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Application of immobilized pH gradient isoelectric focusing to forensic hemogenetics: a survey on a three year experience with the transferrin (TF) and alpha 1-antitrypsin (PI) systems.

The subtypes of transferrin (TF) and alpha 1-antitrypsin (PI), first discovered using isoelectric focusing, are now mostly determined in immobilized pH gradient gels. We report on our experience in the parentage expertise with both polymorphisms over a period of three years. The complexity of the technology was compensated by the fact that most subtypes of TF and PI could be more reliably recognized. The PI alleles PI*M1, M2, M3, S, F, T, and Z and TF alleles TF*C1, C2 and C3, and in addition four further rare TF alleles were observed. The allele frequencies from non-related individuals did not deviate from the Hardy-Weinberg equilibria and corresponded well to known frequencies from West Germany and other Caucasoid populations. With the TF system 36 accused men, and with the PI system 54 were excluded from paternity from a total of 344 (TF) respectively 347 (PI) cases. From the data presented here isoelectric focusing in immobilized pH gradient gels appears to be a major improvement over carrier ampholyte generated pH gradients in the distinction of TF and PI phenotypes.

Alleles↗

Multipoint affected sibpair linkage methods for localizing susceptibility genes of complex diseases.

Recently, Liang et al. ([2001] Hum. Hered. 51:64-78) proposed a general multipoint linkage method for estimating the chromosomal position of a putative susceptibility locus. Their technique is computationally simple and does not require specification of penetrance or a mode of inheritance. In complex genetic diseases, covariate data may be available which reflect etiologic or locus heterogeneity. We developed approaches to incorporating covariates into the method of Liang et al. ([2001] Hum. Hered. 51:64-78) with particular attention to exploiting age-at-onset information. The results of simulation studies, and a worked data example using a family data set ascertained through probands with schizophrenia, suggest that utilizing covariate information can yield substantial efficiency gains in localizing susceptibility genes.

Age of Onset↗

Availability of schizophrenic patients and their families for genetic linkage studies: findings from the Maryland epidemiology sample.

It has been suggested that collections of affected sib pairs, or their nuclear families, may be an efficient method for screening for genetic linkages in schizophrenia. We present the data collected in five years from 15 hospitals in the state of Maryland in an effort to determine if such a collection scheme will be feasible. Probands in our sample were eligible for inclusion in the sample if they were white, were age 16 years or older, and carried a research diagnosis of schizophrenia. Family data are reported for 258 probands. Using the most stringent category of affected (RDC schizophrenia) revealed ten families with two or more affected sibs. The broadest category of affected (any psychotic disorder or psychiatric hospitalization) identified only 36 families with two or more affected sibs. We conclude that, if schizophrenia is a heterogeneous disorder with decreased penetrance, an effort to collect multiplex nuclear families is unlikely to provide enough data to identify genetic linkage. Alternatively, an effort to seek out and collect larger multiplex, multigenerational families rather than a collection of affected sib pairs may be more efficacious.

Adolescent↗

Estimating effects of proband characteristics on familial risk: II. The association between age at onset and familial risk in the Maryland schizophrenia sample.

In this report we apply methods outlined in the companion paper [Liang, Genet Epidemiol 8:329-338, 1991] to study the association between proband age at onset and familial risk among first-degree relatives of 374 schizophrenic probands. The analyses take into consideration the potential problems of censoring and correlation of age at onset within families. All analyses were done by gender of the proband; age at onset was dichotomized. The results of the analyses of the male probands suggest that there is an increased risk of schizophrenia among the relatives of male probands who have an onset prior to age 17 when compared to relatives of male probands who have an onset later than 16. We did not find an association between age at onset and familial risk among the female probands, but this may be due to the smaller number of female probands and the lower power associated with the analyses.

Adolescent↗

Extended latent class approach to the study of familial/sporadic forms of a disease: its application to the study of the heterogeneity of schizophrenia.

