X chromosome inactivation pattern in female carriers of X linked hypophosphataemic rickets.

X linked hypophosphataemia (XLH) results from an abnormality of renal tubular phosphate reabsorption. The disorder is inherited as an X linked dominant trait and the gene has been mapped to Xp22.1-p22.2. A candidate gene (PEX) has recently been isolated. The most striking clinical features are growth retardation and skeletal abnormalities. As expected for X linked dominant disorders, females are less affected. However, such a gene dosage effect does not exist for renal phosphate reabsorption. Preferential X chromosome inactivation has been proposed as a possible explanation for this lack of gene dosage. We have examined the X inactivation pattern in peripheral blood cells from 12 females belonging to seven families with XLH using PCR analysis at the androgen receptor locus. The X inactivation pattern in these patients did not differ significantly from the pattern in 30 healthy females. The X inactivation pattern in peripheral blood cells does not necessarily reflect the X inactivation pattern in renal cells. However, the finding of a normal distribution of X inactivation in peripheral blood cells indicates that the similarity in the renal handling of phosphate in male and female patients is not related to a ubiquitous preferential X inactivation.     

Identification of lymphocyte subsets in peripheral blood and thyroid gland in Graves' disease by alpha-naphthyl-acetate-esterase reaction

T lymphocyte subsets were prospectively examined in the peripheral blood and thyroid aspirates of 10 patients with hyperthyroid Graves' disease before and after treatment with methimazole and attainment of euthyroidism. T lymphocyte subsets were identified with monoclonal antibodies and pattern of alpha-naphthyl-acetate esterase (ANAE) staining pattern in the case of peripheral blood and ANAE staining pattern with thyroid aspirate smears. Before treatment, OKT8+ lymphocytes were significantly decreased (18.4% +/- 4.8) (S. D.) in the patients compared to control (28.8 +/- 6.7%, p less than 0.05), the OKT4/OKT8 ratio was increased (2.92 vs 2.11). Percent OKT8+ lymphocytes were not different from the controls when the ten patients had been rendered euthyroid. ANAE mononuclear cells with a diffuse pattern (presumed suppressor cells) were 4.2% +/- 1.8 before treatment and 8.3 +/- 2.4 (p less than 0.05) after treatment and 11.5% +/- 2.2 in controls. ANAE mononuclear cells with diffuse pattern represented 4.2% +/- 1.8 of the mononuclear cells infiltrating the thyroid gland of untreated patients and rose to 8.3% +/- 2.4 after the patients had become euthyroid. ANAE negative cells (B cells and some T cells) were increased in the thyroid of untreated patients. It is concluded that mononuclear cells with presumed suppressor T cell phenotype are decreased in the blood and thyroid glands of patients with active Graves' disease and that this defect is corrected when euthyroidism has been established.     

Distinct Pattern of Human Vδ/51 γδ/5 T Cells Recognizing MICA

γδ T cells represent one unique recognition pattern, the limited recognition, which distinguishes from the specific recognition for αβ T cells and pattern recognition for macrophages. Vδ1 γδT cell is the major subset of human γδT cells, which predominates in mucosal tissue including the intestinal epithelia. Presently, a few antigens that human Vδ1TCR can recognize have been identified. Among them, MHC class I chain-related molecules A （MICA） have been studied most intensively. Besides Vδ1TCR, MICA is also the ligand of NKG2D, a C-type lectin-like activating immunoreceptor. In human, only Vδ1 cells can simultaneously express both types of receptors of MICA while NK cells, αβ T cells and other subsets of γδT cells likewise express NKG2D. Although the precise mechanisms are still enigmatic, this distinct pattern of Vδ1 cells recognizing MICA predicts unique biological significance of Vδ1 cells in immune defense. Recent years, some progresses have been made in this issue. In this review we summarize the related reports and put forward some novel views based on our group＇s studies.     

