Defective WNT signaling associates with bone marrow fibrosis-a cross-sectional cohort study in a family with WNT1 osteoporosis.

This study explores bone marrow function in patients with defective WNT1 signaling. Bone marrow samples showed increased reticulin and altered granulopoiesis while overall hematopoiesis was normal. Findings did not associate with severity of osteoporosis. These observations provide new insight into the role of WNT signaling in bone marrow homeostasis. INTRODUCTION: WNT signaling regulates bone homeostasis and survival and self-renewal of hematopoietic stem cells. Aberrant activation may lead to osteoporosis and bone marrow pathology. We aimed to explore bone marrow findings in a large family with early-onset osteoporosis due to a heterozygous WNT1 mutation. METHODS: We analyzed peripheral blood samples, and bone marrow aspirates and biopsies from 10 subjects with WNT1 mutation p.C218G. One subject was previously diagnosed with idiopathic myelofibrosis and others had no previously diagnosed hematologic disorders. The findings were correlated with the skeletal phenotype, as evaluated by number of peripheral and spinal fractures and bone mineral density. RESULTS: Peripheral blood samples showed no abnormalities in cell counts, morphology or distributions but mild increase in platelet count. Bone marrow aspirates (from 8/10 subjects) showed mild decrease in bone marrow iron storages in 6 and variation in cell distributions in 5 subjects. Bone marrow biopsies (from 6/10 subjects) showed increased bone marrow reticulin (grade MF-2 in the myelofibrosis subject and grade MF-1 in 4 others), and an increase in overall, and a shift towards early-phase, granulopoiesis. The bone marrow findings did not associate with the severity of skeletal phenotype. CONCLUSIONS: Defective WNT signaling associates with a mild increase in bone marrow reticulin and may predispose to myelofibrosis, while overall hematopoiesis and peripheral blood values are unaltered in individuals with a WNT1 mutation. In this family with WNT1 osteoporosis, bone marrow findings were not related to the severity of osteoporosis.

Degenerative cervical spine changes among early career fighter pilots: a 5-year follow-up.

INTRODUCTION: Degenerative changes of the cervical spine often cause disability and flight duty limitations among Finnish Air Force (FINAF) fighter pilots. We aimed to study the effect of +Gz exposure on degenerative changes in the cervical spine by comparing cervical MRIs of FINAF fighter pilots and controls. METHODS: At baseline, the volunteer study population consisted of 56 20-year-old FINAF male fighter pilots (exposure group) and 56 21-year-old Army and Navy cadets (control group). Both groups underwent MRI of the cervical spine at the baseline and after 5 years. Degenerative changes evaluated using MRI included intervertebral disc (IVD) degeneration (Pfirrmann classification), disc herniations, uncovertebral arthrosis, Schmorl's nodes, Modic changes, spinal canal stenosis, kyphosis and scoliosis. RESULTS: The degree of IVD degeneration in the whole cervical spine increased significantly in both populations with no between-group differences. The prevalence of disc herniations also tended to increase in both populations with no difference in the incidence over the follow-up. However, pilots proved to have more disc herniations at the baseline and at the follow-up. There were virtually no between-group differences in other assessed degenerative changes. DISCUSSION: We found that IVD degeneration and the prevalence of disc herniations increased at a similar rate for fighter pilots and non-flying military students when all cervical levels were summed up. The lack of difference may be explained by the relatively low cumulative +Gz exposure during the first 5 years of a pilots' career.

Proliferative and anti-apoptotic activity of esophageal mucosa in gastroesophageal reflux disease is not affected by fundoplication: a 4-year follow-up study.

