Biotin carboxylases in mitochondria and the cytosol from skeletal and cardiac muscle as detected by avidin binding

Biotin carboxylases in mammalian cells are regulatory enzymes in lipogenesis and gluconeogenesis. In this study, endogenous biotin in skeletal and cardiac muscle was detected using avidin conjugated with alkaline phosphatase and applied in high concentrations to muscle sections. The avidin binding was subsequently visualized by histochemical demonstration of the alkaline phosphatase activity. All cardiac muscle cells showed high affinity for avidin with only the nuclei and the intercalated discs remaining unstained. In skeletal muscle a diffuse reaction could be detected in the sarcoplasm of the muscle fibres. A granular reaction was noted in the same fibres that showed activity for succinic dehydrogenase. The specificity of the coloured reaction product in the muscle sections was investigated and is suggested to be caused by avidin binding to biotin moieties in mitochondria and the cytosol. Mitochondrial and cytosolic preparations of skeletal muscle were electrophoresed in sodium dodecyl sulphate gels. After blotting and incubation with conjugated avidin, two bands with molecular weights of 75 kDa and 130 kDa respectively were evident in the mitochondrial preparation. It is suggested that the 75-kDa band represents comigration of the biotin-containing subunits of propionyl-CoA carboxylase and methylcrotonyl-CoA carboxylase. The 130-kDa band may represent the biotin-containing pyruvate carboxylase. In the cytosolic preparation a 270-kDa band was stained in blots that had been incubated with conjugated avidin; this band is suggested to represent acetyl-CoA carboxylase. A 190-kDa cytosolic band might be a cleavage product of acetyl-CoA carboxylase. We propose that using alkaline phosphatase-conjugated avidin it is possible to detect the mitochondrial and cytosolic biotin-dependent carboxylases in striated muscle.     

Use of nuclear morphometry characteristics to distinguish between normal and abnormal cervical glandular histologies.

This is a methodological study exploring the use of quantitative histopathology applied to the cervix to discriminate between normal and cancerous (consisting of adenocarcinoma and adenocarcinoma in situ) tissue samples. The goal is classifying tissue samples, which are populations of cells, from measurements on the cells. Our method uses one particular feature, the IODs-Index, to create a tissue level feature. The specific goal of this study is to find a threshold for the IODs-Index that is used to create the tissue level feature. The main statistical tool is Receiver Operating Characteristic (ROC) curve analysis. When applied to the data, our method achieved promising results with good estimated sensitivity and specificity for our data set. The optimal threshold for the IODs-Index was found to be 2.12.     

SEASONAL PATTERNS OF MORPHOLOGICAL VARIABILITY IN SARGASSUM POLYCERATIUM (PHAEOPHYTA)1

Temporal variability in certain morphological and taxonomically important features was quantified for Sargassum polyceratium Mont. from a population in the Content Keys, Florida (U.S.A.). Patterns of blade development, senescence, and loss caused pronounced seasonal changes in blade length-width ratios. Blade length and width were maximal early in the growing season (August-November) and decreased as the annual stems matured. Early in the growing season, plants had broader blades with randomly distributed cryptostomata. Late in the growing season (February-April), plants had more linear blades with cryptostomata approximately arranged in two rows, one on each side of the midrib. The length-width ratio of blades increased acropetally along the stems and were directly correlated to the size of the cryptostomatal opening and inversely correlated with the number of cryptostomata. The branching pattern of the annual stems ranged from short spur branches to well-developed, lateral axillary branches. The frequency of bifurcated blades increased significantly late in the growing season. Vesicle shape and size and pedicel length were temporally stable. Alated pedicels and mucronate vesicles occurred in low frequencies. The variability of the morphological features used to delineate species within the genus Sargassum on the tropical eastern coasts of the Americas is poorly understood.     

Marginality: scale determined processes and the conservation of the British butterfly fauna

Marginality describes the impact that environmental and landscape factors have in decreasing the probability of population survival and persistence. It may be imposed by extreme conditions or resource scarcity. Typically, it affects populations at the range edge but can also affect populations within the core of ranges, and produces a number of symptoms: characteristically demographic, but also morphological, physiological, biochemical and genetic. In this paper, the causes and effects of marginality on British butterflies are compared in edge and centre of range populations. Issues of temporal and spatial scales are examined, as is the relevance of marginality to the conservation of single and multiple species populations. The recognition of marginality questions the appropriateness of many so-called spatially realistic models of populations and highlights areas of research which have hitherto been ignored. Projected changes in land use and climate have implications for marginality in core and peripheral populations; in view of this, current scales of mapping are found to be unsuitable for monitoring fragmentation and the increasing marginalization of butterfly species in the British landscape.     

Secondary structure of β-hydroxydecanoyl thiol ester dehydrase, a 39-kDa protein, derived from Hα, Cα, Cβ and CO signal assignments and the Chemical Shift Index: Comparison with the crystal structure

Summary Nearly complete backbone ¹H, ¹⁵N and ¹³C signal assignments are reported for -hydroxydecanoyl thiol ester dehydrase, a 39-kDa homodimer containing 342 amino acids. Although ¹⁵N relaxation data show that the protein has a rotational correlation time of 18 ns, assignments were derived from triple-resonance experiments recorded at 500 MHz and pH 6.8, without deuteration. The Chemical Shift Index, CSI, identified two long helices and numerous -strands in dehydrase. The CSI predictions are in close agreement with the secondary structure identified in the recently derived crystal structure, particularly when one takes account of the numerous bulges in the -strands. The assignment of dehydrase and a large deuterated protein [Yamazaki et al. (1994) J. Am. Chem. Soc., 116, 11655–11666] suggest that assignment of 40–60 kDa proteins is feasible. Hence, further progress in understanding the chemical shift/structure relationship could open the way to determine the structures of such large proteins. Supplementary Material is available on request, comprising Table S1 listing the spectral parameters; Table S2 listing the assignments; Fig. S1 showing the 2D ¹H–¹⁵N HSQC spectrum; Fig. S2 showing sequential NOEs, secondary shifts, H-exchange and ³J_HN data; and Fig. S3 showing plots of the H, C, CO and C Chemical Shift Indexes.To whom correspondence should be addressed.     

