ABSTRACT
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.
Subject(s)
Amnesia, Retrograde/diagnosis , Consciousness Disorders/diagnosis , Insulinoma/diagnosis , Insulinoma/surgery , Pancreatic Neoplasms/diagnosis , Pancreatic Neoplasms/surgery , Personality Disorders/diagnosis , Adult , Amnesia, Retrograde/etiology , Amnesia, Retrograde/prevention & control , Consciousness Disorders/etiology , Consciousness Disorders/prevention & control , Diagnosis, Differential , Humans , Insulinoma/complications , Male , Pancreatectomy , Pancreatic Neoplasms/complications , Personality Disorders/etiology , Personality Disorders/prevention & control , Recurrence , Treatment OutcomeABSTRACT
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.
Subject(s)
Abdominal Pain/etiology , Fever of Unknown Origin/etiology , Peritoneal Neoplasms/complications , Peritoneal Neoplasms/diagnosis , Plasmacytoma/complications , Plasmacytoma/diagnosis , Abdominal Pain/diagnosis , Abdominal Pain/prevention & control , Anti-Inflammatory Agents/therapeutic use , Diagnosis, Differential , Fever of Unknown Origin/diagnosis , Fever of Unknown Origin/prevention & control , Humans , Male , Middle Aged , Peritoneal Neoplasms/drug therapy , Plasmacytoma/drug therapy , Prednisolone/therapeutic use , Treatment OutcomeABSTRACT
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.
Subject(s)
Image Processing, Computer-Assisted/methods , Sheep, Domestic/physiology , Testis/physiology , Ultrasonography/methods , Animals , Male , Testis/chemistry , Testis/diagnostic imaging , Ultrasonography/veterinaryABSTRACT
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.
ABSTRACT
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.
Subject(s)
Rete Testis , Sheep , Spermatogonia/metabolism , Spermatogonia/transplantation , Transduction, Genetic/veterinary , Animals , Cell Survival , Gene Expression , Genetic Vectors , Green Fluorescent Proteins/genetics , Immunohistochemistry/veterinary , Lentivirus/genetics , Lentivirus/growth & development , Male , Seminiferous Tubules/cytology , SpermatogenesisABSTRACT
BACKGROUND AND OBJECTIVE: The gate-keeping role of general practitioners (GPs) is currently the topic of much debate in Germany. Currently it is possible for patients in Germany to see a specialist either through referral by their GP or directly through self-referral. To determine whether the gate-keeping role of GPs has a filtering effect, we compared patients referred by their GPs with self-referred patients presenting with suspected chronic venous insufficiency (CVI) to a specialist practice. PATIENTS AND METHODS: From September to December 2001, we prospectively recruited 316 patients seen for suspected CVI in a specialist practice for vascular surgery and phlebology. Symptoms and clinical findings were recorded using a standardized form. RESULTS: 58.2 % of patients were referred by their GPs. These patients were on average 6 years older and presented at a more advanced stage of disease than self-referred patients. No difference was found between patients with and without referral with respect to the symptoms reported or the therapy recommended by the specialist. CVI was excluded in 7.1 % of patients with a referral and in 6.8 % of those without a referral. CONCLUSIONS: The majority of patients consulting a specialist were referred by their GP. The more advanced disease stage of these patients indicates that a filtering process occurs in referral by GPs. However, the share of patients without referrals in whom CVI could be excluded was low and not significantly different from that of patients with referrals. This indicates that misdiagnosis due to self-referral is relatively modest. A cost reduction effect in a gate-keeper system could therefore only be small.
Subject(s)
Patient Acceptance of Health Care , Physicians, Family , Referral and Consultation/statistics & numerical data , Venous Insufficiency/therapy , Adult , Age Factors , Chronic Disease , Female , Gatekeeping/economics , Gatekeeping/statistics & numerical data , Germany/epidemiology , Health Services Research , Hematology , Humans , Male , Middle Aged , Prospective Studies , Referral and Consultation/classification , Referral and Consultation/economics , Severity of Illness Index , Vascular Surgical Procedures , Venous Insufficiency/economics , Venous Insufficiency/epidemiologyABSTRACT
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.
