Antibodies to metabotropic glutamate receptor 5 in the Ophelia syndrome.

OBJECTIVE: To report the metabotropic glutamate receptor 5 (mGluR5) as the autoantigen of antibodies from 2 patients with Hodgkin lymphoma (HL) and limbic encephalopathy (Ophelia syndrome). METHODS: Immunohistochemistry with brain tissue and cultures of rat hippocampal neurons were used to demonstrate antibodies. Immunoprecipitation, mass spectrometry, and mGluR5-null mice served to identify the antigen. HEK293 cells transfected with mGluR5 or mGluR1 were used to determine immunologic crossreactivity. RESULTS: Both patients developed symptoms consistent with limbic encephalopathy; one had MRI findings typical of this disorder and the other had more extensive radiologic involvement, including parietal and occipital cortex. Patients' sera had antibodies that predominantly reacted with the neuropil of hippocampus and cell surface of live hippocampal neurons. Immunoprecipitation from cultured neurons and mass spectrometry demonstrated that the antigen was mGluR5, a receptor involved in processes of learning and memory. The reactivity of patients' sera was abrogated in brain of mGluR5-null mice, further confirming the antibody specificity. Studies with a large number of controls including 2 patients with cerebellar ataxia and mGluR1 antibodies showed that mGluR5 was only identified by sera of the 2 patients with the Ophelia syndrome, and that despite the homology of this receptor with mGluR1 each autoantigen was specific for a distinct syndrome. CONCLUSIONS: Antibodies to mGluR5 should be considered in patients with symptoms of limbic encephalitis and HL (Ophelia syndrome). Recognition of this disorder is important because it can affect young individuals and is reversible.

Dried plum prevents bone loss in a male osteoporosis model via IGF-I and the RANK pathway.

Previously, dietary supplementation with dried plums, a rich source of polyphenolic compounds with antioxidant and anti-inflammatory properties, has been shown to improve bone density, microstructure and biomechanics in female animal models of osteopenia. We designed this study to determine the extent to which dried plum prevents skeletal deterioration in gonadal hormone deficient male animals and to begin to understand its mechanism of action. Sixty 6-month-old male Sprague-Dawley rats were either sham-operated (Sham = 1 group) or orchidectomized (ORX = 4 groups) and randomly assigned to dietary treatments: standard semi-purified diet (Control) with either LD = 5%, MD = 15%, or HD = 25% (w/w) dried plum for 90 days. At the end of the treatment period, both the MD and HD dried plum completely prevented the ORX-induced decrease in whole body, femur, and lumbar vertebra bone mineral density (BMD). Biomechanical testing indicated that the MD and HD of dried plum prevented the ORX-induced decrease in ultimate load of the cortical bone as well as the compressive force and stiffness of trabecular bone within the vertebrae. Analyses of trabecular microarchitecture of the distal femur metaphysis and vertebral body revealed that HD dried plum protected against the decrease in trabecular bone volume (BV/TV) induced by ORX. In the distal femur, all doses of dried plum improved trabecular number (TbN) and separation (TbSp) compared to the ORX-control group, while MD and HD dried plum prevented the ORX-induced changes in vertebral TbN and TbSp. At the end of the 90-day treatment, no remarkable changes in serum osteocalcin or alkaline phosphatase in any of the treatment groups were observed, while serum insulin-like growth factor (IGF)-I was increased by dried plum. The ORX-induced increase in urinary deoxypyridinoline (DPD) excretion was completely prevented by all doses of dried plum coinciding with down-regulation of gene expression for receptor activator of NFkappa-B ligand (RANKL) and osteoprotegerin (OPG) in the bone. We conclude that dried plum prevents osteopenia in androgen deficient male rats, and these beneficial effects may be attributed in part to a decrease in osteoclastogenesis via down-regulation of RANKL and stimulation of bone formation mediated by IGF-I.

Case selection in macular relocation surgery for age related macular degeneration.

