In vitro method to evaluate virus competition between BVDV-1 and BVDV-2 strains using the PrimeFlow RNA assay.

Bovine viral diarrhea viruses (BVDV), segregated in BVDV-1 and BVDV-2 species, lead to substantial economic losses to the cattle industry worldwide. It has been hypothesized that there could be differences in level of replication, pathogenesis and tissue tropism between BVDV-1 and BVDV-2 strains. Thus, this study developed an in vitro method to evaluate virus competition between BVDV-1 and BVDV-2 strains. To this end the competitive dynamics of BVDV-1a, BVDV-1b, and BVDV-2a strains in cell cultures was evaluated by a PrimeFlow RNA assay. Similar results were observed in this study, as was observed in an earlier in vivo transmission study. Competitive exclusion was observed as the BVDV-2a strains dominated and excluded the BVDV-1a and BVDV-1b strains. The in vitro model developed can be used to identify viral variations that result in differences in frequency of subgenotypes detected in the field, vaccine failure, pathogenesis, and strain dependent variation in immune responses.

Comparison of reproductive performance of primiparous dairy cattle following revaccination with either modified-live or killed multivalent viral vaccines in early lactation.

The objective of this randomized clinical trial was to compare the effect of revaccination in primiparous dairy cows with modified live viral (MLV) or killed viral (KV) vaccines containing bovine viral diarrhea virus (BVDV) and bovine herpesvirus-1 (BoHV-1) on (1) pregnancy rate following estrus synchronization-timed artificial insemination (TAI), (2) serum progesterone concentrations, and (3) serum neutralizing antibody titers at revaccination and at TAI. Primiparous dairy cows (n=692) that had been previously vaccinated with 4 doses of MLV vaccine as calves or heifers were randomized to receive either an MLV or a KV vaccine between 21 and 28 d in milk and 17 d before initiation of a double-Ovsynch-TAI protocol. Serum was collected within the double-Ovsynch protocol for determination of progesterone concentrations, and at vaccination and TAI for serum neutralizing antibody titers. Ultrasound pregnancy determinations were made at 30 and 60 d after TAI. No differences in pregnancy rates were observed between cows receiving MLV vaccine (44%; n=326) or KV vaccine (43%; n=336). No differences were observed in serum progesterone concentrations during a double-Ovsynch-TAI protocol between cows receiving MLV and KV vaccines. No differences were observed in BVDV 1 or BVDV 2 antibody titers at vaccination and TAI between cows receiving MLV or KV vaccine; however, BoHV-1 antibody titers were greater at TAI in cows receiving KV vaccine. Overall response to vaccination-defined as the percent of all individual cows that had any detectable increase in antibody titer from vaccination to TAI-was 39% for BVDV 1, 45% for BVDV 2, and 61% for BoHV-1. In this research, use of an MLV vaccine did not impede reproduction when revaccination was performed between 21 and 28 DIM and just before enrollment in an estrus synchronization-TAI program in primiparous dairy cows; however, response to vaccination as defined by increases in virus-specific antibody titers could be considered less than ideal for this population of cattle.

Distribution of bovine viral diarrhoea virus antigen in persistently infected white-tailed deer (Odocoileus virginianus).

Infection with bovine viral diarrhoea virus (BVDV), analogous to that occurring in cattle, is reported rarely in white-tailed deer (Odocoileus virginianus). This study evaluated the distribution of BVDV antigen in persistently infected (PI) white-tailed deer and compared the findings with those from PI cattle. Six PI fawns (four live-born and two stillborn) from does exposed experimentally to either BVDV-1 or BVDV-2 were evaluated. Distribution and intensity of antigen expression in tissues was evaluated by immunohistochemistry. Data were analyzed in binary fashion with a proportional odds model. Viral antigen was distributed widely and was present in all 11 organ systems. Hepatobiliary, integumentary and reproductive systems were respectively 11.8, 15.4 and 21.6 times more likely to have higher antigen scores than the musculoskeletal system. Pronounced labelling occurred in epithelial tissues, which were 1.9-3.0 times likelier than other tissues to contain BVDV antigen. Antigen was present in >90% of samples of liver and skin, suggesting that skin biopsy samples are appropriate for BVDV diagnosis. Moderate to severe lymphoid depletion was detected and may hamper reliable detection of BVDV in lymphoid organs. Muscle tissue contained little antigen, except for in the cardiovascular system. Antigen was present infrequently in connective tissues. In nervous tissues, antigen expression frequency was 0.3-0.67. In the central nervous system (CNS), antigen was present in neurons and non-neuronal cells, including microglia, emphasizing that the CNS is a primary target for fetal BVDV infection. BVDV antigen distribution in PI white-tailed deer is similar to that in PI cattle.

