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Biomedical subjects

W Rohde

Publications and source records attributed to W Rohde.

At least 91 records · Page 5Linked to original sources

[Cushing's syndrome in CRF-producing mediastinal carcinoid].

A 36-year-old patient developed marked pigmentation, marked myopathy and severe hypokalaemic alkalosis which at first pointed towards an ectopic ACTH syndrome. The dexamethasone test at a high dose indicated cortisol suppression. A mediastinal tumour was seen radiologically, but the sella was of normal size. Computed tomography provided indirect signs of a sellar space-occupying lesion which suggested an ectopic production of corticotropin-releasing factor (CRF) as cause of the Cushing's syndrome. CRF concentration in antecubital venous blood was markedly elevated to 280 ng/l. The mediastinal tumour was excised and proved to be a carcinoid histologically. Postoperatively the CRF concentration fell to 70 ng/l. An extract of the carcinoid contained 15.5 ng/g wet-weight of CRF and 254 ng/g wet-weight of beta-endorphin. The patient died 5 weeks postoperatively of sepsis with bilateral pneumonia. At autopsy the hypophysis was of normal size but showed nodular ACTH-cell hyperplasia. This was thus a case of Cushing's syndrome resulting from ectopic CRF production in a mediastinal carcinoid tumour.

Adult↗

Nucleotide sequence of a circular single-stranded DNA associated with coconut foliar decay virus.

A circular single-stranded (ss) covalently closed (ccc) DNA associated with coconut foliar decay virus (CFDV) was purified, amplified by the polymerase chain reaction, and subcloned and its sequence established by analysis of overlapping subgenomic cDNA clones. The complete CFDV sequence comprised 1291 nucleotides and contained open reading frames for six proteins of molecular weight larger than 5 kDa. One of these (ORF1, 33.4 kDa) codes for a leucine-rich protein with the nucleoside triphosphate-binding motif GXGKS and may possibly participate in virus replication. The putative viral protein encoded by ORF3 (6.4 kDa) is a positively charged arginine-rich protein with homology to the capsid protein of nuclear polyhedrosis virus, and may represent the CFDV coat protein. CFDV DNA can form a stable stem structure of 10 GC base pairs subtending a loop sequence which in one orientation closely resembles the motif TAATATTAC conserved in a similar structural arrangement within the geminivirus group. Otherwise no sequence homology to DNA-containing plant viruses of the gemini- or caulimovirus groups was found. CFDV therefore represents a new taxonomic group of plant viruses.

Amino Acid Sequence↗

Increased cell-mediated cytotoxicity against beta-cells in streptozotocin-treated offspring of mother animals with gestational hyperglycaemia.

The offspring of mother animals with mild gestational hyperglycaemia exhibited basal hyperinsulinism, decreased glucose tolerance and increased susceptibility to streptozotocin diabetes. Following low-dose successive streptozotocin treatment a significantly increased spleen cell cytotoxicity against beta-cells was found in these animals as compared to the offspring of gestational normoglycaemic control mothers. Such increased cell-mediated cytotoxicity, considered as enhanced autoimmune reactivity, was positively correlated to blood glucose levels and negatively correlated to pancreatic insulin contents. Thus, hyperinsulinism, occurring during pre- and neonatal brain organization and produced by gestational hyperglycaemia, is a predisposing teratogenetic risk factor not only for the development of type II, but also of type I diabetes. Thus, it is comprehensible that the prevalence of type I diabetes in children could be decisively reduced by preventing gestational hyperglycaemia in their mothers (Dörner et al., 1985).

Animals↗

Radioimmunoassay of corticotropin-releasing hormone.

A specific double antibody radioimmunoassay (RIA) for human-Corticotropin-Releasing Hormone (hCRF) using an antibody to synthetic hCRF was established. This antibody allowed a usable range of 10 pg to 5 ng of CRF per ml in the assay. Comparing the efficiency of various plasma extraction procedures, the extraction with ice-cold methanol was found to be the most simple and rapid method with an extraction efficiency of more than 80%. The reliability of the radioimmunoassay was shown under physiological and pathophysiological conditions.

Adolescent↗

Early postnatal overfeeding and diabetes susceptibility.

