Isolation and mapping of a polymorphic CA repeat sequence at the human VRK1 locus

VRK1 is a novel human putative serine/threonine kinase, and is located on chromosome 14 at band q32 where an autosomal recessive congenital microphthalmia (CMIC) is mapped. We isolated a polymorphic dinucleotide CA repeat marker from a genomic clone containing the human VRK1 gene. This polymorphism will be useful in genetic studies of disorders localized at the 14q32 region, such as CMIC. Received: October 8, 1998 / Accepted: October 16, 1998     

Histopathologic study of partial hydatidiform moles and DNA triploid placentas

Sixty-two placentas with a triploid DNA content, which were analyzed by flow cytometry using paraffin-embedded tissues, were histologically investigated. These placentas were histologically classified as follows: 51 partial hydatidiform moles (PM), two hydropic abortuses and nine non-hydropic placentas. The DNA indices of the triploid peaks were between 1.41 and 1.60. Histologically, two populations of normal and edematous villi, vesicular villous edema with cistern formation, focal syncytiotrophoblastic hyperplasia with vacuola-tion, and villous scalloping with trophoblastic inclusion were almost always observed in the PM, but no single pathologic feature was specific for PM. The two entities, PM and triploid placenta, overlapped. Not all triploid gestations are PM and not all PM moles are triploid as shown in previously reported diploid or tetraploid PM. Although no patient with triploid PM developed persistent disease in this series, follow up of triploid PM is required as long as its true biological potential remains unclear. Flow cytometry is a reliable aid in the diagnosis of PM.     

Comparison of immunological activities of guinea pig IgG1 and IgG2 antibodies to low-molecular antigen]

The present study was carried out using a low-molecular antigen to clarify which of the two IgG subclasses (IgG1 and IgG2) of guinea pigs is responsible for the immunological assay systems in ordinary antigenicity tests for the development of a novel drug, and to what extent such IgG subclass antibodies contribute to the assay systems in addition to IgM antibody. Guinea pigs were immunized with TNBS plus FCA (3 mg/body), at intervals of 10 days, 3 times in total. The anti-TNBS serum was fractionated into peak I (IgG2), peak II (IgG1) and peak IV (IgM) by DEAE-cellulose column chromatography, and the immunological activities as well as the functions of the above three peaks were estimated by HA, homologous PCA reaction, PSANA and P-Arthus. The IgM and two IgG peaks possessed immunological activities on HA and P-Arthus, and the following degree of activities was shown in both assay systems: peak II (IgG1) greater than peak IV (IgM) greater than peak I (IgG2). In the homologous PCA reaction and PSANA, immunological activities were seen only in peak II (IgG1). It is confirmed that the IgG1 subclass is a homocytotropic antibody involved in the PCA and PSANA assay systems in the case of low-molecular immunogens such as TNBS.     

Coenzyme models, 6. Addition reaction of sulfite ion to monomeric and polymeric nicotinamides. Correlation between equilibria and reaction rates

Rate and association constants (k and K) for the addition of sulfite ions to five nicotinamides substituted at the ring nitrogen, ( 2a–e ), and to poly(1-(4-vinylbenzyl)nicotinamide chloride) (poly{1-[4-(3-aminocarbonylpyridiniomethyl)phenyl]ethylene chloride}) (poly( 1 )) were determined at 30°C in aqueous systems. It was found that the reaction parameters for the addition of SO to poly( 1 ) are markedly enhanced (20500-fold in the k term and 510-fold in the K term) compared with the addition to the corresponding monomeric compound, 3-aminocarbonyl-1-benzylpyridinium chloride ( 2d ), and the enhancements are suppressed with increased ionic strengths. These enhanced reaction parameters for poly( 1 ) are deviated to the upper area by two logarithmic units from a linear log K vs. logk relationship which holds for monomeric nicotinamides. This means that the rate constant is enhanced more effectively that the association constant in the polymeric system. Plots of log K_CN^? vs. log K and of log k_{f, CN}- vs. log k- gave good linear relationships. The plot for poly( 1 ), greatly deviated again to the upper area. The SO ion interacts with poly( 1 ), a cationic polyelectrolyte, more strongly than the CN^? ion.     

Palisading subcutaneous fibrous histiocytoma

A case of palisading subcutaneous fibrous histiocytoma, a very rare variant of fibrous histiocytoma (dermatofibroma), arising in the wrist of a 41-year-old man is presented. An unencapsulated subcutaneous tumor measuring 0.8 x 0.8 x 0.7 cm was histologically characterized by predominant nuclear palisading and a peripheral area with a pattern quite characteristic of conventional fibrous histiocytoma. Immunohistochemically, the tumor cells were strongly positive for vimentin, alpha-smooth muscle, and muscle actin, but negative for S-100 protein, indicating a fibroblastic or myofibroblastic nature. The patient has been well without recurrence for 6 years and 8 months after the excision. This neoplasm should be differentiated from benign and malignant skin or soft tissue tumors with a palisading pattern. Pathologists and clinicians should know of the existence of this type of fibrous histiocytoma and should avoid overdiagnosis and overtreatment.     

