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A clinical implementation of ion-beam radiography (iRad) is envisaged to provide a method for on-couch verification of ion-beam treatment plans. The aim of this work is to introduce and evaluate a method for quantitative water-equivalent thickness (WET) measurements for a specific helium-ion imaging system for WETs that are relevant for imaging thicker body parts in the future.Helium-beam radiographs (αRads) are measured at the Heidelberg Ion-beam Therapy Center with an initial beam energy of 239.5 MeV/u. An imaging system based on three pairs of thin silicon pixel detectors is used for ion path reconstruction and measuring the energy deposition (dE) of each particle behind the object to be imaged. The dE behind homogeneous plastic blocks is related to their well-known WETs between 280.6 and 312.6 mm with a calibration curve that is created by a fit to measured data points. The quality of the quantitative WET measurements is determined by the uncertainty of the measured WET of a single ion (single-ion WET precision) and the deviation of a measured WET value to the well-known WET (WET accuracy). Subsequently, the fitted calibration curve is applied to an energy deposition radiograph of a phantom with a complex geometry. The spatial resolution (modulation transfer function at 10 % -MTFIn the optimal imaging WET-range from ∼280 to 300 mm, the fitted calibration curve reached a mean single-ion WET precision of 1.55Using a beam energy of 239.5 MeV/u and the proposed calibration procedure, quantitative αRads of WETs between ∼280 and 300 mm can be measured and show high potential for clinical use. The proposed approach with the resulting image qualities encourages further investigation toward the clinical application of helium-beam radiography.

Transformation studies of chlorinated paraffins (CPs) and the effects of CP transformation products on humans, biota and environment are rare. The focus here is on hydroxylation reactions. As for polyhalogenated persistent organic pollutants (POPs) in general, hydroxylation reactions convert lipophilic material to more polar compounds with increased mobility. We investigated the in-vitro transformation of single-chain CP-mixtures to hydroxylated products with the dehalogenase LinB from Sphingobium indicum. C11-, C12- and C13-single-chain CP-homologues were exposed to LinB and mono-hydroxylated (CP-ols) and di-hydroxylated (CP-diols) transformation products were formed. Liquid-chromatography coupled to mass-spectrometry (LC-MS) was used to detect hydroxylated products and separate them from the starting material. The presented data can be used to identify these CP-ol and CP-diol homologues in other samples. Hydroxylated products had lower chlorination degrees (nCl) than respective CP-starting-materials. Reactive and persistent CP-material was found in each homologue group. Reactive material is converted within hours by LinB, while more persistent CPs are transformed within days. Homologue-specific kinetic models were established to simulate the stepwise hydroxylation of persistent CPs to mono- and di-hydroxylated products. First-order rate constants for the formation of CP-ols (k1) and CP-diols (k2) were deduced for different homologues. Lower-chlorinated CP-ols did not accumulate to large extent and were transformed quickly to CP-diols, while higher-chlorinated CP-ols and -diols both accumulated. By enzymatic transformation of single-chain CPs with LinB, we synthesized unique sets of mono- and di-hydroxylated materials, which can be used as analytical standards and as starting materials for metabolic, toxicity and environmental fate studies.

Technical chlorinated paraffins (CPs) are produced via radical chlorination of n-alkane feedstocks with different carbon chain-lengths (∼C10–C30). Short-chain CPs (SCCPs, C10–C13) are classified as persistent organic pollutants (POPs) under the Stockholm Convention. This regulation has induced a shift to use longer-chain CPs as substitutes. Consequently, medium-chain (MCCPs, C14–C17) and long-chain (LCCPs, C > 17) CPs have become dominant homologues in recent environmental samples. However, no suitable LCCP-standard materials are available. Herein, we report on the chemical synthesis of single-chain C18-CP materials, starting with pure n-alkane and sulfuryl chloride (SO2Cl2). Fractionation of the crude product by normal-phase liquid-chromatography and pooling of suitable fractions yielded in four C18-CP-materials with different chlorination degrees (mCl,EA = 39–52%). In addition, polar side-products, tentatively identified as sulfite-, sulfate- and bis-sulfate-diesters, were separated from CPs. The new single-chain materials were characterized by LC-MS, 1H NMR and EA. LC-MS provided Relative retention times for different C18-CP homologues and side-products. Mathematical deconvolution of full-scan mass spectra revealed the presence of chloroparaffins (57–93%) and chloroolefins (COs, 7–26%) in the four single-chain C18-CP-materials. Homologue distributions and chlorination degrees were deduced for CPs and COs. 1H NMR revealed chemical shift ranges of mono-chlorinated (δ = 3.2–5.3 ppm) and non-chlorinated (δ = 1.0–3.2 ppm) hydrocarbon moieties. The synthesized C18 single-chain standard materials and respective spectroscopic data are useful to identify and quantify LCCPs in various materials and environmental samples. CP- and CO-distributions resemble the ones of existing SCCP and MCCP reference materials and technical mixtures. Furthermore, these materials now allow specific studies on the environmental fate and the transformation of long-chain chloroparaffins and chloroolefins.

