Capillary growth, ultrastructure remodelling and exercise training in skeletal muscle of essential hypertensive patients

AIM The aim was to elucidate whether essential hypertension is associated with altered capillary morphology and density and to what extent exercise training can normalize these parameters. METHODS To investigate angiogenesis and capillary morphology in essential hypertension, muscle biopsies were obtained from m. vastus lateralis in subjects with essential hypertension (n = 10) and normotensive controls (n = 11) before and after 8 weeks of aerobic exercise training. Morphometry was performed after transmission electron microscopy, and protein levels of several angioregulatory factors were determined. RESULTS At baseline, capillary density and capillary-to-fibre ratio were not different between the two groups. However, the hypertensive subjects had 9% lower capillary area (12.7 ± 0.4 vs. 13.9 ± 0.2 μm(2)) and tended to have thicker capillary basement membranes (399 ± 16 vs. 358 ± 13 nm; P = 0.094) than controls. Protein expression of vascular endothelial growth factor (VEGF), VEGF receptor-2 and thrombospondin-1 were similar in normotensive and hypertensive subjects, but tissue inhibitor of matrix metalloproteinase was 69% lower in the hypertensive group. After training, angiogenesis was evident by 15% increased capillary-to-fibre ratio in the hypertensive subjects only. Capillary area and capillary lumen area were increased by 7 and 15% in the hypertensive patients, whereas capillary basement membrane thickness was decreased by 17% (P < 0.05). VEGF expression after training was increased in both groups, whereas VEGF receptor-2 was decreased by 25% in the hypertensive patients(P < 0.05). CONCLUSION Essential hypertension is associated with decreased lumen area and a tendency for increased basement membrane thickening in capillaries of skeletal muscle. Exercise training may improve the diffusion conditions in essential hypertension by altering capillary structure and capillary number.

STUDIES INTO THE MOLECULAR MECHANISMS REGULATING MUSCULAR ATROPHY RESULTING FROM HINDLIMB IMMOBILIZATION IN THE RAT (PROTEIN, SYNTHESIS, RNA)

The purpose of the work performed in this dissertation was to examine some of the possible regulatory mechanisms involved in the initiation of muscular atrophy during periods of decreased muscle utilization resulting from hindlimb immobilization in the rat. A 37% decrease in the rate of total muscle protein synthesis which has been observed to occur in the first 6 h of immobilization contributes significantly to the observed loss of protein during immobilization.^ The rates of cytochrome c and actin synthesis were determined in adult rat red vastus lateralis and gastrocnemius muscles, respectively, by the constant infusion and incorporation of ('3)H-tyrosine into protein. The fractional synthesis rates of both actin and cytochrome c were significantly decreased (P < 0.05) in the 6th h of hindlimb immobilization.^ RHA was extracted from adult rat gastrocnemius muscle by modification of the phenol: chloroform: SDS extraction procedures commonly used for preparation of RNA for hybridization analysis from other mammalian tissues. RNA content of rat gastrocnemius muscle, as determined by this method of extraction and its subsequent quantification by UV absorbance and orcinol assay, was significantly greater than the RNA content previously determined for adult rat gastrocnemius by other commonly employed methods.^ RNA extracted by this method from gastrocnemius muscles of control and 6h immobilized rats was subjected to "dot blot" hybridization to ('32)P-labelled probe from plasmid p749, containing a cDNA sequence complementary to (alpha)-actin mRNA and from rat skeletal muscle. (alpha)-Actin specific mRNA content as estimated by this procedure is not significantly decreased in rat gastrocnemius following 6h or hindlimb immobilization. However, (alpha)-actin specific mRNA content is significantly decreased (P < 0.05) in adult rat gastrocnemius (alpha)-actin specific mRNA is not decreased in adult rat gastrocnemius muscle following 6h of immobilization, a time when actin synthesis is significantly decreased, it is concluded that a change in (alpha)-actin specific mRNA content is not the initiating event responsible for the early decrease in actin synthesis observed in the 6th h of immobilization. ^