Peripheral primitive neuroectodermal tumour in a lumbar vertebra and the liver of a dromedary camel (Camelus dromedarius).

A 9-year-old castrated male dromedary camel developed weakness and ataxia, progressing to sternal recumbency and hindlimb paralysis. Necropsy revealed multiple liver tumours and a mass in the 3rd lumbar vertebra, compressing the spinal cord. The hepatic and vertebral masses consisted of uniform sheets of primitive cells, with perivascular pseudorosettes and small numbers of neuroblastic Homer-Wright rosettes. Immunohistochemically, the tumour cells were uniformly positive for vimentin and variably positive for neuron-specific enolase and glial fibrillary acidic protein. The histopathological and immunohistochemical findings indicated a peripheral primitive neuroectodermal tumour (pPNET) exhibiting neuroblastic, glial and ependymomatous differentiation, probably reflecting the tumour's primitive multipotential neuroepithelial nature. To the authors' knowledge, this is the first reported case in the camel of a pPNET, presumably intraosseous in origin with hepatic metastasis, and morphologically similar to Ewing's sarcoma in man.

Study of cattle persistently infected with bovine viral diarrhea virus that lack detectable virus in serum.

OBJECTIVE: To determine whether cattle persistently infected with bovine viral diarrhea virus (BVDV) that lack virus detectable in serum by use of the immunoperoxidase microtiter assay (IPMA) can transmit the virus to susceptible herdmates and determine prevalence of these cattle. DESIGN: Clinical trial and serologic survey. SAMPLE POPULATION: 2 cattle and 1,952 blood samples. PROCEDURE: A persistently infected cow in which virus could not be detected in serum was housed with a BVDV-seronegative steer. Blood and nasal swab specimens were tested via virus isolation and serum virus neutralization. Parallel WBC preparations and sera from blood samples of 1,952 adult cows were screened for BVDV by use of IPMA. RESULTS: The steer seroconverted to BVDV within 4 weeks of contact with the cow. Virus was detected in sera and WBC of 5 adult cows that were verified as persistently infected by retest 3 weeks later. Cattle persistently infected with BVDV in which virus could not be detected in both serum and WBC by use of IPMA were not found. CONCLUSION AND CLINICAL RELEVANCE: Cattle persistently infected with BVDV in which virus cannot be detected in serum by use of IPMA may serve as virus reservoirs for infecting susceptible cattle. Persistent infection was detected at a prevalence of 0.26%. Screening adult cattle by use of IPMA on serum samples appears to be a reliable means of detecting persistent infection with BVDV. Prevalence of cattle persistently infected with BVDV that have negative results of IPMA on serum is extremely low.

Relationship between degree of viremia and disease manifestation in calves with experimentally induced bovine viral diarrhea virus infection.

OBJECTIVE: To compare degree of viremia and disease manifestations in calves with type-I and -II bovine viral diarrhea virus (BVDV) infection. ANIMALS: 16 calves. PROCEDURE: Colostrum-deprived calves obtained immediately after birth were assigned to 1 control and 3 treatment groups (4 calves/group). Calves in treatment groups were inoculated (day 0) by intranasal instillation of 10(7) median tissue culture infective dose BVDV 890 (type II), BVDV 7937 (type II), or BVDV TGAN (type I). Blood cell counts and virus isolation from serum and leukocytes were performed daily, whereas degree of viremia was determined immediately before and 4, 6, 8, and 12 days after inoculation. Calves were euthanatized on day 12, and pathologic, virologic, and immunohistochemical examinations were performed. RESULTS: Type-II BVDV 890 induced the highest degree of viremia, and type-I BVDV TGAN induced the lowest. Virus was isolated more frequently and for a longer duration in calves inoculated with BVDV 890. A parallel relationship between degree of viremia and rectal temperature and an inverse relationship between degree of viremia and blood cell counts was observed. Pathologic and immunohistochemical examinations revealed more pronounced lesions and more extensive distribution of viral antigen in calves inoculated with type-II BVDV. CONCLUSIONS AND CLINICAL RELEVANCE: Degree of viremia induced during BVDV infection is associated with severity of clinical disease. Isolates of BVDV that induce a high degree of viremia may be more capable of inducing clinical signs of disease. Strategies (eg, vaccination) that reduce viremia may control clinical signs of acute infection with BVDV.

