[Acute abdominal pain due to a ventriculo-peritoneal shunt metastasis of an intracranial germ cell tumor]. / Akute Nierenkolik aufgrund einer ventrikuloperitonealen Shunt-Metastase eines intrakraniellen Germinoms.

INTRODUCTION: Extracranial metastases of pineal germinomas are rare. CASE REPORT: The case of a metastasis of a primary intracranial germ cell tumor via a temporary-ventriculo-peritoneal shunt as a cause of acute abdominal pain is reported. The germ cell tumor markers were useful in establishing the diagnosis. CONCLUSIONS: With an appropriate clinical history, intra-abdominal metastases via a temporary ventriculo-peritoneal shunt should be considered in the differential diagnosis of acute abdominal pain.

Bladder tumor recurrence after primary surgery for transitional cell carcinoma of the upper urinary tract.

OBJECTIVE: Primary transitional cell carcinoma (TCC) of the upper urinary tract represents 6-8% of all TCC cases. Nephroureterectomy with removal of a bladder cuff is the treatment of choice. The rates of TCC recurrence in the bladder after primary upper urinary tract surgery described in the literature range between 12.5 and 37.5%. In a retrospective analysis we examined the occurrence of TCC after nephroureterectomy for upper tract TCC in patients without a previous history of bladder TCC at the time of surgery. METHODS: Between 1990 and 2002, 29 patients underwent primary nephroureterectomy for upper tract TCC. The mean age of the patients was 69.5 years. In 5 cases upper urinary tract tumors were multilocular, in the remaining cases unilocular in the renal pelvis (n=12) or the ureter (n=12). The follow-up was available for 29 patients with a mean follow-up of 3.37 (0.1-11.2) years. RESULTS: 11/29 (37.9%) patients had TCC recurrence with 9/11 patients having bladder TCC diagnosed within 2.5 years (0.9-6.0) after nephroureterectomy. 13/29 patients are alive without TCC recurrence, 3/29 patients died due to systemic TCC progression and 5/29 died of unrelated causes without evidence of TCC recurrence. CONCLUSION: Our data indicate a high incidence of bladder TCC after nephroureterectomy for primary upper tract TCC of up to 6 years after primary surgery. Because of the high incidence of bladder TCC within the first 3 years of surgery, careful follow-up is needed over at least this period.

Comparative evaluation of the performance of two commercial kits for the detection of central nervous system tissue in meat.

The ScheBo Brainostic test, which detects neuron-specific enolase by Western blotting, and the r-Biopharm Ridascreen Risk Material ELISA test, which detects the presence of glial fibrillary acidic protein, were evaluated using meats containing spinal cord and brain central nervous system (CNS) tissue from ovine and bovine species. The meats were pork, cooked pork sausage, raw minced lamb and cooked minced lamb. Spiking of the CNS tissue ranged from 0.01 to 5%. No false-positives were observed with either test using the manufacturers' analytical protocols. The presence or absence of CNS tissue was correctly determined in 20 of 20 samples using the ScheBo Brainostic test and 18 of 20 samples using the Ridascreen tests. When results were placed in categories according to quantity of CNS tissue detected, 19 of 20 samples were classified correctly using the Brainostic test and 14 of 20 samples using the Ridascreen test. Both kits were considered appropriate for reporting the presence of 1% or more CNS tissue in meat products, but the ScheBo Brainostic test was more consistent at detecting the presence of CNS tissues below the 1% level. Overall, the format of the Ridascreen test was technically easier to use, and the data simpler to interpret.

Manifestation, latency and management of late urological complications after curative radiotherapy for cervical carcinoma.