Inhibitor of cytochrome c messenger RNA -protein interaction in stimulated rat skeletal muscle

The purpose of this work was to examine the possible mechanisms for the regulation of cytochrome c gene expression in response to increased contractile activity in rat skeletal muscle. The working hypothesis was that increased contractile activity enhances cytochrome c gene expression through a cis-element. A 110% increase in cytochrome c mRNA concentration was observed in tibialis anterior (TA) muscle after 9 days of chronic stimulation. Similar difference (120%) exists between soleus (SO) muscle of higher contractile activity and white vastus lateralis (WV) muscle of lower contractile activity. These results suggest that the endogenous cytochrome c gene expression is regulated by contractile activity. Cytochrome c-reporter genes were injected into skeletal muscles to identify the cis-element that is responsible for the regulation. Although the data was inconclusive, part of it suggested the importance of the 3$\sp\prime$-untranslated region (3$\sp\prime$-UTR) in mediating the response to increased contractile activity.^ RNA gel mobility shift (GMSA) and ultraviolet (UV) cross-linking assays revealed specific RNA-protein interaction in a 50-nucleotide region of the 3$\sp\prime$-UTR in unstimulated TA muscle. Computer analysis predicted a stem-loop structure of 17 nucleotides, which provides a structural basis for RNA-protein interaction. These 17 nucleotides are 100% conserved among rat, mouse and human cytochrome c genes and their 13 pseudogenes, suggesting a functional role for this region. The RNA-protein interaction was significantly less in highly active SO muscle than in inactive WV muscle and was dramatically decreased in stimulated TA muscle due to a protein inhibitor(s) associated with ribosome. It is possible that cytochrome c mRNAs undergoing translation are subject to a compartmentalized regulatory influence.^ The conclusion from these results is that increases in contractile activity induce or activate a protein inhibitor(s) associated with ribosome in rat skeletal muscle. The inhibitor decreases RNA-protein interaction in the 3$\sp\prime$-UTR of cytochrome c mRNA, which may result in increased mRNA stability and/or translation. ^

Benthic foraminifers from the Japan Sea

Five holes were drilled at two sites in the Sea of Japan during Ocean Drilling Program (ODP) Leg 128. Site 798 is located on Oki Ridge at a depth of about 900 m. Sediment age at Site 798 ranges from Pliocene to Holocene. Site 799 is located in the Kita-Yamato Trough at depth of 2000 m and below the present calcite compensation depth (CCD); the sediment ranges from Miocene to Holocene in age. Samples from all holes contain benthic foraminifers. Faunal evidence of downslope displacement is frequent in Holes 799A and 799B. The vertical frequency distribution of some dominant species shows that significant faunal changes occur in Holes 798A-C on Oki Ridge. Based on the faunal change and the thickness of sediments, it appears that the Oki Ridge was uplifted more than 1,000 m during last 4 m.y. Benthic foraminifers also demonstrate that the water depth of Site 799 rapidly changed from upper bathyal to lower bathyal during middle Miocene time. The appearance of benthic foraminifer species common to anaerobic environments suggests that the dysaerobic to anaerobic bottom conditions existed during the evolution of the Sea of Japan. Faunal distributions also suggest that the 'Tertiary-type' species recognized in the Neogene strata of the Japan Sea coastal regions disappeared sequentially from the Sea of Japan during Pliocene to late Pleistocene.

Response of deep-sea benthic foraminifera to Late Quaternary climate changes, southeast Indian Ocean, offshore Western Australia

