Spiritual dimensions of a mind-body group for people with severe mental illness.

In a mind-body group designed to address issues of well-being for people with severe mental disorders, experiences with spiritual themes of optimal functioning and ultimate meaning emerged with surprising clarity.

Design of the EPC-9, a computer-controlled patch-clamp amplifier. 2. Software.

The EPC-9 computer-controlled amplifier has no front-panel controls; therefore the user interface to the EPC-9 patch-clamp amplifier is defined entirely by software. This paper describes various user interfaces that have been implemented, including a high-level programming interface, a user interface based on the PostScript language, and graphical user interfaces that control the EPC-9 from data-acquisition programs. Also described are the algorithms used for automatic adjustment of the C-Fast and C-Slow transient cancellation circuitry. An overview of the procedures that perform automatic testing and calibration is given.

[Drug-induced (probably allopurinol) agranulocytosis. Case report and discussion]. / Medikamentöse, wahrscheinlich Allopurinol-induzierte Agranulozytose. Fallbeschreibung und Diskussion.

Agranulocytosis is a rare but sometimes extremely dangerous adverse drug reaction which can be induced by almost any drug. We report the case of a 89-year-old man with a well-documented granulocyte chart, who received allopurinol in addition to preexisting therapy with cardiovascular drugs. Three weeks later agranulocytosis was found which bone-marrow biopsy indicated was drug-induced. After cessation of all drugs, isolation and antibiotic therapy, the leukocyte count returned to normal but the patient died four weeks later from progressive renal failure. A relation between allopurinol therapy and agranulocytosis was presumed. The drugs which might have caused this adverse reaction are discussed. The incidence, signs, symptoms and treatment are summarized and proposals are made concerning the action to be taken in the event of drug-induced agranulocytosis.

Nerve growth factor (NGF) regulates tachykinin gene expression and biosynthesis in rat sensory neurons during early postnatal development.

The regulatory effects of nerve growth factor (NGF) on tachykinin biosynthesis in rat primary sensory neurons during the period of postnatal development were examined under in vivo and in vitro conditions. Administration of NGF to neonatal rats led to a significant increase in protein levels of substance P (SP) and neurokinin A (NKA) in trigeminal and dorsal root ganglia (DRG). In addition, Northern blot analysis revealed that preprotachykinin mRNA was upregulated in sensory ganglia of neonatal animals after treatment with NGF. Using a well-defined in vitro system for neonatal rat DRG and trigeminal ganglia neurons, we found that addition of NGF induced SP and NKA protein levels in a dose-dependent manner. Furthermore, preprotachykinin mRNA was markedly increased in cultured DRG and trigeminal ganglia neurons in the presence of NGF. Thus, our results clearly demonstrate that NGF regulates tachykinin gene expression and biosynthesis both in vivo and in vitro during the developmental period of rat sensory neurons.

[Acral vasospasm in Crohn's disease]. / Akrale Vasospasmen bei M. Crohn.

55 patients with Crohn's disease and 55 sex- and age-matched healthy controls were studied by nailfold capillary microscopy for evidence of microcirculatory abnormalities. Digital capillary blood flow measurements in combination with a local cooling test were assessed by videocapillaroscopy using the flying-sport technique. We found a blood flow stop with cold exposure in 44 of the 55 patients with Crohn's disease (mean duration 39 seconds) but in 5 of the 55 control subjects only (mean duration 19 seconds). There was no significant difference in skin temperature between the two groups. Intravital microscopy of nailfold capillaries revealed a marked reduction in capillary density (p less than 0.001) compared to normal subjects. The association of vasoconstrictive reaction in finger microcirculation of patients with Crohn's disease suggests a vasospastic tendency in this disorder.

The axonally secreted cell adhesion molecule, axonin-1. Primary structure, immunoglobulin-like and fibronectin-type-III-like domains and glycosyl-phosphatidylinositol anchorage.

Axonin-1 is an axon-associated cell adhesion molecule (AxCAM) of the chicken, which promotes neurite outgrowth by interaction with the AxCAM L1(G4) of the neuritic membrane. Here we report the cloning and sequence determination of a cDNA encoding axonin-1. Peptides generated by enzymatic cleavage showed similarity to the AxCAM F11. Degenerated polymerase chain reaction (PCR) primers were designed and an axonin-1 fragment was amplified from mRNA of embryonic retina. Screening of a cDNA library from embryonic brain resulted in the isolation of a 4.0-kb cDNA insert with an open reading frame of 3108 nucleotides. The deduced polypeptide of 1036 amino acids includes a putative hydrophobic N-terminal signal sequence of 23 or 25 amino acids and a C-terminal hydrophobic sequence of 29 amino acids which is suggestive of sequences serving as signal for the attachment of a glycosyl-phosphatidylinositol (glycosyl-PtdIns) anchor. The putative mature form of axonin-1 comprises six immunoglobulin-like repeats, followed by four fibronectin-type III repeats. Axonin-1 exhibits 75% amino acid identity with the AxCAM TAG-1 of the rat, suggesting that it is the chicken homologue of TAG-1. Like TAG-1, axonin-1 is glycosyl-PtdIns-anchored to the neuronal membrane; in contrast to TAG-1, it does not exhibit an Arg-Gly-Asp sequence.

