shown that for carcinoma of the upper gastrointestinal tract the prognosis can be improved markedly if the lymph node ratio is below 0.2. For tumors in the early phase of lymphatic metastasis this can be achieved by extensive lymph node dissection. In practice, this requires as a minimum a lymphadenectomy of compartments I and II of the tumor's lymphatic drainage (D2 lymphadenectomy). The individual compartments are determined by the embryogenesis of the affected organ and defined by the tumor location. In patients with advanced lymphatic metastases, lymphadenectomy does not improve the prognosis and can only result in a reduction of local recurrences. Lymphadenectomy does not increase the risk and morbidity of the surgical procedure, provided it is restricted to the removal of nodes. These basic principles of lymphadenectomy are valid for carcinomas of the esophagus, cardia and stomach.

[Extent of resection in surgery of stomach carcinoma]. / Resektionsausmass in der Chirurgie des Magenkarzinoms.

The extraluminal extent of resection in cases of advanced gastric cancer is controversial. If, however, following meticulous staging--including the detection of free abdominal tumor cells--complete resection seems possible, then multivisceral resection is justified. If complete resection is achieved, the prognosis of these patients can be improved. Left pancreatic resection should be performed only if the tumor invades the pancreas directly. Splenectomy is indicated if the tumor invades the organ directly or if there are locally advanced tumors of the proximal third of the stomach and tumors of the esophageal-gastric junction. However, it has to be kept in mind that splenectomy is an independent negative prognostic factor. The extent of lymphadenectomy (LA) in gastric cancer is still under discussion. According to the 10-year results of the Dutch Gastric Cancer Study, there might be subgroups which have a survival benefit after extended (D2) LA. These include, as the German Gastric Cancer Study corroborated, patients with very early stage II and stage IIIa lymph node metastases. As neither of these stages can at present be diagnosed before or during surgery, D2 lymphadenectomy should be the standard procedure for all patients with gastric cancer. Recent studies have shown that it might be possible with the help of the Sentinel Node Technique to individualize lymphadenectomy in locally gastric cancer as well. The beneficial effects of adjuvant chemoradiation in gastric cancer do not mean, however, that the extent of resection may be reduced. Adjuvant chemoradiation following complete resection and D2 lymphadenectomy should still not be regarded as standard therapy.

[Does the new UICC classification allow better prognostic assessment for ductal pancreatic carcinoma?]. / Erlaubt die neue UICC-Klassifikation eine bessere Prognoseabschätzung für das duktale Pankreascarcinom?

INTRODUCTION: To compare treatment results an international established classification is necessary. In 1997 the TNM classification of 1992 was modified. METHODS: Between 1983 and 1997, 159 patients with a ductal carcinoma of the pancreas underwent resection. All data of the resected specimens were documented in standardized manner prospectively in a protocol that offered ready transfer of the collected data to a new classification. The TNM categories and stage groupings were transferred to the new UICC classification of 1997 and analyzed in comparison to the classification of 1992. RESULTS: The inclusion of a pT4 category equivalent to the other GI tumors made a new stage grouping necessary. Also division into pN1 a and pN1 b was established. According to the clinical experience only few tumors in early stages (pT1/2 and stage I) were observed in the new classification. There was a significant improval in the patient's distribution to the new stage grouping because of the homogeneous groups. In comparison to the 1992 classification the new stage II shows a relevant prognostic value and a significant difference to stage III. CONCLUSION: We conclude that the new UICC classification relates to prognosis better than the old classification.

[Risk of surgical therapy of stomach cancer in Germany. Results of the German 1992 Stomach Cancer Study. German Stomach Cancer Study Group ('92)]. / Risiko der chirurgischen Therapie des Magencarcinoms in Deutschland. Ergebnisse der Deutschen Magencarcinom-Studie 1992. Deutsche Magencarcinom-Studiengruppe (GGCS '92).