Technical chlorinated paraffins (CPs) are produced via radical chlorination of n-alkane feedstocks with different carbon chain-lengths (∼C

Distichodus notospilus was described from the Ogooué River and is considered to occur throughout the Lower Guinea ichthyofaunal province and the western tributaries of the middle and lower Congo River. Recent expeditions in Equatorial Guinea collected D. notospilus specimens in the Mbini River drainage and the Mbia River; a small coastal river that is located between the Ntem and Mbini river drainages. Detailed morphological analyses and multilocus molecular analyses confirm that these two populations are distinct from one another. Topotypic populations of D. notospilus were included in the analyses and demonstrated that populations in the Mbini and Mbia rivers are distinct and these two new species are described herein. Distichodus microps sp. nov. is endemic to the Mbia River drainage and is distinguished from D. notospilus in having more scales along the lateral line (41, rarely 40 versus 37–39, rarely 40), a nearly inferior mouth versus subterminal in D. notospilus, a curved posterolateral margin of the opercle versus straight in D. notospilus, a smaller eye (56.7–80.4 versus 70.1–104.3 % of snout length), and a less prominent elongated spot at the base of the caudal fin. Distichodus mbiniensis sp. nov. is endemic to the upper Mbini River drainage and distinguished from D. notospilus in having more scales along the lateral line (41–42, rarely 40 versus 37–39, rarely 40), a much less prominent elongated dark spot at the base of the caudal fin, and a shorter dorsal fin (21.4–27.2 versus 22.7–34.2% standard length). Distichodus microps is distinguished from D. mbiniensis in having a shallower body (usually six scales from lateral line to the pelvic fin versus seven), fewer anal-fin rays (usually 12 total rays versus 13 or 14), a more inferior mouth, a deeper and longer caudal peduncle, a smaller eye, and differences in several features associated with the head. In addition to the two new species described this study also revealed potential undescribed diversity in the D. notospilus species complex in the Ntem River and Dja River (Congo R. basin) in Cameroon. The biogeography of these fishes in the rivers of Lower Guinea suggests that the Mbini River and smaller coastal rivers are overlooked areas of endemism. Studies of other reported widespread species will likely reveal additional diversity and further elucidate the processes promoting and maintaining freshwater diversity in Central Africa.

Structure, reactivity and physico-chemical properties of polyhalogenated compounds determine their up-take, transport, bio-accumulation, transformation and toxicity and their environmental fate. In technical mixtures of chlorinated paraffins (CPs), these properties are distributed due to the presence of thousands of homologues. We hypothesized that roles of CP dehalogenation reactions, catalyzed by the haloalkane dehalogenase LinB, depend on structural properties of the substrates, e.g. chlorination degree and carbon-chain length. We exposed mixtures of chlorinated undecanes, dodecanes and tridecanes in-vitro to LinB from Sphingobium Indicum bacteria. These single-chain CP-materials also contain small amounts of chlorinated olefins (COs), which can be distinct by mathematical deconvolution of respective mass-spectra. With this procedure, we obtained homologue-specific transformation kinetics of substrates differing in saturation degree, chlorination degree and carbon chain-length. For all homologues, two-stage first-order kinetic models were established, which described the faster conversion of reactive material and the slower transformation of more persistent material. Half-lifes of 0.5–3.2 h and 56–162 h were determined for more reactive and more persistent CP-material. Proportions of persistent material increased steadily from 18 to 67% for lower (Cl6) to higher (Cl11) chlorinated paraffins and olefins. Conversion efficiencies decreased with increasing chlorination degree from 97 to 70%. Carbon-chain length had only minor effects on transformation rates. Hence, the conversion was faster and more efficient for lower-chlorinated material, and slower for higher-chlorinated and longer-chained CPs and COs. Current legislation has banned short-chain chlorinated paraffins (SCCPs) and forced a transition to longer-chain CPs. This may be counterproductive with regard to enzymatic transformation with LinB.

Short-chain chlorinated paraffins (SCCPs) are listed as persistent organic pollutants (POPs) under the Stockholm Convention. Such substances are toxic, bioaccumulating, transported over long distances and degrade slowly in the environment. Certain bacterial strains of the Sphingomonadacea family are able to degrade POPs, such as hexachlorocyclohexanes (HCHs) and hexabromocyclododecanes (HBCDs). The haloalkane dehalogenase LinB, expressed in certain Sphingomonadacea, is able to catalyze the transformation of haloalkanes to hydroxylated compounds. Therefore, LinB is a promising candidate for conversion of SCCPs. Hence, a mixture of chlorinated tridecanes was exposed in vitro to LinB, which was obtained through heterologous expression in Escherichia coli. Liquid chromatography mass spectrometry (LC-MS) was used to analyze chlorinated tridecanes and their transformation products. A chloride-enhanced soft ionization method, which favors the formation of chloride adducts [M+Cl]- without fragmentation, was applied. Mathematical deconvolution was used to distinguish interfering mass spectra of paraffinic, mono-olefinic and di-olefinic compounds. Several mono- and di-hydroxylated products including paraffinic, mono-olefinic and di-olefinic compounds were found after LinB exposure. Mono- (rt = 5.9-6.9 min) and di-hydroxylated (rt = 3.2-4.5 min) compounds were separated from starting material (rt = 7.7-8.5 min) by reversed phase LC. Chlorination degrees of chlorinated tridecanes increased during LinB-exposure from nCl = 8.80 to 9.07, indicating a preferential transformation of lower chlorinated (Cl