BACKGROUND: The role of fundoplication in the prevention of esophageal adenocarcinoma is controversial. Development of cancer is associated with proliferation and anti-apoptosis, for which little data exist regarding their response to fundoplication. METHODS: Ki-67 and Bcl-2 expression was assessed in the esophagogastric junction (EGJ) and the distal and proximal esophagus of 20 patients with gastroesophageal reflux disease (GERD) treated by fundoplication and in 7 controls. Endoscopy was performed preoperatively and 6 (20 patients) and 48 months (16 patients) postoperatively. RESULTS: There were positive correlations between Ki-67 and Bcl-2 levels in the EGJ (p > 0.001) and in the distal (p = 0.001) and proximal esophagus (p = 0.013). Compared to the preoperative level, Ki-67 expression was elevated in the distal (p = 0.012) and proximal (p = 0.007) esophagus at 48 months. In addition, compared to control values, Ki-67 expression was lower at the 6-month follow-up in the EGJ (p = 0.037) and the proximal esophagus (p = 0.003), and higher at the 48-month follow-up in the distal esophagus (p = 0.002). Compared to control values, Bcl-2 was lower at 6 months in the EGJ (p = 0.038). CONCLUSIONS: Proliferative activity after fundoplication increased in the long term in the distal esophagus despite a normal fundic wrap and healing of GERD.

Indoleamine 2,3-dioxygenase expression is associated with chronic rhinosinusitis with nasal polyps and antrochoanal polyps.

Chronic rhinosinusitis without and with nasal polyps (CRSwNP and CRSsNP), and antrochoanal polyps are different phenotypes with different pathomechanisms. Indoleamine 2,3-dioxygenase (IDO) is an enzyme expressed in many cells involved in the catabolism of the essential amino acid tryptophan to kynurenine. IDO might have a role in allergic airway inflammation. The aim was to evaluate if IDO expression is associated with CRSsNP, CRSwNP, or ACP. One hundred fifty specimens from the nasal cavity and sinus mucosa were immunohistochemically stained with mAb anti-IDO. The expression of epithelial and leukocyte IDO was associated with CRSwNP and ACP. The presence of ASA intolerance, asthma, atopy, smoking and use of medication did not significantly change the results. The different expression of IDO could putatively indicate the differences in the pathomechanisms of CRSsNP, CRSwNP and ACP. Further studies on the role of IDO in upper airways pathologies are required.

Detection of enteroviruses in the intestine of type 1 diabetic patients.

Enterovirus infections have been diagnosed more frequently in type 1 diabetic patients than in the healthy population, and enteroviruses have also been found in the pancreas of diabetic patients. Primary replication of the virus occurs in the gut, but there are no previous studies evaluating possible presence of virus in the intestine of diabetic patients. The purpose of this study was to investigate if enteroviruses can be found in small intestinal tissue of type 1 diabetic patients. Formalin-fixed, paraffin-embedded upper intestinal biopsy samples were analysed for the presence of enterovirus using in situ hybridization and immunohistochemistry. Enterovirus was detected by in situ hybridization in six (50%) of the type 1 diabetic patients (n = 12) but in none of the control subjects (n = 10, P = 0.015). Immunohistochemistry identified enterovirus in nine (75%) of the patients and one (10%) control subject (P = 0.004). The presence of the virus was confirmed by reverse transcription-polymerase chain reaction in one of the four patients from whom a frozen and unfixed sample was available. Intestinal morphology was normal in all study subjects. The results suggest that a substantial proportion of type 1 diabetic patients have an ongoing enterovirus infection in gut mucosa, possibly reflecting persistent enterovirus infection. This observation opens new avenues for further studies on the possible role of enteroviruses in human type 1 diabetes.

Effects of nutrients, herbivory, and depth on the macroalgal community in the rocky sublittoral.

We studied the interacting roles of nutrient availability and herbivory in determining the macroalgal community in a rocky littoral environment. We conducted a factorial field experiment where we manipulated nutrient levels and herbivory at two sublittoral depths and measured macroalgal colonization and the following young assemblage during the growing season. At the community level, grazing reduced algal colonization, though the effect varied with depth and its interaction with nutrient availability varied in time. In shallow water, the total density of macroalgae increased in response to nutrient enrichment, but the ability of grazers to reduce macroalgal density also increased with the nutrient enrichment, and thus, the community could not escape from the top-down control. In deep water, the algal density was lower, except in July when nutrient enrichment caused a very dense algal growth. Grazing at the greater depth, though effective, was generally of smaller magnitude, and in July it could not limit algal recruitment and growth. Species richness peaked at the intermediate nutrient level in deep but not in shallow water during most of the growing season. Grazing had no effect on diversity of the algal community at either depth and only a minor effect on species richness at the greater depth. Opportunistic and ephemeral algae benefited from the nutrient enrichment but were also grazed to very low densities. Slowly growing and/or perennial species colonized poorly in the nutrient enriched treatments, and depending on the species, either suffered or indirectly benefited from herbivory. For all species, effects of nutrients on colonization depended on depth; usually both nutrient and herbivory effects were more pronounced at the shallow depth. We conclude that grazers are able to reduce macroalgae over a large range of nutrient availabilities, up to 12-fold nutrient enrichment in the current experiment, and that the sublittoral depth gradient generates variation in the algal community control exerted by both herbivory and nutrient availability. Thus temporal and spatial variability in both top-down and bottom-up control and in their interaction, especially along the depth gradient, may be crucially important for producer diversity and for the successional dynamic in a rocky sublittoral environment.

