The assessment of carotid and vertebral arteries: a comparison of CFM duplex ultrasound with intravenous digital subtraction angiography.

The use of duplex ultrasound with colour flow mapping is compared with that of intravenous digital subtraction angiography (IVDSA) in the investigation of carotid and vertebral artery disease. Ninety-nine carotid and vertebral arteries were examined independently by IVDSA and ultrasound for location and degree of disease in 30 men (mean age 58) and 20 women (mean age 53). For purposes of comparison the carotid tree was divided into six segments. In 46% of cases no disease was detected by either modality. In 36% of cases where disease was found, ultrasound found mild disease in arteries reported as normal by IVDSA. Both modalities detected the six cases of total occlusion found in the study. IVDSA underestimated five cases of major disease found by ultrasound at the bifurcation or in the bulb. Overall agreement in grading vessel segments was good, with 74.5% in complete agreement and 90.3% grading stenoses to within +/- 25% of the other modality. There was good qualitative agreement in the findings for vertebral arteries. In four patients conventional arteriography was also available for comparison.     

Arterial repair after stenting and the effects of GM6001, a matrix metalloproteinase inhibitor

OBJECTIVES: This study compared the extracellular matrix (ECM) and cellular responses after stenting to balloon angioplasty (BA) and to determine the late effects of matrix metalloproteinase (MMP) inhibition on arterial repair after stenting. BACKGROUND: Although stenting is the predominant form of coronary intervention, there is limited understanding of the early and late arterial response. METHODS: In a double-injury rabbit model, adjacent iliac arteries in 87 animals received BA (3.0 mm diameter) or stenting (3.0 mm NIR). Rabbits were treated for 1 week postprocedure with either GM6001 (100 mg/kg per day), an MMP inhibitor or placebo and sacrificed at 1 week or at 10 weeks' postprocedure. Arteries were analyzed for morphometry, collagen content, gelatinase activity, cell proliferation and DNA content.RESULTS: Stented arteries had significant increases in collagen content (2-fold) at 10 weeks compared to BA-treated arteries. At one week, overall gelatinase activity was increased >2-fold in stented arteries, with both 72 kD and 92 kD gelatinase activity. Stented arteries also had increases in both intimal DNA content (1.5-fold) and absolute cell proliferation (4-fold). Compared to placebo, GM6001 significantly inhibited intimal hyperplasia and intimal collagen content, and it increased lumen area in stented arteries without effects on proliferation rates. CONCLUSIONS: Stenting causes a more vigorous ECM and MMP response than BA, which involves all layers of the vessel wall. Inhibition by MMP blocks in-stent intimal hyperplasia and offers a novel approach to prevent in-stent restenosis.     

Growth in vitro and acquisition of meiotic competence after the cryopreservation of isolated mouse primary ovarian follicles.

The growth and acquisition of meiotic competence of oocytes from fresh and frozen-thawed primary follicles collected from 10-day-old mice was compared during culture in collagen gels for 12 days. The oocytes contained in primary follicles have a mean diameter of about 48 microns and do not resume meiosis without further growth and development. During the 12-day culture period the mean diameter of the oocytes increased to over 60 microns. The oocytes were capable of resuming meiosis when isolated from the gel and cultured in the absence of follicular cells in a manner similar to that observed in vivo. Freezing and thawing did not affect oocyte growth or the ability to resume meiosis; this demonstrates the possibility of storing large numbers of female gametes for subsequent development.     

Ovary and ovulation: Follicular development in cryopreserved marmoset ovarian tissue after transplantation

Pieces of marmoset ovary were frozen by slow cooling in 1.5M dimethylsulphoxide. The follicles in fresh and frozen tissuewere counted and examined for morphological appearance in stainedserial sections. The proportion of normal follicles was similarin fresh tissue and frozen tissue examined immediately afterthawing. Follicles at all stages of folliculogenesis up to thesmall antral stage survived freezing and thawing. Fresh andfrozen tissue was transplanted underneath the kidney capsulesof ovariectomized immunodeficient mice. The establishment ofgrafts was similar, and oestrogenic activity (cornificationof the vaginal epithelium) was observed in the recipients 20and 16 days after transplantation of fresh and frozen graftsrespectively. The total number of follicles and the proportionof normal follicles were similar in fresh and frozen grafts.Grafts of frozen tissue recovered between 7 and 15 days aftertransfer contained follicles up to the small antral stage ofdevelopment. Grafts recovered between 21 and 32 days containedfollicles at all stages of folliculogenesis, including largeantral follicles (1–2 mm diameter). Our results suggestthat freezing and thawing do not substantially damage marmosetovarian tissue, and the cryopreserved tissue retains its abilityto support the development of large antral follicles.     