When no method exists for detecting genetic forms of a disorder, epidemiologists classify probands according to the presence or absence of an affected relative (familial or sporadic). Not only is this a surrogate measure but if the risk for the disorder is associated with characteristics such as age and gender, then probands with varied distributions of these characteristics among their relatives are subject to misclassification. A latent class approach is presented which explicitly models the relationship between the affected status of the relatives and the unobservable familial/sporadic status of the proband in order to adjust for these characteristics. Lastly, an approach is introduced to correct for attenuation in measures of association between familial/sporadic status and other variables that could result if probands are misclassified. This approach incorporates the latent class probabilities directly into the regression model without classifying probands. These methods are applied to a study of the heterogeneity of schizophrenia.

Adolescent↗

Serotonin in the developing stomatogastric system of the lobster, Homarus americanus.

We studied the development of the serotonergic modulation of the stomatogastric nervous system of the lobster, Homarus americanus. Although the stomatogastric ganglion (STG) is present early in embryonic development, serotonin immunoreactivity is not visible in the STG until the second larval stage. However, incubation of the STG with exogenous serotonin showed that a serotonin transporter is present in embryonic and early larval stages. Serotonin uptake was blocked by paroxetine and 0% Na(+) saline. The presence of a serotonin transporter in the embryonic STG suggests that hormonally liberated serotonin could be taken up by the STG, and potentially released as a "borrowed transmitter". Consistent with a potential hormonal role, serotonin is found in the pericardial organs, a major neurosecretory structure, by midembryonic development. The rhythmic motor patterns produced by embryonic and larval STGs were decreased in frequency by serotonin. Lateral Pyloric (LP) neuron-evoked excitatory junctional potentials (EJPs) in the embryos and the first larval stage (LI) were larger, slower, and more variable than those in the adult. The amplitude of adult LP neuron-evoked EJPs was increased more than twofold in serotonin, but in embryos and LI preparations this effect was negligible. In embryos and LI preparations, serotonin increased the occurrence of muscle fiber action potentials and altered the EJP wave-form. These data demonstrate that serotonin receptors are present in the stomatogastric nervous system early in development, and suggest that the role of serotonin changes from modulation of muscle fiber excitability early in development to enhancement of neurally evoked EJPs in the adult.

Age Factors↗

Constant amplitude of postsynaptic responses for single presynaptic action potentials but not bursting input during growth of an identified neuromuscular junction in the lobster, Homarus americanus.

As lobsters grow from early juveniles to adults their body size increases more than 20-fold, raising the question of how function is maintained during these ongoing changes in size. To address this question we studied the pyloric 1 (p1) muscle of the stomach of the lobster, Homarus americanus. The p1 muscle receives multiterminal innervation from one motor neuron, the lateral pyloric neuron of the stomatogastric ganglion. Staining with antibodies raised against synaptotagmin showed that as the muscle fibers increased in length, the spacing between the terminal innervation increased proportionally, so the number of synaptic contact regions/muscle fiber did not change. Muscle fibers were electrically coupled in both juveniles and adults. The amplitude of single intracellularly recorded excitatory junctional potentials evoked by motor nerve stimulation was the same in both juveniles and adults. Nonetheless, the peak depolarizations reached in response to ongoing pyloric rhythm activity or in response to high-frequency trains of stimuli similar to those produced during the pyloric rhythm were approximately twofold larger in juveniles than in adults. This suggests that homeostatic regulation of synaptic connections may operate at the level of the amplitude of the single synaptic potential rather than on the summed depolarization evoked during strong rhythmic activity.

Action Potentials↗

A Pilot Study of Mood in Epilepsy Patients Treated with Vagus Nerve Stimulation.

