[A 43-year-old patient with character changes, recurrent impaired consciousness and retrograde amnesia]. / 43-jähriger Patient mit Wesensveränderung, rezidivierenden Bewusstseinsstörungen und retrograder Amnesie.

A 43-year-old male patient with recurring impaired consciousness and retrograde amnesia was admitted to the department of neurology. During the neurological evaluation no pathological findings could initially be revealed but one day the patient was confused again and presented with inadequate behavior: at this time a blood glucose value of 40 mg/dl was measured. For further evaluation the patient was transferred to our department. As the reason for the impaired consciousness was suspected to be of neuroglucopenic origin a rapid adrenocorticotropic hormone (ACTH) stimulation test was first performed to rule out adrenal insufficiency. For further evaluation a fasting test was conducted: after 48 h an episode with neuroglucopenic symptoms occurred again which disappeared after intravenous administration of glucose. The laboratory results of glucose, insulin and c-peptide determined at this point in time led to the diagnosis of an insulinoma. By ultrasound examination a hypoechogenic lesion 1.5 cm in size could be shown in the head of the pancreas and was confirmed by magnetic resonance imaging (MRI). After duodenum-preserving partial pancreatic head resection with enucleation of the insulinoma no further neuroglucopenic symptoms occurred.

[A 55-year-old man with abdominal pain and fever]. / 55-jähriger Patient mit abdominellen Schmerzen und Fieber.

A 55-year-old man was admitted for evaluation of chronic abdominal pain and fever. Computed tomography demonstrated a retroperitoneal inflammatory process involving the mesenteric root. Adipose tissue biopsy showed panniculitis mesenterica with granulomas. Further examinations confirmed the diagnosis of plasmocytoma type IgG kappa. Treatment with steroids (prednisolone), resulted in immediate improvement of pain and fever. Mesenteric panniculitis represents a paraneoplastic syndrome associated with non-Hodgkin lymphoma.

Nanoscale patterning induced strain redistribution in ultrathin strained Si layers on oxide.

We present a comparative study of the influence of the thickness on the strain behavior upon nanoscale patterning of ultrathin strained Si layers directly on oxide. The strained layers were grown on a SiGe virtual substrate and transferred onto a SiO(2)/Si substrate using wafer bonding and hydrogen ion induced exfoliation. The post-patterning strain was evaluated using UV micro-Raman spectroscopy for thin (20 nm) and thick (60 nm) nanostructures with lateral dimensions in the range of 80-400 nm. We found that about 40-50% of the initial strain is maintained in the 20 nm thick nanostructures, whereas this fraction drops significantly to approximately 2-20% for the 60 nm thick ones. This phenomenon of free surface induced relaxation is described using detailed three-dimensional finite element simulations. The simulated strain 3D maps confirm the limited relaxation in thin nanostructures. This result has direct implications for the fabrication and manipulation of strained Si nanodevices.

Transduction and transplantation of spermatogonia into the testis of ram lambs through the extra-testicular rete.

Spermatogonial transplantation will provide a new way to study spermatogenesis in domestic animals, disseminate male genetics and produce transgenic animals, if efficiency can be improved. We evaluated a 'surgical' method for transplanting donor cells into testes of ram lambs, where the head of the epididymis is reflected, and a catheter introduced into the extra-testicular rete testis. We also tested transduction of ram spermatogonia with a lentiviral (LV) vector as a means to identify permanent colonization, and introduce genes into donor cells. Eight ram lambs, 11- to 13-week olds, were the recipients: in five, spermatogonia were injected into one testis, and the contralateral testis was an un-manipulated control: in two, spermatogonia were injected into one testis and the contralateral was sham-injected: in one, both testes were injected. Six lambs received spermatogonia labelled with a cell-tracking dye and these were collected 1 or 2 weeks after transplantation; three lambs received spermatogonia transduced with a LV vector driving the expression of enhanced Green Fluorescence Protein and these were collected after 2 months. Donor cells were detected by immunohistochemistry in tubules of seven of nine recipient testes. Approximately 22% of tubule cross-sections contained donor cells immediately after transplantation, and 0.2% contained virally transduced cells 2 months after transplantation. The onset of spermatogenesis was delayed, and there were lesions in both injected and sham-injected testes. Despite the effects of the surgery, elongated spermatids were present in one recipient testis 2 months after surgery. The results suggest that, after modifying the surgical and transduction techniques, this approach will be a means to produce good colonization by donor spermatogonia in sheep testes.

Insulin-like growth factor binding proteins are transcribed by preimplantation mouse embryos.