Visually defined zones of focal ischemia in the rat brain

A technique to guide brain tissue sampling in experimental focal ischemia is described. This technique allows visual identification for accurate metabolic analysis of regions with variable tissue perfusion. After 2.5 hours of unilateral middle cerebral artery occlusion (MCAO) in the rat, intravenous infusion of the vital dye, neutral red (NR), identified three distinct regions: (a) an intensely stained region consisting of the entire contralateral hemisphere and the medial ipsilateral hemisphere; (b) a blanched region in the lateral ipsilateral hemisphere; and (c) a zone of intermediately stained tissue interposed between the medial and lateral ipsilateral regions. Close regional correlation between NR staining and the iodoantipyrine intensity of adjacent brain slices suggests that NR distribution is a qualitative indicator of brain perfusion. Using the NR staining pattern to guide tissue dissection for metabolite analysis of high energy phosphates (adenosine triphosphate and phosphocreatine) and glucose-related metabolites showed that the blanched region contained ischemic tissue, whereas values from the contralateral cortex resembled those of control tissue. The intermediately stained region exhibited a gradient of metabolic perturbation. Tissue near intensely stained regions resembled control tissue, and tissue near blanched regions resembled ischemic tissue. These results confirm the concept of a metabolic penumbra. When present, it occupies only the most medial area of the intermediate region at 2.5 hours after MCAO.     

Effects of Growth Hormone Administration on Bone Mineral Metabolism,PTH Sensitivity and PTH Secretory Rhythm in Postmenopausal Women With Established Osteoporosis

Introduction : Growth hormone (GH) replacement improves target organ sensitivity to PTH, PTH circadian rhythm, calcium and phosphate metabolism, bone turnover, and BMD in adult GH‐deficient (AGHD) patients. In postmenopausal women with established osteoporosis, GH and insulin like growth factor‐1 (IGF‐1) concentrations are low, and administration of GH has been shown to increase bone turnover and BMD, but the mechanisms remain unclear. We studied the effects of GH administration on PTH sensitivity, PTH circadian rhythm, and bone mineral metabolism in postmenopausal women with established osteoporosis. Materials and Methods : Fourteen postmenopausal women with osteoporosis were compared with 14 healthy premenopausal controls at baseline that then received GH for a period of 12 mo. Patients were hospitalized for 24 h before and 1, 3, 6, and 12 mo after GH administration and half‐hourly blood and 3‐h urine samples were collected. PTH, calcium (Ca), phosphate (PO₄), nephrogenous cyclic AMP (NcAMP), β C‐telopeptide of type 1 collagen (βCTX), procollagen type I amino‐terminal propeptide (PINP), and 1,25‐dihydroxyvitamin D [1,25(OH)₂D] were measured. Circadian rhythm analysis was performed using Chronolab 3.0 and Student's t‐test and general linear model ANOVAs for repeated measures were used where appropriate. Results : IGF‐1 concentration was significantly lower in the women with established osteoporosis compared with controls (101.5 ± 8.9 versus 140.9 ± 10.8 μg/liter; p < 0.05) and increased significantly after 1, 3, 6, and 12 mo of GH administration (p < 0.001). Twenty‐four‐hour mean PTH concentration was higher in the osteoporotic women (5.4 ± 0.1 pM) than in healthy controls (4.4 ± 0.1 pM, p < 0.001) and decreased after 1 (5.2 ± 0.1 pM, p < 0.001), 3 (5.0 ± 0.1 pM, p < 0.001), 6 (4.7 ± 0.1 pM, p < 0.001), and 12 mo (4.9 ± 0.1 pM, p < 0.05) of GH administration compared with baseline. NcAMP was significantly lower in osteoporotic women (17.2 ± 1.2 nM glomerular filtration rate [GFR]) compared with controls (21.4 ± 1.4 nM GFR, p < 0.05) and increased after 1 (24.2 ± 2.5 nM GFR, p < 0.05), 3 (27.3 ± 1.5 nM GFR, p < 0.001), and 6 mo (32.4 ± 2.5 nM GFR, p < 0.001) compared with baseline. PTH secretion was characterized by two peaks in premenopausal women and was altered in postmenopausal women with a sustained increase in PTH concentration. GH administration also restored a normal PTH secretory pattern in the osteoporotic women. The 24‐h mean adjusted serum calcium (ACa) concentration increased at 1 and 3 mo (p < 0.001) and PO₄ at 1, 3, 6, and 12 mo (p < 0.001). 1,25(OH)₂D concentration increased after 3, 6, and 12 mo of GH (p < 0.05). An increase in urine Ca excretion was observed at 3 and 6 mo (p < 0.05), and the renal threshold for maximum tubular phosphate reabsorption rate (TmPO4/GFR) increased after 1, 3, 6, and 12 mo (p < 0.05). βCTX concentration increased progressively from 0.74 ± 0.07 μg/liter at baseline to 0.83 ± 0.07 μg/liter (p < 0.05) at 1 mo and 1.07 ± 0.09 μg/liter (p < 0.01) at 3 mo, with no further increase at 6 or 12 mo. PINP concentration increased progressively from baseline (60 ± 5 μg/liter) to 6 mo (126 ± 11 μg/liter, p < 0.001), with no further increase at 12 mo. The percentage increase in PINP concentration was significantly higher than βCTX (p < 0.05). Conclusions : Our study shows that GH has a regulatory role in bone mineral metabolism. GH administration to postmenopausal osteoporotic women improves target organ sensitivity to PTH and bone mineral metabolism and alters PTH secretory pattern with greater increases in bone formation than resorption. These changes, resulting in a net positive bone balance, may partly explain the mechanism causing the increase in BMD after long‐term administration of GH in postmenopausal women with osteoporosis shown in previous studies and proposes a further component in the development of age‐related postmenopausal osteoporosis.     