The Late Quaternary benthic foraminifera of four deep-sea cores off Western Australia (ODP 122-760A, ODP 122-762B, BMR96GC21 and RC9-150) have been examined for evidence of increased surface productivity to explain the anomalously low sea-surface paleotemperatures inferred by planktic foraminifera for the last and penultimate glaciations. The delta13C trends of Cibicidoides wuellerstorfi, and differences between the delta13C trends of planktics (Globigerinoides sacculifer) and benthics (C. wuellerstorfi) in the four cores indicate that during stage 6 bottom waters were significantly depleted in delta13C, and strong delta13C gradients were established in the water column, while during stage 2 and the Last Glacial Maximum, delta13C trends did not differ greatly from that of the Holocene. Two main assemblages of benthic foraminifera were identified by principal component analyses: one dominated by Uvigerina peregrina, another dominated by U. proboscidea. Abundance of these Uvigerinids, and of taxa preferring an infaunal microhabitat, and of Epistominella exigua and Bulimina aculeata indicate that episodes of high influx of particulate organic matter were established in most sites during glacial episodes, and particularly so during stage 6, while evidence for upwelling during the Last Glacial Maximum is less strong. The Penultimate Glaciation upwellings were established within the areas of low sea-surface paleotemperature indicated by planktic foraminifera. During the Last Interglacial Climax, upwelling appears to have been established in an isolated region offshore from a strengthened Leeuwin Current off North West Cape. Last Glacial Maximum delta13C values of C. wuellerstorfi at waterdepths of less than 2000 m show smaller than global mean glacial-interglacial changes suggesting the development of a deep hydrological front. A similar vertical stratification/bathyal front was also established during the Penultimate Glaciation.

Shallow water foraminifera of ODP Hole 133-821A

Reworked shallow-water foraminifers that settled on the upper slope of the central Great Barrier Reef at Site 821 (water depth, 212.6 m) were used as indicators of the paleoclimatic and paleoenvironmental conditions that have controlled the Pleistocene evolution of the adjacent platform. Throughout the 400-m-thick sequence drilled, the nature, composition, and distribution of the shallow-water foraminiferal assemblages studied indicate that (1) all the species recorded are at present living in diverse tropical, reef-related areas of the Indo-Pacific and Atlantic provinces; (2) the composition of the microfaunal taphocoenoses is almost identical between the different stratigraphic intervals studied and the modern Great Barrier Reef environments; (3) inner-neritic, tropical environments have continued to develop since the middle Pleistocene; (4) high- to moderate-energy platform edges occurred repeatedly throughout Pleistocene time. These factors may suggest that, since the beginning of the Pleistocene, several reef-like tracts have grown successively on the central area of the northeastern Australian shelf edge. These tracts probably had a sufficiently evolved morphological zonation to act as shelters for foraminiferal biocoenoses of high species diversity.

Presence-absence species/sample matrix for the measured sections at the area of Maghara, North Sinai, Egypt

As age-diagnostic fossils are rare in the Middle to Upper Jurassic sedimentary succession of Gebel Maghara, North Sinai, Egypt, and in order to ensure maximal stratigraphic resolution, chronostratigraphic boundaries were determined based on quantitative biostratigraphy. A data matrix comprising 231 macrofaunal taxa in 93 samples from four sections has been processed with the Unitary Association (UA) Method. This led to construction of a sequence of 29 UAs (maximal sets of actually or virtually coexisting taxa), which have been grouped into 14 laterally reproducible association zones. The UA method allowed an in-depth analysis of the stratigraphically conflicting taxa, enabled the biostratigraphic subdivision of the studied interval, and also provided stratigraphic correlation among the measured sections and with the Tethyan ammonite zones.

Occurence of foraminifera and other microfossils in sediment core CRP-2/2A (Table 1)

Sparse to moderately abundant foraminiferal assemblages from Oligocene and Lower Miocene sediments in the CRP-2/2A drillhole contain C.27 genera and 42 species of calcareous benthic foraminifera. No planktic or agglutinated taxa were observed. On the basis of their faunal characteristics, four Foraminiferal Units are defined in drillhole succession: Foraminiferal Unit I (26.91-193.95 mbsf), mostly sparse assemblages with Elphidium magellanicum and Cribroelphidium sp.; Foraminiferal Unit II (193.95-342.42 mbsf), mostly moderately abundant assemblages with Cassidulinoides aequilatera and Eponides bradyi; Foraminiferal Unit III (342.42-486.19 mbsf), moderately abundant to sparse assemblages characterised by Cassidulinoides chapmani and Stainforthia sp.; and Foraminiferal Unit IV, Improverished (486.19-624.15, total depth, mbsf), with mostly barren residues, but with large Milioliidae recorded in situ at various horizons in the drill core. Foraminiferal Units I-IV lack taxa allowing correlation to standard zonal schemes. Inspection of faunal records from CIROS-1 and DSDP 270 indicates that, although the faunas show an overall similarity, CRP-2/2A Foraminiferal Units I-IV are not identifiable at these sites. The units are therefore most likely to reflect local environmental changes, and probably will prove useful for local correlation, but their lateral extent is undetermined. All four assemblages apparently represent various glacially-influenced shelf environments, and appear to reflect a long term deepening trend from Units IV to II, from perhaps inner to mid or outer-shelf depths, followed by a return to shallower, inner shelf, conditios for Unit I.