Context. Antiepileptic drugs (AEDs) are frequently used for their beneficial mood effects.Objective. We sought to determine if there was a quantifiable effect on mood of the vagus nerve stimulator (VNS) when used as an antiseizure treatment.Design. Mood was assessed before and 3 months after VNS implantation in adult epilepsy patients. A group of adult epilepsy patients on stable AED regimens were used as a comparison group. AED regimens were unchanged during the study. The change in mood scale scores across time was assessed by t test (intragroup) and two-factor repeated-measures ANOVA (intergroup).Setting. An epilepsy center in a university hospital was the setting.Subjects. Twenty consecutive adult epilepsy patients undergoing VNS implantation to improve seizure control and twenty adult seizure patients with no intervention were enrolled.Main outcome measures. The mood scales used were the Cornell Dysthymia Rating Scale (CDRS) and the Hamilton Depression (Ham-D), Hamilton Rating Scale for Anxiety (Ham-A), and Beck Depression Inventory (BDI) scales.Results. The VNS group showed a significant decrease in mood scale scores across time (t test CDRS P = 0.001, Ham-D P = 0.017, BDI P = 0.045), indicating a decrease in depressive symptoms. The Ham-A scores in the VNS group and the comparison group scores did not significantly change across time. There were no significant differences between groups across time, although the BDI approached significance at P = 0.07. The VNS group had a significant decrease in seizure frequency compared with the comparison group (P = 0.01). There was no difference in mood scales over time between the VNS treatment responders (defined by >50% decrease in seizure frequency) and nonresponders, suggesting dissociation between seizure frequency reduction and mood change.Conclusion. VNS treatment is associated with mood improvement as measured by multiple scales, but differences in mood scale scores over time between the VNS and a comparison group were not found.

Journal Article↗

Behavior of lymphocyte subsets and expression of activation markers in response to immunotherapy with galactoside-specific lectin from mistletoe in breast cancer patients.

Cellular aspects of the immunomodulating activity of the galactoside-specific lectin from mistletoe (ML-1) were investigated in 10 cancer patients. Regular subcutaneous injections (4 weeks) of the optimal dosis of ML-1 (1 ng per kg body weight, twice a week) yielded notable increases in the apparent numbers of certain lymphocyte subsets [pan T cells; helper T cells; natural killer (NK) cells] which are generally believed to be involved in antitumor immunity. Moreover, ML-1 administration resulted in an increased level of expression of interleukin (IL)2 receptors on lymphatic cells, an indicator of cellular activation. In vitro, the exposure of human lymphocytes to ML-1 resulted in an enhanced expression of receptors for IL-2 (T cells) and HLA-DQ (B cells), which similarly substantiated the capacity of ML-1 to affect immunological parameters within the host defense system. Thorough clinical trials are now required to assess any impact of the application of the lectin on the course of the disease.

Biomarkers↗

The role of hepatic lectins and the activity of the mononuclear phagocyte system in systemic Listeria monocytogenes infection in Balb/c mice.

Hepatic lectin blocking experiments with D-galactose in Balb/c mice showed that parenchymal liver cells are obviously not involved in Listeria monocytogenes infection (strain SLCC 4013, 5 X 10(6) cells i.v.). Using the bacterial immunomodifier Propionibacterium avidum KP-40 the importance of an activated mononuclear phagocyte system in the early stage of Listeria infection could be demonstrated.

Adjuvants, Immunologic↗

Immunostimulating cell surface substance from Staphylococcus epidermidis strain ATCC-31432 prevents metastatic lung colonization in Balb/c-mice.

The antineoplastic activity of cell surface substance from Staphylococcus epidermidis ATCC-31432 (CSS-subfraction 2) in mice and its influence on the activation of human leucocytes was studied. After in vitro incubation with CSS-subfraction 2, human polymorphonuclear leucocytes were evidently stimulated. In vivo (Balb/c-mice) intraperitoneal application of subfraction 2 of CSS induced a considerable splenomegaly and greatly increased IgM levels indicating a strong immuno-stimulating activity. In order to evaluate antineoplastic effects of CSS-subfraction 2, we used sarcoma L-1 cells (Balb/c-mouse origin) which cause heavy tumor colonization of the lung. After single systemic injection of subfraction 2 of CSS, the number of lung tumor-cell colonies drastically decreased. The combination of this immunomodulating therapy with a temporary anticoagulation resulted in a further reduction of tumor colonies in the lungs of Balb/c-mice.

