Identifying pathogenic genetic background of simplex or multiplex retinitis pigmentosa patients: a large scale mutation screening study.

BACKGROUND AND PURPOSE: More than half of the retinitis pigmentosa (RP) cases are genetically simplex or multiplex. To date, 37 causative genes of RP have been identified; however, the elucidation of gene defects in simplex or multiplex RP patients/families remains problematic. The aim of our study was to identify the genetic causes of RP in patients with unknown or non-Mendelian inheritance. METHODS AND RESULTS: Since 2003, 52 simplex RP patients, 151 patients from 141 multiplex RP families, and six sporadic patients with retinal degeneration were studied. A total of 108 exons of 30 RP-causing genes that harboured the reported mutations were screened by an efficient denaturing high performance liquid chromatography (dHPLC) based assay. Aberrant fragments were subsequently analysed by automatic sequencing. Twenty-six mutations, including two frameshift mutations, one single amino acid deletion, and 23 missense mutations, were identified in 28 probands (14.07%). Eighteen mutations have not been reported to date. Three pairs of combined mutations in different genes were identified in two sporadic cases and one multiplex family, indicating the possibility of novel digenic patterns. Of the 23 missense mutations, 21 were predicted as deleterious mutations by computational methods using PolyPhen, SIFT, PANTHER, and PMut programs. CONCLUSION: We elucidated the mutation spectrum in Japanese RP patients and demonstrated the validity of the mutation detection system using dHPLC sequencing for genetic diagnosis in RP patients independent of familial incidence, which may provide a model strategy for identifying genetic causes in other diseases linked to a wide range of genes.

Generation of self HLA-DR-specific CD3+CD4-CD8+ cytotoxic T cells in chronic graft-versus-host disease.

To analyze the mechanism of chronic graft-versus-host disease (GVHD) characteristic of autoimmune disease, we used a cell-mediated lympholysis assay to study the autoreactivity of PBL from two patients after MHC-matched BMT. Our data indicate the induction of CD3+CD4-CD8+ autoreactive cytotoxic T lymphocytes (CTL) in the one patient with chronic GVHD and an important role for allo-non-MHC (minor histocompatibility) antigen-specific CD3+CD4+CD8- helper T cells in this induction. Experiments using HLA-DR gene-transfected mouse L cells as target cells and blocking assays with anti-HLA class I and class II antibodies provided evidence that autoreactive CTL recognized HLA-DR antigen on autologous cells. Analysis of antigen-specific T cell proliferative responses in these patients to examine the effect of self HLA-DR-specific CTL on the antigen presenting cell (APC)-T cell interaction suggested that donor bone marrow-derived self HLA-DR-specific CTL are responsible for the decreased antigen-presenting ability of the patient's APC. These results suggest a new interpretation of the induction mechanism of chronic GVHD and its associated immunosuppression after MHC-matched BMT based on diminished APC function.

Sex differences in proximal humeral outline shape: elliptical Fourier functions.

A method is presented for the numerical analysis of sex differences in size and shape of the proximal humeral outlines using elliptical Fourier functions (EFFs). A skeletal sample consisting of right and left humeri pairs of 69 individuals, 36 males and 33 females, was used. The proximal superior view in the plane of the proximo-distal axis of each humerus was photographed and then 54 boundary points were located on the two-dimensional outline tracings. These points were digitized and used to compute EFFs with 27 harmonics. From the EFFs, a set of expected points on the proximal humeral outline was generated using the centroid as an origin. Superimposition of the male and female outlines on this centroid provided a detailed picture of the relative sex differences in size and shape with respect to that center. The bounded area of the proximal humeral outline showed statistically significant sex differences. Additionally, statistical results of the amplitudes derived from the "area-standardized" EFFs and visual assessments of the mean outline plots indicated significant sex differences in shape of the proximal humeral outlines. Focusing on localized regional differences, the greater tubercle was located more postero-medially and the lesser tubercle was located more anteriorly in the males compared to the females. Sex determinations from the proximal humeri were also examined with discriminant functions based on the amplitudes, which represent shape characteristics of the outline, and the hounded area. Using a cross-validation method, predictions of the percentages of cases correctly classified with the discriminant functions were ranged from 92.8% to 95.7% for the right and left humeral data. These results suggest that differences in size and shape of the proximal humeral outlines may be better predictors of sex when compared with conventional measurements of the humerus.

Sagittal configuration of spinal curvature line in sportsmen using Moire technique.