The German Gastric Cancer Study (GGCS '92) permits a representative pictures of the current situation in surgery for gastric carcinoma at university centers in Germany. The analysis of frequency of resection, complication rate and mortality provides a quality measure for anybody who performs surgical therapy for gastric carcinoma. The results of multivariate analyses will have therapeutic consequences.

[Telemetric impedance analysis of the liver: evaluation of a noninvasive device for diagnosis of acute graft rejection after experimental liver transplantation]. / Telemetrische Impedanzanalyse der Leber: Evaluation eines nichtinvasiven Instrumentariums zur Diagnose akuter Abstossungsreaktionen nach experimenteller Lebertransplantation.

Allograft rejection and its differentiation from other causes of organ dysfunction remains a diagnostic problem in liver transplant patients. Currently, acute rejection can be prevented only by a combination of diagnostic and therapeutic modalities. The diagnostic potential of a novel implantable telemetric rejection monitoring device has been assessed on the basis of the noninvasive impedance analysis in normal and liver transplanted pigs. The electric impedance data were correlated with biochemical and histological parameters. Acute rejection was correctly predicted in n = 4, and correctly excluded in n = 32, biopsy-related impedance recordings (p = 0.004). A correlation between impedance measurements and severity of histological findings r = 0.84; p = 0.0001) was confirmed. Only the biochemical parameters SGLDH and serum bilirubin revealed a comparable correlation. Impedance gradient analysis revealed evidence of a physiological relationship between liver function and the electrical properties of the organ. Telemetric impedance analysis would appear a promising means of assessing acute rejection noninvasively.

[Cecal diverticulitis]. / Die Coecumdivertikulitis.

INTRODUCTION: Cecal diverticulitis is an important differential diagnosis to acute appendicitis. The diagnosis is often difficult to make and the therapeutic procedure is still a point of discussion. PATIENTS AND METHOD: Seven patients (24-77 years old) who underwent surgery for abdominal reasons were investigated retrospectively. RESULTS: Five patients who had not underwent appendectomy before were operated under the leading diagnosis of acute appendicitis. Despite further diagnostic measures, only in one case of the two patients who had already had an appendectomy before, the diagnosis cecal diverticulitis could be made preoperatively. DISCUSSION: Preoperative diagnostics, if conducted at all, only rarely lead to the diagnosis of cecal diverticulitis. In prolonged courses and in patients after appendectomy, laparoscopy can be of diagnostic and therapeutic value. If it is possible to diagnose cecal diverticulitis preoperatively, conservative treatment can be indicated. If the diagnosis is made intraoperatively and malignancy can be securely excluded, we recommend ileocecal resection, but not appendectomy and further conservative treatment. If a carcinoma can not be excluded definitively, a hemicolectomy must be performed.

[A phenocopy of a homocygote Pelger's nuclear anomaly associated with an antibody deficiency (author's transl)]. / Die Phänokopie einer Pelger-Huëtschen Kernanomalie bei idiopathischem Antikörpermangelsyndrom.

A case of a boy is reported with an antibody deficiency syndrome type Bruton leading to a phenocopy of a homozygote Pelger's nuclear anomaly at the age of 10 months. The dominant inheritance and the absence of the anomaly in the peripheral blood of the parents and a sister support the presence of a phenocopy of this leukocyte anomaly. The recurrent bacterial infections are probably the cause of this phenocopy, since the substitution with gammaglobulins did not control the severe antibody deficiency syndrome.

Membrane-associated carbonic anhydrase from the crab gill: purification, characterization, and comparison with mammalian CAs.

