Childhood cerebral X-linked adrenoleukodystrophy with atypical neuroimaging abnormalities and a novel mutation.

Childhood cerebral X-linked adrenoleukodystrophy (XALD) typically manifests with symptoms of adrenocortical insufficiency and a variety of neurocognitive and behavioral abnormalities. A major diagnostic clue is the characteristic neuroinflammatory parieto-occipital white matter lesions on magnetic resonance imaging. This study reports a 5-year 10-month old boy presenting with generalized skin hyperpigmentation since 3 years of age. Over the past 9 months, he had developed right-sided hemiparesis and speech and behavioral abnormalities, which had progressed over 5 months to bilateral hemiparesis. Retrospective analyses of serial brain magnetic resonance images revealed an unusual pattern of lesions involving the internal capsules, corticospinal tracts in the midbrain and brainstem, and cerebellar white matter. The clinical diagnosis of childhood cerebral adrenoleukodystrophy was confirmed by elevated basal levels of adrenocorticotropin hormone and plasma very long chain fatty acid levels. Additionally, sequencing of the ABCD1 gene revealed a novel mutation. The only specific palliative therapy that could be offered after diagnosis was dietary intervention. The patient died within 16 months of onset of neurological symptoms. Awareness that childhood cerebral XALD can present with atypical neuroimaging patterns early in its course may aid diagnosis at a stage when definitive treatment can be attempted and timely genetic counseling be offered to the family.

Transferrin-receptor-mediated iron accumulation controls proliferation and glutamate release in glioma cells.

Transferrin receptors (TfR) are overexpressed in brain tumors, but the pathological relevance has not been fully explored. Here, we show that TfR is an important downstream effector of ets transcription factors that promotes glioma proliferation and increases glioma-evoked neuronal death. TfR mediates iron accumulation and reactive oxygen formation and thereby enhanced proliferation in clonal human glioma lines, as shown by the following experiments: (1) downregulating TfR expression reduced proliferation in vitro and in vivo; (2) forced TfR expression in low-grade glioma accelerated proliferation to the level of high-grade glioma; (3) iron and oxidant chelators attenuated tumor proliferation in vitro and tumor size in vivo. TfR-induced oxidant accumulation modified cellular signaling by inactivating a protein tyrosine phosphatase (low-molecular-weight protein tyrosine phosphatase), activating mitogen-activated protein kinase and Akt and by inactivating p21/cdkn1a and pRB. Inactivation of these cell cycle regulators facilitated S-phase entry. Besides its effect on proliferation, TfR also boosted glutamate release, which caused N-methyl-D-aspartate-receptor-mediated reduction of neuron cell mass. Our results indicate that TfR promotes glioma progression by two mechanisms, an increase in proliferation rate and glutamate production, the latter mechanism providing space for the progressing tumor mass.

[Sonographic diagnosis of parathyroid carcinoma]. / Sonographische Diagnostik eines Nebenschilddrüsenkarzinoms.

Prognosis and survival of patients suffering from parathyroid carcinoma are completely dependent on correct surgical therapy in form of an en-bloc resection of the tumour. A correct preoperative diagnosis of this rare carcinoma is therefore absolutely mandatory in order to perform a curative operation. We demonstrate a case of parathyroid carcinoma in which scintigraphy and CT diagnostics did not produce the right diagnosis. Preoperative high-resolution ultrasound (8 MHz) in combination with power-Doppler-sonography, however, led to the right diagnosis by demonstrating the correct topography, signs of malignancy and the feeding vessels of the tumour. As it could be demonstrated that the inferior thyroid artery was displaced by the tumour and was not a tumour feeding artery, the inferior thyroid artery was an important additional landmark for making the decision between thyroid or parathyroid carcinoma. The use of power-Doppler-sonography for identification of feeding arteries associated with parathyroid carcinoma in addition to high-resolution small parts sonography is of great value for the distinction between thyroid or parathyroid tumour.

Development of symbionts in triatomine bugs and the effects of infections with trypanosomatids.