BACKGROUND: To date there has been no randomised controlled trial demonstrating the safety and efficacy of macular relocation surgery (MRS) for age related macular degeneration (AMD). Vision can be improved in some patients and made worse in others despite successful surgery or because of complications. PURPOSE: To determine which patients would benefit from MRS. METHODS: Twenty nine patients with exudative AMD took part in a prospective, non-comparative, interventional study. Macular relocation surgery involved phacoemulsification, vitrectomy, 360 degrees retinotomy, excision of choroidal neovascular membrane, and macular relocation using an infusion of 5-fluorouracil and low molecular weight heparin as adjuvant to prevent proliferative vitreoretinopathy. Patients underwent protocol refraction preoperatively and six-monthly postoperatively by designated optometrists. Preoperative fundus fluorescein angiograms were read by masked observers and the lesions were classified according to a set protocol. The main outcome measures were visual improvement, final vision of better than 20/400, reading speed, critical print size. Logistic and multiple stepwise linear regressions were used to identify independent factors which predicted the main outcomes. RESULTS: Preoperative visual acuity (20/120 or worse) and lesion type (predominantly classic or submacular haemorrhage) were significantly associated with visual improvement (coefficient of regression B = 26.8, p<0.001 and B = 14.9 with p = 0.045 respectively). There were no significant independent factors which predicted a final distance logMAR visual acuity of 1.3 (20/400) or any arbitrary definition of blindness. CONCLUSIONS: The study showed that it was possible to select cases that were more likely to experience an improvement in vision following MRS.

Sodium currents in vagotomized primary afferent neurones of the rat.

1. Nodose ganglion neurones (NGNs) become less excitable following section of the vagus nerve. To determine the role of sodium currents (I(Na)) in these changes, standard patch-clamp recording techniques were used to measure I(Na) in rat NGNs maintained in vivo for 5-6 days following vagotomy, and then in vitro for 2-9 h. 2. Total I(Na) and I(Na) density in vagotomized NGNs were similar to control values. However, steady-state I(Na) inactivation in vagotomized NGNs was shifted -9 mV relative to control values (V(1/2), -74 +/- 2 vs. -65 +/- 2 mV, P < 0.01) and I(Na) activation was shifted by -7 mV (V(1/2), -21 +/- 2 vs. -14 +/- 2 mV, P < 0.006). I(Na) recovery from inactivation was also slower in vagotomized NGNs (fast time constant, 2.8 +/- 0.4 vs. 1.6 +/- 0.3 ms, P < 0.02). 3. The fraction of I(Na) resistant to 1 microM tetrodotoxin (TTX-R) was halved in vagotomized NGNs (21 +/- 8 vs. 56 +/- 8 % of total I(Na), P < 0.05). This change from TTX-R I(Na) to TTX-sensitive (TTX-S) I(Na) may explain altered I(Na) activation, inactivation and repriming in vagotomized NGNs. 4. The contribution of alterations in I(Na) to NGN firing patterns was assessed by measuring I(Na) evoked by a series of action potential (AP) waveforms. In general, control NGNs produced large, repetitive TTX-R I(Na) while vagotomized NGNs produced smaller TTX-S I(Na) that rapidly inactivated during AP discharge. We conclude that TTX-R I(Na) is important for sustained AP discharge in NGNs, and that its diminution underlies the decreased AP discharge of vagotomized NGNs.

Expression of endogenous lectins (galectins, receptors for ABH-epitopes) and the MIB-1 antigen in esophageal carcinomas and their syntactic structure analysis in relation to post-surgical tumor stage and lymph node involvement.

Squamous cell carcinomas of the esophagus, a disease with poor prognosis, are especially frequent in China and South Africa. To initiate the study of endogenous lectins in this tumor class we employed synthetic neoglycoconjugates and focused on galectins as markers. Histological sections of 43 cases of esophageal carcinomas were analyzed with labeled galectins-1 and -3 and their specific antibodies, neoglycoconjugates exposing chemically prepared histo-blood group A-, B- and H-trisaccharides and the antibody MIB-1 (Ki-67). Features of structural and numerical staining intensities determined quantitatively were correlated to clinical data sets of pTN stages, sex and age of patients. Low tumor stages (pT1/T2) were seen in 10/43 cases (23%) and 65% of the carcinomas surgically treated lacked notable lymph node involvement (pN0). The women were younger than the men (47 years versus 54 years). The proliferation activity of the tumor cells was high and amounted to 75% at average. The presence of galectin-1 and the structural entropy of distribution of staining with carrier-immobilized A-trisaccharide were associated with pN stages. These initial data indicate that distinct glycohistochemical features appear to have prognostic significance in this tumor class, adding to the emerging significance of this marker class in lung cancer.