Oral induction and consolidation of acute myeloid leukemia with etoposide, 6-thioguanine, and idarubicin (ETI) in elderly patients: a randomized comparison with 5-day TAD. Finnish Leukemia Group.

In order to study the efficacy of an oral induction and consolidation regimen in the treatment of acute myeloid leukemia (AML) in elderly patients assessed not to tolerate full-scale intensive chemotherapy, 51 patients over 65 years of age with newly diagnosed AML were randomized to receive two cycles of either totally oral ETI (25 patients) or conventional 5-day TAD (26 patients). The median age of the patients was 73 years, range 65-87 years. Thirty-eight patients had de novo AML and the remaining patients AML subsequent to myelodysplastic syndrome ((n = 11) or treatment related AML (n = 2)). ETI consisted of etoposide 80 mg/m2 and thioguanine 100 mg/m2 twice a day on days 1-5, and idarubicin 15 mg/m2 on days 1-3, all given orally. TAD consisted of oral thioguanine and i.v. cytarabine, both in the dose of 100 mg/m2 twice a day on days 1-5, and daunorubicin 60 mg/m2 on day 5. The maintenance treatment was daily oral mercaptopurine 70 mg/m2 and weekly oral methotrexate 12 mg/m2. In the ETI group complete remission (CR) was achieved in six patients after the first cycle and in nine more patients after the second cycle. The CR rate was 15/25 = 60%. The corresponding figures for the TAD group were four and two remissions, CR rate 6/26 = 23% (p = 0.007). The survival was significantly longer in the ETI arm (p = 0.042). The median survival was 9.9 months in the ETI group and 3.7 months in the TAD group. There were no significant differences in the side effects between the two arms. In conclusion, the totally oral ETI regimen resulted in a significantly higher remission rate and longer survival than the 5-day TAD regimen in elderly patients with AML, with no more toxicity.

Glucose inhibits insulin release induced by Na+ mobilization of intracellular calcium.

45Ca2+ incorporated in response to glucose was selectively mobilized from the beta-cell-rich pancreatic islets of ob/ob-mice after raising the intracellular Na+ by removal of K+ or addition of ouabain or veratridine. Also studies of insulin release indicated opposite effects of glucose and Na+ on the intracellular sequestration of calcium. The fact that glucose inhibits insulin release induced by raised intracellular Na+ indicates that this sugar can lower the cytoplasmic [Ca2+]. The concept of a dual action of glucose on the cytoplasmic [Ca2+]. The concept of a dual action of glucose on the cytoplasmic [Ca2+] might well explain previous observations of an inhibitory component in the glucose action on the 45Ca2+ efflux.

An immunocytochemical study of the olfactory projections in the three-spined stickleback, Gasterosteus aculeatus, L.

The distribution of olfactory fibers in the brain of the three-spined stickleback was visualized by means of immunohistochemistry. The labeling of the olfactory fibers was produced by serum containing antibodies against somatostatin-14. Olfactory fibers were observed entering the olfactory bulbs, where they terminated in the glomerular layer or collected into fascicles that coursed through the bulbs into the telencephalon without participating in the formation of the glomerules. In the telencephalon the fascicles, which belonged to the medial olfactory tract, formed two fiber systems: ventral descending fibers and dorsal descending fibers. The ventral descending fibers could be followed through the ventral telencephalon to the vicinity of the lateral tuberal nucleus. The dorsal descending fibers coursed via the anterior commissure to the posterior part of the telencephalon. Part of the postcommissural fibers of the dorsal descending system coursed to the posterior zone of the area dorsalis telencephali while others left the telencephalon via the medial forebrain bundle and could be followed to the periventricular hypothalamus. Some axons formed synaptic contacts with unlabeled cell bodies in the nucleus of the terminal nerve which, in this species, is situated immediately behind the bulbs. In addition, an extensive terminal field associated with the dorsal descending fibers was found in the ventromedial aspects of the telencephalon. It is unlikely that the labeling represents immunoreactive somatostatin-14 because: 1) the labeling persisted after the absorption of the antiserum with synthetic somatostatin-14; 2) antiserum against somatostatin-14 from another manufacturer did not have this labeling property; and 3) the production of the absorbable labeling depended on the choice of fixative whereas the production of the unabsorbable labeling did not.