Human large granular lymphocytes contain an esterase activity usually considered as specific for the myeloid series

A cytochemical marker such as alpha-naphthyl acetate esterase (ANAE) has been found useful for the morphological identification of the subset of T lymphocytes having receptors for Fcμ (T_M cells). ANAE reaction on T_M cells gives a typical pattern of one to four positive spots, whereas this pattern is not found on T cells with receptors for Fcγ (T_G cells). ANAE is abundant in monocytes but not detectable in granulocytes. Herein another type of esterase activity, naphthol-AS-D chloroacetate esterase (NCAE), is described; it is known to be abundant in granulocytes and was found to give a specific pattern of reactivity with the subpopulation of large granular lymphocytes (LGL). This pattern of fine granular staining was observed not only on LGL present in the T_G cell subpopulation but also in LGL present in the non-T, non-B cells. Fractions of peripheral blood mononuclear cells which were ènriched up to 80% in LGL by Percoll discontinuous density gradient gave a similar percentage of specific NCAE pattern. In addition, among the different fractions from Percoll gradient, there was a good correlation (r = 0.94) between the number of NCAE-positive cells and the natural killer activity against the natural killer susceptible K562 target cells. It will be important to determine whether or not this enzymatic activity plays a role in the cytotoxic activities of LGL.     

Pigeon pattern electroretinogram: A response unaffected by chronic section of the optic nerve

Summary Retinal evoked responses to sinusoidal gratings modulated in counterphase (pattern ERG) have been recorded from the pigeon eye. The pattern ERG amplitude depends upon the temporal frequency of the modulation, the contrast, the spatial frequency and the area of the stimulus. In 8 pigeons the pattern ERG has been recorded at different times after the unilateral section of the optic nerve. It has been found that the pattern ERG has a comparable amplitude in the two eyes as a function of the spatial frequency, 3 and 9 months after the section of the left optic nerve. At these times, histological evidence shows a drastic reduction in the density of the retinal ganglion cells on the operated side in comparison to the control one. These findings suggest that retinal sources other than the ganglion cells are responsible for the generation of the pigeon pattern ERG.     

The morphological and functional observation of the gap junction proteins in the oviduct epithelia in young and adult hamsters

The histological morphology of oviduct epithelia have been described well, however, the expression pattern of the gap junction proteins in the cells, and the function related with the proteins, such as [Ca2+]i dynamics pattern of living oviduct epithelia at different ages have not been clarified. We used immunohistochemistry to compare the expression pattern of gap junction proteins in the cells of the young and adult groups. Moreover, we used real-time confocal microscopy to observe the spontaneous Ca2+ oscillation (spontaneous fluctuation) in freshly isolated epithelia (ciliated cells) in ampulla potion of oviduct from the two groups. The results show as demonstrated by immunohistochemistry the gap junction proteins (Cx26, Cx32 and Cx43) formed a well-regulated expression in the young animals, but not in the adult animals. In addition, the [Ca2+]i dynamics of ciliated cells in freshly oviduct epithelia have a spontaneous fluctuation pattern that occurs without any stimulation in the young animals, but this pattern was not observed in the adult animals. In conclusions, our findings suggest that gap junctions regulate the spontaneous fluctuation of [Ca2+]i dynamics in ciliated cells of oviduct epithelia in young animals.     

Vacuolated cell pattern of pancreatobiliary adenocarcinoma: a clinicopathological analysis of 24 cases of a poorly recognized distinctive morphologic variant important in the differential diagnosis

Pancreatic ductal adenocarcinoma (PDCA) is characterized by well-defined tubular units in the vast majority of the cases; however, variations in this theme do occur. It is important to recognize the morphologic spectrum of PDCA to avoid misdiagnosis especially in small specimens and also in metastatic foci. Here, we document a morphologic variant of PDCA that is characterized by a distinctive pattern of infiltrating cribriform nests in a distinctive “microcystic” or “secretory” pattern. Twenty-four cases of PDCA have been identified in a review of 505 cases diagnosed with PDCA. Histologically, this pattern was characterized by infiltrating nests of tumor cells with large vacuoles and “signet-ring” like appearance imparting a cribriform growth pattern. The vacuoles were one to five cells in size, often merging to form multilocular spaces separated by a thin rim of cell membrane. Many of these spaces contained CA19.9 positive granular secretory material. The nuclei were often pushed to the periphery and compressed in a pattern resembling adipocytes, although the nuclei were often densely hyperchromatic and displayed significant atypia. Especially in biopsies from the peripancreatic fat and peritoneum, these neoplastic cells had been misdiagnosed as degenerating adipocytes, and in the lymph nodes, they had been misinterpreted as lipogranulomas. Clinical findings of the patients were similar to that of conventional PDCA, except higher incidence of history of smoking (83% vs. 60%; p = 0.034). In conclusion, vacuolated cell adenocarcinoma is a distinct morphologic variant of PDCA, and the presence of this peculiar pattern in a metastatic site, although not specific, should raise the suspicion of a PDCA.     