A separate moire photograph of 380 men's back was taken. Using a micro-computer the sagittal configuration of the spinal curvature line was reconstructed based on the moire fringe on the back. Then the sagittal configuration of this line was compared among groups according to their sports: rugby, soccer, kendo, swimming, yachting, four track and field events (sprinting, distance running, jumping, and throwing), body-building as well as non-athletic men. Distance runners and sprinters had a greater degree of thoracic kyphosis and greater degree of lumbar lordosis, while non-athletes had a smaller degree of both kyphosis and lordosis. Swimmers, bodybuilders, rugby and soccer players had partial lordosis. The degree of lumbar lordosis of swimmers and bodybuilders was less than average while the degree of thoracic kyphosis was ordinary and the degree of thoracic kyphosis of rugby and soccer players was less than average while the degree of lumbar lordosis was ordinary.

Fourier analysis of shape changes in the Japanese skull: occipital view.

Photographs (norma occipitalis) of Japanese adult skulls (n = 171) were mathematically described with Fourier analysis to assess shape changes over time. The materials used were adult male skulls excavated from the Kanto District and covered four age-periods; Jomon (2000-1000 BC). Kamakura (14C. AD), Muromachi (15C. AD). and Edo (18C. AD). Mean raw data values, containing both size and shape information of each vector, were compared for the four age-periods. Statistically significant differences among the four age-periods were found using an ANOVA. The form of the Jomon skulls displayed the most difference among the four groups. This trend was especially evident at the parietal margin. Fourier analysis was subsequently undertaken to detect the presence of secular changes in skull form after standardization for size. Once the effect of size was removed, the occipital shape of the Jomon skulls was found to be essentially similar to those of the more recent age-periods.(ABSTRACT TRUNCATED AT 400 WORDS)

Change in facial shape in two cohorts of Japanese adult female students twenty years apart.

INTRODUCTION: Secular changes in the head and body dimensions of the Japanese population were recorded during the last century, but studies on Japanese adult facial shape were poorly documented. This study aimed to document the secular changes in facial shape and their association with craniofacial measures of Japanese adult female students. METHODS: Data were collected from 1998-2001 and 1975-1979 from institutions in Tokyo and Kyoto. The prosopic index (PI) was derived from morphological facial height (n-gn) and bizygomatic breadth (zy-zy). Multiple regression analysis was used to determine the association between PI (facial form) and craniofacial measures. RESULTS: Larger mean values for morphological facial height (n-gn) and for PI, but smaller values for bizygomatic breadth (zy-zy) and bigonial breadth (go-go) were observed in the current sample compared to their predecessors 21 years ago. The most prevalent facial shapes were mesoprosopic (30.53 percent) and euryprosopic (45.14 percent) for the current and previous series, respectively. There was a significant positive relationship between the PI and head length (g-op) (p less than 0.01), head breadth (eu-eu) (p less than 0.01) and head height (v-po) (p less than 0.05), and a negative relationship between the PI and head circumference (g-g) (p less than 0.01) and minimum frontal breadth (ft-ft) (p less than 0.01). CONCLUSION: Our results suggest that a narrowing of facial shape has been occurring for approximately 21 years in adult Japanese females. The best predictor variables for PI were head length, head circumference, head breadth, minimum frontal breadth and head height.

Cranial vault shape in fossil hominids: Fourier descriptors in norma lateralis.

Two major views of human evolution have elicited considerable controversy. These are: [1] the "out of Africa" hypothesis and [2] the "multiregional" hypothesis. This paper is an attempt to try to reconcile these two scenarios using hominid cranial vault data. Elliptical Fourier functions (EFFs) were used to describe, in visual and numerical terms, the shape of the human cranial vault in norma lateralis. Using jpeg images, contours of the cranial vault of a large sample of hominid specimens were pre-processed in Photoshop CS and rotated in 2D space (positional-orientation) so that a line drawn from nasion to porion was horizontal. The cranial vault image was then digitized with 72 closely-spaced points and submitted to a specially written routine that computed EFFs normalized by scaling (size-standardization). This ensured that the representation was invariant with respect to starting point, size and orientation. Statistically significant differences were found between the H. sapiens sample and both the H. erectus and H. neanderthalensis samples. In contrast, there were no statistically significant differences between the H. erectus and H. neanderthalensis groups, leading to three conclusions: [1] the similarity in cranial vault shape between H. erectus and H. neanderthalensis suggests a single gradually evolving lineage; [2] The taxon H. heidelbergensis can be embedded into the H. erectus→H. neanderthalensis line; and [3] H. sapiens seems to be a separate evolutionary development and is considered here either as a separate species or as a possible example of an allopatric semispecies (Grant, 1977). The results here suggest that human evolution over the last 2 Ma may turn out to be neither totally multiregional or simply out of Africa but rather represents a considerably more complicated picture.