Autoimmunity in a rural community

As part of a long term prospective study, sera from 3492 subjects, over 90% of the adult population of the rural Australian town of Busselton, were tested by indirect immunofluorescence for various autoantibodies, and by latex particle agglutination for rheumatoid factor. Autoantibodies were detected in 21·6% of all subjects and the overall prevalence increased with age in both sexes. A dip in the age prevalence curve of autoantibodies amongst subjects aged 75–80 years preceded a terminal sharp rise. The greater prevalence in females, 27·5%, than in males, 13·7%, was accounted for by autoantibodies to nuclei, gastric parietal cells and thyroid epithelial cells; the prevalence of smooth muscle antibody (SMA) and rheumatoid factor (RF) was similar in females and males, and the prevalence of SMA did not increase with age. Thus development of `age-sex dependent' autoantibodies, to gastric and thyroid antigens, may have a considerable genetic component, whereas the non `age-sex dependent' autoantibodies, SMA and RF, may have a considerable extrinsic component, e.g. infection. Autoantibodies were associated in two groups, a `nuclear' group and a `thyrogastric' group; these groups were neither positively nor negatively associated, implying independent genetic or other causes for their appearance.     

Populations of lymphocytes separated from human thymocytes.

Thymocytes from twenty-two human foetal and post-natal thymuses were separated according to their buoyant density. Thymocytes from eight were separated into multiple fractions by means of continuous gradients of bovine serum albumin (BSA), pH 5.1 and iso-osmolar with human cells, and thymocytes from fourteen were separated into two fractions of density less than and greater than 1.068 g/cm3. Fractions were tested for antigen-binding lymphocytes (ABL) to 125I-labelled human thyroglobulin, for response to phytohaemagglutinin (PHA) and for rosette-forming cells (RFC) using sheep red blood cells. For subjects of all ages there was a pronounced enrichment of both ABL and thymocytes responsive to PHA amoung low or 1.064-1.065 g/cm3 density thymocytes. In older subjects there was a second enrichment of ABL among high or 1.072-1.073 g/cm3 density thymocytes. RFC were distributed over a wider range of densities, and although they did not form a discrete subpopulation they predominated among high density thymocytes.