Immune cell characterization in spontaneous autoimmune orchitis in dogs.

With a prevalence of up to 35% in dogs with reproductive problems, azoospermia is one of the most important reasons for male infertility. Non-obstructive azoospermia, without clinical symptoms, but histopathological damage of the testicular tissue and immune cell infiltration is referred to as spontaneous autoimmune orchitis (AIO) in the literature. Published cases in dogs describe immune cell infiltration; however, there is no consent about the involved immune cell types. We aimed to characterize immune cells in testicular biopsies of dogs with AIO (n = 9) and to compare them to those in testicular specimens from healthy control dogs with normospermic ejaculates (CG; n = 5). Immunohistochemistry was performed using specific antibodies against CD3, PAX5, MAC387, IgG and IgM to proof the presence of T lymphocytes, B lymphocytes, macrophages and early and late plasma cells, respectively. Presence of immune cells in healthy testicular tissue was low and restricted to T lymphocytes and macrophages in the interstitium with the latter also being found within the blood vessels. Different to this, AIO samples revealed presence of all investigated immune cells, underlining lymphoplasmacytic nature of chronic asymptomatic immune-mediated orchitis. Canine spontaneous AIO is characterised by a significantly increased number of immune cells, namely ≥33 immune cells/mm2 (sensitivity/specificity: 100% based on our data). The pathogenesis of canine AIO is hypothesized to be as follows: 1. Macrophages initiate AIO via T lymphocyte activation. 2. T lymphocytes lead to a "delayed type immunological response" and development of AIO. 3. Invaded B lymphocytes later differentiating to plasma cells are responsible for the second humoral immunological response and cause progression of AIO. Different to the situation in CG, T lymphocytes and plasma cells were identified within the seminiferous tubules indicating that disruption of spermatogenesis in AIO might be related to invading immune cells. Testicular biopsies provide an essential tool in the diagnosis of spontaneous AIO.

Expression of PTGS2, PGFS and PTGFR during downregulation and restart of spermatogenesis following GnRH agonist treatment in the dog.

Prostaglandins (PGs) and prostaglandin endoperoxide synthase (PTGS) are considered to be relevant for spermatogenesis and steroidogenesis. PTGS2, prostaglandin F synthase (PGFS) and PGF receptor (PTGFR) are investigated in the adult male dog using the model of the GnRH-agonist implant downregulated canine testis and its subsequent restart of spermatogenesis following abolition of treatment (3, 6, 9, 12 weeks after implant removal). On the mRNA level (ratio), PTGS2, PGFS and PTGFR expression did not differ between downregulation, different stages of recovery of spermatogenesis and untreated adult controls (CG). On the protein level, Sertoli and Leydig cells in all samples and some peritubular cells stained immunopositive for PTGS2. In the tubular compartment, the percentage of the PTGS2-immunopositive area (PIA) and the mean PTGS2-staining intensity (gray scale, GS) did not differ between groups but in the interstitial compartment, the PIA (p = 0.0494) and the GS (p < 0.0001) were significantly upregulated during early recrudescence (week 3/6). Comparing downregulation by two GnRH-agonist implants with juvenile controls (JG) and CG, the mRNA expression (ratio) did not differ. In the tubular compartment, the GS (p = 0.0321) was significantly higher at downregulation compared to CG and in the interstitial compartment, the PIA (p = 0.0073) and the GS (p = 0.0097) were significantly higher in JG compared to downregulation/CG. PTGS2, PGFS and PTGFR mRNA and PTGS2 protein are regularly expressed in the adult, juvenile and downregulated canine testis and downregulation and subsequent recrudescence affect PTGS2 protein expression mainly in Leydig cells. PTGS2 expression in the downregulated testis resembles the one in seasonal Syrian hamster but not juvenile canine testis.

Purification of morphologically and functionally intact human basophils to near homogeneity.

Certain studies on basophils require highly purified functionally intact cell preparations. Here, a three-step procedure is described that meets these requirements. The procedure consists of a Ficoll density gradient step, counterflow elutriation and negative selection by magnetic cell sorting (MACS). The mean purity of basophils obtained from 30 donors was 97.6+/-3.96% with a viability of 99.6+/-0.83%. The recovery rate was 49.7+/-15.6%. The cells had a normal morphological appearance as assessed by May-Grünwald-stained cytospins and were functionally intact as shown by their unaltered capacity to release histamine and interleukin 4 (IL-4) following immunological activation. This procedure is a clear improvement over currently available techniques and should facilitate future investigations on basophils.

Extended Electrokinetic Characterization of Flat Solid Surfaces.