Areal growth in the human fetal parietal bone.

The areal growth of the human fetal parietal bone is described using 51 dissected right parietal bones of Japanese fetuses ranging from the fifth month to term. Their shadows were taken on printing paper and analyzed by a sonic digitizer. The absolute growth of the projected area of the fetal parietal bones progresses at a fairly constant rate during the latter half of the fetal period, despite some deceleration in the 9-10th month. By allometric analysis, the allometric coefficients against crown-rump length are 2.201 for males, 2.202 for females, and 2.204 for both sexes, respectively. The allometry is essentially monophasic, showing no inflection point, indicating a change in the slope of the regression line; that is, the growth of the human fetal parietal bone is continuous with a constant specific growth rate. These results are discussed in connection with growth in thickness of the bone.

Developmental changes of the cranial bone thickness in the human fetal period.

The description of the developmental changes of cranial bone thickness in the fetal period is the primary aim of this paper. The materials utilized in this study consisted of the two series. One is those of dry bones of 62 Japanese fetuses ranging from the fifth month to term and the other is those of 56 Japanese fetuses ranging from the fourth month to term. The first series is mainly used for the tests of bilateral difference in the thickness of cranial bones. No statistical significance could be found. Allometry technique was then applied for the analysis in the growth of the thickness of cranial bones and an extremely positive alometric coefficient was obtained. This required us to study another material, i.e., the second series. The materials of the second series were first measured for crown-rump length, head length, head breadth and head circumference. The heads were then carefully dissected and the left side of individual bones of the cranium were removed to take further measurements for the arc length. The thicknesses at the ossification centers and midpoints were successively measured by a micrometer readable to 0.01 mm. The results of the second series were investigated from the viewpoint of absolute and relative growth. By allometric analysis, vertical and transverse arc lengths were found to be isometric with two exceptions, while every measurement of thickness shows an extremely positive allometry. This coincides with the results of the first series.

Prediction of adult stature for Japanese population: a stepwise regression approach.

The longitudinal growth in stature for 509 males and 311 females was characterized from early childhood to adulthood. A triphasic generalized logistic (BTT) model (Bock et al. [1994] Chicago: SSI) was used through the AUXAL software program. Growth parameters were derived from the estimated distance and velocity curves for each individual. A set of estimated growth parameters, including adult stature, was selected to develop equations, through the forward stepwise regression method, for the prediction of adult stature for Japanese boys and girls.

Adolescent growth events in eight decades of Japanese cohort data: sex differences.

The presence of secular trends in sex differences for the onset or takeoff of the adolescent growth spurt and subsequent (adolescent) growth of Japanese boys and girls were investigated using data published in "The Statistical Report of the School Health Survey" of the Ministry of Education, Science, Sports and Culture, Japan. An optimum kernel regression method was used to derive the biological parameters of the adolescent growth curve. The six parameters derived from the regression analysis were age at takeoff (ATO), height at takeoff (HTO), velocity at takeoff (VTO), peak height velocity (PHV), age at PHV (APHV), and height at PHV (HPHV). The sex difference in age at PHV is about 2.1 years. The sex difference in age at takeoff was about 2.1 years starting with the birth year of 1943, but has become almost insignificant with the birth year of 1980. After 1960, the sex difference in mean HPHV varied within 12-13 cm, but the difference in HTO showed a decreasing trend. After the birth year of 1911, the rate of growth at HTO for girls has become markedly greater than for boys. In contrast, the rate of growth at HPHV for girls has become significantly smaller than for boys (P < 0.05). Sex differences in ATO, HTO and VTO were significantly and negatively correlated with differences in APHV and HPHV, but positively correlated with the difference in PHV. The sex difference in PHV had no relationship with the differences in APHV and HPHV.

A factor analysis of cranial base and vault dimensions in children.

Lengths within the cranial base and vault were measured in cephalometric radiographs of 220 boys and 177 girls ranging in age from 0 to 15 years; all these children are participants in The Fels Longitudinal Growth Study. The present study is based on mixed longitudinal data derived from 1640 radiographs for boys and 1260 radiographs for girls. Factor analysis was applied separately for boys and girls for each age group; i.e., 0-3, 4-6, 7-9, 10-12, and 13-15 years. For the 0-3 year age group, two factors were extracted in each sex, whereas four factors were extracted in the rest of the age groups. The factor structures are similar in the three older age groups of boys (7-9, 10-12, and 13-15 years). The first four factors for these groups are labelled, respectively: cranial vault size, posterior cranial base length, presphenoid length, and basisphenoid length. The order of the third and fourth factors is reversed in the 7-9 year olds. For girls, the factors extracted were also the same in both the 7-9 and 10-12 year age groups, even though the order of factors was different between age groups; i.e., anterior cranial base length, cranial vault size, basisphenoid length, and basioccipital length. Differential growth rates among cranial base dimensions probably cause changes in factor patterns. Obliteration of the spheno-occipital synchondrosis is suggested as the mechanism responsible for the change of factor pattern in the girls. closure of this synchondrosis would have occurred too late to affect the patterns in boys.