Milk intake of suckling rats was controlled by adjusting little size at birth to maximum (small litter, overnutrition), ten (normal litter, normal nutrition) or about 19 (large litter, undernutrition) pups. After weaning all animals received the same diet ad libitum. Overnutrition before weaning resulted in an early postnatal hyperinsulinaemia, decreased glucose tolerance and increased susceptibility to streptozotocin-induced diabetes in adulthood. However, the preweaning undernutrition seemed to have no preventive effect on diabetes susceptibility in adulthood. The study suggested that early postnatal overnutrition, which is associated with an early postnatal hyperinsulinemia, can increase diabetes susceptibility in adult life.

Animals↗

Characterization of a potato leafroll luteovirus subgenomic RNA: differential expression by internal translation initiation and UAG suppression.

Northern blot analysis of Solanum tuberosum infected with potato leafroll luteovirus revealed the 6 kb genomic RNA and a major 2.3 kb subgenomic RNA. The 5' end of the subgenomic RNA was located at nucleotide 3653 in an intergenic region located at the centre of the viral genome upstream of three open reading frames (ORFs). Transient expression in tobacco and potato protoplasts of the beta-glucuronidase reporter gene fused to various putative regulatory sequences present in the subgenomic RNA was used to study their influence on expression levels. We observed a suppression of the amber stop codon separating the coat protein (CP) gene from a downstream ORF (56K protein), to a level of 0.9% to 1.3%. Translation initiation at the AUG of an ORF (17K protein) which is nested within the CP gene, exceeds translation of the CP gene itself by a factor of 7.

Base Sequence↗

Nicotiana velutina mosaic virus: evidence for a bipartite genome comprising 3 kb and 8 kb RNAs.

DNA complementary to Nicotiana velutina mosaic virus (NVMV) RNA was cloned and five segments larger than 0.9 kb were used in Northern blot hybridization analysis to identify two virus-specific RNAs, approximately 8 kb (RNA 1) and 3 kb (RNA 2) in size. The clones selected as probes did not hybridize with RNA from various tobamoviruses, or from beet necrotic yellow vein (BNYVV) and peanut clump furoviruses. In an attempt to determine the taxonomic position of the virus, about 75% of the NVMV RNA 2 was sequenced and four open reading frames (ORFs) were identified. ORFs 1, 2 and 3 encode proteins of Mr 20K, 39K and 13K, whereas ORF 4 was incomplete. ORFs 2, 3 and 4 overlapped in an arrangement closely resembling the triple gene block identified in BNYVV RNA 2, barley stripe mosaic virus (BSMV) RNA 2, potato virus X and potato virus M RNA. The presumed coat protein gene of NVMV RNA 2 (ORF 1) is situated to the 5' side of the triple gene block as for BNYVV and BSMV RNA 2. Amino acid homologies were detected among the 13K and 14K proteins of NVMV RNA 2, BNYVV RNA 2 and BSMV RNA 2. Significant homology was also detected between the 39K protein of NVMV RNA 2 and the 42K protein of BNYVV RNA 2, with a motif specific for ATP- and GTP-binding (NTP-binding motif), and a conserved viral DNA polymerase domain. The presence of a triple gene block in NVMV RNA 2 indicates that NVMV has affinities with members of the hordei-, furo-, potex- and carlavirus groups but not with the tobamovirus group. The divided RNA genome of NVMV, and the sizes of the two RNAs suggest that NVMV is most closely allied to the furoviruses, but the unique nature of its different biological properties and lack of any serological relationships with furoviruses lead us to conclude that NVMV has no clear relatedness to any taxonomic group of plant viruses.

Amino Acid Sequence↗

Evidence that desensitization of the negative feedback of oestrogen and priming for the positive feedback of oestrogen depend upon a common mechanism of oestrogen action in rats.