Reduction in myocardial collagen cross-linking parallels left ventricular dilatation in rat models of systolic chamber dysfunction.

BACKGROUND: The transition from compensated left ventricular hypertrophy (LVH) to heart failure is associated with alterations in the myocardial interstitium. We hypothesized that LV dilatation is associated with modifications in collagen cross-linking. METHODS AND RESULTS: We studied 2 rat models of LV dilatation: (1) pressure-overload hypertrophy with heart failure (POH-F) induced by suprarenal abdominal aortic banding and (2) LVH induced by 7 months of isoproterenol (ISO, 0.04 mg x kg(-1) x d(-1)) administration. In POH-F rats and in rats receiving ISO, LV dilatation and a reduced systolic chamber performance were noted. Myocardial hydroxyproline concentrations ([HPRO]) were increased in the POH-F rats, whereas in rats receiving ISO, [HPRO] was decreased. In POH-F rats, the ratio of myocardial collagen type I to type III was increased, but in rats receiving ISO, myocardial collagen I/III was unchanged. In contrast to the diverse changes in myocardial collagen concentrations and phenotypes observed in the 2 models of LV dilatation, the ratio of myocardial insoluble to soluble (relationship between cross-linked and non-cross-linked) collagen was decreased in both the POH-F and ISO groups. Moreover, administration of captopril (0.22 mmol x kg(-1) x d(-1)), which inhibited the ISO-induced reduction in myocardial insoluble/soluble collagen but not the reduction in [HPRO], prevented the ISO-induced alterations in LV dimensions and performance. CONCLUSIONS: Because decreases in the ratio of myocardial insoluble to soluble collagen parallel LV dilatation in rats, reductions in myocardial collagen cross-linking may be an important mechanism contributing to LV dilatation in heart disease.

Vagotomy decreases excitability in primary vagal afferent somata.

Standard patch-clamp and intracellular recording techniques were used to monitor membrane excitability changes in adult inferior vagal ganglion neurons (nodose ganglion neurons, NGNs) 5 days following section of the vagus nerve (vagotomy). NGNs were maintained in vivo for 5 days following vagotomy, and then in vitro for 2-9 h prior to recording. Vagotomy increased action potential (AP) threshold by over 200% (264 +/- 19 pA, mean +/- SE, n = 66) compared with control values (81 +/- 20 pA, n = 68; P < 0.001). The number of APs evoked by a 3 times threshold 750-ms depolarizing current decreased by >70% (from 8.3 to 2.3 APs, P < 0.001) and the number of APs evoked by a standardized series of (0.1-0.9 nA, 750 ms) depolarizing current steps decreased by over 80% (from 16.9 APs to 2.6 APs, P < 0.001) in vagotomized NGNs. Similar decreases in excitability were observed in vagotomized NGNs in intact ganglia in vitro studied with "sharp" microelectrode techniques. Baseline electrophysiological properties and changes following vagotomy were similar in right and left NGNs. A "sham" vagotomy procedure had no effect on NGN properties at 5 days, indicating that changes were due to severing the vagus nerve itself, not surrounding tissue damage. NGNs isolated after being maintained 17 h in vivo following vagotomy revealed no differences in excitability, suggesting that vagotomy-induced changes occur some time from 1-5 days after injury. Decreased excitability was still observed in NGNs isolated after 20-21 days in vivo following vagotomy. These data indicate that, in contrast to many primary sensory neurons that are thought to become hyperexcitable following section of their axons, NGNs undergo a marked decrease in electrical excitability following vagotomy.

Nephroblastoma (Wilms' tumour) diagnosed in the Department of Anatomical Pathology Medunsa/Ga-Rankuwa Hospital for the period 1980 to 1997.