BACKGROUND: Radiotherapy alone or in combination with radical hysterectomy remains a potentially curative treatment for cervical carcinoma. Late urological complications after curative radiotreatment are rare but often present difficult problems of management due to the progressive nature of radiogenic tissue damage. We reviewed all cases of radiogenic urologic complications after radiotherapy for cervical carcinoma treated at our institution over the past 10 years. PATIENTS AND METHODS: In a retrospective analysis, out of all female patients treated for hydronephrosis (n = 543), 93 patients with hydronephrosis occurring after curative treatment for gynecological malignancies were identified. The most frequent cause was progressive disease (n = 55); 30 patients without recurrence had severe radiogenic urological complications (study population). RESULTS: Out of 30 patients with a mean age of 44.9 years at the time of treatment for cervical cancer, 6 had undergone primary radiotherapy and 24 had received adjuvant radiotreatment after radical hysterectomy. The observed late urological complications were: distal ureteral stenoses (bilateral in 4 cases, unilateral in 15 cases), distal ureteral necrosis (n = 1), bilateral ureteral reflux (n = 3), vesico-vaginal fistulae (n = 5), vesico-intestinal fistula (n = 1), severe fibrotic bladder shrinkage (n = 6) and urethral stenosis (n = 2). Marked signs of radiogenic cystitis were observed in almost all patients. The mean latency time between radiotherapy and manifestation of severe radiogenic urological sequelae was 19.4 years (range 0.5-41.5). Primary treatment of urological complications consisted in temporary urinary diversion by percutaneous nephrostomies or insertion of internal ureteral stents. Permanent treatment for radiogenic ureteral stenosis in all patients with sufficient general health consisted of surgical or endourological urinary diversion or reconstruction. However, the majority of patients were managed by permanent endourological measures. Lower tract fistulae were treated by distal ureteral occlusion and diversion. CONCLUSIONS: Although severe late urological sequelae of curative radiotherapy for cervical carcinoma are rare they represent major complications, result in considerable distress for the patient, and often present difficult therapeutic problems for the urologist. As these urological complications can have extremely long latency times, focussed follow-up for early detection may be advisable.

Early changes of oxalate and calcium urine excretion in those with calcium oxalate stone formation after extracorporeal shock wave lithotripsy.

OBJECTIVES: To determine the extent of transient changes of tubular function in idiopathic calcium-oxalate (CaOx) stone-bearing patients after extracorporeal shock wave lithotripsy (ESWL), calcium and oxalate excretion were measured before and after ESWL. METHODS: In 22 patients with renal CaOx stones, the plasma values and urine excretion of creatinine, calcium, oxalate, magnesium, and citrate were measured before and on days 1 and 2 after ESWL under conditions of a standardized diet. Overnight urine collection for an 8-hour period was used to measure the urine excretion, and the values were extrapolated to a 24-hour period. For calculation of the urine ion activity, the AP(CaOx) index EQ(s) and the CaOx risk index were used. RESULTS: After ESWL, hyperoxaluria was noted in 10 patients compared with 2 before ESWL. Hypercalciuria was seen in 11 patients after ESWL compared with 3 before. Combined hyperoxaluria and hypercalciuria was found in 7 patients after ESWL compared with 1 before. Both the oxalate/creatinine and calcium/creatinine ratios were significantly increased after ESWL. The AP(CaOx) index EQ(s) and the CaOx risk index were significantly increased after ESWL in patients with increased post-ESWL calcium and/or oxalate excretion. CONCLUSIONS: Increased calcium and/or oxalate excretion can be seen in patients with CaOx stones early after ESWL. This increased excretion of lithogenic substances represents an increased risk of fragment apposition after ESWL for the group with a significantly increased AP(CaOx) index EQ(s) and CaOx risk index. Therefore, prophylactic measures in patients at risk early after ESWL might be warranted to prevent possible recurrent stone formation.

Measurement of free radicals and NO by chemiluminescence to identify the reperfusion injury in renal transplantation.

Oxygen free radicals are generated during the reperfusion of ischaemic organs. Several experimental studies have demonstrated that the damage produced by reperfusion can be prevented by a scavenger of free radicals. Furthermore, a significantly improved 5 year graft survival rate after cadaveric renal transplantation has been reported in patients treated with scavengers of free radicals (Land et al., 1993). Therefore, a question remains to be answered: whether a routine monitoring of the radical-mediated reperfusion injury with renal transplantation is useful, and whether there is a necessity for a generalized protective treatment in transplant patients. In a prospective trial, we evaluated a group of eight patients during and after renal cadaveric transplantation (three men, five women), using the chemiluminometric measurement of serum free radicals and NO. The serum quantities of free radicals and NO were significantly increased after reperfusion of the transplant kidney (p < 0.02). The mean time of noticeably increased levels of serum free radicals was 4.8 +/- 1.2 h after reperfusion. The results thus showed an increased liberation of free radicals in the peripheral blood of transplant recipients as possible evidence of free radicals-mediated reperfusion injury in renal transplantation. The generation of free radicals measured by chemiluminescence allow a controlled therapy to decrease the generation of free radicals with antioxidants during the early transplantation period e.g. in older recipients.

Food allergy--towards predictive testing for novel foods.