The role of nailbed vasospasm in Crohn's disease.

Local cooling of the nailfold capillaries produces a typical blood flow stop reaction in patients with Raynaud's phenomenon. We applied the same test to 35 patients with Crohn's disease and compared the results with the findings in a control group of 35 healthy subjects matched for age and sex by nailfold videomicroscopy. We found a flow stop with cold exposure in 27 of the 35 patients with Crohn's disease (mean duration 38 s) in contrast to 2 of the 35 control subjects only (mean duration 12 s). There was no significant difference in skin temperature between the two groups. Intravital microscopy of nailfold capillaries revealed an evident reduction of the capillary density (p = 0.001) and crest diameter of the capillary loop (p = 0.02) compared to normal subjects. The strong association of a vasoconstrictive reaction in finger microcirculation in patients with Crohn's disease suggests a vasospastic tendency in this disorder that might manifest in the mesenteric capillaries also. This new finding of microcirculatory disturbance may play a role in the pathophysiology of Crohn's disease and be related to a general vasospastic syndrome.

[Fecal occult blood--does a hemoglobin-specific tests improve the diagnosis of relevant colonic neoplasms?]. / Okkultes Blut im Stuhl--Verbessert ein hämoglobinspezifischer Test die Diagnostik relevanter Kolon-Neoplasien?

The object of this study was to assess whether an immunological test specific for human hemoglobin improves the detection of bleeding from colorectal tumors as compared to a conventional guaiac test for occult fecal blood. We examined one stool sample from 134 patients prior to a complete colonoscopy by the Colo-Immun-Test (CI) as well as the Colo-Rectal-Test (CR). CI proved to have a higher sensitivity for occult bleeding from all colonic lesions, but this difference was not statistically significant. There was, however, no difference in the sensitivity of the two tests in detecting bleeding from relevant colonic neoplasms. In this study the immunological CI did not improve the detection of occult fecal blood loss due to adenomas and carcinomas of the colon in comparison with CR.

Expression of substance P and preprotachykinin mRNA by primary sensory neurons in culture: regulation by factors present in peripheral and central target tissues.

Primary sensory neurons display various neuronal phenotypes which may be influenced by factors present in central or peripheral targets. In the case of DRG cells expressing substance P (SP), the influence of peripheral or central targets was tested on the neuronal expression of this neuropeptide. DRG cells were cultured from chick embryo at E6 or E10 (before or after establishment of functional connections with targets). Preprotachykinin mRNA was visualized in DRG cell cultures by either Northern blot or in situ hybridization using an antisense labeled riboprobe, while the neuropeptide SP was detected by immunostaining with a monoclonal antibody. In DRG cell cultures from E10, only 60% of neurons expressed SP. In contrast, DRG cell cultures performed at E6 showed a significant hybridization signal and SP-like immunoreactivity in virtually all the neurons (98%). The addition of extracts from muscle, skin, brain or spinal cord to DRG cells cultured at E6 reduced by 20% the percentage of neurons which express preprotachykinin mRNA and SP-like immunoreactivity. Our results indicate that factors issued from targets inhibit SP-expression by a subset of primary sensory neurons and act on the transcriptional control of preprotachykinin gene.

Carboxypeptidase H in the hypothalamo-neurohypophysial system: evidence for processing and activation of a prohormone-processing enzyme during axonal transport.

Investigations of peptide precursor processing in nerve cells, including studies on prooxytocin and provasopressin processing in the rat hypothalamo-neurohypophysial system, show that prohormone processing occurs during axonal transport of maturing secretory vesicles. Recent studies (Fricker et al., 1989; Rodriguez et al., 1989) show that carboxypeptidase H (CPH), one of several proteases required for prohormone processing, is synthesized as a proenzyme that presumably requires activation. To determine if pro-CPH, like prohormone precursors, is processed and activated during axonal transport, we have analyzed the molecular forms of CPH present at several levels in the rat hypothalamo-neurohypophysial system. These biochemical and immunochemical studies showed that the supraoptic nucleus (SON), a region enriched in neuronal cell bodies, possesses primarily an inactive 65-kDa species of CPH. The median eminence and pituitary stalk regions that are enriched in axons possess both the inactive 65-kDa and the active 55-kDa forms of CPH, and nerve terminals of the posterior pituitary contain primarily the active 55-kDa CPH. These results support the hypothesis that pro-CPH is processed and activated during axonal transport from neuronal perikarya of SON to nerve terminals of the posterior pituitary. Furthermore, analysis of immunoreactive CPH in the rat and bovine pituitary showed that each tissue possessed different relative amounts of zymogen compared to mature forms of CPH, suggesting that tissue-specific processing of pro-CPH occurs. Thus, the biosynthesis of active peptide hormones requires the simultaneous processing of proenzyme and prohormone.