Carbonic anhydrase (CA) was purified from the gills of the shore crab Carcinus maenas using affinity chromatography and HPLC. The predominantly membrane-bound CA was found to share several features with mammalian CA IV. Its apparent molecular weight of 36 kDa was reduced to 33 kDa by treatment with PNGase F, suggesting that crab CA is a glycoprotein with one N-linked oligosaccharide chain. More than half of the membrane-bound crab CA was released from membranes by treatment with a phosphatidylinositol-specific phospholipase C, indicating that the branchial CA is anchored to membrane surfaces by a phosphatidylinositol-glycan linkage. The enzyme also resembles mammalian CA IV in its relative sensitivity to inhibition by sulfonamides and the resistance to inhibition by halide ions. Amino acid composition of the HPLC-purified crab CA was examined and CNBr cleavage was carried out followed by N-terminal amino acid sequencing. Amino-terminal sequence of the native enzyme differed considerably from those of mammalian isozymes (human CA I and CA II, bovine CA III, human and rat CA IV). However, antisera raised against rat CA IV, CA II, and CA I all cross-reacted weakly with crab CA. Unlike mammalian CA IVs, crab gill CA was sensitive to 0.2% sodium dodecyl sulfate, suggesting that although crab gill CA is like mammalian CA IVs in many ways, it is less stabilized by intramolecular disulfide bonds.

[Prognostic factors of gastrointestinal stromal tumors of the stomach]. / Prognosefaktoren gastrointestinaler Stromatumoren des Magens.

The stomach is the most common gastrointestinal site of mesenchymal tumors which traditionally have been designated as smooth muscle tumors. However, with increasing analytic tools most investigators were unable to demonstrate true myogenic differentiation. Furthermore, the biological behavior of gastrointestinal stromal tumors (GIST) is difficult to predict. The aim of this study was to evaluate MIB-1 and p53 as additional prognostic markers, as well as myogenic differentiation immunohistochemically in GIST. 43 gastric stromal tumors were reviewed, 19 were classified as benign, and 10 as malignant. 14 tumors were considered indeterminate for biological behavior. In addition to MIB-1 and p53, immunohistochemistry was also performed for sm-actin, desmin and S 100-protein (ABC). 41 patients had a clinical follow-up of more than 2.5 years, 5 patients had metastases. Mean proliferation rates defined as percentage of MIB-1 positive tumor cells in 3 HPF were as follows: typical leiomyoma: 0.2%; benign GIST, spindle cell type: 1.8%; benign GIST, epithelioid cell type: 2.4%; borderline GIST, spindle cell type: 2.1%; borderline GIST, epithelioid cell type: 2.5%; malignant GIST, spindle cell type: 4.9%; and malignant GIST, epithelioid cell type: 7.3%. All 5 metastasizing tumors had a proliferation index > 4% (p < 0.0001). 4/5 metastasizing tumors had p53 positive cells (p < 0.05). 36/43 tumors were sm-actin positive, 7 of which were positive for desmin as well. Classification of gastric mesenchymal tumors as GIST is appropriate because only a small percentage show true smooth muscle differentiation. A MIB-1 proliferation index above 4% might indicate a more aggressive course, as well as p53 positivity.

[Value of radical lymph node dissection (compartment I and II) in stomach carcinoma]. / Wertigkeit der radikalen Lymphknotendissektion (Compartment I und II) beim Magenkarzinom.

In a prospective multicentre study of 2394 patients with gastric carcinoma the prognostic relevance of systematic lymph node dissection was evaluated. Of 1654 patients undergoing resection, 558 had a standard lymph node dissection, defined as fewer than 26 nodes in the specimen, and 1096 underwent radical lymphadenectomy, i.e. 26 or more nodes in the specimen. Radical dissection significantly improved the survival rate in patients with Union Internacional Contra la Cancrum (UICC) stages II and IIIA tumors. Multivariate analysis identified radical dissection as an independent prognostic factor in the subgroups of patients with UICC tumor stages II and IIIA. Radical dissection conferred no survival advantage in patients with pN2 tumors. There was no significant difference in morbidity and mortality rates between radical and standard lymph node dissection. Radical lymphadenectomy improves survival in patients with UICC gastric cancer stages II and IIIA, and should be the recommended treatment for such patients.