Oestrogen priming of the central nervous system is required for the positive feedback of oestrogen, and the sensitivity of the negative feedback of oestrogen can be reduced by oestrogen itself. Using adult female and male rats we examined the possibility that these effects depend upon a common mechanism of oestrogen action that is mediated by the medial preoptic area (MPOA). Guide cannulae were implanted in the MPOA of 4-day cyclic rats which were ovariectomized during the evening of day 1 of dioestrus. Glass capillary tubes containing different substances were placed in the cannulae between 09.00 and 12.00 h on the presumptive day 2 of dioestrus. The effectiveness of oestrogen priming was evaluated by examining whether an s.c. implant of oestradiol-17 beta (OE2) induced an LH surge, and the inhibitory effect of oestrogen on tonic LH secretion was investigated by injecting the rats with 3 micrograms oestradiol benzoate (OB)/100 g body weight. The priming effect of an s.c. implant of OE2 could be imitated by the bilateral implantation in the MPOA of a mixture of OB and cholesterol at a ratio of 1:360 for 3 h only. Similar medial preoptic oestrogen implantation also significantly reduced the LH-inhibiting effect of OB. In accord with findings obtained in former studies on desensitization of the negative oestrogen feedback, oestrogen priming resulting from the s.c. administration of OE2 could be suppressed by short-term medial preoptic implantation of clomiphene citrate or apomorphine; bilateral electrical stimulation of the medial amygdaloid nucleus induced an increase in the serum concentration of LH in ovariectomized females implanted with OB in the MPOA, but not in castrated males pretreated and implanted with OB.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Multiple genes are transcribed in Hordeum vulgare and Zea mays that carry the DNA binding domain of the myb oncoproteins.

cDNA clones were isolated from tissue specific cDNA libraries of barley and maize using as a probe the cDNA of the maize gene C1, a regulator of anthocyanin gene expression. C1-related homology for all of the four cDNAs characterized by sequence analysis is restricted to the N-terminal 120 amino acids of the putative proteins. This region shows striking homology to the N-proximal domain of the myb oncoproteins from vertebrates and invertebrates. Within the myb proto-oncogene family this part of the respective gene products functions as a DNA binding domain. Acidic domains are present in the C-proximal protein segments. Conservation of these sequences, together with the genetically defined regulator function of the C1 gene product, suggest that myb-related plant genes code for trans-acting factors which regulate gene expression in a given biosynthetic pathway.

Amino Acid Sequence↗

Cloning of the gene for the capsid protein of potato leafroll virus.

DNA complementary to the RNA of purified potato leafroll virus (PLRV) was synthesized and cloned into the lambda insertion vector NM1149. Overlapping PLRV-specific cDNA clones were isolated that represent some 80% of the viral genome. Sequences coding for the capsid protein were identified by subcloning size-selected cDNAs into the lambda expression vector gt11 and screening with PLRV-specific antisera. The gene for the viral capsid protein was shown to reside in the 3' terminal half of the genomic RNA. Sequence comparisons with the recently published genomes of the beet western yellows virus (BWYV) and the barely yellow dwarf virus (BYDV) reveal some 65% protein sequence homology between the capsid proteins of BWYV and PLRV and some 45% homology between BYDV and PLRV. Furthermore, it is evident that the structural organization of the PLRV genome in the CP gene region is similar to that of BWYV and BYDV.

Amino Acid Sequence↗

Cloning of the potato virus Y genes encoding the capsid protein CP and the nuclear inclusion protein NIb.

DNA was synthesized complementary to the RNA genome of potato virus Y (PVY; strain GO 16) and cloned into lambda vectors. The size of PVY-specific Eco RI-restricted cDNA ranged from 0.3 to approximately 22kb. Two of the cDNA clones each of which contained some 4kb of cDNA sequence starting from the 3'-polyadenylated terminus were characterized by sequence analysis. Presence of a single open reading frame suggests that PVY-specific proteins are synthesized as a polyprotein precursor. As with other sequenced potyvirus RNAs the gene for the PVY capsid protein CP is located next to the 3'-untranslated region followed by the genes for the putative RNA polymerase (nuclear inclusion protein NIb) and the virus-specific protease (nuclear inclusion protein NIa). The 3'-trailing sequence of the PVY strains cloned is highly homologous to the corresponding region of pepper mottle virus (PeMV) and suggests that PeMV is not a distinct member of the potyvirus group, but another strain of PVY.

Amino Acid Sequence↗

Sex-specific effects on the fetal neuroendocrine system during acute stress in late pregnancy of rat and the influence of a simultaneous treatment by tyrosine.