Intermediate filament aggregates in mitoses of primary cutaneous neuroendocrine (Merkel cell) carcinoma

Primary cutaneous neuroendocrine carcinomas express different kinds of intermediate filaments and frequently in a 'paranuclear globular' pattern. We have observed the same pattern not only in interphase but also in mitotic cells, which are very frequent in these tumours. We report a quantitative and morphological study of eight primary cutaneous neuroendocrine carcinomas stained with different antibodies against cytokeratins (CAM 5.2 and anti-cytokeratin 20), neurofilaments (70 kDa and 200 kDa) and peripherin. We have found a predominance of CAM 5.2 expression in interphase cells and of neurofilament proteins in mitotic cells; 87.02% of the interphase cells were positive with CAM 5.2 whereas only 6.08% were positive for neurofilaments ( P <0.01); 35.41% of the mitotic cells were positive with CAM 5.2, whereas 50% were positive for neurofilaments ( P <0.01). A correlation between a globular pattern of intermediate filament proteins and prognosis has not been found. We describe for the first time the division of neoplastic cells with a globular pattern; the presence of intermediate filament proteins with a globular pattern in all mitotic stages; and the uneven distribution of this formation between the two daughter cells.     

Extracellular matrix environment influences chondrogenic pattern formation in limb bud micromass culture: experimental verification of theoretical models

Various theoretical models have been proposed to explain the periodicity in the pattern of limb chondrogenesis, but experimental comparison of these models have seldom been performed properly. In the present study, micromass culture of limb bud mesenchyme cells was undertaken to test the validity of three theoretical models: the reaction-diffusion model, the cell sorting model, and the mechanochemical model. Computer simulations were undertaken to predict the factors that can affect the coarseness of the chondrogenic pattern. According to the predictions, we performed micromass culture of limb mesenchyme in collagen and agarose gel. Then we carried out time-lapse observation to analyze the cell movement during pattern formation. From computer simulations it was theoretically predicted that changes in the surrounding extracellular matrix should alter the periodicity of the chondrogenic pattern in vitro, and we found that pattern changes actually occurred under different culture conditions. When compared with the culture in a liquid medium, the chondrogenic pattern became less coarse when the cells were cultured in collagen or agarose gel, and the pattern change appeared to be independent of the cell differentiation. Time-lapse observation revealed a decrease in cell motility when the cells were cultured in gel. It was found that both the reaction-diffusion and cell sorting models fit the pattern change produced and that the mechanochemical model is not primarily important in the chondrogenic pattern formation in vitro.     

Cytological features of lung adenocarcinoma with micropapillary pattern in the pleural or pericardial effusion: analysis of 5 cases

Micropapillary pattern is a distinct histopathological pattern, and usually shows a high frequency of lymphatic invasion and lymph node metastases. This pattern is also reported in lung adenocarcinoma, however, only one cytological report of lung adenocarcinoma with micropapillary pattern has been reported. In this study, we analyzed the cytological features of this type of carcinoma in the pleural or pericardial effusion. This study was comprised of 5 consecutive cases of lung adenocarcinoma with micropapillary pattern, in which the tumor cells were present in the pleural or pericardial effusion and whose diagnoses were histopathologically confirmed. The characteristic cytological findings in the pleural or pericardial effusion were as follows: i) tightly cohesive small nests of tumor cells showing papillary structure without fibrovascular core, ii) these nests were comprised of approximately 5-20 tumor cells, iii) cauliflower-like and acinar-like structures were also observed, iv) intracytoplasmic vacuoles were observed in 40% of the cases, and v) the neoplastic cells had large round to oval nuclei containing coarse chromatin and occasional conspicuous nucleoli. It has been reported that the presence of micropapillary structure and intracytoplasmic vacuolation are also characteristic cytological features of micropapillary carcinoma of the urinary bladder, therefore, they are thought to be common cytological features of carcinomas with micropapillary pattern. Consequently, detection of these features can lead to a cytodiagnosis of lung adenocarcinoma with micropapillary pattern in the pleural or pericardial effusion. Recognition of these features is important because this type of tumor shows an aggressive clinical course.     

