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T Boehm

Publications and source records attributed to T Boehm.

At least 37 records · Page 2Linked to original sources
PubMed2001

A point mutation in the IL-12R beta 2 gene underlies the IL-12 unresponsiveness of Lps-defective C57BL/10ScCr mice.

Lps-defective C57BL/10ScCr (Cr) mice are homozygous for a deletion encompassing Toll-like receptor 4 that makes them refractory to the biological activity of LPS. In addition, these mice exhibit an inherited IL-12 unresponsiveness resulting in impaired IFN-gamma responses to different microorganisms. By positional cloning methods, we show here that this second defect of Cr mice is due to a mutation in a single gene located on mouse chromosome 6, in close proximity to the Igkappa locus. The gene is IL-12Rbeta2. Cr mice carry a point mutation creating a stop codon that is predicted to cause premature termination of the translated IL-12Rbeta2 after a lysine residue at position 777. The truncated beta2 chain can still form a heterodimeric IL-12R that allows phosphorylation of Janus kinase 2, but, unlike the wild-type IL-12R, can no longer mediate phosphorylation of STAT4. Because the phosphorylation of STAT4 is a prerequisite for the IL-12-mediated induction of IFN-gamma, its absence in Cr mice is responsible for their defective IFN-gamma response to microorganisms.

Animals↗
PubMed2001

Laser capture microdissection-based expression profiling identifies PD1-ligand as a target of the nude locus gene product.

T cell development requires the interaction of developing thymocytes with thymic epithelial cells. Thymic epithelial cells acquire their unique phenotype under the control of the winged-helix transcription factor Whn, which is lacking in the nude mouse. Whn-dependent genes may therefore be important regulators of lympho-epithelial interactions. To identify Whn target genes we isolated RNA populations of wild-type and nude thymic anlagen from embryonic day 12.5 embryos by laser capture microdissection and compared them by gene expression profiling on microarrays representing 22,000 transcripts. All cDNA with expression differences confirmed by quantitative RT-PCR using RNA from individual anlagen and by in situ hybridization were found to be present in wild-type and absent in nude samples. Three of eight confirmed transcripts were of hematopoietic origin; these transcripts emanate from hematopoietic precursors which have just entered the thymic anlage. Five transcripts were of epithelial origin; one of these corresponds to the recently identified PD-1 ligand (PD-L1), the receptor of which is known to modulate positive selection and to play a role in the control of autoimmunity, and the remaining transcripts code for novel genes. The presented results support our prediction that this systematic approach by gene expression profiling yields regulators of thymopoiesis.

Algorithms↗
PubMed2001

Abnormal bone marrow stroma in mice deficient for nemo-like kinase, Nlk.

The stromal compartment of the bone marrow is composed of various cell types that provide trophic and instructive signals for hematopoiesis. The mesenchymal stem cell is believed to give rise to all major cellular components of the bone marrow microenvironment. Nemo-like kinase, Nlk, is a serine-threonine kinase that connects MAP kinase and Wnt signaling pathways; its in vivo function in mouse is unknown. We have generated mice with a targeted disruption of Nlk and find that the complex phenotype significantly varies with the genetic background. Whereas C57BL/6 mice lacking Nlk die during the third trimester of pregnancy, the 129/Sv background supports survival into adolescence; such mice are growth retarded and suffer from various neurological abnormalities. We show here that the Nlk deficiency syndrome includes aberrant differentiation of bone marrow stromal cells. Varying degrees of morphological abnormality, such as increased numbers of adipocytes, large blood sinuses and absence of bone-lining cells are observed in the bone marrow of mutant mice. Nlk deficient mice thus provide a novel model to study the genetic requirements for bone marrow stromal differentiation.

Animals↗
PubMed2001

Expression domains in the skin of genes affected by the nude mutation and identified by gene expression profiling.

