First histological observations on the incorporation of a novel calcium phosphate bone substitute material in human cancellous bone

Calcium phosphates are frequently used as bone substitute materials because of their similarity to the mineral phase of bone, absence of antigenicity, and excellent osteoconductivity. However, in most currently available mineral substitutes, resorption occurs slowly if at all. In contrast, calcium phosphate cements have shown rapid resorption and remodeling in animal studies. In two prospective studies, a novel amorphous calcium phosphate cement (Biobon) was implanted in human patients for the first time. After 2-12 months, ten biopsies were obtained from nine individuals during secondary surgical interventions, for example, for implant removal. In all specimens, partial replacement of the material by new bone was observed, while residues of the cement were still visible. Undecalcified sections revealed extensive bone formation in immediate contact to the cement without fibrous interface. Polynucleated cells and superficial lacunae were indicative of resorptive activity, but inflammatory tissue response was absent. The new bone displayed regular trabecular and osteonal patterns. The histologic findings are in accordance with the excellent biocompatibility observed in the clinical follow-up. Though still incomplete, the resorbability of this cement appears superior to sintered calcium phosphates in these biopsy specimens. Presumably this is due to its amorphous crystalline structure. Biobon merits further studies as a promising substance for bone defect reconstruction in non-stress-bearing areas.     

Preliminary observations on polar body extrusion and pronuclear formation in human oocytes using time-lapse video cinematography

In this study, we have used time-lapse video cinematography to study fertilization in 50 human oocytes that had undergone intracytoplasmic sperm injection (ICSI). Time-lapse recording commenced shortly after ICSI and proceeded for 17-20 h. Oocytes were cultured in an environmental chamber which was maintained under standard culture conditions. Overall, 38 oocytes (76%) were fertilized normally, and the fertilization rate and embryo quality were not significantly different from 487 sibling oocytes cultured in a conventional incubator. Normal fertilization followed a defined course of events, although the timing of these events varied markedly between oocytes. In 35 of the 38 fertilized oocytes (92%), there were circular waves of granulation within the ooplasm which had a periodicity of 20-53 min. The sperm head decondensed during this granulation phase. The second polar body was then extruded, and this was followed by the central formation of the male pronucleus. The female pronucleus formed in the cytoplasm adjacent to the second polar body at the same time as, or slightly after, the male pronucleus, and was subsequently drawn towards the male pronucleus until the two abutted. Both pronuclei then increased in size, the nucleoli moved around within the pronuclei and some nucleoli coalesced. During pronuclear growth, the organelles contracted from the cortex towards the centre of the oocyte, leaving a clear cortical zone. The oocyte decreased in diameter from 112 to 106 microm (P < 0.0001) during the course of the observation period. The female pronucleus was significantly smaller in diameter than the male pronucleus (24.1 and 22.4 microm respectively, P = 0.008) and contained fewer nucleoli (4.2 and 7.0 respectively, P < 0.0001). After time-lapse recording, oocytes were cultured for 48 h prior to embryo transfer or cryopreservation. Embryo quality was related to fertilization events and periodicity of the cytoplasmic wave, and it was found that good quality embryos arose from oocytes that had more uniform timing from injection to pronuclear abuttal and tended to have a longer cytoplasmic wave. In conclusion, we have shown that time-lapse video cinematography is an excellent tool for studying fertilization and early embryo development, and have demonstrated that human fertilization comprises numerous complex dynamic events.      

Immobilization of antibodies on a new solid phase for use in ELISA

Antibodies to myoglobin were immobilized by covalent linkage to polyester film for use in a solid-phase ELISA. The covalent linkage of antibody to this new solid phase was accomplished by partial acid hydrolysis of the film followed by periodate oxidation. About 60 ng of protein could be immobilized per cm2 of film and the binding was stable. Antimyoglobin IgM immobilized on the films was used in the ELISA technique to detect myoglobin within the range 0.25-10 ng. The assay procedure was found to be very accurate and the coefficient of variation of each concentration ranged from 0.63 to 2.1%. Furthermore the immobilized film could be re-used after dissociating the antigen antibody complexes.     

