Biomedical subjects

Solveig Schulz

Publications and source records attributed to Solveig Schulz.

14 recordsLinked to original sources

PubMed2006

Immunohistochemical detection of bombesin receptor subtypes GRP-R and BRS-3 in human tumors using novel antipeptide antibodies.

Bombesin (BN)-like peptides can stimulate cancer cell growth through binding to their specific G protein-coupled receptors. It is well established that BN receptors are being overexpressed in a subset of human tumors; however, little is known about the cellular and subcellular localization of individual BN receptor subtypes in these tissues. In this study, we developed and characterized novel antipeptide antibodies to the carboxy terminal regions of the gastrin-releasing peptide-preferring bombesin receptor (GRP-R) and the bombesin receptor subtype-3 (BRS-3). Specificity of the antisera was demonstrated by (1) detection of broad bands migrating at Mr 50,000-70,000 in Western blots of membranes from receptor-expressing tissues; (2) cell surface staining of transfected cells; (3) translocation of GRP-R receptor immunostaining after BN exposure; and (4) abolition of tissue immunostaining by preadsorbtion of the antibodies with their immunizing peptides. The distribution of BN receptors was investigated in 74 formalin-fixed, paraffin-embedded human tumors. GRP-R receptors were most frequently detected in breast and prostate carcinomas. BRS-3 receptors were often detected in prostate and pancreatic carcinomas and in pituitary adenomas. Immunoreactive GRP-R and BRS-3 receptors were in many cases predominantly confined to the plasma membrane and uniformly present on nearly all tumor cells. The development of these novel antipeptide antibodies will facilitate the identification of those tumors, which may be targets for diagnostic or radiotherapeutic application of subtype-selective BN analogs.

Amino Acid Sequence↗

PubMed2006

Immunocytochemical localisation of plasma membrane GHRH receptors in human tumours using a novel anti-peptide antibody.

Antagonists of growth hormone releasing hormone (GHRH) directly inhibit the growth of a variety of human neoplasms. However, the plasma membrane receptor mediating these effects has not been immunocytochemically visualised in primary tumour cells. Given that previous attempts using an antibody to the amino-terminal region did not result in the visualisation of plasma membrane receptors, we have developed and characterised an anti-peptide antibody to the carboxy-terminal region 403-422 of the human pituitary GHRH receptor. This sequence is identical to residues 339-358 of splice variant 1 (SV1) of tumoural GHRH receptors. Specificity of the antibody was demonstrated by (1) immunocytochemical staining of GHRH receptor-transfected cells, (2) detection of a broad glycosylated protein band migrating at Mr 50,000-60,000 in Western blots of membranes from human pituitary, and (3) abolition of tissue immunostaining by preadsorbtion of the antibody with its immunising peptide. The distribution of GHRH receptors was investigated in 69 formalin-fixed, paraffin-embedded human tumours showing that GHRH receptors were frequently expressed in breast, ovarian and prostate carcinomas. Immunoreactive GHRH receptors were clearly confined to the plasma membrane and uniformly present on nearly all tumour cells. In Western blots of membranes prepared from human tumours, the anti-GHRH receptor antibody detected a non-glycosylated protein band migrating at Mr 40,000, which corresponds to the expected molecular weight of splice variant 1 of tumoural GHRH receptors. Together, our findings provide direct evidence for the presence of GHRH receptor protein on the plasma membrane of primary human tumour cells. The GHRH receptor visualisation could be of value for a rapid immunohistochemical identification of those tumours which could be a target for diagnostic or therapeutic intervention using GHRH analogues.

Antibodies↗

PubMed2006

Immunolocalization of full-length NK1 tachykinin receptors in human tumors.