Although several genes affected by the nude mutation inactivating the transcription factor Whn have recently been identified, a comprehensive molecular analysis of the nude phenotype is still missing. Gene expression profiling of wildtype and nude mice back skin reveals several so far unknown differences in mRNA levels and demonstrates that microarray hybridization is ideal to identify even quantitative changes in expression. Some genes are upregulated in the absence of Whn. Most of the differentially expressed genes are downregulated in nude skin. Our results identify metallothionein IV among these genes. This is the first report on metallothionein IV expression in the murine hair follicle; its expression domain almost completely overlaps that of Whn.

Animals↗
PubMed2001

Vacuum-assisted resection of malignant tumors with and without subsequent radiofrequency ablation: feasibility of complete tumor treatment tested in an animal model.

PURPOSE: To evaluate the feasibility of vacuum-assisted tumor excision with and without RF ablation for the minimally invasive treatment of small tumors. MATERIALS AND METHODS: Twenty VX2 tumors were implanted bilaterally into the spine muscle of 10 rabbits. Tumor excision was performed after tumor sizes reached 10 mm (12-27 d incubation) with use of a vacuum-assisted biopsy device. Three or four directed vacuum-assisted biopsies were performed in angle steps of 30 degrees. In 10 tumors, ultrasound (US)-guided radiofrequency (RF) ablation (8 min, 60 W) was subsequently performed with use of a cooled-tip electrode system. Follow-up US was performed at 3-4-day intervals for as long as 3 weeks after excision/RF ablation. Autopsy and histopathologic analysis were performed. RESULTS: The duration of vacuum excision ranged from 12 to 45 minutes (25 min +/- 7). Histologically tumor-free margins in the outer round of the core biopsy specimens were found in only four of 20 cases (20%). Maximum lesion sizes during RF ablation ranged from 18 to 25 mm (20 mm +/- 2.6). Histologic examination of the excision specimens documented tumor-free margins in only three tumors (30%) among the excision-only group and only one (10%) among the combined excision/ablation group. Local recurrences occurred in eight of 10 cases (80%) after vacuum excision alone, whereas recurrence after combined excision and RF ablation occurred only in two of 10 cases (20%; P <.05). CONCLUSIONS: Local tumor resection with use of vacuum-assisted biopsy is feasible and promising as a minimally invasive therapy for the treatment of small focal breast neoplasms. Combined excision and RF ablation techniques may reduce the rate of local recurrence considerably.

Animals↗
PubMed2001

Differentiation of mammographically suspicious lesions: evaluation of breast ultrasound, MRI mammography and electrical impedance scanning as adjunctive technologies in breast cancer detection.

AIM: Various modalities are used as an adjunct to mammography for differentiation of potentially suspicious breast lesions. Electrical impedance scanning (EIS) is a new technique based upon the principle that cancer cells exhibit altered local dielectric properties and thus show measurably higher conductivity values. The accuracy of differentiation of benign and malignant breast lesions was evaluated to determine whether EIS duplicates or supplements the results obtainable from ultrasound (US) or magnetic resonance imaging (MRI). MATERIALS AND METHODS: One hundred mammographically suspicious lesions were examined using US, MRI and EIS. Definitive histology was acquired through either lesion biopsy or surgical excision. RESULTS: Fifty of 62 malignant lesions were correctly identified using EIS (81% overall sensitivity), 24/38 benign lesions were correctly identified as benign (63% specificity). Negative predictive value and positive predictive value of 67 and 78% were observed, respectively. kappa-factor evaluation revealed a value of 0.82 between MRI and EIS and 0.62 between US and EIS. CONCLUSIONS: EIS may be a valuable adjunct for differentiation of suspicious mammographic lesions. Based upon the calculated kappa-factor, EIS results supplement US examinations. Artifacts (superficial skin lesions, poor contact, air bubbles) currently result in the high false-positive rate of EIS.

Breast Neoplasms↗
PubMed2001

[Cost-effectiveness of MR-imaging in the preoperative work-up of suspicious breast lesions].