Plio-Pleistocene benthic foraminifera of the Tyrrhenian Sea

Benthic foraminifers from Site 652, Site 653 (Hole 653A), and Site 654 of Leg 107 (Tyrrhenian Sea, Western Mediterranean), which penetrated with more or less good recovery the Plio-Pleistocene stratigraphic interval, were studied in a total of 699 close-spaced samples. A total number of 269 species have been classified and their quantitative distribution in each sample is reported. The benthic foraminifers assemblage is more diversified in Site 654, less diversified in Site 652. Less than a half of the benthic foraminifers species listed from Plio-Pleistocene Italian land sections are present in the coeval deep-sea Tyrrhenian record, in which shallow water species are missing and Nodosarids are poorly represented. A very few species have comparable stratigraphic distribution in the three deep-sea sequences and in Italian land sections when compared against calcareous plankton biostratigraphy. In the same three sites, the first appearance levels of several species are younger and younger, and last appearance levels are earlier and earlier from Site 654 to Site 653 and Site 652. Five biostratigraphic events, biochronologically evaluated and occurring at the same level in the deepsea Tyrrhenian record and in several land sections, have been selected as zonal boundaries of the proposed benthic foraminifers biostratigraphic scheme. The Plio-Pleistocene interval has been subdivided into four biozones and one subzone, recognizable both in the deep-sea and land-based sequences. The Cibicidoides (?) italicus assemblage zone stretches from the base of the Pliocene to the extinction level of the zonal marker, biochronologically evaluated at 2.9 Ma. The Cibicidoides robertsonianus interval zone stretches from the Cibicidoides (?) italicus extinction level to the Pliocene Mediterranean FO of Gyroidinoides altiformis, evaluated at 2.4 Ma. The Gyroidinoides altiformis interval zone stretches from the Mediterranean Pliocene FO of the zonal marker to the appearance level of Articulina tubulosa, evaluated at 1.62 Ma. The Articulina tubulosa assemblage zone stretches from the appearance level of the zonal marker to the Recent. In the Articulina tubulosa biozone, the Hyalinea baltica subzone is proposed. The appearance level of Hyalinea baltica is evaluated at 1.35 Ma, well above the Plio-Pleistocene boundary as defined in the Vrica stratotype section.

Benthic foraminifera in sapropels of the eastern Mediterranean Sea

High-resolution benthic foraminiferal and geochemical investigations were carried out across sapropels S5 and S6 from two sediment cores in the Levantine Sea to evaluate the impact of climatic and environmental changes on benthic ecosystems during times of sapropel formation. The faunal successions indicate that eutrophication and/or oxygen reduction started several thousand years prior to the onset of sapropel formation, suggesting an early response of the bathyal ecosystems to climatic changes. Severest oxygen depletions appear in the early phases of sapropel formation. The initial reduction of deep-water ventilation is caused by a warming and fresh water-induced stratification of Eastern Mediterranean surface waters. During the late phase of S5 formation improved oxygenation is restricted to middle bathyal ecosystems, indicating that at least some formation of subsurface water took place. During S6 formation oxygen depletions and eutrophication were less severe and more variable than during S5 formation. Estimated oxygen contents were low dysoxic at middle bathyal to anoxic at lower bathyal depths during the early phase of S6 formation but never dropped to anoxic values in its late phase. The high benthic ecosystem variability during S6 formation suggests that water column stratification at deep-water formation sites was in a very unstable mode and susceptible to minor temperature fluctuations at a millennial time-scale.