Introduction: Given that primary care has been found to be the most likely point of entry into the health care system for concussion patients, diagnosis and management of concussions are vital to the education of family medicine physicians. Studies of primary care residencies reveal a self-perceived deficiency in sports medicine education. This project was designed to determine the effect of a concussion management clinic on family medicine residents’ confidence in and personal value placed on diagnosing and managing concussions. Methods: This project was completed in a family medicine residency as a retrospective evaluation of an educational improvement project during the 2016-2017 academic year. The intervention involved implementation of a structured clinical experience focused on management of concussions. Residents were surveyed preintervention, as well as 5 and 11 months postintervention, measuring resident exposure to and confidence in diagnosing and managing concussions. Results: Residents demonstrated significantly increased confidence in diagnosing concussions and managing complicated concussions following the intervention (both F [2, 84]=3.56, P=0.03). Post hoc analysis indicated the statistical difference was found between preintervention measures and 1 academic year later. The number of concussions seen positively correlated with resident confidence in both diagnosing and managing concussions while personal value remained high. Conclusions: This project is the first to evaluate the impact of a concussion clinic on resident education, demonstrating significant improvement in multiple areas of concussion education. Replicating the project in other family medicine residencies could be beneficial in determining if this clinic education model improves resident outcomes in other residencies.

Background and Objectives: The transition to clerkships is one of the most challenging times during medical school. To help students better cope, many schools have established transition-to-clerkship curricula. Such curricula may optimally prepare students through increasing their self-efficacy and response efficacy. We hypothesized that a small-group, near-peer-led format would be ideally suited to help students achieve these outcomes. Methods: During process improvement for a transition-to-clerkship curriculum, we conducted an informal focus group and subsequent survey of postclerkship students to guide curricular innovation, including incorporation of third- and fourth-year students as near-peer instructors in a seminar format. Seminars included three sequential small-group discussions focused on discrete topic areas and concluded with a large-group session highlighting salient discussion points. To evaluate the impact of this educational strategy, near-peer learners were surveyed before and after the seminars. Results: Junior student participants reported feeling more prepared to integrate into the health care team, develop a clerkship study plan, and access applicable, valuable study materials, both immediately following the seminars and 6 months later, demonstrating increased self-efficacy. These students placed equal or greater value on these topics as compared to students in previous year groups, demonstrating similar response efficacy. Conclusions: This study demonstrated an increase in student self-efficacy that persisted 6 months postintervention, in addition to similar response efficacy. Future research could be directed toward: (1) investigating whether improvements in self-efficacy among students transitioning to clerkships are associated with improved clerkship performance and (2) studying outcomes for near-peer teachers.

Die chronische Kniegelenkluxation ist ein seltenes Ereignis und stellt eine Herausforderung in der Therapie dar. Ein stufenweises Behandlungskonzept ist notwendig, um die aus mehreren Komponenten bestehende Pathologie zu behandeln. In diesem Artikel wird aufgrund eines Fallberichts ein Algorithmus fur die Therapie der chronischen Kniegelenkluxation gegeben.

Malignant melanoma derives from pigmented cells of neuroectodermal origin. Melanoma cells express estrogen receptors. An influence of steroid hormones on the proliferation rate of melanoma cells was demonstrated in some animal experiments. Thus it is of concern whether or not women with malignant melanoma are candidates for hormone replacement therapy (HRT). Until today, no prospective randomized trial on the association between HRT and malignant melanoma has been published. However, from the data available from in vitro-studies, in vivo-studies, and epidemiological reports, we can draw the following conclusions: there is no convincing evidence that the intake of sexual steroids increases the risk for the development of malignant melanoma and there is no evidence that HRT, taken after a diagnosis of malignant melanoma, negatively influences the outcome of the disease.

The diagnosis of severe combined immunodeficiency complicated by chronic graft-vs-host disease affecting liver and skin in association with engraftment of maternal T cells was established in a 5-mo-old boy. Detailed immunologic and molecular genetic studies were performed because a unique T cell phenotype was identified on initial evaluation. A major proportion of the patient's peripheral T cells expressed a CD8+ and TCR-gamma/delta+ phenotype while CD4+ T cells were virtually absent. Southern blot analysis of cell subpopulations isolated by fluorescence activated cell sorting indicated that approximately 50% of CD8+/TCR-gamma/delta+ cells were clonally related. Immunophenotyping and -genotyping also identified a clonal TCR-gamma/delta+ cell population in the child's mother. Clonal identity of these T cell populations in mother and child was demonstrated by studies using a clonspecific TCR-delta probe generated by polymerase chain reaction as well DNA sequence analysis. HLA typing and DNA fingerprinting confirmed that the child had acquired this clone diaplacentally from the mother. According to immunohistology and DNA analysis the clone was found to be virtually absent in the liver tissue suggesting that this clonal T cell population plays a minor role, if any, in the pathogenesis of the liver abnormalities in the patient. In the mother the CD8+/TCR-gamma/delta+ clone spontaneously declined to a level around 1% of PBMC several months later and has remained at this level since. We conclude that 1) a clonal expansion of TCR-gamma/delta T cells, triggered by yet unknown stimuli, may occur in otherwise healthy individuals, 2) respective T cells are able to cross the placental barrier, and 3) in an microenvironment precluding rejection, i.e., in severely immunocompromised patients, these cells may persist and even represent a significant proportion of circulating T cells.