Immunohistochemical characterization of p57(KIP2) expression in early hydatidiform moles

The differentiation of complete mole (CM), an aberrant androgenetic conceptus, from partial mole (PM) and hydropic abortion (HA) in early gestations is very important for patient management. In this study, 10 diploid voluntary artificial abortions (ABs), 20 diploid HAs, 20 triploid PMs, and 44 diploid CMs (including 4 persistent diseases), all of which were in the first trimester, were evaluated by immunohistochemistry of formalin-fixed tissues using a monoclonal antibody against p57(KIP2) protein (p57), a putative paternally imprinted inhibitor gene. DNA ploidy in all cases was analyzed by flow cytometry. In all ABs, nuclear p57 was strongly expressed in cytotrophoblasts, intermediate trophoblasts, villous stromal cells, and decidual stromal cells but was absent in syncytiotrophoblast. In diploid CMs, p57 expression in cytotrophoblasts and villous stromal cells was either absent (37 cases) or very low (7 cases). Villous intermediate trophoblasts stained for p57 in 12 cases of CM. On the other hand, 16 HAs and 19 PMs showed p57 levels comparable to those observed in ABs. Decidual stromal cells provided a reliable internal control in all cases. These findings support the hypothesis that misexpression of p57 is involved in the abnormal development of androgenetic CMs. This immunohistochemical analysis is a useful tool for the differential diagnosis of CMs.     

Interaction of rat ascites hepatoma cells with cultured mesothelial cell layers: a model for tumor invasion

Interactions of rat ascites hepatoma cells with primary cultured layers of rat mesentery-derived cells were studied. The mesentery-derived cells were isolated from rat mesentery and cultured in Eagle's minimum essential medium with a 2-fold concentration of amino acids and vitamins supplemented with 10% calf serum. The primary cultured cells, consisting mainly of mesothelial cells in polygonal shape, forms a "paving stone" sheet. Upon seeding the tumor cells on the mesentery-derived cell layers, three different types of tumor cell growth were observed. Type 1 was the formation of piled-up tumor cell nests on mesothelial cell layers. Type 2 was the formation of flattened tumor cell islands underneath mesothelial cell layers. This island formation was clearly observed under a phase contrast microscope 2 days after the tumor cell seeding. Protrusion of cellular processes of the tumor cells beneath mesothelial cells was occasionally seen. Type 3 was the growth of tumor cells in suspension. These types of tumor cell growth closely resemble those in the peritoneal cavity observed after i.p. implantation of the tumor cells. When the tumor cells recovered from the blood of tumor-bearing rats were seeded, flattened tumor cell islands were formed 15 times more frequently than when the tumor cells isolated from host peritoneal cavity were seeded. Shortly after the appearance of small flattened tumor cell islands, a distinct morphological change of mesothelial cells from polygonal to spindle shape was seen preferentially at the marginal area of the cell layers (a partial retraction of cell edges). The retraction of mesothelial cells was induced not only by seeding the tumor cells but by adding the tumor ascites fluid or the medium conditioned by the tumor cell culture. The morphological change was reversed by changing the culture medium to remove the effectors. These results indicate that the system described in this study can provide a useful model to study tumor cell invasion.     

The anti-proliferative effect of proliferating cell nuclear antigen-specific antisense oligonucleotides on human gastric cancer cell lines

The proliferating cell nuclear antigen (PCNA) is a nuclear protein that leads DNA synthesis by the DNA polymerase delta. As the PCNA gene is strongly expressed in invasive gastric cancer cells with high proliferative activity, PCNA is suspected of playing an important role in the proliferation and advancement of gastric cancer. Thus, the effects of antisense oligonucleotides specific for PCNA mRNA were examined in seven gastric cancer cell lines. It was found that treatment with antisense oligonucleotides at concentrations of 10–40 M dose-dependently inhibited the growth of all cell lines; however, random sequence oligonucleotides did not modify the proliferation of any type of cells. These results indicate that PCNA is essential for cell proliferation in gastric cancer cells, and that the growth inhibitory effect results from the inhibition of PCNA gene expression. Therefore, PCNA-specific antisense oligonucleotides may be effective in the treatment of gastric cancer.     

A study on how a 6-month aerobic exercise program can modify coronary risk factors depending on their severity in middle-aged sedentary women

It is well known that physical exercise can reduce coronary risk factors. But how an aerobic exercise modifies coronary risk factors in relation to severity and physical fitness is still controversial. Fifty-four middle-aged women (mean age, 55 years) completed a 6-month on-site and home-based anaerobic threshold-level exercise program. The changes in coronary risk factor profiles were observed during the pre-intervention and intervention periods. Before the intervention (during control period), most coronary risk factors showed a rather unfavorable trend. After the program, their mean body weight decreased from 56.7 to 55.7 kg (p>0.05) and the proportion of body fat from 30.9 to 27.9% (p>0.05) without any reduction in lean body mass. Systolic blood pressure (SBP) decreased from 129.0 to 125.0 mm Hg (p>0.05) and diastolic blood pressure from 79.5 to 76.6 mm Hg (p>0.05). Fasting plasma glucose (FPG) declined from 109.6 to 103.4 mg/dl (p>0.05). Changes in SBP and FPG were most remarkable in their respective worst tertile. Serum lipids improved only modestly. Maximum oxygen uptake increased from 23.6 to 26.1 ml/kg/min (p>0.01). However, no significant correlations were found between changes in coronary risk factors and those in physical fitness. We conclude that the 6-month aerobic exercise program would modify women’s coronary risk factors depending on their initial values, probably independently of the changes in physical fitness.