Platelet aggregation responses and virus isolation from platelets in calves experimentally infected with type I or type II bovine viral diarrhea virus.

Altered platelet function has been reported in calves experimentally infected with type II bovine viral diarrhea virus (BVDV). The purpose of the present study was to further evaluate the ability of BVDV isolates to alter platelet function and to examine for the presence of a virus-platelet interaction during BVDV infection. Colostrum-deprived Holstein calves were obtained immediately after birth, housed in isolation, and assigned to 1 of 4 groups (1 control and 3 treatment groups). Control calves (n = 4) were sham inoculated, while calves in the infected groups (n = 4 for each group) were inoculated by intranasal instillation with 10(7) TCID50 of either BVDV 890 (type II), BVDV 7937 (type II), or BVDV TGAN (type I). Whole blood was collected prior to inoculation (day 0) and on days 4, 6, 8, 10, and 12 after inoculation for platelet function testing by optical aggregometry by using adenosine diphosphate and platelet activating factor. The maximum percentage aggregation and the slope of the aggregation curve decreased over time in BVDV-infected calves; however, statistically significant differences (Freidman repeated measures ANOVA on ranks, P < 0.05) were only observed in calves infected with the type II BVDV isolates. Bovine viral diarrhea virus was not isolated from control calves, but was isolated from all calves infected with both type II BVDV isolates from days 4 through 12 after inoculation. In calves infected with type I BVDV, virus was isolated from 1 of 4 calves on days 4 and 12 after inoculation and from all calves on days 6 and 8 after inoculation. Altered platelet function was observed in calves infected with both type II BVDV isolates, but was not observed in calves infected with type I BVDV. Altered platelet function may be important as a difference in virulence between type I and type II BVDV infection.

Effect of experimentally induced type II bovine viral diarrhea virus infection on platelet function in calves.

OBJECTIVE: To evaluate platelet aggregation responses in calves experimentally infected with a thrombocytopenia-inducing type II bovine viral diarrhea virus (BVDV) isolate (BVDV 890). ANIMALS: 9 neonatal male Holstein calves. PROCEDURE: 5 calves were inoculated with BVDV 890, and 4 were used as controls. Platelet aggregation studies and attempts to isolate BVDV from platelets were performed 2 days before, the day of, and every 2 days for 12 days after inoculation. Platelet function was assessed by means of optical aggregometry, using adenosine diphosphate and platelet-activating factor as agonists. Bovine viral diarrhea virus was isolated from purified platelet preparations by use of an immunoperoxidase monolayer assay. RESULTS: Maximum percentage aggregation and slope of the aggregation curve decreased over time in calves infected with BVDV. Bovine viral diarrhea virus was not isolated from platelets from control calves, but it was isolated from infected calves from 4 through 12 days after inoculation. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that platelet function may be depressed in calves infected with type II BVDV. Although the mechanism for altered platelet function was not determined, it likely involved an increase in the percentage of aged platelets in the circulation, a direct virus-platelet interaction, or an indirect virus-platelet interaction. Platelet dysfunction, in addition to thrombocytopenia, may contribute to the hemorrhagic syndrome associated with acute type II BVDV infection in calves.

Differential uptake of estramustine phosphate metabolites and its correlation with the levels of estramustine binding protein in prostate tumor tissue.