UNLABELLED: Cost-effectiveness of MR-imaging in the preoperative work-up of suspicious breast lesions. PURPOSE: To assess the clinical and economic consequences of a preoperative magnetic resonance mammography (MRM) in patients with suspicious breast lesions. PATIENTS AND METHODS: 483 out of 2426 patients operated because of breast neoplasms at the University Hospital of Jena between August 1995 and March 1999 underwent preoperative Gd-enhanced breast MRI in addition to mammography and breast ultrasound because of suspicious breast lesions. The histological assessment was correlated with the results of the diagnostic imaging work-up in 447 cases. The costs of surgery for malignant lesions were calculated based on case- dependent billing agreements, costs of surgery of benign lesions were calculated based on current billing procedure. In 36 cases no surgery was performed due to the exclusion of malignancy by breast MRI. 140 benign lesions were treated surgically in spite of negative breast MRI diagnosis. The hospitalization time related costs caused by these cases were compared to the costs of preoperative MR-imaging. RESULTS: The total costs for MR mammography in all 483 patients amounted to DM 499,857. In 11 cases false positive results in MRM led to additional costs of DM 32,565. Assuming that there is no need for surgical treatment of benign breast lesions, total savings of DM 521,048 due to avoided hospitalization in 176 cases could have been possible. The savings per patient amount in these 176 cases are DM 2,960.50 compared to DM 1,034.90 costs for breast MRI. CONCLUSION: Despite the currently still high costs of breast MRI a major cost reduction can be achieved by prevention of unnecessary surgery, avoidance of additional surgery and associated trauma for patients.

Breast Neoplasms↗
PubMed2001

Percutaneous radiofrequency (RF) thermal ablation of rabbit tumors embedded in fat: a model for RF ablation of breast tumors.

RATIONALE AND OBJECTIVES: To develop an experimental tumor model for RF ablation of breast tumors surrounded by fat and to investigate the minimally invasive treatment of such tumors by saline-enhanced RF ablation monitored by ultrasound (US). METHODS: Twenty-eight VX2 tumors were implanted into the retroperitoneum of 14 rabbits and monitored by B-mode US at regular intervals of 2 to 3 days. Saline-enhanced RF ablation (25-mm tip length) was performed 16 days after tumor implantation (10-minute treatment time, 28 W, 15 mL/h infusion of 0.9% NaCl, which was increased to 30 mL/h in cases of an impedance increase). Thermal lesion growth was monitored by B-mode US. Treatment was considered complete if no relapse was detectable histopathologically after a follow-up period of up to 3 weeks. RESULTS: All tumor implantations were successful, reaching sizes from 5 to 38 mm 16 days after implantation. Tumors >20 mm showed central necrosis on US. Smaller tumors without signs of necrosis required greater needle perfusion to maintain constant needle-to-tissue impedance. Local relapses occurred in 14 of 27 tumors (51.8%), all with tumors >20 mm. In 12 cases, they were already detected sonographically. In 2 cases, "islands" of vital tumor were detected only during histopathological assessment. CONCLUSIONS: VX2 tumors are implantable into hypovascular adipose tissue with a high success rate and may be used as a model for RF ablation of breast tumors. B-mode US is not suitable for guiding RF ablation of tumors embedded in fat. The method appears to be more successful with smaller tumors.

Adipose Tissue↗
PubMed2001

Contrast-enhanced near-infrared laser mammography with a prototype breast scanner: feasibility study with tissue phantoms and preliminary results of imaging experimental tumors.