Preimplantation embryos have been reported to synthesize insulin-like growth factors (IGF) and their receptors, and reproductive tract fluid has been found to contain insulin and IGF I. In this communication, we report that all stages of preimplantation mouse embryos transcribe IGF binding proteins (IGF-BP) 2, 3 and 4, and that blastocysts also transcribe IGF-BP6. IGF-BP5 was not detected at any preimplantation stage. Reproductive tract cells in proximity to preimplantation mouse embryos transcribe all of IGF-BP2 through 6. Thus studies of the mechanisms of IGF action on preimplantation mouse development must consider the IGF-BP.

Inhibitor profiles of alkaline phosphatases in bovine preattachment embryos and adult tissues.

The alkaline phosphatases are a small family of isozymes. Bovine preattachment embryos transcribe mRNA for two tissue-specific alkaline phosphatases (TSAP2 and TSAP3) beginning at the 4- and 8-cell stages. Whereas no mRNA has been detected in oocytes, there is maternally inherited alkaline phosphatase activity. It is not known which isozyme(s) is responsible for the maternal activity or when TSAP2 and TSAP3 form functional protein. No antibodies are available that recognize the relevant bovine alkaline phosphatases. Therefore, sensitivity to heat and chemical inhibition was used to separate the different isozymes. By screening tissues, it was determined that the bovine tissue-nonspecific alkaline phosphatase (TNAP) is inactivated by low temperatures (65C) and low concentrations of levamisole (<1 mM), whereas bovine tissue-specific isozymes require higher temperatures (90C) and levamisole concentrations (>5 mM). Inhibition by L-homoarginine and L-phenylalanine was less informative. Cumulus cells transcribe two isozymes and the pattern of inhibition suggested heterodimer formation. Inhibition of alkaline phosphatase in bovine embryos before the 8-cell stage indicated the presence of only TNAP. At the 16-cell stage the pattern was consistent with TNAP plus TSAP2 or -3 activity, and in morulae and blastocysts the pattern indicated that the maternal TNAP is fully supplanted by TSAP2 or TSAP3.

The distribution of two cell surface determinants of mouse embryonal carcinoma and early embryonic cells.

This study examines the distribution of two carbohydrate determinants recognized by monoclonal antibodies EMA-1 and EMA-6 on cells in culture, adult mouse tissues and day 0-7 embryos, and provides partial biochemical characterization of the molecules carrying the determinants for EMA-1 on embryonal carcinoma cells. Both antigens are present on primordial germ cells of day 8-day 10 mouse embryos (Hahnel and Eddy, 1986), and are expressed recurrently on pluripotent cells during earlier embryogenesis. Both antigens are also present on mouse embryonal carcinoma cells and the same set of adult tissue types. However, the distribution of the EMA-6 determinant on adult urogenital epithelia and during embryogenesis is more restricted than that of EMA-1. The determinants are detected only on lumenal surfaces of epithelial cells, suggesting that they are either not involved in processes of cell-cell or cell-substrate interaction, or are masked or altered during such interactions. Present biochemical evidence suggests that EMA-1 determinant on Nulli SCC1 cells may be on a large glycoprotein or a glycoprotein complex that is firmly attached to the cell membrane.

Syngeneic antiserum to Nulli SCC1 embryonal carcinoma cells recognizing surface antigens of embryonic cells.

A syngeneic antiserum was prepared in mice against the nullipotent embryonal carcinoma cell line Nulli SCC1 to determine whether cell surface determinants are expressed commonly by this cell line and early embryos. After antibodies to calf serum components were removed from the antiserum by affinity purification, the antiserum was tested on preimplantation and 6 1/2-day postimplantation mouse embryos, embryonal carcinoma cells, tumor cells, and somatic cells. It recognizes antigens which are stage and tissue specific on early embryos and have a restricted distribution in adults. These antigens are first detected on 8-cell embryos and display a polarized distribution on blastomeres corresponding to the distribution on microvilli.

An analysis of the uterine lymphocyte-derived hybridoma cell line GWM 1-2 for expression of receptors for estrogen, progesterone and interleukin 2.