It is well-known that prenatal chronic intermittent stress affects the reproductive system of both sexes. Investigating the effects of an acute maternal stress on the fetal neuroendocrine system, parameters such as hypothalamic catecholamines. CRF, GRF, LH-RH, beta-endorphin, hypophysial beta-endorphin and beta-LPH as well as plasma LH, corticosterone and androstenedione were measured. Pregnant rats of Wistar strain were exposed to restraint stress at day 22 of gestation or to forced immobilization at day 20 of gestation, respectively, and were sacrificed before stress and 10, 30, 60, and 120 min after starting stress. A decrease of fetal hypothalamic catecholamines and an increase of LH-RH content of the hypothalamus as well as of plasma catecholamines were observed under stress on day 22 of gestation. On day 20 of gestation hypothalamic beta-endorphin was depleted in male and unchanged in female fetuses under stress. A depletion of hypothalamic CRF was observed in male fetuses, whereas female fetuses showed an increase of hypothalamic CRF. An increase of GRF was found in fetuses of both sexes. Pituitary opioid content increased in fetuses of both sexes initially, but was depleted secondarily in male fetuses. The LH plasma level was markedly reduced in male, the corticosterone level was elevated in fetuses of both sexes as well as the androstenedione level in female fetuses. A simultaneous treatment of mother animals with tyrosine--a catecholamine precursor--prevented the depletion of hypothalamic and pituitary beta-endorphin as well as in part the reduction of plasma LH levels in male fetuses. Hypothalamic GRF content does not increase under tyrosine treatment in male fetuses, whereas in female fetuses the stress-induced increase of GRF content was rather pronounced under tyrosine than attenuated. These results indicate that fetal hypothalamic neurotransmitters and neurohormones (such as LH-RH, CRF, GRF and opioids) are involved in changing circulating hypophysial and adrenal hormones in fetuses exposed to maternal stress in late pregnancy, whereby sex-specific different pathways might be effective in fetal stress processing. The prenatal administration of tyrosine prevented at least in part--those neurohormonal changes which are affecting the sex-specific brain differentiation.

Androstenedione↗

Oestrogen priming for the positive oestrogen feedback: site of action.

The significance of oestrogen priming for efficacy of the positive, ovulation-inducing oestrogen feedback has been known for more than 15 years, but the site and mechanism of oestrogen action in the priming effect have not yet been elucidated. Long-term ovariectomized adult female rats were injected once or twice with 20 micrograms oestradiol benzoate (OB), and the serum LH concentration was estimated. Whereas a single injection of OB induced significant inhibition of LH secretion, high circulating LH levels were recorded in rats injected twice with the hormone at an interval of 48 h. This increase was prevented in ovariectomized females fitted with guide cannulae, if the antioestrogen clomiphene citrate was implanted into the medial preoptic area (MPOA) before the first injection of oestrogen and removed prior to the second. On the other hand, replacement of the first oestrogen administration by the implantation of a very low dose of OB into the MPOA resulted in stimulation of LH secretion. OB implants placed into the hypothalamic ventromedial-arcuate region were ineffective in this regard. Taken together, the findings suggest that the priming effect of oestrogen is mediated in rats, at least in part, by the MPOA.

Animals↗

Paradoxical response of plasma beta-endorphin to combined administration of TRH and GnRH in adrenal disorders.

The combined intravenous injection of TRH and GnRH elicited paradoxical responses of plasma beta-endorphin in active and successfully treated pituitary dependent Cushing's disease as well as in ectopic ACTH syndrome and in congenital adrenal hyperplasia. No response was observed in Cushing's syndrome due to adrenal tumours. It is concluded that an abnormal response to inappropriate releasing hormones cannot verify the existence of a pituitary corticotrophic microadenoma.

Adenoma↗

[The immunohistochemistry of pituitary adenomas].

A great deal of progress has been achieved in recent years in the field of immunohistochemistry of pituitary adenoma. Continued use of more new antisera and monoclonal antibodies against numerous hormones in the adenohypophysis have resulted in new approaches to classification of pituitary adenoma. However, new problems have been discovered, on the other hand, by large-scale studies in recent years. The great number of multihormonal pituitary adenomas and possible change of the immunohistochemically detectable hormone status in cases of recurrent tumours have particularly re-emphasised the need for new thinking about patterns of classification. It would appear somewhat problematic, in this context, to uncritically accept terms, such as ACTH cellular adenoma or GH cellular adenoma. Reference is also made to the distribution pattern of cell and tissue markers in pituitary adenomas. The paper is based on thorough literature screening as well as on experience obtained by the authors from 450 cases of pituitary adenoma of which 260 had been analysed by immunohistochemistry, 131 by morphometry, and 80 by electron microscopy.

Adenoma↗