Development and evaluation of a phage typing scheme for Vibrio cholerae O139

The scenario of cholera that existed previously changed in 1992 and 1993 with the emergence of toxigenic Vibrio cholerae O139 in India. The genesis of the new serogroup formed the impetus to search for O139 phages in and around the country. A total of five newly isolated phages lytic to V. cholerae O139 strains were used for the development of this phage typing scheme. These phages differed from each other and also differed from the existing O1 phages in their lytic patterns, morphologies, restriction endonuclease digestion profiles, and immunological criteria. With this scheme, 500 V. cholerae O139 strains were evaluated for their phage types, and almost all strains were found to be typeable. The strains clustered into 10 different phage types, of which type 1 (38.2%) was the dominant type, followed by type 2 (22.4%) and type 3 (18%). Additionally, a comparative study of phage types in 1993 and 1994 versus those from 1996 to 1998 for O139 strains showed a higher percentage of phage type 1 (40.5%), followed by type 3 (18.8%) during the period between 1993 and 1994, whereas phage type 2 (32. 1%) was the next major type during the period from 1996 to 1998. This scheme comprising five newly isolated phages would be another useful tool in the study of the epidemiology of cholera caused by V. cholerae O139.     

Effect of lycoriside,an acylglucosyloxy alkaloid,on mast cells

The effect of lycoriside, an acylglucosyloxy alkaloid from Crinum asiaticum Linn, (family Amaryllidaceae), with or without sitosterol-3-O--D-glucoside, was studied on the rate of degranulation of peritoneal mast cells of albino rats. Lycoriside, at lower concentrations (1–20 µg/ml), in vitro, produced statistically significant protection against Tween 80-induced degranulation, as also to sensitized mast cells challenged with an antigen (horse serum). It also provided protection against compound 48/80-induced degranulation of mast cells when administered in vivo (1–5 mg/kg, po). At higher concentrations (100 µg/ml and above), in vitro, however, it had a mast-cell degranulation effect per se. The addition of sitosterol-3-O--D-glucoside to lycoriside did not modify the effect of the latter compound. The mechanism of the dual response elicited by lycoriside is appraised in view of a concentration-dependent anti- or prerelease effect on mast-cell mediators.     

Management of Neglected Upper Cervical Spine Injuries

BackgroundInjuries involving upper cervical spine are serious and fatal injuries which are associated with alteration of normal occipital–cervical anatomy. These injuries may result in permanent neurologic deficits or neck deformity if not treated in a timely and appropriate manner.ObjectiveTo evaluate the outcomes of neglected upper cervical spine injuries treated by various methods.Study designRetrospective study.Materials and methodsTwelve patients attending ER or OPD with a history of neck trauma and who were diagnosed with fractures and fracture dislocations C1 and C2 were included in the study. Fresh injuries sustained within a week were excluded from study. The outcomes were measured in terms of improvement in VAS, ODI Scores and correction of the neck deformity. Surgical parameters like duration of surgery and blood loss were also observed.ResultsEleven males and one female. The mean age was 40.9 ± 16.9 (07–67 years). Eleven patients underwent posterior instrumentation, while one patient was treated anteriorly. The mean delay in presentation was 28 ± 8.67 days (15–42 days). The mean duration of surgery was 188.3 ± 34.35 min (120–240 min), average blood loss was 350 ± 111.8 ml (150–600 ml). The mean VAS improved from 8.45 ± 0.89 to 3.9 ± 0.51 (p < 0.05). The mean ODI Pre-operatively was 88.45 ± 5.89 which improved to 31.9 ± 4.01 (p < 0.05). The neck deformity/torticollis was corrected in all the patients.ConclusionsNeglected upper cervical spine injuries are difficult to treat and a posterior approach is helpful in reducing the subluxations indirectly and to obtain a posterior fusion.     

