Developmental changes in mucosubstances revealed by immunostaining with antimucus monoclonal antibodies and lectin staining in the epithelium lining the segment from gizzard to duodenum of the chick embryo

The mucosubstances in the epithelium lining the segment from gizzard to duodenum during development of the chick embryo was studied histochemically using monoclonal antibodies against gizzard mucus and lectins, with attention to the regional differentiation of the epithelium in this segment. The anterior limit of epithelial CdxA mRNA expression detected by in situ hybridisation, which served as the position of the gizzard-duodenal boundary, was clearly found from d 3. Granules positive for some antibodies or lectins were found in the region ranging from the posterior part of the gizzard to the duodenum at d 3, which was followed by an increase in the number of granules and a gradual enlargement of the granule-positive area to the anterior part of the gizzard over 4–6 d. From d 4, the epithelia of the gizzard body and of the pyloric or duodenal region came to be differently stained with some antibodies or lectins. From d 10, each region showed a specific pattern of staining. The epithelia of the gizzard body and pyloric region contained abundant mucus granules with a different staining pattern. In the duodenum the number of stained granules was low except in occasional goblet cells. Thus the epithelia of the gizzard body, pyloric region and duodenum may produce different mucosubstances and the regional differentiation in these epithelia may start at rather early stages soon after the formation of digestive tube.     

Granulocyte-colony stimulating factor-induced proliferation of primary adult T-cell leukaemia cells

Granulocyte-colony stimulating factor (G-CSF) is known to induce proliferation and differentiation of granulocyte progenitors, and is widely used to treat neutropenia induced by intensive chemotherapy for malignant lymphoma or adult T-cell leukaemia/lymphoma (ATL). G-CSF is thought not to stimulate malignant lymphoid cells. In the present study we examined the ability of G-CSF to induce in vitro growth of primary ATL cells from 14 patients (nine acute-type, two chronic-type and three lymphoma-type), and we analysed the in vivo counts of ATL cells in patients who received G-CSF for neutropenia. FACS analysis using phycoerythrin-labelled recombinant G-CSF demonstrated that ATL cells from 11/14 patients express some G-CSF receptor (G-CSFR), with a range between 5.4% and 87.3%. Cells expressing G-CSFR also expressed CD4. Reverse polymerase chain reaction (PCR) analysis demonstrated expression of G-CSFR messenger RNA in G-CSFR expressing cells. Leukaemic cells derived from seven (four acute-type, one chronic-type and two lymphoma-type) of the 14 patients proliferated in vitro in response to G-CSF, as measured by [³H]thymidine incorporation; maximum responses were at G-CSF concentrations of 10–100 ng/ml. Nine of 14 patients receiving rG-CSF for neutropenia were analysed retrospectively for ATL cell numbers. Four patients whose primary tumour cells proliferated in response to rG-CSF in vitro showed a significant increase in ATL cell count after administration of rG-CSF ( P  =0.038), whereas five patients whose leukaemic cells did not proliferate in vitro showed no significant increase in ATL cell count. G-CSF can stimulate proliferation of ATL cells which may complicate therapy for this disease.     

A possible mechanism for Inv22‐related F8 large deletions in severe hemophilia A patients with high responding factor VIII inhibitors

Summary. Background: Intron 22 inversion (Inv22) of the coagulation factor (F)VIII gene (F8) is a frequent cause of severe hemophilia A. In addition to Inv22, a variety of F8 mutations (1492 unique mutations) causing hemophilia A have been reported, of which 171 involve deletions of over 50 bp (HAMSTeRs database; http://hadb.org.uk/ ). However, only 10% of these large deletions have been fully characterized at the nucleotide level. Patients and methods: We investigated gene abnormalities in three unrelated severe hemophilia A patients with high titer FVIII inhibitors. They had previously been shown to carry large deletions of the F8, but the precise gene abnormalities remain to be elucidated. Results: Inverse shifting‐PCR (IS‐PCR) Inv22 diagnostic tests revealed that these patients carried either type I or II Inv22. However, they showed a wild‐type (WT) pattern in the IS‐PCR Inv22 complementary tests. We further analyzed their X chromosomes to account for the puzzling results, and found that they had different centromeric breakpoints in the Inv22 X chromosomes, adjacent to the palindromic regions containing int22h‐2 or ‐3, and their spacer region, respectively. The connections appeared to be shifted towards the telomere of the WT F8 Xq28, resulting in a new telomere with an additional intact int22h copy. Conclusions: These gene rearrangements might result from double‐strand breaks in the most distal regions of the long arms of the Inv22 X chromosomes, followed by DNA restorations using the WT F8 Xq28 by non‐homologous end joining or break‐induced replication; thus leading to large F8 deletions in severe hemophilia A patients.     