In the intestinal tract of fifth instars of the hematophagous reduviid bugs Rhodnius prolixus and Triatoma infestans blood ingestion induced an initial decrease of the concentration of the respective symbiotic bacteria Rhodococcus rhodnii and Nocardia sp. and then within 10 days a 15- or 18-fold increase of the total population/bug to about 0.8 x 10(9) colony-forming units in R. prolixus and 1.8 x 10(9) colony-forming units in T. infestans. About 95-99% of the total populations of both symbionts developed in the anterior midgut regions, i.e., cardia and stomach. The passage from the blood-storing stomach to the digesting small intestine caused a considerable breakdown of symbiont populations, and only about 0.01% of the total population was present in the rectum. These were excreted mainly within 4 h after a blood meal. After infection with three species of trypanosomatids, R. rhodnii, the symbiont of R. prolixus, was affected by Trypanosoma rangeli, but not by Blastocrithidia triatomae or Trypanosoma cruzi. On the other hand, in T. infestans the concentration of Nocardia sp. was reduced after infection with B. triatomae, but not by T. rangeli nor T. cruzi. In long-term B. triatomae-infected T. infestans, this reduction and a reduced diuretic activity after feeding synergistically lowered the symbiont concentration in the singly deposited feces/urine drops drastically compared to uninfected controls. These data strongly support the theory of the mechanisms of pathogenicity of T. rangeli and B. triatomae for R. prolixus and T. infestans, respectively, that the primal point of attack is the host-specific symbiont, R. rhodnii and Nocardia sp., respectively.

Bilateral human fetal striatal transplantation in Huntington's disease.

BACKGROUND: Transplanted striatal cells have been demonstrated to survive, grow, establish afferent and efferent connections, and improve behavioral signs in animal models of Huntington's disease (HD). OBJECTIVE: To evaluate feasibility and safety and to provide preliminary information regarding the efficacy of bilateral human fetal striatal transplantation in HD. METHODS: Seven symptomatic patients with genetically confirmed HD underwent bilateral stereotactic transplantation of two to eight fetal striata per side in two staged procedures. Tissue was dissected from the lateral half of the lateral ventricular eminence of donors 8 to 9 weeks postconception. Subjects received cyclosporine for 6 months. RESULTS: Three subjects developed subdural hemorrhages (SDHs) and two required surgical drainage. One subject died 18 months after surgery from probable cardiac arrhythmia secondary to severe atherosclerotic cardiac disease. Autopsy demonstrated clearly demarcated grafts of typical developing striatal morphology, with host-derived dopaminergic fibers extending into the grafts and no evidence of immune rejection. Other adverse events were generally mild and transient. Mean Unified HD Rating Scale (UHDRS) motor scores were 32.9 plus minus 6.2 at baseline and 29.7 plus minus 7.5 12 months after surgery (p = 0.24). Post-hoc analysis, excluding one subject who experienced cognitive and motor deterioration after the development of symptomatic bilateral SDHs, found that UHDRS motor scores were 33.8 plus minus 6.2 at baseline and 27.5 plus minus 5.2 at 12 months (p = 0.03). CONCLUSIONS: Transplantation of human fetal striatal cells is feasible and survival of transplanted cells was demonstrated. Patients with moderately advanced HD are at risk for SDH after transplantation surgery.

Software based digital signal processing and spectrum deconvolution in X-ray spectroscopy.

Approaches for software based digital signal processing and numerical deconvolution of measured signals which overcome limitations of state-of-the-art systems are described. The basic technical equipment for digital signal processing consists of an energy resolving detector with a preamplifier followed by a fast sampling analogue-to-digital converter (ADC). The main idea is the numerical decomposition of the measured signal into contributions caused by single photon absorption using standard pulses. The latter can be obtained by measurements under definite conditions. The maximum pulse rate is then limited only by the ratio of sampling time to the time between two pulses which should be attributed to single events. Thus pulse overlaps do not require pulse rejection. At sampling rates of 10(8) samples per second theoretically a comparable photon rate can be detected at throughputs of 100%. Beyond that it is outlined that in a comparable manner a numerical deconvolution of measured energy spectra (statistic distribution functions of single events) into combinations of standard spectra, which can likewise be determined by measurement, offers outstanding possibilities, too. On the one hand the energy resolution attainable for individual events for a given detector can be improved drastically by the statistical treatment of spectra. On the other hand an energy resolving work principle becomes possible for certain detectors, which do not permit this conventionally due to their poor signal to noise ratio.