Characterisation of murine hybridoma cell lines derived from the fusion of lymphocytes migrating from explant cultures of early, pregnancy-associated metrial glands (days 6-8 of gestation) to SP 2/0 cells, has been extended (van den Heuvel et al., J. Reprod. Immunol., 27 (1994) 13-36). These hybridomas have been grown in culture for over 2 years and are thought to represent the only immortalized lines of murine pregnancy-associated, uterine natural killer (uNK) cells. Previous studies had shown that these hybridomas, known as GWM cells, lack uNK cell surface markers, but share with uNK cells the expression of the lytic protein perforin and the ability to lyse YAC cells, a natural killer cell target (van den Heuvel et al., J. Reprod. Immunol., 27 (1994) 13-36). We report here, the evaluation of the transcription and expression of genes encoding the estrogen receptor (ER), the progesterone receptor (PR) and the interleukin 2 receptor complex (IL 2R alpha, beta and gamma) by uNK cells at day 8 of gestation and by GWM 1-2 cells and SP 2/0 cells. Our investigations indicate that expression of these genes divides day 8 uNK cells into subsets, with the predominant population being ER+, PR-, IL 2R alpha +, IL 2R beta + and IL 2R gamma +. Like day 8 uNK cells, most GWM 1-2 cells expressed all three chains of the IL 2R complex. In addition, GWM 1-2 cells expressed the ER but the PR was not detected on this cell line. Only the IL 2R alpha was detected on the SP 2/0 myeloma cell line. These studies further validate the use of GWM hybridomas as models for pregnancy-associated uNK cells.

Using temporal information to construct, update, and retrieve situation models of narratives.

Four experiments explored how readers use temporal information to construct and update situation models and retrieve them from memory. In Experiment 1, readers spontaneously constructed temporal and spatial situation models of single sentences. In Experiment 2, temporal inconsistencies caused problems in updating situation models similar to those observed previously for other dimensions of situation models. In Experiment 3, merely implied temporal order information was inferred from narratives, affecting comprehension of later sentences like explicitly stated order information. Moreover, inconsistent temporal order information prevented the creation and storage in memory of an integrated situation model. In Experiment 4, a temporal inconsistency increased processing time even if readers were unable to report the inconsistency. These results confirm the significance of the temporal dimension of situation models.

The metrics of spatial situation models.

The authors investigated the metrics of spatial distance represented in situation models of narratives. In 3 experiments, a spatial gradient of accessibility in situation models was observed: The accessibility of objects contained in the situation model decreased with increasing spatial distance between the object and the reader's focus of attention. The first 2 experiments demonstrated that this effect of spatial distance was purely categorical rather than Euclidean: Accessibility depended on the number of rooms located between the object and the focus of attention, not on the size of the rooms. Experiment 3 revealed, however, that participants were able to use information about Euclidean distance in a secondary task when necessary. The implications of these results for theories of narrative comprehension and hierarchical versus nonhierarchical theories of spatial memory are discussed.

Comparisons of oocyte maturation times and of three methods of sperm preparation for their effects on the production of goat embryos in vitro.

Various times of in vitro maturation of oocytes, and three methods of separating spermatozoa from frozen-thawed semen (Percoll density-gradient centrifugation, swim-up, and glass-wool filtration), were compared for their effects on goat embryo production in vitro. Cumulus-oocyte-complexes (COCs) from abattoir ovaries were matured in M199 supplemented with 10% fetal calf serum and hormones. In Experiment 1, COCs were fixed at 4 h intervals from 0 to 27 h of culture to assess oocyte nuclear maturation. A higher proportion cultured for 27 h than for 24 h were in Metaphase II (27/37, 73% vs. 18/33, 55%, P < 0.05). In Experiment 2, the effects of separation methods on total numbers and numbers of membrane-intact spermatozoa, and the acrosome reaction were compared. Total numbers after Percoll density-gradient centrifugation were approximately 4 times higher than after swim-up and approximately 2 times higher than after glass-wool filtration (P < 0.001). Progression of the acrosome reaction was not affected differentially. In Experiments 3 and 4, after 27 h of culture the COCs were inseminated with sperm isolated by the three methods. In Experiment 3, presumptive zygotes were examined for pronucleus (PN) formation at 6, 12, 18 and 24 h post-insemination. At 12 h, male PN formation rate from Percoll-treated spermatozoa was higher than from sperm subjected to swim-up and glass-wool treatments (20/37, 54% vs. 6/37, 16% and 6/38, 16%, respectively; P < 0.001). In Experiment 4, embryos were compared for cleavage at 48 h and development into blastocysts, hatching rates and cell number at 192 h. The rates of cleavage and blastocyst formation in the Percoll-treated group were higher (P < 0.05) than in the swim-up and glass-wool groups (62% and 18% vs. 50% and 11%, and 45% and 8%, respectively). Similarly, the mean cell number in the Percoll group was higher (P < 0.05) than in the swim-up and glass-wool groups (167 +/- 5 vs. 149 +/- 4 and 126 +/- 4, respectively). We conclude that Percoll density-gradient centrifugation is superior to the other two methods for separating goat spermatozoa from frozen-thawed semen in preparation for IVF.