RATIONALE AND OBJECTIVES: Near-infrared (NIR) optical mammography without contrast has a low specificity. The application of optical contrast medium may improve the performance. The concentration-dependent detectability of a new NIR contrast medium was determined with a prototype optical breast scanner. In vivo imaging of experimental tumors was performed. METHODS: The NIR contrast agent NIR96010 is a newly synthesized, hydrophilic contrast agent for NIR mammography. A concentration-dependent contrast resolution was determined for tissue phantoms consisting of whole milk powder and gelatin. A central part of the phantoms measuring 2 x 2 cm2 without contrast was replaced with phantom material containing 1 micromol/L to 25 nmol/L NIR96010. The composite phantoms were measured with a prototype NIR breast scanner with lasers of lambda1 = 785 nm and lambda2 = 850 nm wavelength. Intensity profiles and standard deviations of the transmission signal in areas with and without contrast were determined by linear fit procedures. Signal-to-noise ratios and spatial resolution as a function of contrast concentration were determined. Near-infrared imaging of five tumor-bearing SCID mice (MX1 breast adenocarcinoma, tumor diameter 5-10 mm) was performed before and after intravenous application of 2 micromol/kg NIR96010. RESULTS: Spectrometry showed an absorption maximum of the contrast agent at 755 nm. No spectral shifts occurred in protein-containing solution. Signal-to-noise ratio in the transmission intensity profiles ranged from 1.1 at 25 nmol/L contrast to 28 at 1 micromol/L. At concentrations <40 nmol/L, no differentiation from the background was possible. The transitional area between the contrast-free edge of the phantom and the central contrast-containing part appeared in the profiles as a steep increase with a width of 4.2 +/- 1.8 mm. The experimental tumors were detectable in nonenhanced images as well as contrast-enhanced images, with better delineation after contrast administration. In postcontrast absorption profiles, a 44.1% +/- 11.3% greater absorption increase was seen in tumor tissue compared with normal tissue. CONCLUSIONS: The laser wavelength lambda1 of the prototype laser mammography device was not situated at maximum absorption of the contrast agent NIR96010 but on the descending shoulder of the absorption spectrum. This implies a 20% signal loss for contrast detection. Despite the nonideal measurement conditions, concentrations as low as 40 nmol/L were detectable in vitro. In vivo, all tumors were detectable in color-coded nonenhanced scans as well as in contrast-enhanced scans, with better delineation after contrast administration.

Alkenes↗
PubMed2001

Electrical impedance scanning: a new technique in the diagnosis of lymph nodes in which malignancy is suspected on ultrasound.

Differentiation between inflammatory and malignant lymph nodes by ultrasound is difficult. Electrical impedance scanning (EIS) is a new diagnostic tool, so far used primarily for the identification of malignant breast lesions. Cancer cells have altered dielectric properties compared with normal cells, thereby distorting the local electrical field. The induced changes in capacitance and conductivity are measurable using EIS. We evaluated EIS in demonstrating the cause of lymph node enlargement. 51 lymph nodes that were suspicious for malignancy on ultrasound (32 patients, mean age 32 years), with a mean size of 18 mm x 12 mm x 10 mm, were examined. The following lymph node locations were included in the study: cervical, inframandibular, axillary, paraaortic and inguinal. EIS results were compared with histopathological and follow-up findings. 30/34 malignant lymph nodes were correctly detected using EIS, while 14/17 inflammatory or benign lymph nodes were correctly identified as benign by EIS; thus, there were 4/51 false negative and 3/51 false positive cases. The sensitivity was 88.2% and the specificity was 82.4%. Corresponding negative and positive predictive values were 77.8% and 90.9%, respectively. Results from this initial study suggest the potential usefulness of EIS as an adjunctive imaging modality in the differentiation of lymphadenopathy that is equivocal on ultrasound. The best accuracy was obtained in the cervical, axillary and inguinal regions. Owing to technical restrictions of the present system, examination of inframandibular and paraaortic lymph nodes should be limited to special cases.

Adult↗
PubMed2000

Formation of regulator/target gene relationships during evolution.

The Foxn1-like forkhead/winged-helix transcription factor genes have been maintained in single copy throughout chordate evolution. Among other functions, Foxn1 (formerly known as Whn) regulates the expression of hair keratin genes in the hair follicle, which represents an evolutionarily novel organ characteristic of mammals. We show here that fish and mouse Foxn1-like genes are functionally equivalent in hair keratin gene activation, suggesting the absence of functionally relevant changes over the course of several hundred million years of vertebrate evolution. In contrast, the Foxn1-like gene from the cephalochordate Branchiostoma lanceolatum is inactive in this assay because of changes in the region located N-terminal to DNA binding and transcriptional activation domains of the protein. Our results indicate that functionally relevant changes in cis-regulatory regions are not necessarily accompanied by corresponding changes in transcription factor proteins in the formation of evolutionarily novel regulator/target gene relationships.