The risks associated with IgE-mediated food allergy highlight the need for methods to screen for potential food allergens. Clinical and immunological tests are available for the diagnosis of food allergy to known food allergens, but this does not extend to the evaluation, or prediction of allergenicity in novel foods. This category, includes foods produced using novel processes genetically modified (GM) foods, and foods that might be used as alternatives to traditional foods. Through the collation and analysis of the protein sequences of known allergens and their epitopes, it is possible to identify related groups which correlate with observed clinical cross-reactivities. 3-D modelling extends the use of sequence data and can be used to display eptiopes on the surface of a molecule. Experimental models support sequence analysis and 3-D modelling. Observed cross-reactivities can be examined by Western blots prepared from native 2-D gels of a whole food preparation (e.g. hazelnut, peanut), and common proteins identified. IgEs to novel proteins can be raised in Brown Norway rat (a high IgE responder strain) and the proteins tested in simulated digest to determine epitope stability. Using the CSL serum bank, epitope binding can be examined through the ability of an allergen to cross-link the high affinity IgE receptor and thereby release mediators using in vitro cell-based models. This range of methods, in combination with data mining, provides a variety of screening options for testing the potential of a novel food to be allergenic, which does not involve prior exposure to the consumer.

An In Vitro Cell Based Model for Assessing the Potential of Allergens to Release Mediators Through the Cross-linking of IgE.

The aim of this study was to examine the capacity of an in vitro model to test the potential of an allergen to cause cross-linking of IgE bound to the surface of mast cells. The model involved the passive sensitization of murine mast cells, with zearalenone IgE and subsequent exposure to anti IgE, zearalenone, zearalenone-BSA (with up to nine bound zearalenone molecules) or peanut lectin. The extent of cross-linking was determined by measuring the release of IgE mediators TNFalpha and histamine. Release of TNFalpha from IgE sensitized cells increased following exposure to zearalenone-BSA, but not following exposure to zearalenone alone or to peanut lectin. Histamine release could not be quantified against background. The results suggested that the model could be used to test allergenic potential through the availability of epitopes to bind and cross-link IgE on the surface of mast cells. As IgE is species specific, the model was adapted for use with a human cell system employing mast cells in lung fragments. TNFalpha release was measured, and the system was calibrated with the inhalant allergen from Timothy Grass and Timothy Grass specific IgE.

Use of a flow cell bioreactor as a chronic toxicity model system.

We describe the use of a model system to mimic chronic toxin exposure, similar to that which might be found in a human situation, where exposure to dietary or environmental toxins occurs at a low level for an extended period of time. This is in contrast to the acute, immediately toxic dose effect usually observed in flask tissue culture. The apparatus used was a flow cell bioreactor in which cells can be cultured for lengthy periods of time as a continuous viable population. The compound used as the toxic agent was 3-nitropropanoic acid (3-NPA), a fungal neurotoxin which acts as a suicide inhibitor of succinate dehydrogenase. The secondary human cell line, Int 407, was tested in conventional flask culture for periods of up to 72 hours to determine the no-observed-effect level (NOEL) for 3-NPA. Cell populations established in the bioreactor were continuously exposed to levels of 3-NPA below that of the NOEL for periods of up to 4 weeks and the viability of the population determined using MTT, trypan blue and ATP assays.

Producing a P1 bacteriophage library from pine: isolation and cloning of very high molecular weight DNA.

We have generated a genomic P1 bacteriophage library using Monterey pine (Pinus radiata) DNA. We first developed a method for isolating from pine tissue the very high molecular weight DNA necessary for the preparation of libraries requiring large inserts. The method involves protoplasting the cells, isolating nuclei and lysis in a high concentration of detergent. Fragments of greater than two megabases in size are produced in solution. Modifications introduced to the protocol for library preparation and for P1 plasmid isolation are described.

Barley anther culture: The effect of position on pollen development in vivo and in vitro.

Locule structure and organization were studied in vivo and in vitro to determine whether the disposition of pollen within barley anthers affected the response of pollen in culture. Following release from the meiotic tetrads, juvenile barley microspores become peripherally organized around the locule, with the single pollen pore oriented towards the tapetum. Scanning electron micrographs of transverse sections from freeze fractured anthers showed that some microspores failed to locate the tapetal surface and occupied a position in the centre of the locule where they continued to develop as small, abnormal pollen grains (dimorphic pollen). Previous evidence has suggested that in some species dimorphic pollen could be the source of embryonic pollen in vitro. Cultured anthers frequently dehisced to reveal a mass of dividing pollen grains, however those anthers that remained intact retained the original locule structure and could be freeze fractured permitting examination of the developing pollen in situ. This showed that pollen embryogenesis was not restricted to dimorphic pollen, and that any grain could become Embryogenic irrespective of position.