An experimental setup has been developed and applied for the combined determination of the electrokinetic potential and the surface conductivity of flat surfaces. The key feature of the new device (designated as microslit electrokinetic setup) is the variability of the distance between two parallel flat sample surfaces (10 mm x 20 mm) forming a slit channel. The setup allows us to decrease this distance down to about 1 µm keeping the surfaces parallel. In consequence, streaming potential measurements can be performed at a given solid/liquid interface both at conditions where surface conductivity is negligible and at conditions where surface conductivity significantly contributes to the total channel conductivity. The zeta potential is calculated at different channel geometries based on streaming potential and channel conductivity data and, alternatively, based on streaming current measurements and the dimensions of the cross section of the slit channel. The results obtained were found to agree well if correct conductivity values for the calculation of the zeta potential based on the streaming potential data are used. The surface conductivity is determined from the extrapolation of the channel conductance values gained at a number of sufficiently small distances between the parallel sample surfaces to the distance zero. An additional feature of the developed microslit electrokinetic setup is the assessability of the hydrodynamic thickness of adsorbed layers of macromolecules or particles at the investigated flat surface. In a series of measurements a plasma-deposited fluoropolymer (PDFP) layer on top of a glass carrier and an adsorption layer of the blood protein fibrinogen on top of the PDFP layer were characterized by zeta potential and surface conductivity measurements in different aqueous electrolyte solutions (KCl, KOH, HCl). For the PDFP/solution interfaces zeta potential up to -100 mV were obtained in solutions of neutral pH exclusively due to preferential ion adsorption. After adsorption of fibrinogen the zeta potential is considerably reduced. For the PDFP/solution interfaces surface conductivities were determined in the range of (1-2) x 10(-9) S. The contribution of the diffuse layer to the surface conductivity has been calculated from the zeta potential according to the approach of Bikerman (Kolloid Z. 72, 100 (1935)) and compared with the experimentally determined surface conductivity. Based on this comparison ions in hydrodynamically immobile interfacial layers were concluded to contribute considerably to the surface conductivity in all investigated cases. This so-called additional surface conductivity is attributed to the accumulation of hydroxide and hydronium ions in the Stern layer. Both the high specific mobility of these ions (as compared to the potassium and the chloride ions) and the conductivity of the charge determining species may contribute to the experimental observations. After adsorption of fibrinogen onto the PDFP surface the additional surface conductivity is increased by about an order of magnitude. The latter fact is assumed to be caused by the presence of mobile ions in the interfacial volume of the adsorbed protein layer. In addition to the electrochemical characterization of the adsorbed protein layer its hydrodynamic thickness has been determined by means of liquid flow measurements with the microslit electrokinetic setup. The obtained value of 48 +/- 5 nm correlates well with the protein dimensions given in the literature and is in the order of magnitude of the optical layer extension determined by ellipsometry. Copyright 1998 Academic Press.

[Percutaneous transhepatic cholangioscopy (PTCS)--an important supplement in diagnosis and therapy of biliary tract diseases (indications, technique and results)]. / Die perkutan transhepatische Cholangioskopie (PTCS)--Eine wichtige Ergänzung in der Diagnostik und Therapie von Gallenwegserkrankungen (Indikationen, Technik und Ergebnisse).

In 39 patients with biliary disease inappropriate for a transpapillary access (21 m, 18 f, mean age 62 y.) 61 percutaneous transhepatic cholangioscopies (PTCS) were performed. In 28 cases (71.8%) diagnostic PTCS was done in order to investigate the etiology of a biliary stenosis. Nine of these patients had histological as well as clinical outcome indicating a benign lesion. Histology was positive for malignancy in 14 (82%) out of 17 patients with clinically assumed malignant stenosis. The etiology of the stenosis remained obscure from a clinical point of view in two cases with negative histology for malignancy. In twelve patients (30.7%) we performed PTCS for endoscopically controlled laser or electrohydraulic lithotripsy of bile duct stones. The procedure was successful in all patients and stone fragments were delivered to the duodenum by saline lavage. Seven of these patients had stones combined with a benign stenosis and were additionally treated by long-term drainage (Yamakawa drain) for three to twelve months. Two of the seven patients had no relaps of the biliary stricture after a follow-up of ten and twelve months respectively. In five cases (12.8%) PTCS controlled insertion of a guide-wire was attempted after failure of the radiologically guided internalization of the percutaneous drainage. The procedure was successful in three of these patients.

Intraoperative recording of parietal SEP can miss hemodynamic infarction during carotid endarterectomy: a case study.