Amino Acid Sequence↗
PubMed2000

Gene structure, chromosomal localization, and expression pattern of Capn12, a new member of the calpain large subunit gene family.

We report the identification of mouse Capn12, a new member of the calpain large subunit gene family. It possesses potential protease and calcium-binding domains, features typical of the classical calpains. In situ hybridization and Northern blot analysis demonstrate that during the anagen phase of the hair cycle the cortex of the hair follicle is the major expression site of Capn12. The gene was sequenced in its entirety and consists of 21 exons spanning 13 kb with an exon-intron structure typical of the calpain gene family. The last exon of the mouse Actn4 gene overlaps the 3' end of Capn12 but in the opposite orientation. This overlap between the two genes is conserved in the human genome. Three versions of the Capn12 mRNA transcript were identified. They occur as a result of alternative splicing, and two of these encode a protein lacking the C-terminal calmodulin-like domain. Radiation hybrid mapping localized Capn12 to mouse chromosome 7, closely linked to a marker positioned at 10.4 cM. Refined mapping of Capn5, also previously localized to chromosome 7, indicated that it was not closely linked to Capn12, mapping tightly linked to a marker positioned at 48.5 cM.

Actinin↗
PubMed2000

Evolutionary conservation of gene structures of the Pax1/9 gene family.

Based on amino acid sequence comparisons, Pax1 and Pax9 genes are considered to form a subgroup of vertebrate Pax genes. We show here that the gene structures of mouse Pax1, human PAX9 genes are similar to that of a single Pax1/9 related gene in Branchiostoma lanceolatum, AmphiPax1. This supports the hypothesis that Pax1 and Pax9 genes were derived from a single ancestral gene. A refined protein alignment of AmphiPax1, mouse Pax1 and human PAX9 proteins based on the determined exon boundaries indicates that sequence divergence at the C-termini may be related to the unique functions of the Pax1 and Pax9 genes in vertebrates. AmphiPax1 is expressed in adult amphioxus in the pharyngeal endoderm.

Amino Acid Sequence↗
PubMed2000

Cytohesin-1 regulates beta-2 integrin-mediated adhesion through both ARF-GEF function and interaction with LFA-1.

Intracellular signaling pathways, which regulate the interactions of integrins with their ligands, affect a wide variety of biological functions. Here we provide evidence of how cytohesin-1, an integrin-binding protein and guanine-nucleotide exchange factor (GEF) for ARF GTPases, regulates cell adhesion. Mutational analyses of the beta-2 cytoplasmic domain revealed that the adhesive function of LFA-1 depends on its interaction with cytohesin-1, unless the integrin is activated by exogenous divalent cations. Secondly, cytohesin-1 induces expression of an extracellular activation epitope of LFA-1, and the exchange factor function is not essential for this activity. In contrast, LFA-1-mediated cell adhesion and spreading on intercellular cell adhesion molecule 1 is strongly inhibited by a cytohesin-1 mutant, which fails to catalyze ARF GDP-GTP exchange in vitro. Thus, cytohesin-1 is involved in the activation of LFA-1, most probably through direct interaction with the integrin, and induces cell spreading by its ARF-GEF activity. We therefore propose that both direct regulation of the integrin and concomitant changes in the membrane topology of adherent T cells are modulated by dissectable functions of cytohesin-1.

ADP-Ribosylation Factors↗
PubMed2000

Forkhead/winged-helix transcription factor Whn regulates hair keratin gene expression: molecular analysis of the nude skin phenotype.