Reduced number of CD1a+ cells in cutaneous B-cell lymphoma.

Cutaneous B-cell lymphoma is difficult to distinguish from pseudolymphoma. The histologic pattern and monoclonal restriction (immunohistochemical analysis and molecular biology) are the criteria used for differentiating these entities. CD1a+ dendritic cells have been observed in the infiltrates of T-cell lymphoma, but the presence of these CD1a+ cells has not been compared in B-cell lymphoma and pseudolymphoma. We studied the presence of CD1a+ cells on frozen sections of 23 B-cell lymphomas, 13 pseudolymphomas, and 17 T-cell lymphomas by immunohistochemical analysis. We found abundant CD1a+ dendritic cells in only 1 (4%) of 23 B-cell lymphomas, whereas in 8 (62%) of 13 pseudolymphomas and 17 (100%) of 17 T-cell lymphomas, strong CD1a staining was present. Our study demonstrates a distinct pattern of CD1a staining in the infiltrates of B-cell lymphoma and pseudolymphoma that may be of value in the differential diagnosis of these skin disorders.     

Replication status in leukocytes of treated and untreated patients with polycythemia vera and essential thrombocytosis

The replication status of malignant cells is usually asynchronous. However, to date the pattern of replication has not been studied in myeloproliferative disorders nor has the effect of chemotherapy been systematically evaluated. Therefore, we used fluorescence in situ hybridization to interphase nuclei in PHA-stimulated peripheral blood lymphocytes to examine replication timing of three alleles associated with the malignant process. The study group comprised hydroxyurea treated and untreated patients with essential thrombocytosis (ET) or polycythemia vera (PV). A significantly higher rate of the asynchronous pattern of replication in both treated and untreated patients was found as compared to healthy controls. The highest rate of asynchronous replication was observed in untreated patients. Also, the frequency of the two doublets pattern was significantly higher in the untreated group compared to the treated patients and to the control groups. In conclusion, patients with PV and ET have a higher rate of asynchronous pattern of replication. A possible correlation between disease activity and the pattern of replication is suggested. The effect of hydroxyurea on the pattern of replication is variable.     

Chromosomally abnormal cells are not selected for the extra-embryonic compartment of the human preimplantation embryo at the blastocyst stage

BACKGROUND: Although well defined for embryos at cleavage stages, the occurrence and frequency of chromosomal aberrations in human blastocysts is relatively unknown. It has been reported that only one in four blastocysts is comprised totally of chromosomally normal cells. One of the selection mechanisms for the embryo proper to become free of these chromosomally abnormal cells would be to sequester them to the extra-embryonic compartment during development. The study aim was to investigate whether such a mechanism of selection exists in human preimplantation embryos. METHODS: Inner cell mass (ICM)/trophectoderm (TE) differentiation was performed, followed by fluorescence in-situ hybridization (FISH), to study the chromosomal distribution in both populations of cells. RESULTS: Of the 94 successfully analysed blastocysts, 68.8 +/- 1.5% of all analysable nuclei per blastocyst showed a disomic chromosomal content. Only 22.6% of blastocysts analysed were classified as normal. Of the embryos classified as abnormal at the blastocyst stage, 11.9% showed a simple mosaic pattern and 32.1% a complex mosaic pattern. An equally large group of blastocysts showed either a chaotic pattern (16.7%), or the chromosomal pattern could not be classified. The average degree of normal cells in the ICM (67.9%) was similar to the degree observed in the TE (69.5%). CONCLUSIONS: These findings indicate that chromosomally abnormal cells are not preferentially segregating to the extra-embryonic compartment of the human preimplantation embryo at the blastocyst stage. Hence, other mechanisms should be responsible for an absence of chromosomally abnormal cells in the embryo proper at later stages of development. One possible mechanism might be the elimination of the chromosomally abnormal cells by selective cell death activation.     