Biological effects of substance P (SP) are mediated by the neurokinin-1 (NK1) receptor that exists as a full-length and as a carboxy-terminally truncated isoform in humans. Although NK1 receptor mRNA and binding sites have been detected in certain malignancies, little is known about the cellular and subcellular localization of NK1 receptor protein in human neoplastic tissues. We developed and characterized a novel anti-peptide antibody to the carboxy-terminal region of the human full-length NK1 receptor. Specificity of the antiserum was demonstrated by (1) detection of a broad band migrating at molecular mass 70,000-90,000 Da in Western blots of membranes from NK1-expressing tissues; (2) cell-surface staining of NK1-transfected cells; (3) translocation of NK1 receptor immunostaining after SP exposure; and (4) abolition of tissue immunostaining by preadsorption of the antibody with its immunizing peptide. Distribution of NK1 receptors was investigated in 72 formalin-fixed, paraffin-embedded human tumors showing that NK1 receptors were frequently expressed in glioblastomas and breast and pancreatic carcinomas. Immunoreactive NK1 receptors were clearly confined to the plasma membrane and uniformly present on nearly all tumor cells. Development of this novel NK1 receptor antibody allows the efficient localization of NK1 receptor protein in human formalin-fixed, paraffin-embedded tissues. NK1 receptor visualization with this simple and rapid immunohistochemical method will facilitate identification of tumors with a sufficient receptor overexpression for diagnostic or therapeutic intervention using SP analogs.

Antibodies↗

PubMed2006

Increased frequency of cystic fibrosis transmembrane conductance regulator gene mutations in infertile males.

OBJECTIVE: To investigate the frequency of mutations of the cystic fibrosis transmembrane regulator (CFTR) gene in males with reduced sperm quality before intracytoplasmic sperm injection (ICSI). DESIGN: The nine most frequent cystic-fibrosis-causing mutations in the German population and IVS8T alleles were analyzed. SETTING: University-based centers for reproductive medicine and clinical genetics. PATIENT(S): An unselected group of 597 males with oligo-, astheno-, terato-, crypto-, oligoasthenoteratozoospermia, or azoospermia, which underwent pre-ICSI genetic counseling over a 5-year period. INTERVENTION(S): Blood samples were collected from the patients during genetic counseling. MAIN OUTCOME MEASURE(S): Frequency of mutations of CFTR gene in infertile males. RESULT(S): A heterozygous CFTR mutation was observed in 34 of 597 patients (5.70%). None of the patients had two CFTR mutations. Given that our mutation panel recognizes about 82% of heterozygotes, it can be assumed that the frequency of CFTR heterozygotes in our cohort is about 6.94%. The frequency of CFTR mutations in our cohort did not correlate with a reduced sperm count. CONCLUSION(S): The frequency of cystic fibrosis in the German population is 1:3300. Thus, a CFTR heterozygosity of 3.42% can be estimated. This indicates that in our cohort of infertile males, the frequency of CFTR heterozygosity is twofold higher than in the general population (P<.0001).

Cohort Studies↗

PubMed2006

Immunocytochemical identification of low-affinity NTS2 neurotensin receptors in parietal cells of human gastric mucosa.

The biological effects of neurotensin (NT) are mediated by two distinct G protein-coupled receptors, NTS(1) and NTS(2). Although it is well established that neurotensin inhibits gastric acid secretion in man, the plasma membrane receptor mediating these effects has not been visualized yet. We developed and characterized a novel antipeptide antibody to the carboxy-terminal region of the human NTS(2) receptor. The cellular and subcellular distribution of NTS(2) receptors was evaluated in various human gastrointestinal tissues. Specificity of the antiserum was demonstrated by (1) detection of a broadband migrating at M(r) 90 000-100 000 in Western blots of membranes from NTS(2)-expressing tissues; (2) cell-surface staining of NTS(2)-transfected cells; (3) translocation of NTS(2) receptor immunostaining after agonist exposure; and (4) abolition of tissue immunostaining by preadsorbtion of the antibody with its immunizing peptide. In the gastrointestinal tract, NTS(2) receptor immunoreactivity was highly abundant in parietal cells of the gastric mucosa, in neuroendocrine cells of the stomach small and large intestine, and in cells of the exocrine pancreas. NTS(2) receptors were clearly located in the plasma membrane and uniformly present on nearly all target cells. The presence of NTS(2) receptors was rarely detected in human tumors. This is the first localization of NTS(2) receptors in human formalin-fixed, paraffin-embedded tissues at the cellular level. The abundant expression of low-affinity NTS(2) receptors on the plasma membrane of human parietal cells provides a morphological substrate for the direct inhibition of gastric acid secretion observed after i.v. administration of neurotensin.