Development of tyrosine hydroxylase-, dopamine- and dopamine beta-hydroxylase-immunoreactive neurons in a teleost, the three-spined stickleback.

The development of catecholaminergic neuronal systems in the brain of a teleost, the three-spined stickleback, was studied through embryonic to early larval stages by immunocytochemistry using specific antibodies against dopamine, tyrosine hydroxylase and dopamine beta-hydroxylase. By analysing the spatiotemporal patterns of development for the catecholaminergic nuclei, possible homologies with nuclei in amniote brains have been identified. The noradrenergic neurons in the isthmus region of the rostral rhombencephalon originate in the same manner as the A4-A7 + subcoeruleus group in mammals. Their developmental characteristics show the largest similarities with the subcoeruleus group of birds and mammals, although some features are shared with developing A6 (locus coeruleus) neurons. Catecholaminergic neurons never appear during development in the ventral mesencephalon of the three-spined stickleback. A group of large dopaminergic neurons that accompany the cerebrospinal fluid (CSF)-contacting neurons follows the border between the hypothalamus and the ventral thalamus into the caudal hypothalamus, where they are continuous with the dopaminergic neurons in the posterior tuberculum. They are thus topologically comparable with the dopaminergic neurons of the zona incerta in mammals. The dopaminergic CSF-contacting neurons that line the median, lateral and posterior recesses of the third ventricle do not contain tyrosine hydroxylase-immunoreactivity at any developmental stage. This indicates that they take up and accumulate exogenous dopamine or L-dihydroxyphenylalanine, and do not synthesize dopamine from tyrosine at any developmental stage. Tyrosine hydroxylase-immunoreactive neurons appear in the pineal organ on the day of hatching (120 h post-fertilization). They were still observed in 240-h-old larvae, but are absent in the pineal organ of adult sticklebacks. The initial appearance and subsequent differentiation of catecholaminergic neurons in the stickleback embryo follow essentially the same spatial and temporal pattern as in amphibian, avian and mammalian embryos. This observation supports the hypothesis that morphologically, topologically and chemically similar monoaminergic neurons in different vertebrate classes are homologous.

Distribution of dopamine-immunoreactive neuronal perikarya and fibres in the brain of a teleost, Gasterosteus aculeatus L. comparison with tyrosine hydroxylase- and dopamine-beta-hydroxylase-immunoreactive neurons.

The distribution of dopamine in the brain of the teleost Gasterosteus aculeatus L. was demonstrated with the indirect peroxidase-antiperoxidase immunohistochemical method using highly specific antibodies against a dopamine-glutaraldehyde-thyroglobulin conjugate. Dopamine-immunoreactive (DAir) neuronal somata were observed in all main brain regions. In the forebrain, DAir neurons were located in a continuous cell column extending from the caudal part of the olfactory bulbs to the preoptic area. The neurons lie lateral to the dorsal (and caudally to the subcommissural) portion of the ventral telencephalic area, and ventromedial to the central nuclei of the dorsal area. In the diencephalon, cerebrospinal fluid-contacting neurons were located in the paraventricular organ and in the subependymal layers of the dorsal and caudal zones of the periventricular hypothalamus. Small DAir neurons were observed in the suprachiasmatic nucleus, in the parvocellular preoptic nucleus and in the ventromedial thalamic nucleus, while large perikarya were observed dorsolateral to the dorsal zone of the periventricular hypothalamus ('PVO-accompanying cells'), in the posterior tuberal nucleus and in the most rostral portion of the mammillary bodies. Numerous small DAir neurons were located in the periventricular pretectal nucleus. In the brainstem, DAir neurons were observed in the isthmus region, in the dorsal raphe nucleus and in the lateral parts of the nucleus of the solitary tract. DAir perikarya were also observed in the area postrema. Direct comparison with the distribution of tyrosine hydroxylase- and dopamine-beta-hydroxylase-immunoreactivity (THir and DBHir) gave the following results: THir neurons were found in all areas where DAir neurons were located, except for the paraventricular organ and the dorsal and caudal zones of the periventricular hypothalamus, which were devoid of THir. DBHir (putatively noradrenergic or adrenergic) neurons were observed in the lateral parts of the nucleus of the solitary tract, and in the isthmus region. The DBHir neurons in the isthmus region, which have previously been shown to be noradrenergic, appeared to be identical with the THir and DAir neurons of the same area. DAir axons were found in high numbers in most parts of the brain. Especially dense innervation was found in the ventrolateral and posterior parts of the dorsal telencephalic area, the region surrounding the lateral recesses of the third ventricle, the interpeduncular nucleus, the dorsal and median raphe nuclei (the rostral raphe nuclei), and in the nucleus of the solitary tract.(ABSTRACT TRUNCATED AT 400 WORDS)