Suspected acute encephalopathy with symmetrical abnormal signal areas in the basal ganglia,thalamus, midbrain and pons diagnosed by magnetic resonance imaging

A 4-year-old boy was admitted with disturbed consciousness following a convulsion. He developed bilateral pyramidal tract signs and showed a decerebrate posture. Laboratory findings revealed severe liver dysfunction and disseminated intravascular coagulation. On the eighth day eight in hospital he developed parkinsonism. However, 5 months from onset he had recovered almost completely. Brain CT on admission showed low density areas in the basal ganglia, thalamus, midbrain and pons. A T2-weighted scan in magnetic resonance imaging (MRI) showed almost symmetrical high signal intensities in both basal ganglia (including putamen, caudate nucleus, globus pallidus), external capsule, internal capsule, thalamus, midbrain, pons and white matter of the peribasal ganglia; but a T1-weighted scan showed low signal intensities in the same regions during all phases. Therefore hemorrhagic lesions or the presence of thalamic methemoglobin were excluded. It was considered that the pathogenesis may be cytotoxic cellular edema due to cytotoxic agents or acute metabolic factors. Clinical presentation, laboratory findings and radiological findings were most suggestive of acute necrotizing encephalopathy. As differential diagnoses, acute disseminated encephalomyelitis and brainstem encephalitis were considered.     

Cytotoxicity Tests against Cultured Human Lung Cancer Cells With Autologous Lymphocytes Activated in vitro by Mitomycin C-Treated Tumor Monolayers in the Presence of T-Cell Growth Factor

Human peripheral blood or lymph node lymphocytes, obtained frompatients with a variety of lung cancer, were incubated in vitrowith mitomycin C-treated tumor monolayers in the presence ofT-cell growth factor. The cytotoxicity of these lymphocytesfor autologous tumor cells (autologous killer activity) wasassessed by a 4-hr ⁵¹Cr-release assay. Cytotoxic activity wasobserved in 14 out of a total of 20 cases. Lymphocytes frompatients with squamous cell carcinoma, large cell carcinomaand carcinoid exhibited positive activity levels of 11.1 ±1.8, 16.3 and 23.9% respectively. Nine out of 13 patients withadenocarcinoma exhibited positive activity with a mean valueof 8.8 ± 6.8%. No lymphocyte activity against small cellcarcinoma was observed. Natural killer (NK) activity did not always correlate with autologouskiller (AK) activity. Treatment of lymphocytes with monoclonalanti-human lymphocyte antibody revealed differences in effectorcell populations concerning these two activities; AK activitywas abrogated only by treatment with anti-human Lyt 3 antibodyand complement, whereas NK activity was abrogated by anti-humanLyt 1, 2 and 3 and partially by anti-human la antibody. Theseresults indicate that AK activity is mediated exclusively byT cells, but that NK activity is mediated by several subpopulationsof lymphocytes such as T cells, null cells and others.     

APOPTOSIS OF CRYPT EPITHELIAL CELLS IN ULCERATIVE COLITIS

In the colon of ulcerative colitis (UC) patients, apoptotic bodies have been recognized in routine histopathological preparations. To investigate the extent of the apoptosis, colonic biopsies were examined from involved and uninvolved areas of untreated active UC and from normal areas in patients with colonic polyps, utilizing various markers of apoptosis. The markers included DNA breaks detected by TUNEL, Fas (CD95/APO-1) and Fas ligand (Fas-L) localized by immunohistochemistry, electron microscopic features of apoptosis, and laddering of extracted DNA. Apoptosis marker positive cells were found mainly on the luminal epithelium of the normal colon and were present in active UC in crypts of involved and uninvolved areas of the colon, in addition to the luminal epithelium. The DNA extracted from active UC colon electrophoresed as a ladder. These findings suggest that the loss of epithelial cells in active UC occurs mainly by apoptosis in crypts of involved and adjacent uninvolved areas and that the Fas/Fas-L interaction is a mediator of the apoptosis.     