Antibodies, Monoclonal↗

PubMed2005

Immunohistochemical localization of CCK1 cholecystokinin receptors in normal and neoplastic human tissues.

BACKGROUND: The biological effects of cholecystokinin (CCK) are mediated by two distinct G protein-coupled receptors, CCK1 and CCK2. Although it is well established that CCK receptors are widely distributed throughout the normal gastrointestinal tract, little is known about their cellular and subcellular localization in human normal and neoplastic tissues. METHODS: We developed and characterized a novel antipeptide antibody to the carboxyl-terminal region of the human CCK1 receptor. Specificity of the antiserum was demonstrated by 1) detection of a broad band migrating at a relative molecular mass of 85,000-95,000 in Western blots of membranes from CCK1-expressing tumors and CCK1-transfected cells, 2) cell surface staining of CCK1-transfected cells, 3) translocation of CCK1 receptor immunostaining after agonist exposure, and 4) abolition of tissue immunostaining by preadsorption of the antibody with its immunizing peptide. The distribution of CCK1 receptors was investigated in 74 human tumors and their tissues of origin. RESULTS: The presence of CCK1 receptors was rarely detected in human tumors except for carcinoids, insulinomas, pituitary adenomas, and meningiomas. CCK1 receptors were clearly located at the plasma membrane and uniformly present on nearly all tumor cells. In the gastrointestinal tract, CCK1 receptor immunoreactivity was highly abundant in chief cells of the gastric mucosa, in myenteric ganglion cells, and in myenteric nerve fibers. CONCLUSION: This is the first localization of CCK1 receptors in human formalin-fixed, paraffin-embedded tissues at the cellular level. The overexpression of CCK1 receptors in a subset of human neuroendocrine tumors may provide a molecular basis for efficient targeting of these tumors with radiolabeled CCK analogs.

Amino Acid Sequence↗

PubMed2005

Clinical and mutational spectrum of Mowat-Wilson syndrome.

Mowat-Wilson Syndrome is a recently delineated mental retardation syndrome usually associated with multiple malformations and a recognizable facial phenotype caused by defects of the transcriptional repressor ZFHX1B. To address the question of clinical and mutational variability, we analysed a large number of patients with suspected Mowat-Wilson Syndrome (MWS). Without prior knowledge of their mutational status, 70 patients were classified into "typical MWS", "ambiguous" and "atypical" groups according to their facial phenotype. Using FISH, qPCR and sequencing, ZFHX1B deletions, splice site or truncating mutations were detected in all 28 patients classified as typical MWS. No ZFHX1B defect was apparent in the remaining 15 cases with ambiguous facial features or in the 27 atypical patients. Genotype-phenotype analysis confirmed that ZFHX1B deletions and stop mutations result in a recognizable facial dysmorphism with associated severe mental retardation and variable malformations such as Hirschsprung disease and congenital heart defects. Our findings indicate that structural eye anomalies such as microphthalmia should be considered as part of the MWS spectrum. We also show that agenesis of the corpus callosum and urogenital anomalies (especially hypospadias) are significant positive predictors of a ZFHX1B defect. Based on our observation of affected siblings and the number of MWS cases previously reported, we suggest a recurrence risk of around 1%. The lack of missense mutations in MWS and MWS-like patients suggests there may be other, as yet unrecognized phenotypes, associated with missense mutations of this transcription factor.