The molecular characteristics of the nude phenotype (alopecia and thymic aplasia) in humans and rodents are unknown. The nude locus encodes Whn, a transcription factor of the forkhead/winged-helix class. Expression of Whn in HeLa cells induced expression of human hair keratin genes Ha3-II and Hb5. Correspondingly, in nude mice, which are homozygous for a loss-of-function mutation of Whn, expression of mouse mHa3 and mHb5 hair keratin genes is severely reduced. Characterization of a previously identified nude allele, nu(Y), revealed a mis-sense mutation (R320C) in the DNA binding domain of Whn. This mutant protein is unable to activate hair keratin gene expression in HeLa cells. When the Whn transcription factor was expressed in two parts, one containing the N-terminal DNA binding domain and the other the C-terminal activation domain, no activation of hair keratin genes in HeLa cells was observed. However, when these two proteins were noncovalently linked by means of synthetic dimerizers, hair keratin gene expression was induced. This finding suggests that target gene activation by Whn depends on the structural integrity and physical proximity of DNA binding and activation domains, providing a molecular framework to explain the loss-of-function phenotypes of all previously characterized nude mutations. Our results implicate Whn as a transcriptional regulator of hair keratin genes and reveal the nude phenotype as the first example of an inherited skin disorder that is caused by loss of expression rather than mutation of keratin genes.

Animals↗
PubMed2000

Genetically separable determinants of hair keratin gene expression.

The nude locus encodes Whn, a transcription factor of the forkhead/winged-helix class. Mutations in Whn cause failure of differentiation of thymic epithelium with a corresponding lack of intrathymic T-cell development; in the skin, differentiation of follicular keratinocytes is disturbed resulting, in the formation of fragile hair shafts. Here, we describe the identification and characterization of a novel nude allele, nu(StL). nu(StL) encodes a truncated Whn transcription factor protein, designated Whn(StL), lacking the activation domain but retaining the characteristic DNA binding domain. In contrast, the previously described Whn(nu) mutant protein lacks both domains. nu(StL)/nu(StL) mice show an alymphoid thymic rudiment and lack of peripheral T cells, similar to nu/nu mice. In the skin, impaired expression of hair keratin genes mHa1, mHa2, mHa3 and mHa4, mHb3, mHb4, mHb5, and mHb6 is observed in a pattern that parallels that of nu/nu mice: both mutant alleles behave as hypomorphs with respect to the expression of these hair keratin genes. However, a significant difference between these two alleles exists for mHa5 expression, which is reduced in nu(StL)/nu(StL) but not in nu/nu mice. We show that the mutant Whn protein in nu/nu mice cannot enter the nucleus, whereas the mutant Whn protein in nu(StL)/nu(StL) mice is present in the nucleus. The antimorphic characteristic of the activation-deficient Whn(StL) protein with respect to mHa5 expression is therefore most likely caused by its non-productive interaction with other proteins at cis-regulatory regions of the mHa5 gene. Our results indicate that the molecular consequences of mutations of the Whn gene can be different and demonstrate an unexpected complexity of transcriptional control mechanisms of hair keratin genes.

Alleles↗
PubMed2000

Chemokines define distinct microenvironments in the developing thymus.

During thymus development, prothymocytes home to the thymus where they migrate as maturing thymocytes from the cortex to the medulla. Chemotaxis assays show that developing T cells of newborn mice respond to certain chemokines depending on their differentiation state. In situ expression analyses indicate that the same chemokines are expressed in distinct microenvironments within the thymic stroma. Expression of chemokines is regulated temporally during embryogenesis; in the alymphoid early thymic anlage, only TECK, SDF-1 and SLC but not ELC, MDC or TARC are expressed. Fetal blood prothymocytes destined to colonize the thymus respond to the embryonic chemokines TECK and SDF-1 in chemotaxis assays with high efficacy. The in vivo significance of this finding is demonstrated by studies in the nude mouse where the thymic anlage lacks TECK and SDF-1 expression and prothymocytes home to the parathyroid anlage rather than to the thymic anlage. Developing thymocytes respond to chemokines expressed in distinct microenvironments within the thymic stroma in a way that correlates well with the previously observed migration pattern from cortex to medulla. The complexity of these chemokine-defined microenvironments increases as the thymic anlage develops to a mature thymus.

Animals↗