Estracyt (EMP) has been used for the treatment of hormone refractory prostate cancer for many years. Recently, new data from combination studies have given rise to new interest in this old drug. Explanations for the synergy found in the clinic are many, but one major factor may be the previous indication that the drug accumulates in the prostate tumor. We have, therefore, examined the level of the four metabolites, estromustine (EoM), estramustine (EaM), estrone, and estradiol in the tumor and serum of 14 patients with T2 and T3 prostate cancer receiving a single i.v. dose of 600 mg of EMP, about 12 h before radical prostatectomy. Because it has been suggested that the uptake into the prostate tumor is due to binding to the estramustine binding protein (EMBP), we have in addition measured the level of EMBP in the prostate tumor tissue. The main serum and tissue metabolite in all patients was EoM followed by EaM, estrone, and estradiol. The levels for EoM ranged from 63.8-162.8 ng/ml in the serum and from 64.8-1209 ng/ml in the prostate tumor, resulting in a mean ratio for serum to tumor of 1:5. The levels for EaM ranged from 8.3-51.4 ng/ml in the serum and 73.9-563.4 ng/ml in the tumor, giving a mean ratio for serum to tumor of 1:13. The levels of EMBP were higher in T3 tumors than in T2 tumors, 54.1 and 40.7 ng/g tissue, respectively. A significant correlation was found between the levels of EaM (r = 0.60) and the levels of EMBP in the tumor. These data demonstrate that 12 h after a single i.v. dose of 600 mg of EMP the levels of the cytotoxic metabolites EoM and EaM are substantially higher in the tumor than in the serum of the same patient and that a correlation exists between the levels of EaM in the tumor and the levels of EMBP. Thus, this supports the hypothesis that the EMBP is responsible for the retention of EoM and EaM in the prostate tumor.

Paclitaxel-based second-line therapy for patients with advanced chemotherapy-resistant bladder carcinoma (M1): a clinical Phase II study.

BACKGROUND: In a previous clinical trial, the authors disclosed that the expression of tumor cell motility factor gp78hAMFR correlates with tumor progression in patients with bladder carcinoma. This study was initiated to evaluate whether the combination of cytostatic drugs with an antimotility factor has an effect on chemotherapy-resistant bladder carcinoma. METHODS: In a Phase II trial, the authors evaluated the influence of paclitaxel, carboplatin, and an antimotility factor (acellular pertussis vaccine [APV]) in 18 patients with cisplatin- and methotrexate-resistant metastatic bladder carcinoma. Intramuscular injection of APV 3 times in the first week, on Days 1, 4, and 7, was followed by paclitaxel 135 mg/m2 and carboplatin 400 mg/m2. After an interval of 1 week APV was given again on Days 15 and 19. Each cycle lasted 3 weeks. On Day 22 the cycle was repeated. RESULTS: Four of 18 patients had objective responses (2 had complete remissions and 2 had partial remissions). After a median number of 2.5 cycles, side effects did not exceed World Health Organization Grade 4. CONCLUSIONS: The results of this clinical Phase II study demonstrate that the combination of paclitaxel-based therapy causes complete remissions previously not obtained with second-line chemotherapy, although no conclusions can be drawn as to the effectiveness of the individual substances. Further trials have to be evaluated with regard to the individual components.

[Differentiated metastasis of mature testicular teratoma as differential kidney and adrenal tumor diagnosis]. / Differenzierte Metastase eines reifen Hodenteratoms als Differentialdiagnose eines Nieren-bzw. Nebennierentumors.

We report on the eighth known case of a mature metastasis of a mature teratoma of the testis. In a 19-year-old patient who had undergone a pyeloplasty of the left renal pelvis 6 years before, a mature metastasis of an occult mature teratoma of the testis was found at the former operation site. This case--in addition to those published previously--emphasizes the fact that a mature teratoma of the testis has to be regarded as a malignant tumor in adults. It is supposed that the mature teratoma originates, as other testicular tumors, from carcinoma-in-situ-cells, which are responsible for metastasize and are not detected after lysis.

[A combination of spermatocytic and classic seminoma, mature teratoma and carcinoma in situ of the testis. An attempt at an etiologic explanation]. / Eine Kombination von spermatozytischem und klassischem Seminom, reifem Teratom und Carcinoma in situ des Hodens. Versuch einer ätiologischen Zuordnung.

We report of a 39 years old patient with a large testicular tumor found during an examination for infertility. The tumor consisted of a spermatocytic seminoma (SS) and a differentiated teratoma (TD). Furthermore, two small foci of seminoma were seen in the surrounding testicular tissue, several testicular tubuli contained carcinoma in situ (CIS). The diagnosis was based on the results obtained with various immunohistochemical markers: keratin, vimentin, desmin, LCA, CD3, CD20, CD45R, ferritin, PLAP, AFP. On the basis of the macroscopic and histopathological features, we propose the following etiology: CIS progressed in an earlier phase to the (larger) TD and later to the (smaller) classical seminoma; likewise, in an earlier phase, SS developed from a still unknown precursor stage. Our case of a mixed tumor as well as other cases reported in the last years do not allow the explanation of a differing etiology for SS. On the contrary, it may be presumed that the origins of seminoma and teratoma on the one hand and SS on the other hand are less divergent than hitherto thought.