Circulating maternal T lymphocytes were noted in the peripheral blood of six patients with severe combined immunodeficiency. Phenotypical analyses revealed the presence of both CD4 and CD8 subsets in some but not all cases. The maternal T cells could be stimulated by anti-TCR/CD3 mAb +/- rIL-2, but were virtually silent in the MLR and against the recall Ag purified protein derivative of tuberculin and tetanus toxoid, even in immunized patients engrafted with T cells from a responding mother. Using a panel of mAb against TCR V region gene encoded epitopes including V beta 5, V beta 6, V beta 8, V beta 12, and V alpha 2, we show that maternal T cells displayed a profoundly reduced TCR diversity, characterized by a lack of one or even several TCR V subsets in all six cases and a dramatic (5- to 25-fold) expansion of other TCR V subsets in three cases. In one patient analyzed, limited TCR diversity was also seen in T cells cultured from bone marrow and skin; restimulation experiments of these cells against cells expressing host MHC Ag were unsuccessful, as were attempts to exclusively allocate anti-host proliferative responses of maternal control T cells to the TCR V subsets that had undergone expansion in vivo. We conclude that a severely reduced TCR diversity is a common feature of maternal T cells engrafted in severe combined immunodeficiency patients. These novel findings provide a structural basis to understand the failure of these cells to protect the host from infections and may also help to understand their relative inefficiency to induce lethal, multi-organ, graft vs host disease. Moreover, as an experiment of nature, the reported phenomenon clearly illustrates the functional consequences in vivo of an insufficient TCR diversity.

Human severe combined immunodeficiency (SCID) patients were analyzed by a polymerase chain reaction assay for their recombination capability at the DHQ52-JH region of the immunoglobulin heavy chain locus. Five patients with B cells (B+ SCID) exhibited a recombination pattern also observed in healthy persons. In contrast, six patients lacking B cells (B- SCID) showed a grossly altered rearrangement pattern characterized by the (partial) absence of regular DHQ52-JH recombinations and the presence of abnormal rearrangements. These events were caused by deletions surpassing the boundaries of immunoglobulin coding elements and thus resemble the pattern of deletional recombinations previously described in SCID mice.

Feature extraction is a crucial part of classification procedures. In this paper we present an approach to utilize feature extraction criteria to predict the potential efficiency of a neural network classifier. Statistical and geometrical criteria are introduced for analysis. The complete system of our research consists of a class of generalized Hough-transformations for feature extraction and a subsequent neural network. The neural network performs the classification based on respective features. For an example we concentrated on a pattern recognition problem-the classification of handwritten numerals. As a result of our work we assign two feature extraction criteria to the employed network for a significant estimation of its efficiency.

Presented here is the case of a paraneoplastic cerebral degeneration (PCD) in a female patient with breast cancer and the indication of anti-Yo antibodies in the cerebrospinal fluid (CSF) and serum. The patient's primary indications were dizziness and a severe gait ataxia. The indication of anti-Yo antibodies led to the conclusion of the existence of a paraneoplastic cerebral degeneration. The antibodies in question are anti-Purkinje-cell autoantibodies acting against the antigens common to tumor and Purkinje cells which occur in association with a certain percentage of breast or ovarian cancers. The diagnosis of the primary tumor, that is clinically undetectable with conventional imaging processes, is performed with the aid of positron emission tomography (PET) to detect the presence of axillary lymph node metastases. The micro-invasive mammary carcinoma was able to be localized with the aid of MR mammography and, after MR mammography marking, was removed. The patient subsequently received adjuvant treatment with epirubicine and cyclophosphamide. This treatment failed to influence the paraneoplastic neurological symptoms.

The study was designed to investigate the role of ovarian volume, as assessed by three-dimensional (3D) sonography, in predicting conception in an in-vitro fertilization-embryo transfer (IVF-ET) programme. Transvaginal 3D sonography was performed in 152 cycles before initiation of ovarian stimulation (day 1) and on the day of oocyte retrieval. Ovarian volume showed no significant correlation with IVF outcome. On the contrary, all ovarian measurements were lower, albeit non-significantly, in the conception group. Fifteen patients (15/152, 9.9%) had a minimum unilateral ovarian volume of ≤3 mL (1 SD below the mean) on day 1 of the stimulation cycle. In this subgroup, the likelihood of conception was 6.7% (1/15) versus 21.9% (30/137) in patients with an initial minimum ovarian volume of >3 mL. This difference did not reach statistical significance. In both groups, cancellation rates due to poor ovarian response or lack of fertilization were similar. In conclusion, ovarian volumetry as assessed by three-dimensional ultrasound failed to predict conception in women undergoing IVF treatment.