Cenozoic benthic foraminifers from ODP Leg 134 holes

This paper discusses the Paleobathymetric and paleoenvironmental history of the New Hebrides Island Arc and North d'Entrecasteaux Ridge during Cenozoic time based on benthic foraminiferal and sedimentological data. Oligocene and Pliocene to Pleistocene benthic foraminiferal assemblages from Sites 827, 828, 829, and 832 of Ocean Drilling Program (ODP) Leg 134 (Vanuatu) are examined by means of Q-mode factor analysis. The results of this analysis recognize the following bathymetrically significant benthic foraminiferal biofacies: (1) Globocassidulina subglobosa biofacies and Bulimina aculeata-Bolivinita quadrilatera biofacies representing the upper bathyal zone (600-1500 m); (2) Gavelinopsis praegeri-Cibicides wuellerstorfi biofacies, indicating the Pacific Intermediate Water (water depth between 1500 and 2400 m); (3) Tosaia hanzawai-Globocassidulina muloccensis biofacies, Valvulineria gunjii biofacies, and the Melonis barleeanus-Melonis sphaeroides biofacies, which characterize the lower bathyal zone; (4) the Nuttallides umbonifera biofacies, which characterizes the interval between the lysocline (approximately 3500 m) and the carbonate compensation depth (approximately 4500 m); and (5) the Rhabdammina abyssorum biofacies representing the abyssal zone below the carbonate compensation depth. Benthic foraminiferal patterns are used to construct Paleobathymetric and paleogeographic profiles of the New Hebrides Island Arc and North d'Entrecasteaux Ridge for the following age boundaries: late Miocene/Pliocene, early/late Pliocene, Pliocene/Pleistocene, and Pleistocene/Holocene.

Benthic foraminiferal response to the emergence of the Isthmus of Panama and coincident paleoceanographic changes

Late Cenozoic benthic foraminiferal faunas from the Caribbean Deep Sea Drilling Project (DSDP) Site 502 (3052 m) and East Pacific DSDP Site 503 (3572 m) were analyzed to interpret bottom-water masses and paleoceanographic changes occurring as the Isthmus of Panama emerged. Major changes during the past 7 Myr occur at 6.7-6.2, 3.4, 2.0, and 1.1 Ma in the Caribbean and 6.7-6.4, 4.0-3.2, 2.1, 1.4, and 0.7 Ma in the Pacific. Prior to 6.7 Ma, benthic foraminiferal faunas at both sites indicate the presence of Antarctic Bottom Water (AABW). After 6.7 Ma benthic foraminiferal faunas indicate a shift to warmer water masses: North Atlantic Deep Water (NADW) in the Caribbean and Pacific Deep Water (PDW) in the Pacific. Flow of NADW may have continued across the rising sill between the Caribbean and Pacific until 5.6 Ma when the Pacific benthic foraminiferal faunas suggest a decrease in bottom-water temperatures. After 5.6 Ma deep-water to intermediate-water flow across the sill appears to have stopped as the bottom-water masses on either side of the sill diverge. The second change recorded by benthic foraminiferal faunas occurs at 3.4 Ma in the Caribbean and 4.0-3.2 Ma in the Pacific. At this time the Caribbean is flooded with cold AABW, which is either gradually warmed or is replaced by Glacial Bottom Water (GBW) at 2.0 Ma and by NADW at 1.1 Ma. These changes are related to global climatic events and to the depth of the sill between the Caribbean and Atlantic rather than the rising Isthmus of Panama. Benthic foraminiferal faunas at East Pacific Site 503 indicate a gradual change from cold PDW to warmer PDW between 4.0 and 3.2 Ma. The PDW is replaced by the warmer, poorly oxygenated PIW at 2.1 Ma. Although the PDW affects the faunas during colder intervals between 1.4 and 0.7 Ma, the PIW remains the principal bottom-water mass in the Guatemala Basin of the East Pacific.