DNA synthesis of human XYY cells

The autoradiographic YY labelling pattern of the DNA replication in 47, XYY cells of two patients has been studied. Both Y chromosomes began DNA replication simultaneously but later than the rest of the chromosomes. During later stages the YY labelling pattern in both patients was correlated to the position of the cells in S, as indicated by the cell grain count: in cells with more than approximately 500 grains the synchronous labelling of the Y chromosomes was the rule; whereas during later stages in cells with approximately 100–400 grains, synchronous as well as asynchronous labelling in the same patient was found. In cells with less than about 100 grains, both Y chromosomes usually had completed replication. No obvious difference between the labelling pattern of the two Y chromosomes was found when cells from the 2 patients were compared at the same stage of S. Our results demonstrate the importance of analysing cells of comparable developmental stages of S when comparing DNA labelling patterns of different XYY individuals in search of a possible correlation between labelling pattern and phenotypic expression of the syndrome.
In one of the patients the QM fluorescence pattern as well as the autoradiographic pattern of the same cells was studied. The characteristic intense fluorescence pattern of both Y chromosomes was found even in cells where the autoradiographic labelling pattern indicated distinct asynchrony between the two Y chromosomes. The significance of these findings is discussed.     

Discharge patterns of cat pontine brain stem neurons during desynchronized sleep.

1. Discharge pattern has been characterized by autocorrelation analysis of stationary portions of extracellularly recorded discharge trains of cat pontine brain stem neurons during spontaneously occurring desynchronized sleep episodes. 2. Neurons localized to the area implicated in control of the desynchronized phase of sleep, the gigantocellular tegmental field (FTG), show the most phasic or clustered discharge pattern, as evinced by initial peaks in the autocorrelations. At the peak, the FTG population average discharge probability is 3 times that expected had the discharges been evenly distributed over time. The initial peak extends beyond a lag of 3 s, indicating runs of clustered discharge extending beyond this duration. Neurons in other reticular tegmental fields, the tegmental reticular nucleus and pontine gray, show a more sustained or tonic discharge pattern. 3. Discharge patterns of a given cell are consistent from one desynchronized sleep episode to the next; units with phasic discharge patterns remain phasic, and tonic patterns remain tonic. 4. There is a three-way correlation among FTG units recorded at sites with many giant cells, units with high discharge rate increases on transition to desynchronized sleep, and units with a markedly phasic discharge pattern. This implicates the giant cells as the source of both the distinctive discharge rate and pattern changes of neurons during desynchronized sleep. 5. Stereotyped, regular discharge patterns are not characteristic of FTG or other units, suggesting they are not pacemakers and that endogenous activation of pacemaker cells is unlikely to be a mechanism for generation of the marked discharge rate increases on transition to desynchronized sleep that are found in FTG units. The irregular, clustered discharge pattern of FTG is more compatible with generation of discharge rate increases through interaction with other cells. The markedly phasic discharge of FTG units is also consistent with a driving role in generation of the phasic electrophysiologic events of desynchronized sleep.     

Antigen-specific regulatory T cells are detected in Peyer's patches after the interaction between T cells and dendritic cells loaded with orally administered antigen

Systemic immune tolerance is induced for orally administered antigen, and this phenomenon is called oral tolerance. However, the mechanism of oral tolerance has not been completely elucidated. It has been suggested that antigen presentation and generation of regulatory T cells in Peyer's patches (PPs) are important for induction of oral tolerance. Hence, we orally administered fluorescence-labelled antigen to mice and examined kinetics of the antigen and interaction between antigen-loaded dendritic cells and T cells. It was visualized that dendritic cells in PP rapidly take up antigen. We next transferred antigen-specific naïve T cells from T cell receptor transgenic mice and administered the antigen orally. Antigen-specific T cells accumulated in IFR in PP and DCs that have ingested antigen come in contact with antigen-specific T cells in IFR. The accumulated T cells were then collected and analyzed for the pattern of gene expression by real-time PCR, which revealed a gene expression pattern similar to that of FoxP3-positive regulatory T (T_reg) cells. CCR9, an intestinal homing marker, was also strongly expressed. These results suggest that DCs that have captured oral antigens in PPs locally induce antigen-specific naïve T cells to differentiate into T_reg cells with the intestinal homing phenotype.     