This study was designed to detect the expression of CD44v splicing variants in primary ovarian carcinomas and their lymph node metastases, in order to evaluate the possible role in intraperitoneal and lymphogenic metastasization.Paraffin-embedded tissue of 50 patients with ovarian cancer was available from both the primary tumors and from the lymph node metastases. The expression of the CD44 standard from (CD44s) and of the variant isoforms CD44v5, CD44v6, CD44v7 and 8, and CD44v10 was investigated in the tumor cell regions by immunohistochemistry.45/50 (90%) primary ovarian carcinomas were found to be immunohistochemically positive for the CD44v5 splicing variant; 10 (20%) tumors expressed both CD44v5 and CD44v6. Apart from an additional v7 stain, all of the ovarian carcinomas studied were negative for the splicing variants v7, v7-8 and v10. No differences were revealed in the pattern of CD44 variants expression investigated in primary tumors and in the lymph node metastases.In ovarian cancer the expression of CD44v5 as a possible first step, as well as CD44v6, probably plays an important role in intraperitoneal implantation. In view of the comparatively heterogeneous expression patterns of CD44v in individual organs, it may be assumed that there are organ-specific differences in this respect, and that the invasion potential is influenced by both the expression pattern and the extent of expression.

Bone marrow transplantation (BMT) plays a central role in the management of congenital immunodeficiencies characterized by profound abnormalities of the T cell system [10, 14, 19]. This heterogeneous group of disorders may be corrected by the establishment of a new, donor-dependent immmune system. BMT in these disorders also represents a unique model to study and to define cellular interactions which are operative in the functional reconstitution of the lymphohemopoietic system in man.

We report on 9 infants with severe combined immunodeficiency (SCID), who additionally showed signs of Omenn syndrome with an exfoliative dermatopathy, alopecia, enlarged lymph nodes, a hepatomegalia and a striking blood eosinophilia. The immunological evaluation revealed the characteristic abnormalities of SCID with cellular and humoral immunodeficiency. All patients however had the unusual finding of mature T cells in the peripheral blood. By HLA typing these cells were noted to be of maternal origin in 5 patients. In the other 4 patients the T cells were of host origin. We asked for additional differences between both patient groups.Both patient groups were analyzed and compared with regard to case histories, clinical, laboratory and histopathological parameters.No clinical or laboratory differences could be detected. The histomorphologic analysis of patients with or without maternal T cells was identical. The skin biopsies showed dense cell infiltrations of lymphocytes, histiocytes and eosinophils, in the enlarged lymph nodes the latter two cell types predominated. Therefore the only difference between the 2 patient groups was the presence or absence of maternal T cells.Since the Omenn syndrome is found in association with maternal as well as patient derived T cells, we postulate that the peculiar symptoms of this syndrome are the result of a T cell induced inflammatory reaction, similar but not identical to a graft versus host reaction, occurring on the basis of an inborn SCID.

We have studied T cell receptor (TCR) diversity in a group of six patients with severe combined immunodeficiency (SCID) previously treated by HLA-haploidentical bone marrow transplantation (BMT). At the time of study, all patients had developed stable T cell chimerism and full reconstitution of T cell functions in the absence of acute or chronic graft-versus-host disease. Peripheral blood lymphocytes (PBL) were analysed by immunofluorescence using a panel of monoclonal antibodies (MoAb) against TCR variable (V) region epitopes including V beta 5, V beta 6, V beta 8, V beta 12, and V alpha 2. Our results showed that in each patient studied a low but significant portion of PBL reacted with each anti-TCR V region epitope MoAb used, in a manner that was, on statistical grounds, indistinguishable from results obtained with PBL from healthy controls. We conclude that within the experimental resolution of a limited number of anti-TCR V region epitope MoAb, T cell reconstitution following BMT for SCID, even when performed across a full HLA-haplotype barrier, leads to an apparently normal TCR diversity. These novel findings may be relevant in the evaluation of functional capacities of T cells that have differentiated from transplanted precursor cells in an HLA-haplodifferent environment.

We have studied a recently described method to purge human marrow of T cells using magnetic beads conjugated to various anti-T cell monoclonal antibodies. Using anti-CD2 and anti-T cell receptor/CD3 monoclonal antibodies, we show that up to 2.76 log T cell purge can be achieved while recovery of hemopoietic activity is 82%. As a control, our clinically established T cell depletion protocol, namely soybean agglutination combined with E-rosetting, provided 3.39 log T cell purge and 59% recovery of hemopoietic activity. The impact of these results on T cell depletion strategies in HLA-nonidentical bone marrow transplantation is discussed.

Reticular Dysgenesis (RD) basically represents a lymphopenic severe combined immunodeficiency (SCID) in association with congenital agranulocytosis. It is presumed that RD results from a primary defect of pluripotent hematopoietic stem cells (HSC). Alternatively RD might be due to an alloreaction induced by T-cells derived from intrauterine transfusion of maternal cells into an immunoincompetent host. In the past 15 years, among 49 newborns with SCID taken care of in the University Hospital of Ulm, 5 children (4 boys, 1 girl) exhibited the characteristics of RD. In 3 of 4 cases studied, maternal T-cells were detected by HLA-typing. All 3 children showed signs of graft-versus-host-disease (GvHD), confirmed histologically. However, 9 of 45 newborns with SCID without congenital agranulocytosis also disclosed maternal T-cell-engraftment; 5 of the 9 had signs of GvHD. Therefore, it is unlikely that RD is caused by GvHD secondary to maternofetal transfusion. The fact that erythropoiesis, thrombopoiesis and the monocyte-macrophage-system basically are intact argues against a global maturation defect of HSC in RD.