Adjuvants, Immunologic↗

Immunochemotherapy of breast cancer with Propionibacterium granulosum.

Eighty-two patients with metastatic breast cancer were treated with adriamycin, cyclophosphamide, and 5-fluorouracil (FAC). These patients were divided by randomization into two groups. One was receiving cytostatic drugs only, while another received the identical cytostatic medication plus additional therapy consisting of IV infusions of gamma-radiation-sterilized cells of Propionibacterium granulosum strain KP-45. Metastases were localized in six different sites and/or organs. Multiple parameters for monitoring changes in metastatic lesions and laboratory tests were performed serially in all patients. The addition of P. granulosum to standard (FAC) chemotherapy of metastatic breast cancer influenced metastases in the liver. It also resulted in increased survival time within the observation period. Possible mechanisms of immunomodulating activity of P. granulosum in this disease are discussed.

Adult↗

The influence of killed Propionibacterium granulosum on experimental infection with Escherichia coli.

Mice infected subcutaneously with 5 X 10(3) viable cells of Escherichia coli ATCC 25922 incorporated in liquified agar developed a systemic infection. Increasing bacterial numbers could be recovered from the liver at several days following infection. Ultimately, the animals died 6 days after infection. Treatment of mice with 1 mg killed Propionibacterium granulosum KP-45 lead to an increased spleen weight at day 7 after intraperitoneal injection. Hence, the animals were highly susceptible to the lethal action of endotoxin. They were, however, markedly protected against infection with E. coli, since definitely lower bacterial counts were found in the liver of pretreated mice in comparison to controls.

Animals↗

Inhibition of Lewis lung carcinoma in mice by local microwave hyperthermia combined with immunomodulating Propionibacterium granulosum KP-45.

C-57 BL/6 mice with Lewis lung carcinoma were treated 2 weeks after tumor implantation with local microwave hyperthermia (2450 MHz, tumor temperature 43.5 degrees C, 30 min) and/or intratumoral or intraperitoneal injection of 1 mg cell walls of Propionibacterium granulosum KP-45. Tumor growth, survival time of the animals, and the delayed skin hypersensitivity to oxazolone were followed up, as well as the 3H-thymidine uptake of tumorous tissues and the number of lung metastases. The combined treatment of microwave hyperthermia with immunomodulating P. granulosum KP-45 resulted in significantly stronger inhibition of tumor growth than with each of these methods alone. The number of lung metastases could be significantly lowered, and the skin reactivity to oxazolone remained enhanced during the whole observation period (over 70 days after tumor implantation). The implications of the test observation are discussed.

Animals↗

Effects of cefotaxime, clindamycin, mezlocillin, and piperacillin on mouse sarcoma L-1 tumor.

For the study described in the paper, the effects of 10 days' chemotherapy with cefotaxime, clindamycin, mezlocillin, and piperacillin on local tumor growth and on spontaneous or artificial metastatic spread into the lungs were studied. For the animal tumor model Balb/c mice and the mouse sarcoma L-1 tumor were used. Chemotherapy was administered before, immediately after, or some time after the injection of tumor cells. The antibiotic dosage given to mice was calculated on a body weight basis from the doses recommended for humans. Cefotaxime and clindamycin did not influence the animal tumor model, whereas mezlocillin and piperacillin showed positive or negative effects depending on the chemotherapy schedule. In vitro none of the four antibiotics caused cytotoxic activity in cell cultures of mouse sarcoma L-1, human lung cancer E-14, or human malignant melanoma MEW.

Animals↗

The role of capsule as a barrier to bacteriophage adsorption in an encapsulated Staphylococcus simulans strain.

The polyvalent staphylococcal bacteriophage U16 failed to adsorb to an encapsulated Staphylococcus simulans strain. Partially purified cell wall and teichoic acid of this strain could, however, inactivate bacteriophage U16 to a great extent, indicating the presence of the phage receptor. It is concluded that the capsule of Staphylococcus simulans acts as a barrier for the interaction of the phage with its receptor in the bacterial cell wall.

Abscess↗