Subject(s)
Alkylating Agents/pharmacology , Busulfan/pharmacology , Spermatogonia/drug effects , Spermatogonia/transplantation , Animals , Cell Death , DNA-Binding Proteins/genetics , Dose-Response Relationship, Drug , Female , Infertility, Male/therapy , Male , Mice , Mice, Knockout , Models, Animal , Pregnancy , Seminiferous Tubules , Sperm Count , Testis/embryology , Transplantation Conditioning , Transplantation, HomologousABSTRACT
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.
Subject(s)
Alkaline Phosphatase/antagonists & inhibitors , Cattle/embryology , Embryo, Mammalian/enzymology , Enzyme Inhibitors/pharmacology , Isoenzymes/antagonists & inhibitors , Alkaline Phosphatase/analysis , Alkaline Phosphatase/genetics , Animals , Blastocyst/enzymology , Dimerization , Fallopian Tubes/enzymology , Female , Hot Temperature , Isoenzymes/analysis , Isoenzymes/genetics , Kidney/enzymology , Levamisole/pharmacology , Liver/enzymology , Morula/enzymology , Oocytes/enzymology , Ovary/enzymology , Reverse Transcriptase Polymerase Chain Reaction , Spleen/enzymology , Time FactorsABSTRACT
The development rate of bovine chimeric embryos reconstituted at the 4-cell stage is relatively low. If chimerism is to be used as an approach in producing transgenic livestock, it is important to investigate whether this rate is affected by the sex of the blastomeres being combined and if all blastomeres survive equally well. In Experiment 1, blastomeres from 4-cell stage embryos were inserted into surrogate zonae pellucidae either in pairs to reconstitute 4-cell chimeras, or as the original sets of four to make handled controls. The development of chimeras with one pair of blastomeres labelled with PKH26-GL was also investigated. The rate of development into blastocysts was similar in chimeras with unlabelled blastomeres (23%) and in those in which one pair of blastomeres was labelled (26%) and was lower (P < 0.001) than in the handled and IVF control groups (43 and 58%, respectively). Labelled cells were distributed approximately evenly between ICM and trophoblast. In Experiment 2, the effect of sex differences between pairs of blastomeres in chimeras was investigated; chimeras were reconstituted from pairs of blastomeres taken from 4-cell embryos in which the remaining pair was sexed by PCR. No significant differences according to the sex of constituent blastomeres were detectable (mixed sex, 27%; males, 24%; females, 21%; P > 0.05). These results suggest that, in addition to the negative effects of micromanipulation, factors other than the sex of the blastomeres are involved in the reduced rate of development of chimeric bovine embryos. They also confirm the usefulness of PKH26-GL labelling for tracking the progeny of cleaving bovine blastomeres at least to the blastocyst stage.
Subject(s)
Blastomeres/physiology , Chimera/physiology , Embryonic and Fetal Development , Organic Chemicals , Animals , Animals, Genetically Modified , Cattle , Cells, Cultured , Female , Fluorescent Dyes/metabolism , Male , Micromanipulation , Sex Differentiation/physiologyABSTRACT
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.
Subject(s)
Fertilization in Vitro/veterinary , Goats/physiology , Oocytes/physiology , Spermatozoa/physiology , Acrosome/physiology , Animals , Centrifugation, Density Gradient/veterinary , Coculture Techniques/veterinary , Female , Fertilization in Vitro/methods , Filtration , Immunohistochemistry/veterinary , Male , Microscopy, Confocal/veterinary , Microscopy, Fluorescence/veterinary , Pregnancy , Time FactorsABSTRACT
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.
Subject(s)
Memory , Reading , Serial Learning , Space Perception , Time Perception , Adult , Cognition , Cues , Female , Humans , Male , Models, Psychological , Time FactorsABSTRACT
Embryonic alkaline phosphatase (EAP) is expressed during the preimplantation period of mouse development; however, its function is unknown. To determine whether the absence of an EAP gene affects development of preimplantation embryos, we studied mice homozygous for the disrupted EAP gene (EAP.ko mice). Time to reach morphologically definedpreimplantation stages, preimplantation loss, cell count, gestation length, and litter size were monitored, and it was found that EAP.ko embryos have slower development and higher rates of degeneration during in vitro preimplantation development. In vivo, EAP.ko mice had a longergestation, smaller litter size, and fewer cells at 93 hr after human chorionic gonadotropin injection. Furthermore, there was no compensation for the absence of EAP gene in EAP.ko embryos by other isozymes of alkaline phosphatase. We conclude that the presence of an active EAP gene is beneficial for preimplantation development of the mouse embryo, and its absence leads to fewer blastocysts in vitro, delayed parturition, and reduced litter size in vivo.