Cleavage of recombinant enkephalin precursor by endoproteolytic activity in bovine chromaffin granules.

To identify endoproteolytic activity that processes the enkephalin precursor, a novel approach was undertaken for the production of model substrate in the form of recombinant 35S-(Met)-preproenkephalin (35S-(Met)-PPE), generated by in vitro transcription and translation of the rat PPE cDNA. Endoproteolytic activity in bovine chromaffin granules cleaved 35S-(Met)-PPE with a pH optimum of 4.5 and generated multiple products containing the NH2-terminal segment of the precursor. Processing of 35S-(Met)-PPE, as well as endogenous enkephalin intermediates, was inhibited by pepstatin A and stimulated by DTT. These results suggest that aspartyl and thiol proteolytic activity(ies) are involved in cleaving the enkephalin precursor.

Identification of zymogen and mature forms of human carboxypeptidase H. A processing enzyme for the synthesis of peptide hormones.

Carboxypeptidase H (CPH) is one of several processing enzymes required for the conversion of peptide hormone precursors into their smaller active forms. In this study, high levels of CPH activity was found in a liver metastasis of a human ileal carcinoid which expresses beta-preprotachykinin mRNA and the tachykinin neuropeptides, substance P and substance K. This human CPH showed properties of a zinc-metallopeptidase that is structurally similar to bovine and rat CPH. Immunoblots of the human ileal carcinoma with anti-bovine CPH showed that CPH activity is represented by two proteins of apparent molecular masses 57 and 55 kDa. Cell-free translation of poly(A)+ RNA followed by immunoprecipitation with anti-bovine CPH showed that human CPH mRNA encodes a precursor protein of apparent molecular mass 75 kDa. These data demonstrate that human CPH is synthesized as a zymogen, prepro-CPH, which must be cleaved to form catalytically active CPH.

[Crohn's disease-- 10 years' experience at a city hospital]. / Morbus Crohn--Erfahrungen eines Stadtspitals über 10 Jahre.

Between 1977 and 1986, 92 patients with Crohn's disease (53 females and 39 males, average age 32 years at diagnosis) were admitted for a total of 158 hospital stays at the St. Claraspital, Basel. In 71 cases the diagnosis was based on one or more of radiologic and endoscopic evaluation and/or operative findings. In 4 cases diagnosis was based solely on clinical findings and in the remaining 17 the original diagnostic workup could not be evaluated retrospectively. 65% of cases were histologically proven, while the rest of the patients were either not biopsied or biopsy yielded inconclusive results. Crohn's disease was located simultaneously in the small and large bowels in 41 patients, in the small bowel only in 23 (25%) and in the large bowel only in 20 (22%). The remaining localizations were either proximal GI tract or anal region only. Surgery was performed a total of 65 times in 44 of the 92 patients (48%), the most frequent procedures being ileocecal resection and surgical treatment of perianal fistulas and abscesses. Accordingly, the main indications for surgery were perianal complications (37%) and stenotic bowel lesions (29%). The resection rate was higher in ileal (35%) than in colorectal (25%) involvement. The 3 rectal amputations eventually necessary were all in patients with anorectal involvement.

[Campylobacter pylori, gastritis and ulcer disease. Microbiological, histological and serological studies]. / Campylobacter pylori, Gastritis und Ulkuskrankheit. Mikrobiologische, histologische und serologische Untersuchungen.

In a prospective study of 70 patients with epigastric pain, gastroduodenoscopy revealed gastric and/or duodenal ulcers in a total of 41 and no ulceration in the remainder. Biopsies were taken to assess the severity of gastritis and the presence of Campylobacter pylori (CP) by histology and culture. Gastritis was found in 54 patients. CP was detected in 78% of the ulcer patients and 52% of the patients without ulcer (p less than 0.05). CP was demonstrated in 83% of the histologically diagnosed cases of gastritis (all grades) but no CP was detectable in patients with normal gastric mucosa. Among the ulcer patients, CP was more frequent in those with no history of medication with nonsteroidal antiinflammatory drugs (p less than 0.01). Sera from CP-positive ulcer and gastritis patients have significantly higher levels of IgG antibodies to CP than sera from those found to be free of ulcer or gastritis. In 200 blood donors an increasing percentage of elevated CP-antibody titres were found with advancing age (50% over 60 years of age). Quantification of circulating CP antibodies, would thus seem a valuable adjunct in the diagnosis of gastritis and probably also of peptic ulcer. The data presented furnish further evidence of the high rate of association of CP and the gastritis-peptic ulcer complex.