Abnormalities, Multiple↗

PubMed2004

Immunocytochemical identification of VPAC1, VPAC2, and PAC1 receptors in normal and neoplastic human tissues with subtype-specific antibodies.

Human tumors frequently overexpress receptors for vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating peptide (PACAP). However, none of the VIP/PACAP receptor proteins has been visualized individually in human tumors. Here, we developed and characterized a panel of antipeptide antibodies to the carboxyl-terminal regions of the VIP/PACAP receptor subtypes vasoactive intestinal peptide receptor (VPAC)1, VPAC2, and pituitary adenylate cyclase-activating peptide receptor (PAC)1. Specificity of the antisera was shown by the following: (1) detection of broad bands migrating at Mr 50,000 to 70,000 in Western blots of membranes from receptor-expressing tumors and receptor-transfected cells; (2) cell surface staining of VIP/PACAP receptor-transfected cells; (3) translocation of VIP/PACAP receptor immunostaining in transfected cells after agonist exposure; and (4) abolition of tissue immunostaining by preadsorbtion of the antibodies with their immunizing peptides. The distribution of VIP/PACAP receptors was investigated in 98 human tumors and their tissues of origin. VPAC1, VPAC2, and PAC1 receptors were clearly located at the plasma membrane of the tumor cells in a variety of human neoplasms. In the gastrointestinal tract, VPAC1 receptor immunoreactivity was abundant in the mucosa and myenteric neurons; VPAC2 receptor immunoreactivity was detected in neuroendocrine cells, blood vessels, and smooth muscle; and PAC1 receptor immunoreactivity was found in myenteric neurons. This is the first localization of all of the VIP/PACAP receptor subtypes in human formalin-fixed, paraffin-embedded tissues. VIP/PACAP receptor visualization with this simple and rapid immunohistochemical method will facilitate identification of tumors with a sufficient receptor overexpression for diagnostic or therapeutic intervention.

Animals↗

PubMed2004

Differential expression of sst1, sst2A, and sst3 somatostatin receptor proteins in low-grade and high-grade astrocytomas.

We have previously reported that sst2A somatostatin receptors are frequently overexpressed in human meningiomas. Initial clinical observations suggest that somatostatin analogues may also be of value for imaging and treatment of other human intracranial tumors, including astrocytomas. However, contradictory results have been reported regarding the expression of somatostatin receptors in low-grade and high-grade astrocytomas. Therefore, we determined the precise pattern of somatostatin receptor protein expression in 8 diffuse astrocytoma (DA), 10 anaplastic astrocytomas (AA), and 32 glioblastoma multiforme (GBM) using immunohistochemistry and Western blot analysis. sst1 and sst2A somatostatin receptors were not present in DA and only occasionally detected in AA. In GBM, sst1 was present in 66%, and sst2A was found in 44% of the tumors. sst3 receptors were present in 38% of DA, 40% of AA, and 84% of GBM. Thus, loss of differentiation was significantly associated with increased expression of sst1, sst2A, and sst3 somatostatin receptors. In contrast, sst4 and sst5 receptors were found in 80% and 25% of all cases, respectively, in a manner independent of histological grade. No significant correlation was found between somatostatin receptor expression and the proliferation rate of the tumors as determined by MIB-I immunostaining. Furthermore, the presence or absence of the 5 somatostatin receptor subtypes did not significantly influence survival time in 14 GBM patients.

Astrocytoma↗

PubMed2003

Frequent expression of immunoreactive somatostatin receptors in cervical and endometrial cancer.