Subject(s)
Alkaline Phosphatase/genetics , Embryonic Development/physiology , Alkaline Phosphatase/metabolism , Animals , Cell Count , Female , Genitalia, Female/metabolism , Humans , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Pregnancy , Time FactorsABSTRACT
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.
Subject(s)
Attention , Models, Psychological , Reading , Humans , Memory , Problem Solving , Psycholinguistics , Space Perception , Time PerceptionABSTRACT
Expression of the X inactive-specific transcript (Xist) is thought to be essential for the initiation of X chromosome inactivation and dosage compensation during female embryo development. In the present study, we analyzed the patterns of Xist transcription and the onset of X chromosome inactivation in bovine preattachment embryos. Reverse transcription-polymerase chain reaction (RT-PCR) revealed the presence of Xist transcripts in all adult female somatic tissues evaluated. In contrast, among the male tissues examined, Xist expression was detected only in testis. No evidence for Xist transcription was observed after a single round of RT-PCR from pools of in vitro-derived embryos at the 2- to 4-cell stage. Xist transcripts were detected as a faint amplicon at the 8-cell stage initially, and consistently thereafter in all stages examined up to and including the expanded blastocyst stage. Xist transcripts, however, were subsequently detected from the 2-cell stage onward after nested RT-PCR. Preferential [3H]thymidine labeling indicative of late replication of one of the X chromosomes was noted in female embryos of different developmental ages as follows: 2 of 7 (28.5%) early blastocysts, 6 of 13 (46.1%) blastocysts, 8 of 11 (72.1%) expanded blastocysts, and 14 of 17 (77.7%) hatched blastocysts. These results suggest that Xist expression precedes the onset of late replication in the bovine embryo, in a pattern compatible with a possible role of bovine Xist in the initiation of X chromosome inactivation.
Subject(s)
Cattle/embryology , Dosage Compensation, Genetic , Gene Expression , RNA, Untranslated , Transcription Factors/genetics , Animals , Blastocyst/metabolism , DNA/analysis , Embryonic Development , Escherichia coli/genetics , Female , Male , Pregnancy , RNA/chemistry , RNA, Long Noncoding , RNA, Messenger/analysis , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology , Sex Characteristics , Testis/chemistry , Tissue DistributionABSTRACT
We report the cloning and partial sequences of two novel bovine tissue-specific alkaline phosphatase (AP) isozymes (TSAP2 and TSAP3) from in vitro-produced bovine blastocysts. Using a reverse-transcribed polymerase chain reaction (RT-PCR)-based assay for mRNA expression and in vitro-produced preattachment bovine embryos, TSAP2 mRNA was detected first at the four-cell stage prior to the major burst of embryonic transcription in cattle and TSAP3 at the eight-cell stage with the major burst in transcription. Furthermore, the transcription of TSAP2 and TSAP3 displays a curious "on-off" pattern during early cleavages between 40 and 120 hr after insemination. Activity of bovine AP, measured by an azo-dye coupling technique, indicates that at least one AP isozyme is functional in oocytes and embryos throughout bovine preattachment development. However, maternal and embryonic-derived AP activity may have different cell-surface distributions. This novel expression pattern of the bovine AP isozymes could provide a useful tool for identifying and clarifying the events controlling transcription and gene expression during early embryo development.