Preliminary assessment of the quantitative relationships between testicular tissue composition and ultrasonographic image attributes in the ram.

This study examined the relationships between gross chemical composition and ultrasonographic characteristics of the ram testes. Ten testes from sexually mature Karakul rams were scanned ex situ with an 8-MHz linear-array transducer, in a transverse and longitudinal plane. All ultrasonograms were saved as digital images and subjected to computerized analyses. Crude protein content was determined by the Kjeldahl method, moisture was determined with an oven-drying method, and fat was measured by the Soxhlet extraction of dried samples. Mean pixel values (r=-0.64, P=0.04), pixel heterogeneity (standard deviation of pixel values; r=-0.64, P=0.04) and maximum pixel intensity (r=-0.76, P=0.01) were all negatively correlated with parenchymal protein content. Pixel heterogeneity correlated directly with extractable lipids (r=0.66, P=0.02). The quantitative correlations between echotextural and biochemical parameters found in the present experiment confirm the utility of ultrasonographic imaging combined with computer-assisted image analysis for determining changes in testicular histophysiology.

[Strategic focusing of attention during narrative reading]. / Strategische Fokussierung der Aufmerksamkeit beim Lesen narrativer Texte.

In 2 experiments based on the constructionist view of text comprehension, we investigated whether readers are able to strategically use, in a problem solving matter, spatial and temporal information that varies in its relevance to the goals of the protagonists. In order to accomplish this, we varied the specificity of the protagonists' goal descriptions. Both experiments consisted of a reading section, a test, and a questionnaire, and differed only in respect to when the questionnaire was administered. The results of both experiments showed that readers were able to use spatial and temporal information of narrative texts in a strategic matter, and they even did so without explicit instruction. This focus of attention, however, was not uniformly reflected in the reading times. Memory data showed a clear disadvantage for temporal information as compared to spatial information. This was the case even though both types of information had been equally well identified in the questionnaire as crucial to the problem solving process of the protagonist.

[Clinical value of antibody titers to Borrelia burgdorferi and titer course in neurologic disease pictures]. / Klinische Wertigkeit von Antikörpertitern gegen Borrelia burgdorferi und Titerverläufe bei neurologischen Krankheitsbildern.

Over a period of 3 years, antibody titres against Borrelia burgdorferi in serum were determined for 492 patients with a wide spectrum of neurological diseases. Using the ELISA technique, we found elevated titres against Borrelia burgdorferi in about 20% of these patients. Cranial nerve paresis was often the leading symptom of an acute Neuroborreliosis. In a number of cases the diagnosis was indicated only by an elevated IgG titre in the patient's serum, or a decrease in the titre level following antibiotic treatment. The IgG titres are, however, unsuitable for control of therapy. Non-specific parameters of inflammation such as blood sedimentation rate (BSC), C-reactive protein (CRP), serum-electrophoresis or leucocyte count are also unsuitable in evaluating the therapeutic effect or for confirming the diagnosis. The most important diagnostic criterion is the demonstration of Borrelian antibodies, synthesised locally in cerebrospinal fluid (CSF). A spirochaete index above 2 suggests autochthonous intrathecal antibody production. This procedure corresponds to the determination of intrathecally produced Treponema pallidum antibodies in neurolues from quantitative TPHA values and total IgG in serum and CSF (ITpA Index according to Prange).

Immunohistochemical localization of two monoclonal antibody-defined carbohydrate antigens during early murine embryogenesis.

Monoclonal antibodies designated "C6" and "A5" identify cell surface carbohydrates shared by embryonal carcinoma cells and early mouse embryos. The binding of both antibodies to F9 embryonal carcinoma cells was inhibited by N-acetyllactosamine. While antibody C6 did not agglutinate human erythrocytes, antibody A5 agglutinated adult, but not fetal, erythrocytes of both type A and O, suggesting partial specificity for branched polylactosamine structures. Antibodies C6 and A5 did not label preimplantation stage embryos; however, labeling with both antibodies was observed following treatment of embryos with neuraminidase. In paraffin sections of postimplantation stage embryos, C6 and A5 exhibited similar yet distinct patterns of labeling, restricted primarily to the luminal surfaces of ectodermal and visceral endodermal epithelia. Neuraminidase treatment was found to expose additional patterns of C6 and A5 labeling within the ectoderm and mesoderm of the postimplantation embryo, not restricted to periluminal surfaces. These results suggest that cell surface carbohydrates are modified during early embryogenesis, in part, by selective patterns of sialylation.