OBJECTIVE: A variety of human cancers has been shown to express somatostatin receptors which can be utilized for in vivo tumor targeting. Although the somatostatin receptor status of mammary and ovarian cancer has been studied in detail, somatostatin receptors have not been examined in other gynecological malignancies such as cervical or endometrial carcinoma. METHODS: We have previously developed a panel of somatostatin receptor subtype-specific antibodies that effectively stain formalin-fixed, paraffin-embedded human tumor tissue. In the present study, we have used these antibodies to determine the somatostatin receptor status of 21 randomly selected cervical carcinomas and 28 randomly selected endometrial carcinomas. RESULTS: In cervical carcinoma, the incidence of somatostatin receptor expression was sst2A (12/21, approximately 57%) > sst3 (9/21, approximately 43%) > sst1 (8/21, approximately 38%) > sst4 (0/21, 0%) = sst5 (0/21, 0%). In endometrial carcinoma, the incidence of somatostatin receptor expression was sst3 (12/28, approximately 43%) > sst2A (11/28, approximately 39%) > sst1 (9/28, approximately 32%) > sst4 (1/28, approximately 4%) = sst5 (1/28, approximately 4%). Somatostatin receptor-like immunoreactivity was in most cases located at the plasma membrane and completely blocked with antigenic peptide. The expression of somatostatin receptor subtypes was independent of patient age, diagnosis, tumor stage, and histological grade. CONCLUSION: Our findings reveal a previously unappreciated high incidence of sst1, sst2A, and sst3 somatostatin receptors in human cervical and endometrial cancers and suggest that a subgroup of receptor-positive uterine carcinomas may be a potential target for diagnostic and therapeutic intervention with stable somatostatin analogs.

Adult↗

PubMed2003

Somatostatin receptor subtypes in human pheochromocytoma: subcellular expression pattern and functional relevance for octreotide scintigraphy.

The stable somatostatin analog octreotide has been successfully used for imaging and treatment of a variety of human tumors. In pheochromocytoma, data on somatostatin receptor subtyping have thus far been sparse. Pheochromocytomas often express more than one somatostatin receptor, and it is uncertain by which receptor subtype the functional responses of octreotide are mediated. Here, we have examined somatostatin receptor protein expression in a panel of 52 pheochromocytomas from 35 randomly selected patients by immunostaining with specific polyclonal anti-sst(1-5) and monoclonal mouse anti-SS-14 antibodies. Staining pattern, distribution and subcellular localization of somatostatin receptor subtypes were investigated. Seventeen patients received (111)In-octreotide scintigraphy. Although the vast majority of tumors (90%) showed positive immunohistochemical staining for sst(3), immunoreactive sst(2A) receptors were only seen in 13 tumors (25%). All other somatostatin receptor subtypes were less frequently detected. Interestingly, among sst(3)-positive tumors strikingly different subcellular distributions of immunoreactive sst(3) receptors were observed. In most cases, immunoreactive sst(3) receptors were distributed throughout the cytosol. Scintigraphic localization of tumors larger than 1 cm in diameter was always successful in the presence of immunoreactive sst(2A) receptors. In the absence of sst(2A), true-positive octreotide scintigraphy was only seen in the presence of membrane-associated sst(3) immunoreactivity. Our findings suggest that selective expression of functional membrane-associated sst(3) receptors is sufficient for high tracer uptake during octreotide scintigraphy in a subgroup of human pheochromocytomas. These tumors may represent a potential target treatment with somatostatin receptor agonists with improved sst(3) activity.

Adrenal Gland Neoplasms↗

PubMed2002

Immunohistochemical detection of somatostatin receptors in human ovarian tumors.