Bacterial symbiosis and paratransgenic control of vector-borne Chagas disease.

The triatomine vectors of Chagas disease are obligate haematophagous insects, feeding on vertebrate blood throughout their entire developmental cycle. As a result of obtaining their nutrition from a single food source, their diet is devoid of certain vitamins and nutrients. Consequently, these insects harbour populations of bacterial symbionts within their intestinal tract, which provide the required nutrients that are lacking from their diet. We have isolated and characterised symbiont cultures from various triatomine species and developed a method for genetically transforming them. We can then reintroduce them into their original host species, thereby producing stable paratransgenic insects in which we are able to express heterologous gene products. Using this methodology, we have generated paratransgenic Rhodnius prolixus that are refractory for infection with Trypanosoma cruzi. Two examples of potentially refractory genes are currently being expressed in paratransgenic insects. These include the insect immune peptide cecropin A and active single chain antibody fragments. We have also developed an approach that would allow introduction of genetically modified bacterial symbionts into natural populations of Chagas disease vectors. This approach utilises the coprophagic behaviour of these insects, which is the way in which the symbionts are transmitted among bug populations in nature. The production and ultimate release of transgenic or paratransgenic insects for public health applications is potentially very promising but also worthy of much careful consideration with respect to environmental, political, and human safety concerns.

[Content of minerals (Na,K,Ca,Mg) and of trace elements (Fe,Cu and Zn) in 13 different tissues of fallow deer (Dama dama L.) of various ages]. / Der Gehalt an Mineralstoffen (Na, K, Ca, Mg) und Spurenelementen (Fe, Cu, Zn) in 13 verschiedenen Geweben vom Damwild (Dama dama L.) unterschiedlichen Alters.

At male deer in the age of 15 and 27 months, at experimentally induced cryptorchids and at castrated animals analyses of the content of Na, K, Ca, Mg, Fe, Cu and Zn in 13 different tissues were performed. In the liver of newborn animals solely the content of Fe, Cu and Zn was determined. The significance of the content of the mentioned elements in the various tissues is described. In the liver of the newborn animals stores of Fe, Cu an Zn were identified. The values can be used as a basis for analyses at deer with a growth inhibition caused by a lack of sodium chloride Fe, Cu and Zn.

A recombinant virus assay using full-length envelope sequences to detect changes in HIV-1 co-receptor usage.

The clinical management of HIV-1 infection has benefited enormously from molecular characterization of drug resistance as well as determination of the viral phenotype in vitro. HIV-1 infected individuals on HAART are currently monitored for the development of drug resistance variants allowing clinicians to redesign drug regimens. An understanding of the molecular basis of the evolution of drug resistance in vivo allows the improvement of the drugs as well as in vitro evaluation of new antiviral compounds alone or in combination with those currently approved. New findings suggest that viral envelopes could be a target to inhibit infection and replication. Therefore the generation of a recombinant virus assay (RVA) to allow the phenotypic determination of drug resistance against entry inhibitors (EI) is anticipated. We constructed an env-deleted clone of HIV-1 using the molecular clone NL-4.3. PCR amplified complete envelope genes (NL-4.3, BaL, primary envelope-genes) were ligated in vitro with a deletion clone (pNL-deltaK) and PM1-cells, supporting the replication of R5- and X4-tropic viruses, were transfected. Determination of co-receptor usage of the harvested recombinant virus-swarm revealed no difference compared to the molecular clones derived individually from three different patients. These results clearly show that an envelope-based RVA is practicable to monitor HIV-co-receptor usage at a given time point. Furthermore, this assay will allow to monitor resistance development against existing and future entry inhibitors and will aid to improve the management of HIV-therapy.