Complete nucleotide sequence of mouse mammary tumor virus from jyg chinese wild mice: Absence of bacterial insertion sequences in the cloned viral gag gene

Mammary tumors of a newly isolated strain of Chinese wild mouse (JYG mouse) harbor exogenous mouse mammary tumor virus (MMTV). The complete nucleotide sequence of exogenous JYG-MMTV was determined on the proviral 5' long terminal repeat (LTR)(partial)-gag-pol-env-3' LTR (partial) fragment cloned into a plasmid vector and the cDNA sequence from JYG-MMTV producing cells. Similarly to the other MMTV species the LTR of JYG-MMTV contains an open reading frame (ORF). The amino acid sequence of the JYG-MMTV ORF resembles that of SW-MMTV (92% identity) and endogenous Mtv-7 (93% identity) especially at the C-terminal region. Thus, a functional similarity in T-cell receptor V beta recognition as a superantigen is implicated among these MMTV species. Analysis of the viral gag nucleotide sequence revealed that this gene is not disrupted by the bacterial insertion sequence IS1 or IS2, which have been reported to be present in the majority of the plasmids containing the gag region. Comparison of amino acid sequences of JYG-MMTV with those of BR6-MMTV showed that over 96% of the amino acids of gag, pol, protease and env products are identical. These results suggest the intact nature of the nucleotide sequence of the near full-length MMTV genome cloned in the plasmid.     

Estrogen augments hypothalamicβ-endorphin secretion and activates an inhibitoryβ-endorphin short-loop feedback system

The role of estrogen in the regulation of hypothalamicβ-endorphin hormone secretion is studied by determiningβ-endorphin concentration in pituitary portal plasma of ovariectomized rats in the presence or absence of this steroid and/or the opioid antagonist naloxone. Twenty-six hours following s.c. injection of 10 /μg estradiol benzoate (estrogen) or oil, rats anesthetized with Saffan (alphaxolone/alphadolone) underwent pituitary stalk exposure and hypophysectomy, after which pituitary portal blood was continuously collected and stored in 15 min aliquots from 1100-1400 h. At 1100 h, animals were given an initial bolus iv injection of naloxone or saline (naloxone, 2 mg/ kg, or saline, 0.1 ml) and then infused (iv) continuously with naloxone (2 mg/kg/h) or saline (0.8 ml/h) until 1400 h. Plasma samples were extracted and assayed by radioimmunoassay forβ-endorphin. Treatment with estrogen increased the meanβ-endorphin levels twofold as compared to oil-treated controls. Naloxone potentiated estrogen action ofβ-endorphin secretion, but did not affect basalβ-endorphin secretion. These results suggest that estrogen enhancedβ-endorphin secretion from the hypothalamus. Furthermore, the hypersecretion ofβ-endorphin induced by naloxone with, but not without, estrogen supports the existence of an estrogen-activated short-loop negative feedback mechanism regulatingβ-endorphin secretion.     

Potentiation of the mitogenic effect of estrogen on the pituitary-gland by alcohol-consumption

The effect of alcohol on estrogen-regulated lactotropic cell proliferation was examined in Fisher 344 rats. Alcohol was administered for 2 weeks using liquid diet containing 8.7% ethanol (v/v) and 37% ethanol-derived calories. The control group was pair-fed with an isocaloric diet minus the ethanol or adlib-fed with normal diet. Ethanol-treated rats showed mean blood ethanol concentrations between 60-90 mg/dl. Alcohol treatment did not effect the body growth rate, but increased the DNA synthesis in lactotropes and reduced the levels of lactotropic growth inhibitory transforming growth factor-beta 1 (TGF-beta 1) protein and mRNA in the pituitary. These results suggest that alcohol promotes estrogen-induced lactotropic proliferation, possibly by down regulating the inhibitory TGF-beta 1 control of lactotropic function.