Aim To investigate the role of 3D-ultrasound in estimating the likelihood of conception in an IVF programme. Materials and methods We studied a total of 152 women during their stimulation cycle. Apart from conventional 2D-parameters we assessed endometrial and uterine volume (sub-)endometrial blood flow (sub-)endometrial vessel density and ovarian volume by 3D-ultrasound. Results With regard to subendometrial blood flow on day 1 of the stimulation cycle, all 3D-indices were significantly lower in the conception vs. non-conception group. On the day of oocyte retrieval, any difference between the two groups failed to reach statistical significance. The same applied to endometrial volumes. The latter showed a significant corrrelation with uterine size. Ovarian volumes ≤ 3 mm (1 SD below the mean) on day 1 of the stimulation cycle were associated with a nonsignificant lower conception rate. Conclusion Quantitative assessment of spiral artery blood flow and vessel density may be an early predictor of IVF success.

The diagnosis of severe combined immunodeficiency (SCID) was established in a 5 months old boy based on typical clinical and laboratory findings. Analysis of surface markers, however, revealed a remarkable pattern: CD3 86%, CD4 5%, CD8 62%, TCR-1 38%, TCR-2 47%, CD8+/TCR-1+ 21%, CD8+/TCR-2+ 24%. Thus, the CD8+/TCR-1+ population almost absent in normal blood was extremely elevated to 21% of PBMC. To further characterize these cells and the complementary fractions they were double labelled using commercial monoclonal antibodies to CD8 and TCR-1 and sorted by fluorescence activated cell sorting (FACSTAR PLUS). DNA from sorted cells was next analyzed by Southern Slot using a common delta-specific probe. A monoclonal rearrangemnt (Vδ1 to Jδ1) was exclusively found in the abnormally elevated cells but not in any other population analyzed. Using PCR we next generated a clonospecific probe directed against the NDN sequences of the rearranged fragment. Dot blot hybridization with this probe confirmed Southern blot results. The mother's PBMC contained 5% T cells with CD8+/TCR-1+ phenotype which had an identical rearrangement on Southern blot and reacted with the child's clonospecific probe, in addition, the NDN nucleotide sequences were identical. Because of signs of GvHR skin and liver biopsies were taken. Southern blots of DNA extracted from the skin revealed the presence of the rearranged fragment while only a minimal reaction could be observed with liver DNA. This case shows that monoclonal maternal cells can metastasize diaplacentally into the child if they are not destroyed by the child's immune system and that CD8+/TCR-1+ cells may have a rule in the skin but not in the liver.

Δ 9 -THC significantly influenced the acute lethal effects (24 hr after a single dose) in mice of several drugs which alter cholinergic, adrenergic and serotoninergic neurotransmission. The LD50 values of carbachol chloride, arecoline HCl, physostigmine salicylate and neostigmine methylsalicylate were all significantly potentiated after pretreatment with 20.0 mg/kg of Δ 9 -THC. Paradoxically, lethality produced by methacholine and exogenously administered acetylcholine were attenuated. Evidence for an α-adrenergic receptor blocking effect of Δ 9 -THC was noted by the significantly larger LD50 values for 1-norepinephrine bitartrate and phenylephrine HCl when each was given in combination with Δ 9 -THC. Moreover, bretylium tosylate and dl -α-methyl- p -tyrosine toxicities were also attenuated by pretreatment with Δ 9 -THC. Finally, the combination of Δ 9 -THC and cyproheptadine HCl was more toxic than the latter compound alone, suggesting an additive antiserotonin effect.

The effects of RX 781094, a new and potent alpha 2-adrenoceptor antagonist, on locus coeruleus (LC) unit activity were examined. Low doses of RX 781094 produced suppression of spontaneous LC unit activity which could be reversed with yohimbine. The increase in LC firing produced by WB 4101 could also be reversed with a low dose of RX 781094. Thus, at low doses, RX 781094 has clonidine-like alpha 2-agonist activity. At higher doses, RX 781094 reversed the effects of clonidine and markedly shifted the dose of clonidine required to suppress LC unit activity. These data suggest that at high doses RX 781094 has alpha 2-antagonist properties. It is concluded that RX 781094 may be a partial agonist at alpha 2-adrenoceptors in the CNS.

The anti-edema activity of THC has been directly compared to that of hydrocortisone and aspirin in rat paw edemas produced by eleven different phlogistic agents. A significant effective dose for THC could only be achieved in carrageenan, dextran, formalin, kaolin and Na urate induced edemas, while hydrocortisone and aspirin were active against all edemas except that produced by serotonin. Based on these data it is suggested that the anti-edema effects of THC most likely occur by a different mode of action than either the classic steroid or non-steroidal anti-inflammatory agents.

Acute rejection of allogeneic and semiallogeneic marrow grafts has long been considered to be a function of the natural immune system because it shares many features with NK activity in mice. With the use of a recently developed in vivo adoptive transfer assay in which spleen cells are transferred from mice able to reject a particular marrow graft into mice that fail to do so, we show that the cells responsible for induction of marrow graft rejection indeed display the phenotype of NK cells: they lack the T cell Ag CD4 and CD8 but express the NK Ag NK1 and ASGM1. The rejection induced by adoptively transferred cells is exquisitely specific--a feature that points to a specific recognition process by the transferred cells. To elucidate what the recognition structure on these cells may be we found that they express CD3 and most likely the beta-chain of the TCR. Highly purified responder cells with the NK1+, CD3+, CD4-, CD8- phenotype, when transferred into nonresponder recipients, cause specific marrow graft rejection. We conclude that the acute rejection of bone marrow grafts is caused by a cell that expresses NK phenotype but is of T cell lineage. This may suggest the specificity of acute marrow graft rejection is caused by a specific recognition process that involves TCR.