Serial recording of median nerve somatosensory evoked potentials (SEPs) provides reliable intraoperative information on critical lowering of brain perfusion which compares favorably with other monitoring methods. However, single-channel parietal SEP recording may not detect hemodynamic cerebral infarctions affecting the pre-rolandic area. Following left carotid endarterectomy for high-grade symptomatic stenosis in a 66-year-old male, predominantly motor hemiparesis and aphasia were detected which were largely reversible. Pre- and intraoperative SEPs were normal. Postoperative SEPs showed significant interhemispheric amplitude differences. Postoperative CT examinations demonstrated hemodynamic watershed-type infarctions. This rare complication of carotid endarterectomy was not detected by intraoperative single-channel parietal SEP monitoring.

Right proatlantal artery type I, right internal carotid occlusion, and left internal carotid stenosis: case report and review of the literature.

A 66-year-old man presented with right cerebellar infarction and ischemic lesions in the left dorsal thalamus and right upper parietal lobe. Angiography showed occlusion of the right internal carotid artery proximal to an ipsilateral proatlantal artery type I, 70% stenosis of the left internal carotid artery, and aplasia of both posterior communicating arteries. The carotid occlusion was successfully treated by thrombendarterectomy. Persistence of a proatlantal artery is a rare condition. In relation to the 38 literature reports on proatlantal arteries, this case demonstrates the clinical significance of a persistent proatlantal artery in the evolution of atypical ischemic cerebrovascular disease.

Estimation of extracellular space and blood volume using bioelectrical impedance measurements.

The bromide-82 dilution space (extracellular space, ECS) and blood volume (BV) were measured in 21 patients with esophageal and gastric cancer and in 27 patients 18-96 months after total gastrectomy. Resistance (R) and reactance (Xc) from bioelectrical impedance measurements were used to obtain multiple regression equations for ECS and BV. The variables weight, gender, and height 2/Xc were independent predictors of ECS (r = 0.767; P less than 0.0001). Height 2/R and gender were predictors of blood volume (r = 0.856; P less than 0.0001). The mean difference between the Br space and the ECS predicted from impedance measurements was 0 +/- 1.54 (mean +/- SD). The limits of agreement (+/- 2 SD) were therefore +/- 3.08 l or 19.6% of the mean Br space of 15.7 l. The limits of agreement for BV were +/- 789 ml or +/- 19.7% of the average BV of 4008 ml. It is concluded that bioelectrical impedance plethysmography using a single frequency can be used for the estimation of ECS and BV. The wide limits of agreement, however, may limit its used in clinical practice.

T-DNA gene 5 of Agrobacterium modulates auxin response by autoregulated synthesis of a growth hormone antagonist in plants.

Oncogenes carried by the transferred DNA (T-DNA) of Agrobacterium Ti plasmids encode the synthesis of plant growth factors, auxin and cytokinin, and induce tumour development in plants. Other T-DNA genes regulate the tumorous growth in ways that are not yet understood. To determine the function of T-DNA gene 5, its coding region was expressed in Escherichia coli. Synthesis of the gene 5 encoded protein (26 kDa) correlated with a 28-fold increase in conversion of tryptophan to indole-3-lactate (ILA), an auxin analogue. Expression of chimeric gene 5 constructs in transgenic tobacco resulted in overproduction of ILA that enhanced shoot formation in undifferentiated tissues and increased the tolerance of germinating seedlings to the inhibitory effect of externally supplied auxin. Promoter analysis of gene 5 in plants revealed that its expression was inducible by auxin and confined to the vascular phloem cells. cis-regulatory elements required for auxin regulation and phloem specific expression of gene 5 were mapped to a 90 bp promoter region that carried DNA sequence motifs common to several auxin induced plant promoters, as well as a binding site for a nuclear factor, Ax-1. ILA was found to inhibit the auxin induction of the gene 5 promoter and to compete with indole-3-acetic acid (IAA) for in vitro binding to purified cellular auxin binding proteins. It is suggested therefore that ILA autoregulates its own synthesis and thereby modulates a number of auxin responses in plants.

[Radiographic studies of ossification of the hoof cartilage of horses and its clinical evaluation]. / Röntgenologische Untersuchung der Hufknorpelverknöcherung des Pferdes und ihre klinische Beurteilung.

Radiographic examination is a valuable means for the identification of ossified lateral cartilage. The extent and stages of development of the ossification can be determined. The active stage of development of ossified lateral cartilage, which by itself can provoke lameness, can be defined. By means of the X-ray it is tried to differentiate the three kinds of ossification: the enchondral, the peri- and parachondral ossification of the lateral cartilage. Centres of ossification and fractures of the ossified lateral cartilage are described.