OBJECTIVE: The aim of the analysis was to describe the clinicopathological features of Wilms' Tumour (WT) diagnosed in our Department and compare results to other WT Registers. DESIGN: All cases of WT for the period 1980 to 1997 were retrieved from the Register of Renal Tumours of Childhood. SETTING: The Medical University of Southern Africa, Department of Anatomical Pathology and Department of Paediatrics, Ga-Rankuwa Hospital. SUBJECTS: A total of 171 cases of WT (97% of all renal tumours) were the subject of the analysis. RESULTS: The age of the patients ranged from four to 216 months. The two sexes were represented equally. Tumours were solid in 73.9%, cystic in 20% and mixed in 6.1%. There were as many tumours involving the right kidney as those involving the left kidney. In nine cases WT was bilateral. Only 2.4% of tumours were in stage I and II. The mass of the kidney with tumour ranged from 50 g to 5,400 g and in diameter from three to 28 cm. Histopathologically classic, blastemal and stromal type were nearly equally represented. Follow up was very inadequate and in 66% of cases the fate of the patient remains unknown. CONCLUSION: Occurrence of WT is similar to that reported from other regions of the world. Cases are in a more advanced stage than reported by SIOP and NWTS. Follow up is highly inadequate.

Lack of normal pattern of cerebral asymmetry in familial schizophrenic patients and their relatives--The Maudsley Family Study.

Lack of the normal cerebral asymmetry has been reported in schizophrenia. We wished to test the hypothesis that this lack of the normal pattern of asymmetry is familial and that it can be found in both schizophrenic and non-schizophrenic family members. In particular, we wanted to know whether those relatives who appear to be transmitting liability to the illness also demonstrate the loss of normal asymmetry. We studied families with several members affected with schizophrenia. We carried out volumetric measurements of prefrontal, premotor, sensorimotor and occipitoparietal regions in each hemisphere using 3D reconstructed MRI images in 29 schizophrenic patients, 55 of their first degree relatives, and 39 unrelated control subjects on contiguous thin slices of the brain. Nine of the unaffected relatives appeared to be transmitting the liability for schizophrenia (e.g. the mother of a schizophrenic patient who, although not psychotic herself, had a schizophrenic parent or sibling). We termed them presumed obligate carriers and the remaining 46 relatives presumed non-obligate carriers. The healthy control subjects showed larger right than left prefrontal regions and larger left than right sensorimotor and occipitoparietal regions. The schizophrenic patients showed lack of this normal brain asymmetry in the prefrontal, sensorimotor and occipitoparietal cortical regions. The presumed obligate carriers were similar to the schizophrenic patients in exhibiting lack of asymmetries in these cortical regions, while the presumed non-obligate relatives showed lack of asymmetry only in the occipitoparietal region. There was no overall reduction in total or regional brain volumes among the groups. Our findings indicate that lack of the normal pattern of frontal and occipital asymmetry is a marker for genetic liability to schizophrenia in families multiply affected with schizophrenia.

Carcinoma of the uterine cervix: results of Ka-Ngwane screening programme and comparison between the results obtained from urban and other unscreened rural communities.

OBJECTIVE: The Ka-Ngwane screening programme was initiated by the Department of Anatomical Pathology and the Department of Community Health at Medunsa to establish the incidence of cervical intraepithelial neoplasia (CIN) and cervical carcinoma (CaCx) in previously unscreened rural population in our catchment area. STUDY DESIGN: Ten thousand consecutive PAP smears from Ka-Ngwane (Mpumalanga) area are the subject of the analysis. The incidence of CIN I-CIN III and CaCx is calculated in the screened material. The changes are related to age, age of first pregnancy and parity of the patients. The incidence of cervical abnormalities is compared with 20,000 consecutive cases from Pretoria and 18,000 consecutive cases from previously unscreened rural population of Transkei. RESULTS: Positive cases consisted of three per cent in Ka-Ngwane, five per cent in Greater Pretoria area and of more than six per cent in Transkei. CaCx constituted 12.35% of all positive cases for Ka-Ngwane, 4.8% for urban population and 26% of Transkei positive cases. More than 75% of positive cases in Ka-Ngwane were below 40 years of age and five cases of CaCx were found in the 21-30 year age group. In Pretoria more than 80% of positive cases were younger than 41 years. In the material from Transkei similarly 80% of the positive cases were younger than 41 years. CONCLUSION: The screening programme confirmed the high incidence of CIN and CaCx in previously unscreened population and an urgent need to develop educational programme which will facilitate early detection, proper treatment and follow up of the cases with cervical pathology. CaCx was found at a younger age than in the developed countries which indicated that screening programmes in our environment should involve all adult females from the age of 25 years.

