1,2,3-Trichlorobenzene (TCB) was ground with calcium oxide (CaO) powder in air with a planetary ball mill. A mechanochemical reaction was induced, resulting in the decomposition of TCB through dechlorination from the benzene nucleus. The mechanochemical dechlorination was monitored by a suite of analytical methods including X-ray diffraction (XRD), thermogravimetry (TG), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, electron spin resonance (ESR), gas chromatography/mass spectrometry (GC/MS), and ion chromatography. With the increase in grinding time, the remaining amount of TCB decreased and reached 0.02% at 360 min grinding. On the other hand, the water-soluble amount of chlorine increased correspondingly and reached 95%, indicating the successful transformation of chlorine from organics into inorganic chloride. Additionally, the other final products detected after grinding were carbon and some minor methane and ethane, although the formation of some chlorine-containing intermediate phases such as dichlorobenzene and tetrachlorobenzene was observed in the early stage of grinding (for 1 h). A possible decomposition pathway based on dechlorination or dehydrochlorination is compared.

The growth plates of the femoral head of Japanese white rabbits aged 5, 10, 15 and 20 weeks were stained for apoptotic and proliferating chondrocytes using the TUNEL and PCNA antibody staining techniques. Both TUNEL-and PCNA-positive chondrocytes were detected in all of the specimens. The positive ratios of both stainings were calculated for the whole plate and for the resting, proliferating and hypertrophic zones. The highest ratios in both stainings occurred in the hypertrophic zone in all age groups. With growth, the TUNEL-positive ratio increased whereas the proliferating ratio decreased.We suggest that the increase in chondrocytic death by apoptosis and the decrease in cell proliferation potential led to closure of the growth plate.J Bone Joint Surg [Br] 1997;79-B:483-6. Received 3 September 1996; Accepted after revision 22 November 1996 In tissues, homeostasis is achieved by the balance between cell death and proliferation. When a population of cells is changing, however, the proportion of proliferating to dying cells may be expected to vary, and should occur in chondrocytes during closure of the growth plate in endochondral ossification. It has been suggested that apoptosis 1 is closely related to closure and endochondral ossification. 2-5 Our aim was to elucidate the kinetics of apoptosis in chondrocytes in closing growth plates by comparing the ratios of dying to proliferating chondrocytes using markers for apoptosis and proliferation. MATERIALS AND METHODSWe used eight Japanese white rabbits, two of which were killed by intravenous injection of pentobarbital sodium at each of 5, 10, 15 and 20 weeks of age. The epiphyseal growth plate with metaphyseal and epiphyseal bones was obtained from the middle of the femoral head of each rabbit.The growth plates were sectioned coronally into blocks 5 mm thick. These were fixed in 10% formalin for three days, decalcified by EDTA at 4°C for ten days, embedded in paraffin and cut into slices 2.5 m thick.For in situ visualisation of apoptotic cells, we performed terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP)-biotin nick end labelling (TUNEL). 6 To show the proliferative portion of the tissues, we used immunostaining for proliferating cell nuclear antigen (PCNA), an auxiliary protein of DNA polymerase. After removal of the paraffin and dehydration, the first slice from each specimen was stained with haematoxylin and eosin for histological differentiation of the three zones of the growth plate: resting, proliferating and hypertrophic (Fig. 1). The second slice was stained by the TUNEL method. The reaction time was 20 minutes for protease K and 100 minutes for both TdT and biotin-dUTP. We performed PCNA immunostaining on the third slice using the labelled streptavidin-biotin (LSAB) method with a reaction time of 60 minutes for anti-PCNA antibody (PC-10, Code No. M879, DAKO, Denmark) at a dilution of 1:50. Counterstaining was carried out by methyl green in both instances. Some sections used for TUNEL staining were pretreated wit...

UC placement is simple, helping to prevent ureteral injury during laparoscopic hysterectomy and enhancing safety of this procedure.

No abstract

Human sperm nucleoproteins consist of protamines and histones. Changes in composition of these proteins are thought to correlate with spermatogenesis and may be involved in some instances of male infertility. We sought to separate sperm nucleoproteins including variants of protamine using an improved two-dimensional electrophoretic method, with the aim of comprehensively analysing all sperm nucleoprotein constituents. After extracting nuclear basic proteins from the sperm of normal volunteers, we analysed these proteins on a gel sheet by a radical free, highly reducing method based on Kaltschmidt and Whittmann's two-dimensional electrophoresis. Basic proteins from sperm nuclei were separated clearly into 12 spots. By amino acid sequence analysis, these spots corresponded to protamine 1 (P1)- (five spots), protamine 2 (P2)-related proteins (six spots) and testis-specific histone H2B (one spot). The N-terminal amino acid sequences of the six P2-related proteins were compatible with those of HPI1, HPI2, HPS1, HPS2, HP2 and HP3, and quantitative comparison could be performed. In conclusion, human sperm nucleoproteins including all P2-related variants could be analysed quantitatively with high resolution on a single electrophoretic gel.