Expression of the major heat shock protein (hsp 70) family during early mouse embryo development.

Stress or heat shock proteins (hsp) are synthesized by most cells in response to adverse environmental conditions. In mammalian cells, the major proteins synthesized in response to stress have relative molecular weights (Mr) in the range of 68 to 74 kilodaltons (kD) and are encoded by a small multi-gene family collectively referred to as hsp 70 genes. In unfertilized mouse eggs, no members of the hsp 70 family appear to be synthesized under normal or stressful (heat shock) conditions. At the two-cell stage, two proteins with Mr = 74 kD and Mr = 70 kD are expressed as a consequence of developmentally activated transcription of these hsp 70 genes. No stress-induced synthesis in response to heat shock is observed at this stage. At the eight-cell stage, constitutive synthesis of the 70-kD protein continues, but, as in the two-cell embryo, no heat shock induced synthesis of a novel heat shock protein is observed. By the blastocyst stage, however, an inducible protein with Mr = 68 kD is synthesized in response to heat shock in addition to constitutive synthesis of the 70-kD protein. The constitutively synthesized cognate proteins are coded for by a set of mRNAs about 2,000 nucleotides in length. The induced hsp 68 proteins are coded for by mRNAs of larger size (about 2,600 nucleotides). Only the smaller mRNA class is detectable on Northern blots of RNA extracted from control or heat shock cleavage stage embryos and control blastocysts. As predicted from protein synthetic studies, both classes are resolved in RNA preparations derived from heat-shocked blastocysts.

Effective treatment of chronically progressive multiple sclerosis with low-dose cyclophosphamide with minor side-effects.

Twenty-one multiple sclerosis (MS) patients with a chronically progressive course were treated with a low dose of cyclophosphamide (CY). The control group consisted of 21 MS patients with a chronically progressive course who received the standard treatment (ACTH or cortisone). The control group consisted of patients who preferred the standard therapy because of its beneficial effects. In contrast, the patients of the CY group wanted to try a new therapy because the standard therapy was not effective. Thus before starting the study the progression of the disease was faster in the CY group than in the standard therapy group. As regards age, sex and degree of disability, the two groups were comparable. For 20 of the 21 patients in the CY group the degree of disability (Kurtzke scale) remained stable over 1 year; for 2 of the 20 stable patients there was even an improvement. In the standard therapy group, 7 out of 21 patients were stable over 1 year, while 14 showed progressive disability. A quantitative neurological score at the beginning and 1 year after the therapy showed a nearly identical difference between the CY group and the control group. The changes of the patients' abilities in daily-life activities (which were observed and recorded by the nurses) were similar to the Kurtzke scale data obtained by the physicians. The beneficial effect of CY in chronically progressive MS was thus highly significant (P less than 0.001). The side-effects of low-dose CY were fewer than those of ACTH.

Dose-response of RAG2-/-/gammac-/- mice to busulfan in preparation for spermatogonial transplantation.

Practical applications of spermatogonial transplantation require good rates of colonization by the donor cells. Recipient testes are usually depleted of competing endogenous spermatogonia by administration of 32-44 mg busulfan kg(-1) body weight before transplantation. However, it is not clear that this is the optimum dose, especially for immunodeficient mice. In the present study, the response of adult RAG2(-/-)/gamma(c)(-/-) (RAG2) male mice to treatment with 10-50 mg busulfan kg(-1) body weight was determined in terms of mortality rates, testicular masses and histology, and colonization of seminiferous tubules by transplanted spermatogonia. Mortality increased from 0 to 50% at doses between 20 mg busulfan kg(-1) and 40 mg busulfan kg(-1), whereas the maximum effects on testicular mass and histology were observed at 20 mg busulfan kg(-1). Colonization of testes by genetically marked spermatogonia after treatment of mice with 20 mg busulfan kg(-1) was equivalent to rates previously reported in recipients treated with 32-44 mg busulfan kg(-1). Thus, 20 mg busulfan kg(-1) appears to be the optimum dose for preparing RAG2 mice for spermatogonial transplantation. However, because the steepness of the dose-response curves indicates that direct administration of busulfan is not ideal for this purpose, 15 mg busulfan kg(-1) was administered to pregnant females at various times between day 10.5 and day 16.5 of gestation to determine whether this would deplete the number of germ cells in male offspring. Although there were large variations in testicular mass and histology, no mortality was observed and administration of busulfan at day 10.5 or 12.5 after mating delayed initiation of spermatogenesis, indicating that prenatal administration of busulfan combined with neonatal transplantation might be an effective method for further increasing rates of colonization by donor spermatogonia.