Histological variety of floral variant of follicular lymphoma

To further clarify the histopathological findings of the floral variant of follicular lymphoma (FVFL), we studied 13 Japanese cases. Two histological subtypes of neoplastic follicles of FVFL have been described: (i) A macrogerminal center pattern where the mantle zone lymphocytes were invaginated into the neoplastic germinal center, often reminiscent of a floral design. (ii) A microgerminal center pattern where the massive invasion of mantle zone lymphocytes resulted in almost complete breakage of the neoplastic follicles. In the former pattern, the neoplastic germinal center usually contained large clusters of tumor cells, whereas in the latter, small clusters of up to 20 tumor cells or isolated tumor cells were observed in the neoplastic germinal centers. Moreover, occasional tumor cells showed a lymphocytic and/or histiocytic Reed-Sternberg cell (L&H cells)-like morphology. Both types of neoplastic follicles were observed to a varying degree in most cases. The macrogerminal center pattern was predominant in nine cases (70%), whilst the microgerminal center pattern was predominant in only four cases (30%). Three lesions (23%) had a marginal zone component. Immunohistochemistry showed that atypical follicular center cells, including L&H cells, were CD3-, CD5-, CD10+, CD20+, CD43-, bcl-2+, cyclinD1-. The overall histological findings of the macrogerminal center are similar to those of florid progressive transformation of germinal center (PTGC), whilst the microgerminal center pattern is similar to that of nodular lymphocyte-predominant Hodgkin lymphoma. Initially, the differential diagnosis between FVFL and florid PTGC was emphasized. However, the present study indicates that nodal marginal zone B-cell lymphoma possessing floral follicles and nodular lymphocyte-predominant Hodgkin lymphoma should be added to the differential diagnosis of FVFL. The germinal center B-cell nature of FVFL is most clearly recognizable by immunohistochemistry, though histological appearance alone may cause some diagnostic problems.     

Abnormalities in the expression of the leucocyte-common antigen in chronic lymphocytic leukaemia.

The surface glycoproteins of lymphocytes isolated from patients suffering from B cell chronic lymphocytic leukaemia (B-CLL) have been studied by radioactive labelling with impermeable probes and with MoAb. Several features not found in normal B cells have been observed. The abnormalities found in the expression of polypeptides of the leucocyte common (L-C) antigen, identified by appropriate MoAb, have been examined in detail. It has been shown by both biochemical analysis and MoAb binding that this group of polypeptides can, within a panel of B-CLL patients, range from a typical B cell pattern to the pattern resembling that normally found in T cells. The T lymphocyte profile is correlated with a poor prognosis (MVA C rating) and in the one patient where a change in the glycoprotein profile towards that of the T cell was observed, the change coincided with a clinical deterioration. The biological significance of the molecular diversity is discussed.     

Metabolic Patterns (NAD(P)H) in Rat Basophilic Leukemia (RBL-2H3) Cells and Human Hepatocellular Carcinoma (Hep G2) Cells with Autofluorescence Imaging

Although the spatial and temporal distributions of cellular NAD(P)H concentrations have been theoretically predicted as typical patterns of the metabolism in living cells, so far such a pattern was observed only in neutrophils. In this work, the dynamic NAD(P)H distributions in rat basophilic leukemia (RBL-2H3) and human hepatocellular carcinoma (Hep G2) cells were studied by imaging the autofluorescence of cellular NAD(P)H with a sensitive CCD detector in a confocal microscope. The typical pattern of the cytoplasmic NAD(P)H wave traveling along the long axis of the elongated cell with a velocity of 2.2±0.6?μm/s was detected in RBL-2H3 cells. While in the case of Hep G2 cells, only the oscillation of the mitochondrial NAD(P)H was observed because the NAD(P)H mainly localized in mitochondria of Hep G2 cells. These results confirm the metabolic pattern of NAD(P)H in living cells and suggest that the expression of the metabolic pattern probably differs in different cell lines.     

The ANAE reaction pattern of reticular stromal cells of lymphatic organs from normal and hydrocortisone--treated mice

The pattern of alpha-naphthyl-acetate esterase (ANAE) activity in stromal and lymphatic cells has been examined in lymphatic organs from normal and hydrocortisone (HC)-treated mice. The distribution of ANAE-positive lymphocytes as well as stromal cells in peripheral lymphatic organs was different in T- and B-dependent regions, while in the thymus there was the difference between cortex and medulla. The HC application evoked a marked changes in the pattern of ANAE-positive cells distribution of the thymus, producing only a slight ones in the remaining organs. The influence of HC on the ANAE reaction intensity and distribution in the examined tissue sections is discussed.