Diazepam (5 mg/kg, ip) and tracazolate (40 mg/kg, ip), a nonbenzodiazepine anxiolytic, blocked electrically-induced head-turning without producing sedation. Bicuculline and picrotoxin, GABA antagonists, at doses not affecting head-turning (2 mg/kg, ip) antagonized the effects of diazepam and tracazolate on head-turning. However, at the same dose, bicuculline was more effective as an antagonist of diazepam whereas picrotoxin was more effective as an antagonist of tracazolate. These results suggest that benzodiazepine as well as nonbenzodiazepine anxiolytics possess GABAmimetic activity. The difference in potency between bicuculline and picrotoxin as antagonists of diazepam and tracazolate may be related to their reported differences as GABA antagonists (e.g., site of receptor interaction).

The effects of acute and subacute administration of the tricyclic antidepressants imipramine and amitriptyline, and the atypical antidepressants mianserin and iprindole, on seizures kindled from the amygdala and the cortex were examined. Whereas amitriptyline selectively antagonized seizures kindled from the amygdala after a single dose, neither amitriptyline nor imipramine was any more effective in antagonizing seizures kindled from the amygdala than from the cortex following subacute treatment. Both acute and subacute administration of iprindole failed to significantly alter seizures kindled from either site. Although only the highest acute dose of mianserin tested selectively attenuated amygdaloid seizures, a lower dose that was ineffective when given acutely, was selective when given subacutely. In contrast to an earlier report, the present findings suggest that kindling may not be a particularly useful model for the evaluation of potential antidepressant agents.

(C57BL/6 x DBA/2)F1 mice transplanted with parental C57BL/6 spleen cells become splenic chimeras, show donor antihost cytotoxic T cell activity, and lose their T cell-mediated, humoral, and natural immunity. Injection of anti-asialo-GM1 (ASGM1) into transplanted mice strongly suppresses splenic cytotoxic activity and causes a significant reduction of spleen cells expressing ASGM1, Thy-1, and Lyt-2. In vitro treatment of spleen cells from transplanted mice with antibody and complement shows that the cytotoxic effector cells are ASGM1+, Thy-1+, Lyt-2+, L3T4-, NK1.1-, and H-2d-, hence of donor origin. The cytotoxic effector cells are specific for H-2d targets and lack NK activity. In an attempt to explore whether in vivo elimination of the cytotoxic effector cells has any influence on splenic chimerism or humoral immunity, F1 mice injected with parental splenocytes were treated with anti-ASGM 1. Results show that this treatment eliminates a substantial proportion of cytotoxic effector cells but has no effect on splenic chimerism or restoration of humoral immunity. It therefore appears that cytotoxic effector cells are not primarily responsible for induction of chimerism or suppression of humoral immunity. In support of this injection of parental spleen cells with the nu/nu mutation induces killer cells in F1 mice but fails to induce splenic chimerism or immunosuppression. In contrast, injection of parental spleen cells with the bg/bg mutation generates both splenic chimerism and suppression of humoral immunity although their ability to generate cytotoxic effector cells in F1 hosts is seriously impaired and comparable to the cytotoxic potential of C57BL/6 nu/nu cells. It is concluded that the ASGM1 + cytotoxic T cells are not primarily responsible for splenic chimerism and suppression of humoral immunity and that the two effects are likely caused by parental cells with a different phenotype and function.

The activity of gold sodium thiomalate (GST) given i.m. to adjuvant-induced polyarthritic rats was studied alone or in combination with active doses of aspirin, indomethacin and hydrocortisone. In addition to paw volume and body weight changes, erythrocyte sedimentation rate, serum albumin/globulin and gold levels as well as plasma activities of beta-glucuronidase, acid phosphatase, lysozyme and lactic acid dehydrogenase were measured. In prophylactic studies the beneficial activity of GST was unaffected by aspirin, suggesting a positive drug interaction, but additive with indomethacin or hydrocortisone for the 1st but not 2nd lesion of the disease. These results were closely correlated with increased serum gold levels. Similar clinical findings were observed in therapeutic studies except that a positive drug interaction occurred between GST and hydrocortisone. Unlike in the prophylactic experiments, serum gold levels were unaffected by any of the agents tested in the therapeutic studies.