The effects of aposymbiosis and of an infection with Blastocrithidia triatomae (Trypanosomatidae) on the tracheal system of the reduviid bugs Rhodnius prolixus and Triatoma infestans.

We have investigated the effects of the absence of symbionts and of infection with a trypanosome on the tracheal supply to different organs in larvae of the blood-sucking bugs, Rhodnius prolixus and Triatoma infestans. In bugs grown without symbionts there were extensive reductions in the tracheal supply to all the internal organs examined. These bugs also excreted less fluid after blood meals, perhaps because of a reduction in the oxygen supply to the Malpighian tubules. Inclusion of vitamin B in the diet of these affected insects reversed both the adverse effects on diuresis and on the extent of the respiratory supply to the different internal organs. The results of these studies suggest that vitamin B may play a key role in the development and maintenance of an adequate tracheal supply to the tissues in these insects. Infection with the trypanosome Blastocrithidia triatomae also greatly reduced the density of tracheoles supplying the rectum, small intestine and Malpighian tubules in infected T. infestans, but not in R. prolixus. It is possible, therefore, that the parasite exerts at least part of its pathogenic effect by causing a vitamin B deficiency, that in turn affects oxygen supply to the tissues.

Effect of once daily milking and concurrent somatotropin on mammary tight junction permeability and yield of cows.

Six pairs of monozygous Friesian twin cows during late lactation were used to assess the effect of once daily milking and bST treatment on yields and tight junction permeability of mammary epithelial cells. During the first 7 d, all cows were milked twice daily; on d 8 through d 21, all cows were milked once daily, but one cow of each twin pair was treated daily with 20 mg of bST on d 13 through 21; and, finally, during d 22 through 28, all cows were again milked twice daily. Once daily milking, a common management practice in New Zealand, resulted in a small (7%) but significant decrease in milk yield. Treatment with bST increased milk yield by 19%, thereby exceeding the milk yield loss from once daily milking. The integrity of mammary tight junctions was assessed indirectly by measuring concentrations of plasma lactose and milk BSA. Once daily milking temporarily disrupted tight junction integrity, based on a 4- to 5-fold increase in plasma lactose and a 42 to 55% increase in the concentrations of milk BSA. In the present study, bST did not affect the permeability of mammary tight junctions.

The relationship between cardiovascular reactivity and heartbeat detection.

This experiment tested the hypothesis that inotropic cardiovascular reactivity to stress is related to performance on heartbeat discrimination tasks. The experiment also compared the efficacy of a specific modification of two popular heartbeat discrimination paradigms, Whitehead's and Katkin's. Subjects were 48 male undergraduates who performed both discrimination tasks and then were subjected to mental arithmetic stress. Results indicated that high cardiovascular reactors were better detectors than low reactors. Results also indicated that subjects performed better on the modified Whitehead task than on the modified Katkin task.

Effect of colostrum intake on alpha-lactalbumin concentrations in serum of calves.

Seven Friesian calves were fed colostrum for four days beginning within 24 hours of birth, and milk thereafter. The concentration of alpha-lactalbumin in serum was measured by specific radioimmunoassay and compared to IgG assayed by electroimmunodiffusion. Serum concentrations of alpha-lactalbumin peaked at 387 +/- 85 ng ml-1 within eight hours of initial intake of colostrum, declining to 12 +/- 3 ng ml-1 by day 6. IgG rose steadily to 17 mg ml-1 by 48 hours of birth and remained relatively constant thereafter. The temporal pattern of alpha-lactalbumin in serum following colostrum intake confirms previous studies suggesting reduced absorption of colostral proteins between 24 and 36 hours. The presence of variable amounts of alpha-lactalbumin in serum even after 17 days, however, indicates limited transfer of milk-derived proteins across the gut at this time. The data further show that cessation of maximal gut transfer does not relate to molecular weight of transferred protein.