[Gigantic pyohydronephrosis and contralateral ureteral bud. Problems of differential diagnosis]. / Gigantische Pyohydronephrose und kontralaterale Ureterknospe. Differentialdiagnostische Probleme.

Both giant hydronephrosis and blind-ending ureteral bud are rare conditions. We report on a patient with an impacted ureteral stone leading to giant pyohydronephrosis (6.51) on the right side, which in turn caused displacement of the ureter containing the stone to the left. An asymptomatic blind-ending ureteral bud initially complicated the diagnosis. After nephrectomy, a unsuspected transitional cell carcinoma of the right renal pelvis and the ureter was found.

Influence of prostatic disease and prostatic manipulations on the concentration of prostate-specific antigen.

We examined the influence of different factors [benign prostatic hyperplasia (BPH), prostatic carcinoma (PCA), organ volume, weight of resected tissue, transurethral catheter] on the serum prostate-specific antigen (PSA) levels in 253 patients with BPH (n = 138; 54%) and PCA (n = 115; 46%). Only in 57.2% of the BPH patients, PSA values were < 4 ng/ml, in 74.6% < 7 ng/ml. In 108 patients with BPH, a transurethral prostatectomy was performed. PSA values correlated significantly with the sonographically determined prostatic volumes and less precisely with the weight of the resected tissue. The PSA concentration per milliliter of prostatic volume was 0.12 ng/ml, per gram of resected tissue it was 0.21 ng/ml. An incidental PCA was found in 12/108 patients (11%). The PSA values were identical with those of the total collective in regard to volume and tissue weight. In 11 patients, we examined possible alterations of the PSA values before and until 24 h after prostatic massage. Only insignificant alterations were seen, a massive increase was not found in any patient. Searching for an absolutely valid 'normal value' appears hardly appropriate. However, the usefulness of PSA is increased when the sonographically determined prostatic volume is included. A rectal examination of the prostate has no influence on the PSA value.

Ultrasound examination of bladder and prostate.

Transabdominal (suprapubic) sonography of the bladder and prostate is not limited to special indications but should be considered an integrated part of the urologic sonographic evaluation of the genitourinary tract. Information about many pathological conditions (e.g. residual urine, stone, tumor, diverticulum, enlarged prostate) is obtained without delay, without need of instrumentation or exposure to X-rays. Transurethral sonography has proven useful in the differentiation of superficial bladder tumors from deep infiltrating ones. Transvaginal or transrectal examination of the bladder neck has developed into a new modality for urodynamic examination. More than any other diagnostic modality, transrectal ultrasound of the prostate has become a valuable tool for the more correct detection and staging of prostatic carcinoma.

Primary megaureter in adults.

The terminology of megaureters, their histological findings and etiology vary greatly in the literature. Primary obstructed megaureters must be clearly differentiated from megaureters caused by infravesical obstruction. The characterization and therapeutical consequences of the two types also differ. In a retrospective study, 43 adult patients with 54 primary obstructed megaureters (male:female = 30:13) were studied. This diagnosis excluded infravesical obstruction, reflux and neurogenic disorders. The main symptoms were flank pain, infection and hematuria. An operation was performed in 25 patients; 29 were treated conservatively. The criteria for surgery are discussed in relation to symptoms, functional reserves of the kidney, transportation capacity of the upper urinary tract, and histological changes of the narrow segment of the ureter. Operative treatment should be limited to well-defined complications, such as deterioration of kidney function or recurrent infection, to avoid its discredit as merely a cosmetic procedure.

[Radical extirpation of a left-sided hypernephroma with tumor thrombus in the vena cava inferior (author's transl)]. / Radikale Exstirpation eines linksseitigen Hypernephroms mit Tumorzapfen in der Vena cava inferior.

A radical nephrectomy of a left-sided renal cell carcinoma with extension of the tumor thrombus into the vena cava as far up as the right atrium is described. The right kidney has no venous collateral system, therefore ligation of the right renal vein and resection of the tumor-invaded vena cava is not possible. A left radical extended nephrectomy, if necessary, with interposition of a vessel prosthesis, increases the prognosis and must be considered the therapy of choice.