FMRFamide-like immunoreactive neurons of the nervus terminalis of teleosts innervate both retina and pineal organ.

The tetrapeptide FMRFamide (Phe-Met-Arg-Phe-NH2) was first isolated from molluscan ganglia. Subsequently, it has become clear that vertebrate brains also contain endogenous FMRFamide-like substances. In teleosts, the neurons of the nervus terminalis contain an FMRFamide-like substance, and provide a direct innervation to the retina (Proc. Natl. Acad. Sci. U.S.A., 81 [1984] 940-944). Here we report the presence of FMRFamide-immunoreactive axonal bundles in the pineal organ of Coho salmon and three-spined sticklebacks. The largest numbers of axons were observed proximal to the brain, in the pineal stalk, while the distal part of the pineal organ contained only few axons. No FMRFamide-like-immunoreactive (IR) cell bodies were observed in the pineal organ. In adult fish it was not possible to determine the origin of these axons, due to the large numbers of FMRFamide-like IR axons in the teleost brain. However, by following the development of FMRFamide-like IR neurons in the embryonic and larval stickleback brain, it was possible to conclude that, at least in newly hatched fish, FMRFamide-like IR axons that originate in the nucleus nervus terminalis reach the pineal organ. Thus, it seems there is a direct connection between a specialized part of the chemosensory system and both the retina and the pineal organ in teleost fish.

An immunocytochemical study of the development of the olfactory system in the three-spined stickleback (Gasterosteus aculeatus L., Teleostei).

Antisera against a variety of substances have been found to produce an identical immunoreaction in the developing olfactory system of a teleost, the three-spined stickleback (Gasterosteus aculeatus). The label is localized in the olfactory placode, the olfactory nerve and those parts of the secondary olfactory tracts which constitute the dorsal descending fascicles and the ventral descending fibers of the medial olfactory tract. The label was first detected 3 days after fertilization (3D) in the olfactory placode where labeled supporting cells were observed. At 4D, the label was observed at the site of the developing olfactory bulbs. At 7D, the olfactory placode lost the direct contact with the brain and the labeled olfactory nerve became visible. At the same time, the medial olfactory tract emerged from the bulbs, and contacts with cells in the nucleus of the terminal nerve were observed. The development of the medial olfactory tract proceeded caudally, and by the end of 10D, the olfactory tract reached the periventricular hypothalamus. Pre-absorption of the antisera with the respective antigens did not abolish the capacity of the antisera to produce the label. The immunoreaction is thus not specific for the antigens against which the antisera have been raised. Yet the label produced by the immunoreaction is an extremely reliable marker for the primary olfactory tract, and the only existing marker by which secondary olfactory tracts can be visualized.

The CYP3A4 inhibitor intraconazole does not affect the pharmacokinetics of a new calcium-sensitizing drug levosimendan.