Subject(s)
Alkaline Phosphatase/genetics , Blastocyst/enzymology , Isoenzymes/genetics , Alkaline Phosphatase/biosynthesis , Animals , Base Sequence , Cattle , DNA, Complementary , Gene Expression , Isoenzymes/biosynthesis , Molecular Sequence Data , Sequence Analysis, DNA , Sequence Homology, Nucleic Acid , Species Specificity , Transcription, GeneticABSTRACT
PROBLEM: Natural Killer lymphocytes (NK cells) from the pregnant uterus and from other tissues in pregnant and nonpregnant mammals can be stimulated by interleukin-2 (IL-2) during culture to become Lymphokine Activated Killer (LAK) cells. The susceptibility of cultured trophoblast cells to lysis by LAK cells raises the enigma of why uterine (u) NK cells that are characterized by morphology and by surface phenotyping as "activated," and thus potentially damaging to the placenta, become localized to implantation sites during normal rodent gestation. METHOD: uNK cells migrating from explant cultures of the metrial gland were assessed for expression (mRNA and protein) of each chain of the IL-2 receptor (IL-2R). Implantation sites from transgenic mice lacking a functional IL-2 gene were examined histologically for the differentiation of mature, granulated uNK cells. RESULTS AND CONCLUSION: Early post-implantation, mRNA from migrating uNK cells contains transcripts for all three chains of the IL-2R. Only IL-2Rgamma was expressed at day 12 of gestation; expression of this gene was also lost by day 16. Loss of IL-2R transcription did not result in loss of protein expression; however, it did coincide with loss of uNK cell viability in vivo. Apparently normal differentiation of uNK cells occurred in IL-2(-/-) mice and in doubly mutant IL-2(-/-).beta2m(-/-) mice. Thus, despite uNK cell expression of the full IL-2R at day 8 of gestation, IL-2 is not required for the maturation of uNK cells to their fully granulated form or for normal placental development.
Subject(s)
Interleukin-2/physiology , Killer Cells, Natural/cytology , Uterus/cytology , Animals , Cell Differentiation , Cell Movement , Cytoplasmic Granules/ultrastructure , Embryo Implantation , Endometrium/cytology , Endometrium/immunology , Female , Gene Expression Regulation, Developmental , Gestational Age , Killer Cells, Natural/immunology , Killer Cells, Natural/metabolism , Killer Cells, Natural/ultrastructure , Lymphocyte Activation , Mice , Mice, Transgenic , Pregnancy , RNA, Messenger/analysis , Receptors, Interleukin-2/biosynthesis , Receptors, Interleukin-2/genetics , Uterus/immunologyABSTRACT
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.
Subject(s)
Memory/physiology , Reading , Spatial Behavior/physiology , Adult , Female , Humans , Male , Models, Psychological , Neuropsychological TestsABSTRACT
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.
Subject(s)
Killer Cells, Natural/immunology , Killer Cells, Natural/metabolism , Receptors, Estrogen/biosynthesis , Receptors, Interleukin-2/biosynthesis , Receptors, Progesterone/biosynthesis , Uterus/cytology , Uterus/immunology , Animals , Cells, Cultured , Female , Hybridomas , Mice , Mice, Inbred Strains , PregnancyABSTRACT
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.
Subject(s)
Carrier Proteins/genetics , Embryonic Development , Transcription, Genetic , Animals , Cattle , Embryonic and Fetal Development , Fallopian Tubes/metabolism , Female , Genes , Insulin-Like Growth Factor Binding Proteins , Mice/embryology , Polymerase Chain Reaction , Pregnancy , RNA, Messenger/metabolism , Somatomedins/genetics , Uterus/metabolismABSTRACT
The temporal patterns of expression of genes encoding insulin-like growth factor (IGF) ligands and receptors during very early development have been investigated in several laboratories in several different mammalian species. Both reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemical techniques have been used to identify the time of appearance of gene transcripts or end-products. In preimplantation mouse embryos, IGF-II ligand and receptor gene activity is detectable as early as at the two-cell stage, the time when transcription from the embryonic genome is activated, but receptors for insulin and IGF-I are not detectable until the compacted eight-cell stage. Transcripts for insulin or IGF-I are not detectable in preimplantation mouse embryos, although the ligands are present in the reproductive tract. The pattern of IGF gene expression is not, however, identical in all mammalian species. In cow embryos, for example, transcripts for IGF-I and IGF-II ligands and receptors and insulin receptors have been detected at all stages of preimplantation development from mature oocyte to blastocyst (Watson et al., 1992). Attempts to quantitate transcript abundance in these early embryos are in progress in our laboratory. In the preimplantation mouse embryo, transcripts for several different IGF-binding proteins (IGFBP-2, -3, -4, and -6) have been detected by RT-PCR procedures. In addition, transcripts for IGFBPs have been identified in RNA derived from cumulus cells, the ovary, the oviduct, the uterus, and the decidua. These findings suggest that the interactions of IGF ligands and receptors in preimplantation development might, indeed, be modulated by IGFPs.(ABSTRACT TRUNCATED AT 250 WORDS)