Granulocyte-macrophage colony-stimulating factor abrogates transforming growth factor-beta 1-mediated cell cycle arrest by up-regulating cyclin D2/Cdk6.

The role of positive and negative cytokine interactions in G1 cell cycle regulation of haemopoietic cells was analysed by determination of the expression patterns of D-type cyclins and cyclin-dependent kinases (cdks) in SKM-1 myelodysplastic syndrome (MDS) cells incubated with granulocyte-macrophage colony-stimulating factor (GM-CSF) and/or transforming growth factor-beta 1 (TGF-beta 1). TGF-beta 1 inhibited SKM-1 cell proliferation due to the cell cycle arrest in G1 phase. GM-CSF abrogated the TGF-beta 1-mediated G1 arrest in these cells. Reverse transcription-polymerase chain reaction (RT-PCR) analysis indicated that TGF-beta 1-mediated G1 arrest correlated with the down-regulation of cdk4, cdk6 and cyclin D2, and that abrogation of TGF-beta 1-mediated G1 arrest by GM-CSF correlated with the constitutive over-expression of cyclin D2 and cdk6 but not cdk4. These results suggest the importance of cyclin D2/cdk6 levels in abrogating G1 arrest in cells exposed to TGF-beta 1, and raise the possibility that the GM-CSF-mediated up-regulatory pathway of signal transduction through cyclin D2/cdk6 differs from the TGF-beta 1-cdk4-mediated pathway in SKM-1 cells. This signal transduction pathway through cyclin D2/cdk6 might play an important role in haemopoietic regulation by the cytokine network.

Sibling correlations for cranial measurements from serial radiographs.

Sibling correlations are presented for nine craniofacial measurements from lateral cephalometric radiographs of participants in the Fels Longitudinal Study of Growth and their siblings. Because the data were collected serially, measurements on sibs at the same age can be compared at one-year intervals. Sibling correlations are generally lowest during the first year of life and highest as adulthood is approached, but no interpretable pattern is established during the intervening years. Most of the correlations are significantly greater than zero, and many exceed 0.5, the value expected for a completely heritable polygenic trait. This suggests that common sibling environment contributes to sibling resemblance for craniofacial dimensions in our sample.

Lymphoblastic transformation of chronic myelomonocytic leukemia in an infant.

A 10-month-old infant with chronic myelomonocytic leukemia (CMML) of 5 months' duration, who had been treated only with transfusion, displayed leukemic transformation characterized by lymphoid morphology, PAS positivity, and myeloperoxidase negativity. Surface marker analysis of blast cells revealed expression of lymphoid-associated antigens (CD10 and CD19) but not myeloid-associated antigens (CD13, CD14, and CD33). These findings suggest that some cases of infantile CMML are clonal disorders arising in a pluripotent stem cell that can also differentiate along the lymphoid cell lineage.

Standard RUS skeletal maturation of Tokyo children.

A total of 704 girls and 753 boys, all healthy, from 3 to 18 years of age, from Tokyo and its suburbs, were radiographed on the left hand and wrist in 1986. Their RUS (TW2) skeletal maturity was estimated, the 50th-centile skeletal maturity scores were obtained, and the smoothed RUS maturity curves were determined applying the cubic spline function to the 50th-centile scores. On this maturity curve the score at each 0.1 year of chronological age was obtained and allocated as a given RUS skeletal age. This set of scores and ages we termed the TW2-J RUS, i.e. the Japanese TW2 RUS maturity standard. Comparing this RUS standard with the British standard, the Belgian, the southern Chinese, and the northern Indian, it became clear that Japanese children's RUS skeletal maturity progresses rapidly during puberty (after ages 9 in girls and 11 in boys), and that the maximum score difference between neighbouring age groups was observed at ages 12.5 in girls and 14.5 in boys on the spline-smoothed curve. Japanese children attain the adult stage 1 or 2 years earlier than other groups of children (at ages 15 in girls and 16 in boys).