Synergistic interactions between XPC and p53 mutations in double-mutant mice: neural tube abnormalities and accelerated UV radiation-induced skin cancer

The significance of DNA repair to human health has been well documented by studies on xeroderma pigmentosum (XP) patients, who suffer a dramatically increased risk of cancer in sun-exposed areas of their skin [1] and [2]. This autosomal recessive disorder has been directly associated with a defect in nucleotide excision–repair (NER) [1] and [2]. Like human XP individuals, mice carrying homozygous mutations in XP genes manifest a predisposition to skin carcinogenesis following exposure to ultraviolet (UV) radiation [3], [4] and [5]. Recent studies have suggested that, in addition to roles in apoptosis [6] and cell-cycle checkpoint control [7] in response to DNA damage, p53 protein may modulate NER [8]. Mutations in the p53 gene have been observed in 50% of all human tumors [9] and have been implicated in both the early [10] and late [11] stages of skin cancer. To examine the consequences of a combined deficiency of the XPC and the p53 proteins in mice, we generated double-mutant animals. We document a spectrum of neural tube defects in XPC p53 mutant embryos. Additionally, we show that, following exposure to UV-B radiation, XPC p53 mutant mice have more severe solar keratosis and suffer accelerated skin cancer compared with XPC mutant mice that are wild-type with respect to p53.     

One-dimensional numerical simulation of primary production: Lagrangian and Eulerian formulations

It has been argued (Wolf and Woods, in Toward a Theory On Biological-PhysicalInteractions in the World Ocean, Rothschild, (ed.), 1988) (WW88)that phytoplankton growth models are sensitive to Lagrangianeffects because populations at a given depth and time containa wide distribution of photoadaptive properties. On the otherhand, Lande and Lewis (Deep-Sea Research. 36. 1161–1175.1989) (LL89) have claimed that for a different photosyntheticmodel, this distribution of properties can be adequately representedby a mean value in a much simpler and more efficient Eulerianformulation. This study compares Lagrangian and Eulerian integrationsof these two different models of photosynthesis under two mixingregimes. For relatively weak mixing, the growth rate predictedby the different formulations of the two models is small (     

The Distribution and Biochemical Properties of a Cdc2-Related Kinase, KKIALRE, in Normal and Alzheimer Brains

Abstract: The biochemical properties and distribution of a Cdc2-related kinase, KKIALRE, were studied in brain tissues and cultured cells with antibodies to a subregion of KKIALRE protein deduced from cDNA. In adult human brain, the KKIALRE-immunoreactive protein consisted of four or five isoforms having a molecular size of 40–52 kDa, whereas in fetal brain, there was one protein of ∼48 kDa. Cultured astrocytes, neuroblastoma cells, and mouse brains contained the fetal form of KKIALRE protein. KKIALRE-immunoreactive proteins were capable of phosphorylating histone and synthetic peptides with the X-Ser-Pro-X motif, indicating that these proteins belong to the proline-directed Ser/Thr protein kinase family. The KKIALRE immunoreactivity was detected primarily in fibrous astrocytes in white matter and perivascular and subpial spaces, as well as in Bergmann glia in the cerebellum. In fetal brains radial glia were weakly immunoreactive. Reactive astrocytes were more intensely labeled than other glia. Neurons in normal brains and brains with Alzheimer's disease (AD) displayed no KKIALRE immunoreactivity. KKIALRE immunoreactivity was similar in neurons with and without neurofibrillary tangles. The results indicate that in CNS, the KKIALRE protein is mainly a glial protein that is up-regulated in gliosis and that it probably plays no role in the hyperphosphorylation of τ in AD brains.     

A permanent prosthesis for converting in situ muscle contractions into hydraulic power for cardiac assist

Trumble, Dennis R., and James A. Magovern. A permanentprosthesis for converting in situ muscle contractions into hydraulic power for cardiac assist. J. Appl.Physiol. 82(5): 1704-1711, 1997.The key toutilizing muscle power for circulatory support lies with thedevelopment of a practical scheme by which contractile energy may becollected and efficiently delivered to the bloodstream. This workdescribes initial in vitro testing of a prototype muscle energyconverter (MEC) designed to transform the power of in situ musclecontractions into hydraulic form. The MEC resembles a simple pistonpump and is designed for implant beneath the humeral insertion of thelatissimus dorsi muscle. Bench tests were conducted to measurecomponent function and to characterize device performance under varioushydraulic loads. Under simulated muscle-pull conditions, MEC energytransfer capacity was found to be 170 mJ/stroke while operating at peakefficiencies (i.e., >98% of input power converted into hydraulicenergy and preload work). Transfer efficiencies dropped from 96 to 38%as mean generated pressures increased from 23 to 36 N/cm² due to metal bellowsflexion. These results demonstrate that a significant amount ofcontractile energy can be efficiently transformed to hydraulic powervia this mechanism.