Itraconazole is a potent inhibitor of CYP3A4 isoenzyme and it can cause clinically significant interactions with some other drugs. Levosimendan is a new calcium-sensitizing drug intended for congestive heart failure. We aimed to study possible interactions of itraconazole with levosimendan in healthy volunteers. Twelve healthy male volunteers were included into a randomized, double-blind, two-phase crossover study. A wash-out period of 4 weeks was held between the phases. The subjects were given orally itraconazole 200 mg or placebo daily for 5 days. On the fifth day, they received a single oral dose of 2 mg of levosimendan. Levosimendan plasma concentrations were determined up to 12 hours and ECG, heart rate, and blood pressure followed-up to 8 hours after intake of levosimendan. Itraconazole had no significant effects on the pharmacokinetic parameters of levosimendan. Neither were there any differences in heart rate, PQ-, QTc- or QRS intervals between the placebo and itraconazole phases. The systolic blood pressure was decreased slightly more (p < 0.05) during the itraconazole phase than during the placebo phase. In conclusion, because the potent CYP3A4 inhibitor itraconazole had no significant pharmacokinetic interaction with levosimendan, interactions with CYP3A4 inhibitor, and oral levosimendan are unlikely.

Venous thrombosis: a controlled study on the performance of scintigraphy with 99Tcm-HMPAO-labelled platelets versus venography.

Thirty-three patients with symptoms and signs of a deep venous thrombosis (DVT) were examined by contrast venography and radionuclide imaging with 99Tcm-hexamethylpropyleneamineoxime (99Tcm-HMPAO)-labelled autologous platelets. There were 13 patients on heparin therapy and 20 without anticoagulation during the scintigraphy. Scintigraphy consisted of blood pool imaging at 5 to 20 min and accumulation imaging at 2, 4-6 and 18-24 h. In scintigraphy a positive finding was either a defect of radioactivity in the immediate blood pool phase or a hot spot indicative of accumulation of platelets in later phases. Fifteen out of 23 patients positive by venography were also positive by scintigraphy. Five of the eight false negative patients were on heparin treatment, two probably had DVT which were not quite fresh. The venography negative patients were also negative on scintigraphy. Nine out of 12 patients without anticoagulation had positive platelet accumulation compared with two out of 11 patients on heparin therapy. This difference was statistically significant (P less than 0.025). The sensitivity and specificity of platelet scintigraphy were 65 and 100%, respectively, in all patients and 83 and 100% in patients without anticoagulation. Our results suggest that scintigraphy with 99Tcm-HMPAO-labelled platelets is a useful alternative in diagnosing DVT in patients in whom a standard contrast X-ray venograph is contraindicated or otherwise unsuccessful.

Altered expression of HSP27 and HSP70 in distal oesophageal mucosa in patients with gastro-oesophageal reflux disease subjected to fundoplication.

BACKGROUND: Gastro-oesophageal reflux disease (GERD) is a risk factor for oesophageal adenocarcinoma. Although fundoplication cures reflux symptoms and oesophagitis, it remains controversial whether it is capable of preventing the development of oesophageal adenocarcinoma. Hsp27 and Hsp70 are associated with the development of cancer, whereas the effect of fundoplication on them is not known. METHODS: The expression of Hsp27 and Hsp70 was assessed semiquantitatively from biopsies of oesophageal mucosa for a prospective cohort of 19 patients with GERD treated with fundoplication and 7 controls without GERD. Upper gastrointestinal endoscopy with biopsies from the oesophagogastric junction (EGJ) and the distal and proximal oesophagus were performed preoperatively (19 patients) and after recovery from GERD at 6 (19 patients) and 48 months (16 patients) postoperatively. RESULTS: The expressions of both Hsp27 (p = 0.001) and Hsp70 (p = 0.002) in the distal oesophagus were lower in patients preoperatively and at 48 months postoperatively (p < 0.001 for both) than in controls. The patients' Hsp27 and Hsp70 levels were lower preoperatively in the proximal oesophagus (p = 0.048 for both) than in controls. Both Hsp27 (p = 0.002) and Hsp70 (p = 0.003) were lower in the distal oesophagus preoperatively and at 48 months postoperatively (p = 0.003 for Hsp27, p = 0.004 for Hsp70) than in the proximal oesophagus. CONCLUSIONS: Our results indicate that there may be some factor interfering with the mucosal defence system of the distal oesophagus in GERD that is uninfluenced by fundoplication and not associated with the acid-reflux-normalizing effect.