B6D2F1 mice acutely reject parental C57Bl/6 bone marrow grafts, a phenomenon that is known as hybrid resistance. Injection of C57Bl/6 splenocytes into B6D2F1 recipients prior to bone marrow transplantation had previously been shown to facilitate growth of C57Bl/6 marrow grafts. We show in this report that for this effect to occur, radiation-sensitive T cells have to be present in the splenocyte inoculum. Loss of hybrid resistance following injection of C57Bl/6 splenocytes into B6D2F1 mice coincides with the appearance of T killer cells of C57Bl/6 origin that are specific for H-2 histocompatibility antigens of DBA/2. The parental T killer cells in unresponsive B6D2F1 mice express in vitro cytotoxic activity on H-2d targets and appear to be responsible for the acquired in vivo rejection of H-2d bone marrow grafts. Appearance of donor-derived T killer cells coincides with marked suppression of host immunity: lymphocytes from unresponsive B6D2F1 mice do not proliferate in mixed lymphocyte reactions and fail to respond to sheep erythrocytes in vitro, nor do they express natural killer (NK) activity. Concomitant with the suppression of NK activity, hybrid resistance to C57B1/6 marrow grafts disappears. This loss of resistance to bone marrow transplants is unspecific since third-party SJL marrow grafts are not rejected. It is concluded that suppression of hybrid resistance by injection of parental splenocytes into B6D2F1 mice is caused by a severe nonspecific suppression of host immune responsiveness by parental T cells that recognize disparate histocompatibility antigens in the host.

The effects of intraperitoneally injected detla9-tetrahydrocannabinol (THC), cannabinol (CBN) and cannabidiol (CBD) were compared to d-amphetamine sulfate (d-AMP) on food and water consumption and intake of two different concentrations of sucrose solutions. Three groups of rats were given the following dietary regimens within a 6-hr feed period day: 1 - water and dry food; 2 - water, dry food and five percent sucrose solution; 3 - water, dry food and 20% sucrose solution. Food and water consumption were dramatically reduced by each test drug at feeding periods immediately following and in some instances up to 4 days after dosing in all 3 groups. However, sucrose consumption was much less affected by each cannabinoid, inidcating a preference for sweet calories, whereas d-AMP had an equal anorexic action on both food and sucrose consumption. These data suggest for the first time in rats that a preference for sweet calories occurs during an overall anorexic effect of THC, CBN and CBD.

The susceptibility of female and male C57Bl/KsJ-mice to multiple low-dose streptozotocin applications was investigated. Both sexes of the strains studied developed hyperglycemia. In the early phase of streptozotocin-applications we failed to observe a strong parallelism between increase of plasma glucose and decrease of pancreatic insulin content.

The aqueous extract (marijuana tea) and volatile oil prepared from marijuana were compared with (−)-trans-∆9-te- trahydrocannabinol for their effect on hexobarbital sleeping time and analgesic action in mice. All three substances prolonged hexobarbital sleeping time with an order of potency of (−)-trans-∆9- tetrahydrocannabinol > aqueous extract > volatile oil. Each agent produced significant analgesic activity. However, the potencies of the aqueous extract and the volatile oil were similar to each other but only 1/200 that of (−)-trans-∆9-tetrahydrocannabinol.

The analgesic effectiveness of delta-9-tetrahydrocannabinol (THC), a crude marihuana extract (CME), cannabinol (CBN), cannabidiol (CBD), morphine SO-4 and aspirin following oral administration was directly compared in mice using the acetic-induced writhing and hot plate tests and the Randall-Selitto paw pressure test in rats. THC and morphine were equipotent in all tests except that morphine was significantly more potent in elevating pain threshold in the uninflamed rat hind paw. In terms of THC content, CME was nearly equipotent in the hot plate and Randall-Selitto tests, but was 3 times more potent in the acetic acid writhing test. On the other hand, CBN, like aspirin, was only effective in reducing writhing frequency in mice (3 times more potent than aspirin) and raising pain threshold of the inflamed hind paw of the rat (equipotent with aspirin). CBD did not display a significantly analgesic effect in any of the test systems used. The results of this investigation seem to suggest that both THC and CME possess narcotic-like analgesic activity similar to morphine, while CBN appears to be a non-narcotic type analgesic like aspirin.

Orally administered delta9-tetrahydrocannabinol (THC) produced a dose-dependent increase in urine output in hydrated rats similar in mg/kg potency and magnitude of effect to hydrochlorothiazide (HCT). Whereas HCT promoted marked excretion of Na+, K+ and Cl- and an increase in the urinary Na+/K+ at all diuretic doses (1.25-20.0 mg/kg), THC had only a slight effect on Na+ and K+ excretion but not Cl- even after the highest dose tested (20.0 mg/kg). Hypophysectomy and adrenalectomy abolished the diuretic effect of THC, thus suggesting both central and peripheral sites of action for the diuretic effect of THC. Tolerance to the effect on urine output by THC developed after 15 days of repeated dosing, while urine output and electrolyte excretion remained significantly elevated after 25 days of HCT administration.

The behavioral consequences of beta-adrenoceptor subsensitivity were investigated by determining whether a physiological response that is mediated by beta-receptors, isoproterenol-induced drinking (IID), would be reduced by subacute antidepressant/alpha 2-antagonist treatment. The coadministration of typical (e.g., imipramine) or atypical (e.g., mianserin) antidepressants with yohimbine or piperoxan twice daily for four consecutive days reduced IID. Both the time course as well as the magnitude of beta-adrenoceptor subsensitivity could be behaviorally demonstrated. In addition, the reduction in IID observed after coadministration of imipramine with yohimbine was a centrally mediated effect since it was observed after systemic (subcutaneous) and central (intraventricular) administration of isoproterenol. These results provide evidence that IID is an appropriate behavioral model to demonstrate beta-adrenoceptor subsensitivity following subacute antidepressant/alpha 2-antagonist treatment.