Spectroscopic analysis of fire suppressants and refrigerants by laser-induced breakdown spectroscopy.

Laser-induced breakdown spectroscopy is evaluated as a means of detecting the fire suppressants CF(3)Br, C(3)F(7)H, and CF(4) and the refrigerant C(2)F(4)H(2). The feasibility of employing laser-induced breakdown spectroscopy for time- and space-resolved measurement of these agents during use, storage, and recharge is discussed. Data are presented that demonstrate the conditions necessary for optimal detection of these chemicals.

Brain changes in schizophrenia. Volumetric MRI study of families multiply affected with schizophrenia--the Maudsley Family Study 5.

BACKGROUND: Structural brain abnormalities have been reported in schizophrenia. We tested the hypothesis that these abnormalities represented a marker for the genetic liability to schizophrenia in a sample of people with schizophrenia and their relatives from families multiply affected with the disorder. METHOD: We compared 31 people with schizophrenia, 57 relatives and 39 unrelated control subjects. Volumetric measurement of brain structures was carried out using stereological principles from three-dimensional reconstructed magnetic resonance images. RESULTS: Subjects with schizophrenia had larger lateral ventricles than their relatives and the normal control subjects. Relatives who were 'presumed obligate carriers' had larger left lateral ventricles than other relatives and the control subjects. Subjects with schizophrenia showed smaller whole brain and cerebellar volumes and larger lateral ventricles than their age- and gender-matched unaffected siblings. CONCLUSIONS: In families multiply affected with schizophrenia lateral ventricular enlargement distinguishes people with schizophrenia and presumed obligate carriers from other relatives and unrelated control subjects. These changes may be a marker for a genetic liability to schizophrenia.

Bilateral juvenile granulosa--cell tumour with multiple metastases. A case report.

We represent the case of premenarchal 14 year old girl complaining of a four month history of painful enlargement of both breasts. The patient died 10 days after admission due to respiratory distress. The post mortem diagnosis: bilateral juvenile granulosa-cell tumour with widespread metastases.

Site of opioid action in the human brain: mu and kappa agonists' subjective and cerebral blood flow effects.

OBJECTIVE: Humans experience the subjective effects of mu and kappa opioid agonists differently: mu agonists produce mainly euphoria, while kappa agonists are more likely to produce dysphoria. This study tested the hypothesis that these subjective effects would be associated with anatomically distinct changes in regional cerebral blood flow (CBF) relative to baseline as assessed with single photon emission computed tomography (SPECT). METHOD: Nine nondependent opioid abusers participated in the study. In the first phase of the study, the participants were acclimated to effects of the study drugs. In the second phase they underwent repeat challenges with the study drugs followed by an assessment of CBF with use of the SPECT tracer [99mTc]HMPAO. Medications tested were the prototypic mu agonist hydromorphone, the mixed agonist/antagonist butorphanol (which has a kappa agonist component of activity), and saline placebo. RESULTS: Subjective effects of the drugs were distinctly different. Hydromorphone produced increased ratings of "good effects," while butorphanol led to more "bad effects." Hydromorphone significantly increased regional CBF in the anterior cingulate cortex, both amygdalae, and the thalamus--all structures belonging to the limbic system. Butorphanol caused a less distinct picture of regional CBF increases, mainly in the area of both temporal lobes. CONCLUSIONS: This study demonstrates that opioids with different subjective effects also produce statistically significant patterns of change in regional CBF from baseline, and the regions of statistical significance appear in different brain regions. In addition, these results demonstrate the applicability of SPECT functional neuroimaging in the study of medications with potential abuse liability.