Extracellular space, blood volume, and the early dumping syndrome after total gastrectomy.

Extracellular space and blood volume were measured using 82Br dilution and 51Cr-tagged erythrocytes in 24 tumor-free patients after total gastrectomy. Eleven of the patients suffered from early dumping. Age, blood volume, and extracellular space were significantly smaller in dumpers (P less than 0.05). The dumping score could be predicted by a multiple regression model considering blood volume per lean body mass and extracellular space (r = 0.637; P = 0.0039). Rapid (t1/2 less than 360 seconds) emptying of the gastric substitute, assessed using a 99Tc-labeled solid test meal, was significantly associated with dumping in addition to extracellular space and blood volume (r = 0.876; P = 0.0018). Both rapid emptying and a narrow extracellular space seem to contribute to the early dumping syndrome.

Regional cerebral blood flow in patients with leuko-araiosis and atherosclerotic carotid artery disease.

The relation between white-matter lesions (WMLs), demonstrated with magnetic resonance imaging, and regional cerebral blood flow (CBF), measured with dynamic positron emission tomography and [18F] fluoromethane, was investigated in 20 patients with atherosclerotic disease of the internal carotid artery. There was no correlation between the extent of small patchy WMLs and hemispheric CBF, but hemispheric CBF was significantly reduced in 5 patients with multiple large or confluent lesions. Distinct focal cortical CBF reductions were observed when large WMLs (greater than 5 mm) were located directly beneath the cortex, whereas large WMLs in deeper white matter were associated with a more diffuse decrease of cortical perfusion. There was no evidence of preferential CBF reduction in vascular border zones with increasing severity of WMLs or stenosis of the internal carotid artery. The side of previous transient ischemic symptoms correlated significantly with hemispheric CBF asymmetries, but not with asymmetries of WMLs and internal carotid artery stenosis. It can be concluded from these results that the presence of small patchy WMLs shown by magnetic resonance imaging cannot be used as evidence of impaired cerebral perfusion, while large lesions indicate clinically relevant cerebrovascular disease affecting cortical blood flow.

High-frequency T-DNA-mediated gene tagging in plants.

An insertion element [transferred DNA (T-DNA)], transferred by soil agrobacteria into the nuclear genome of plants, was used for induction of gene fusions in Arabidopsis thaliana, Nicotiana tabacum, and Nicotiana plumbaginifolia. A promoterless aph(3')II (aminoglycoside phosphotransferase II) reporter gene was linked to the right end of the T-DNA and transformed into plants along with a plasmid replicon and a selectable hygromycin-resistance gene. Transcriptional and translational reporter gene fusions were identified by screening for APH(3')II enzyme activity in diverse tissues of transgenic plants. The frequency of gene fusions, estimated by determination of the copy number of T-DNA insertions, showed that on average 30% of T-DNA inserts induced gene fusions in Arabidopsis and Nicotiana. Gene fusions were rescued from plants by transformation of the T-DNA-linked plasmid and flanking plant DNA into Escherichia coli. By dissection of gene fusions and construction of chimeric genes, callus- and root-specific promoters were identified that showed an altered tissue specificity in the presence of a 3'-downstream-located 35S promoter. Transcript mapping of a gene fusion and expression of a non-frame transcriptional fusion of bacterial luciferase luxA and luxB genes demonstrated that dicistronic transcripts are translated in tobacco.

[Health-related evaluation of plant protective agents in drinking water]. / Gesundheitliche Bewertung von Pflanzenschutzmitteln im Trinkwasser.

Crop protection products are anthropogenous substances that should have no place in drinking water. Some of them are long-lived, can accumulate and are poorly biodegradable and many of them are not so harmless to human health as would be desirable or necessary. Accordingly, Federal German legislation provides for limit values of crop protection products close to the identification limit. Although it does not appear justified to close down waterworks effective 1 October 1989 if these limit values are exceeded, it must be carefully considered up to which concentration--depending on the individual substance--special permissions can be granted for a limited period of time. It seems that such concentrations should preferably be in accordance with general food legislation. Recommendations will be shortly issued by the Federal Health Ministry. The fact that special permissions are at all under consideration to prevent complete closing down of water supply reservoirs should sound a note of warning to all concerned, including crop protection product users and the relevant Government agencies issuing special permits.

Embolization therapy in patients with severe arterial bleeding after hysterectomy.

The case history of a patient aged 41 years, who suffered recurrent hemorrhages following vaginal hysterectomy is reported. After several surgical interventions complete hemostasis was finally achieved by selective embolization of the internal iliacal artery. The application of this procedure is recommended in case of recurrent bleeding complications caused by pelvic surgery or neoplasms.