OBJECTIVE: It is well established that many human tumors can express somatostatin receptors. This is the molecular basis for the application of long-acting somatostatin analogues for therapeutic and diagnostic purposes. However, there are discrepancies in earlier studies regarding the prevalence of somatostatin receptor expression in human ovarian tumors detected using autoradiographic binding studies or reverse transcription-polymerase chain reaction. METHODS: We have previously developed a panel of somatostatin receptor subtype-specific antibodies that effectively stain formalin-fixed, paraffin-embedded human tumor tissue. In the present study, we have used these antibodies to determine the somatostatin receptor status of 47 randomly selected human ovarian tumors. RESULTS: Somatostatin receptor-like immunoreactivity was in most cases located at the plasma membrane as well as in the cytoplasm of the tumor cells and was completely blocked with antigenic peptide. The pattern of expression of somatostatin receptor subtypes varied greatly between individual tumors. Of the 47 ovarian tumor specimens analyzed, 9 tumors (approximately 19%) were positive for sst(1), 13 (approximately 28%) revealed immunoreactive sst(2A) receptors, 20 (approximately 42%) showed sst(3)-like immunoreactivity, 8 (approximately 17%) revealed immunoreactive sst(4) receptors, and 10 (approximately 21%) were positive for sst(5). Whereas 18 tumors (approximately 38%) revealed no somatostatin receptor immunoreactive staining, 14 (approximately 30%) expressed more than one somatostatin receptor subtype. The expression of somatostatin receptor subtypes was independent of patient age, diagnosis, and histological grade. CONCLUSION: We provide direct evidence of somatostatin receptor protein expression in human ovarian tumors. Our findings suggest that a subgroup of receptor-positive ovarian tumors may be a potential target for treatment with somatostatin receptor subtype-selective analogs. However, due to the wide variations in the somatostatin receptor status of similar cases, each tumor specimen must be tested individually for the presence of particular somatostatin receptor subtypes.

Adolescent↗

PubMed2002

True positive somatostatin receptor scintigraphy in primary breast cancer correlates with expression of sst2A and sst5.

UNLABELLED: [111In-DTPA-D-Phe1]-octreotide scintigraphy has been shown to reveal somatostatin receptor-positive lesions in the majority of primary breast cancers. We have recently developed a panel of somatostatin receptor subtype-specific antibodies that effectively stain formalin-fixed, paraffin-embedded breast cancer tissue. However, it is uncertain to what extend somatostatin receptors detected during immunohistochemical staining represent functional binding sites responsible for high tracer uptake during somatostatin receptor scintigraphy. PATIENTS AND METHODS: We, therefore, conducted a prospective study in which 23 patients with suspected breast tumors were included. All patients received [111In]-pentetreotide scintigraphy. After surgical removal of the tumor, the somatostatin receptor status was determined by immunohistochemistry. RESULTS: Among 20 pathologically proven malignant tumors (14 ductal and six lobular carcinomas), 13 (approximately 65%) were scintigraphically visible. Of the 20 primary breast cancer specimens analyzed, three tumors (approximately 15%) were positive for sst1, nine (approximately 45%) revealed immunoreactive sst2A receptors, eight (approximately 40%) showed sst3-like immunoreactivity, and 14 (approximately 70%) were positive for sst5. There was an excellent correlation between the outcome of somatostatin receptor scintigraphy and expression of sst2A (P = 0.025) as well as sst5 (P < 0.001) but not expression of either sst1 (P = 0.343) or sst3 (P = 0.400). CONCLUSION: Both sst2A and sst5 can be responsible for high tracer uptake during [111In]-pentetreotide scintigraphy in primary breast cancer. Thus, somatostatin receptor scintigraphy may possibly be of value in the detection of proven somatostatin receptor sst2A- and/or sst5-positive lesions in metastatic breast cancer.

Adult↗

PubMedDate not supplied

Pseudotrisomy 13: clinical findings and genetic implications.

The combination of holoprosencephaly, postaxial polydactyly, and normal karyotype has been termed pseudotrisomy 13 syndrome. Here, we report the prenatal diagnosis of pseudotrisomy 13 in three siblings suggesting autosomal recessive inheritance of this syndrome. Clinical overlap with hydrolethalus syndrome, Smith-Lemli-Opitz syndrome, Meckel syndrome, and Pallister-Hall syndrome is discussed.

Diploidy↗