A cDNA encoding the precursor of the rat neuropeptide, neurokinin B.

We have isolated a cDNA clone from a rat cerebral cortex library which encodes the 116 amino acid precursor of the neuropeptide, neurokinin B. The precursor has 68% amino acid homology to the bovine precursor and encodes a single peptide of the tachykinin family. Except for possible small variations at both ends of the message, there appears to be only a single species of neurokinin B mRNA in rat cerebral cortex. In situ hybridization histochemistry indicates that the message is widely distributed in the rat brain in a pattern distinct from that of substance P message.

Comparative distribution of mRNAs for glutamic acid decarboxylase, tyrosine hydroxylase, and tachykinins in the basal ganglia: an in situ hybridization study in the rodent brain.

Neurotransmitter-related messenger RNAs were detected by in situ hybridization in sections of rat and mouse brains by using 35S-radiolabelled RNA probes transcribed from cDNAs cloned in SP6 promoter-containing vectors. The distribution of messenger RNAs for glutamic acid decarboxylase, tachykinins (substance P and K), and tyrosine hydroxylase was examined in the striatum, pallidum, and substantia nigra. Dense clusters of silver grains were observed with the RNA probe complementary of the cellular messenger RNA for glutamic acid decarboxylase (antisense RNA) over most large neurons in the substantia nigra pars reticulata and medium-sized to large neurons in all pallidal subdivisions. A few very densely and numerous lightly labelled medium-sized neurons were present in the striatum. Among the areas examined, only the striatum contained neurons labelled with the antisense tachykinin RNA. Most of these neurons were of medium size, and a few were large. With the antisense tyrosine hydroxylase RNA, silver grains were found over neurons of the substantia nigra pars compacta and adjacent A10 and A8 dopaminergic cell groups. No signal was observed with RNAs identical to the cellular messenger RNA for glutamic acid decarboxylase or tachykinin (sense RNA). These results show a good correlation with immunohistochemical studies, suggesting that documented differences in the distribution and the level of glutamic acid decarboxylase, tyrosine hydroxylase, and substance P immunoreactivities in neurons of the basal ganglia are related to differences in the level of expression of the corresponding genes rather than to translation accessibility, stability, or transport of the gene products.

Preprotachykinin messenger RNA detected by in situ hybridization in striatal neurons of the human brain.

Sections from postmortem human brain were processed for in situ hybridization histochemistry using a 35S-labelled RNA probe transcribed from a cDNA coding for the human preprotachykinin which contains both substance P and K. Labelled neurons were observed in the caudate nucleus and the putamen but not the cerebellum. The labelled cells were of medium size and their distribution and morphology were compatible with previous data on substance P-like immunoreactivity in the human brain. The results confirm the presence of preprotachykinin mRNA in a subpopulation of striatal neurons in the human and show that in situ hybridization can be used to detect specific neurotransmitter-related mRNAs in postmortem tissue from normal and diseased humans.

Hormone receptor regulated proopiomelanocortin gene expression.

The activity of the POMC gene is regulated by both stimulatory and inhibitory hormones. Presumably, some balance exists in the influence of these hormones in order to maintain a steady-state level of activity of this gene. Physiological insults, such as stress, may upset this balance and change the rate of production of POMC and its biologically active peptides. The relative strength of these different hormones may therefore determine the long-term expression of this gene. Chronic administration of CRF to rats, primates and humans produces prolonged increases in plasma ACTH levels. This long-term effect is most likely due to an activation of the POMC gene. Interestingly, chronic treatment of anterior pituitary cells with CRF desensitizes CRF receptors. Thus, despite the corticotrophs becoming refractory to the acute stimulatory actions of CRF, the POMC gene remains stimulated. These findings suggest that corticotrophs do not have to be continuously activated by CRF to maintain a long-term increase in POMC gene expression. This contrasts with the actions of glucocorticoids whose effects are abruptly terminated following their removal from the target tissue. The molecular basis of this form of cellular memory to the actions of CRF may involve cAMP regulatory phosphoproteins binding to and activating the POMC gene. If this phenomenon is shown to occur and the phosphorylation state of these nuclear proteins is found to govern their level of interaction with POMC gene, then it would represent a novel mechanism of gene regulation. Proof for this mechanism and the elucidation of how other second messengers such as protein kinase C and calcium regulate the POMC gene will greatly aid our understanding of the precise molecular mechanisms controlling opioid peptide expression.