Proliferative activity of normal and neoplastic colonic mucosa in population groups with high and low risk for colorectal carcinoma.

Europeans have a high incidence of colorectal cancer in comparison to Africans. Lack of detectable sequence adenoma-colorectal carcinoma in Africans may suggest the development of adenocarcinoma is de novo. The aim of this study is to assess colonic mucosal proliferative activity in various pathological conditions of diverse population groups. Materials included routinely processed tissue specimens from consecutively resected well- and moderately differentiated colorectal adenocarcinomas from 32 rural Africans (South Africa) and 27 urban Europeans (Poland) and from apparently normal rectal mucous membrane from the age and sex matched each population group (28 and 25 samples respectively). In addition, 32 resected adenomatous polyps were examined in Europeans as well. The MIB 1 monoclonal antibody was used to assay the expression of Ki67 antigen in routinely processed tissue specimens. Proliferative activity in colonic carcinomas was scored by the percentage of positively stained cells. Labelling indices were estimated in 5 crypt compartments in apparently normal colonic mucosa adjacent and distant to the tumour, in mucosal samples of controls from both population groups and in adenomatous polyps from Europeans. The mean age of African patients with adenocarcinoma was markedly lower than in European counterparts (48.6 yrs vs. 66.4 yrs). The overall proliferative activity in cancerous tumours of Africans was higher than in Europeans. The labelling indices were lower in all compartments in normal colonic mucosa in Africans. Overall increase of the labelling indices in adjacent and distant to the tumour mucosa noted when compared to the mucosa of healthy individuals. No such differences were detected between indices in the mucosa adjacent and the mucosa distant to the tumour. Proliferative activity in the mucosa adjacent to adenoma was also higher than in normal mucosa from healthy individuals. Adenomas with marked dysplasia showed higher and diffuse proliferative activity, when regular adenomas shown superficial labelling only. Relatively young age, lack of detectable evidence of adenoma-carcinoma sequence and low proliferative activity in all compartments of mucosa from healthy individuals indicate different etiopathogenesis of colorectal carcinoma in rural Africans.

Distribution and activation of intracellular Ca2+ stores in cultured olfactory bulb neurons.

The presence and distribution of intracellular Ca2+ release pathways in olfactory bulb neurons were studied in dissociated cell cultures. Histochemical techniques and imaging of Ca2+ fluxes were used to identify two major intracellular Ca2+ release mechanisms: inositol 1, 4,5-triphosphate receptor (IP3R)-mediated release, and ryanodine receptor-mediated release. Cultured neurons were identified by immunocytochemistry for the neuron-specificmarker beta-tubulin III. Morphometric analyses and immunocytochemistry for glutamic acid-decarboxylase revealed a heterogeneous population of cultured neurons with phenotypes corresponding to both projection (mitral/tufted) and intrinsic (periglomerular/granule) neurons of the in vivo olfactory bulb. Immunocytochemistry for the IP3R, and labeling with fluorescent-tagged ryanodine, revealed that, irrespective of cell type, almost all cultured neurons express IP3R and ryanodine binding sites in both somata and dendrites. Functional imaging revealed that intracellular Ca2+ fluxes can be generated in the absence of external Ca2+, using agonists specific to each of the intracellular release pathways. Local pressure application of glutamate or quisqualate evoked Ca2+ fluxes in both somata and dendrites in nominally Ca2+ free extracellular solutions, suggesting the presence of IP3-dependent Ca2+ release. These fluxes were blocked by preincubation with thapsigargin and persisted in the presence of the glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione. Local application of caffeine, a ryanodine receptor agonist, also evoked intracellular Ca2+ fluxes in the absence of extracellular Ca2+. These Ca2+ fluxes were suppressed by preincubation with ryanodine. In all neurons, both IP3- and ryanodine-dependent release pathways coexisted, suggesting that they interact to modulate intracellular Ca2+ concentrations.