Usefulness of the Adenosine Triphosphate with a Sufficient Observation Period for Detecting Reconduction after Pulmonary Vein Isolation

Background: Although reconduction after pulmonary vein (PV) isolation is considered to play a key role in the recurrence of paroxysmal atrial fibrillation (AF), there have been few reports regarding the precise time course of early reconduction. Several studies have suggested that transient PV reconduction facilitated by adenosine may predict long-term AF recurrence. This study was designed to clarify the incidence and time course of early reconduction after PVI during the procedure and to confirm whether the use of ATP after a certain observation period was useful to detect early reconduction after PVI.
Methods: In 21 patients (18 males, 56 ± 11 years) with drug refractory AF, radiofrequency circumferential PV antrum ablation was performed in all 4 PVs. After the completion of isolation, electrograms in each PV were repeatedly recorded (1.98 ± 0.57 times per PV) using a circular mapping catheter for an observation period of 87 ± 29 minutes. After isolation of all 4 PVs, 30 mg of adenosine triphosphate (ATP) was administered during isoproterenol infusion.
Results: PV electrical isolation was initially achieved in all 81 PVs. During the observation period, 12 (15%) PVs in 10 (48%) of 21 patients exhibited spontaneous reconduction. Among the remaining 69 PVs, 8 (12%) additional PVs had reconduction with the use of ATP. All PV reconduction was successfully eliminated by 4.5 ± 2.2 additional radiofrequency applications.
Conclusion: A sufficient observation period and the use of ATP are useful to detect early reconduction after PV isolation.     

Early diagnosis of Alzheimer's type dementia with special reference to the clinicopathology of mild cognitive impairment

A considerable number of patients with Alzheimer‐type pathology begin with various types of psychiatric symptoms. In discussing mild cognitive impairment (MCI) and Alzheimer‐type dementia (ATD), psychiatric symptoms other than memory impairment must be considered. There is no obvious neuropathological boundary that distinguishes dementia from non‐dementia. In fact, although Alzheimer‐type pathology of MCI corresponds mainly to Braak's stage I or II, it has a wide range extending from Braak's stage I to V or VI. Difference in compensatory ability and environmental factors among individuals seem to affect the mental state more extensively than previously thought. MCI manifests most frequently in Braak's stage I and II (transentorhinal stage) and its Alzheimer‐type pathology is almost confined to the hippocampal region. Although the degree and distribution of senile plaques vary in each patient, the cerebral isocortex is virtually free from degeneration. There is discrepancy between the corresponding neuropathology and hypo‐metabolism in the posterior cingulated cortex (PCC), which is regarded as an important finding for the early diagnosis of ATD. In PCC, hypo‐metabolism seems to precede that morphological change especially in its premorbid or early period of ATD. The state of MCI could also occur in other diseases causing dementia, among which tangle‐only dementia is particularly notable. The characteristic of tangle‐only dementia is the gradually progressive, persistent pathological memory impairment with relative preservation of personality and cognitive function. These characteristics meet the criteria for MCI. Thus, tangle‐only dementia should be taken into account in patients who exhibit MCI but do not progress to the development of dementia over a prolonged period.     

New visual rating system for medial temporal lobe atrophy: a simple diagnostic tool for routine examinations

Aim: We estimated the usefulness of our new scale to rate medial temporal atrophy with short inversion time inversion recovery images. Methods: Alzheimer's disease (AD) subjects (n= 34) and non‐demented subjects (n= 19) were recruited for this study. First, coronal short inversion time inversion recovery images were scanned vertical to the long axis of hippocampus. Next, the single image in which peduncles appeared widest was adopted for estimation. The parahippocampal cerebrospinal fluid space was divided into three parts: the outer, upper and inner parts. The hippocampus was defined as a structure being of equal intensity to grey matter. Two radiologists compared each part of the parahippocampal cerebrospinal fluid space with the hippocampus and rated them on a 0–3 scale. Interrater and intrarater kappa statistics and sensitivity/specificity for the diagnosis of AD were calculated using the scores of the right, left and both sides combined. Results: There were no significant differences between AD and ND subjects with regards to sex. AD subjects had lower Mini‐Mental State Examination scores and were older than non‐demented subjects. Interrater and intrarater kappa statistics were 0.52–0.68 and 0.76–0.83, respectively. Sensitivity was 88.2% using the scores of both sides. Conclusions: Interrater and intrarater agreements were fair to good and good to excellent, respectively. Our new visual rating method detected medial temporal atrophy in AD patients at a highly sensitive rate. As such, we conclude that this visual rating scale is useful for judging medial temporal atrophy simply and objectively in clinical use, and it is helpful in establishing an AD diagnosis.