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Biomedical subjects

Z Yang

Publications and source records attributed to Z Yang.

At least 451 records · Page 25Linked to original sources

Utility of adenoviral-mediated Fas ligand gene transfer to modulate islet allograft survival.

BACKGROUND: One of the best-defined mechanisms for the induction of apoptosis involves signaling via the cell surface molecule Fas, after binding of Fas ligand. Expression of Fas ligand is tightly regulated, being expressed primarily by T cells after activation, where it serves as a self-regulatory mechanism for immune responses. Fas ligand has also been found to be expressed constitutively at sites of immune privilege such as the testes and the anterior chamber of the eye. Recently, co-transplantation of Fas ligand-transfected myoblasts in association with islet cell allografts was shown to prolong islet allograft survival but only rarely led to indefinite graft survival. Graft rejection was associated with loss of Fas ligand on the myoblasts, suggesting that direct expression of the transgene on the islets might be more effective. METHODS: A replication-defective adenoviral construct containing murine Fas ligand (Ad/MFL) was prepared by homologous recombination. NIH 3T3 cells, rodent splenocytes, and murine islets were infected with Ad/MFL and examined in vitro for functional murine Fas ligand expression. Survival of Ad/MFL-infected islets was subsequently evaluated in vivo in both syngeneic and allogeneic islet transplantation models. RESULTS: Cell lines and islet allografts transfected with Ad/MFL expressed a functional Fas ligand, capable of inducing apoptosis (confirmed by three distinct assays for DNA fragmentation) in Fas+ targets, but not in Fas- controls. Furthermore, Ad/MFL was able to modify allogeneic immune responses in vitro, as addition of this virus, but not a control adenovirus, significantly reduced proliferation in a mixed lymphocyte reaction. Surprisingly, however, transplantation of islet allografts transfected with Ad/MFL resulted in long-term allograft survival in only 1 of 30 recipients. Moreover, adenoviral-mediated Fas ligand gene transfer was complicated by transient, dose-dependent islet dysfunction, perhaps contributing to the lack of long-term engraftment. CONCLUSION: These data suggest that adenoviral-mediated Fas ligand expression may impair normal islet function in vivo, and indicate that alternative strategies for Fas ligand transgene delivery may be required in this setting.

Adenoviridae↗

Studies on diffusion maximum in x-ray diffraction patterns of plasma-sprayed hydroxyapatite coatings.

Study of an amorphous phase in plasma-sprayed hydroxyapatite (HA) coatings is important owing to its unique characteristics and nonnegligible amount of the amorphous phase compared to crystalline HA. However, little is known about the component parts of an amorphous phase. It is known that amorphous phase usually appears as the diffusion maximum (Dmax) in X-ray diffraction (XRD) patterns. Analyzing Dmax, including the position (Pmax) and area of Dmax, we can indicate the component parts of an amorphous phase and their transitions. In this study, the variation of Dmax in XRD patterns of the coatings during plasma spraying, in postheating, and in dissolving in vitro was studied with the aid of XRD. It was found that component parts of the amorphous phase in the coating varied with increasing thickness, consisting of two part represented by Dmax1, located between 29.4 and 29.8 degrees (2 theta), and Dmax2, located between 31.0 and 31.4 degrees (2 theta). It was concluded that Dmax3, located between 32.0 and 32.4 degrees (2 theta), should be referred to as nanocrystals of HA. In addition, the particle size of the starting powder may affect the component parts of the amorphous phase in the coating in addition to thickness. With vacuum heating (650 degrees C) and water vapor treatment at a low temperature (125 degrees C) in a saturated vaporic atmosphere, transition of the amorphous components was not as efficient as that at 490 degrees C with water vapor. The reason might be that the amorphous-to-crystalline HA conversion is dependent on both temperature and water vapor pressure. It was found that amorphous components were transformed completely into crystalline HA after heating at 490 degrees C with a partial water vapor pressure of 0.01 MPa for 2 h. It was concluded that the unstable amorphous components (Dmax1, Dmax2) converted into more stable nanocrystals of HA (Dmax3). Degradation in vitro showed that Dmax3 was more stable than Dmax1 and Dmax2. It was concluded that nucleation of apatite in vitro should be attributed to nanocrystals of HA (Dmax3) except for the amorphous components. It is recommended that the optimal phasic contents of the plasma-sprayed HA coating be mainly composed of crystalline HA and nanocrystals of HA (Dmax3) in terms of the stability and biocompatibility of the coating.

Air↗

Mass screening for lung cancer with mobile spiral computed tomography scanner.

BACKGROUND: The incidence of and mortality from lung cancer have increased steadily. Most lung cancers are not localised when first detected, but early detection is mandatory to improve prognosis. Since curable early cases are hard to visualise with conventional chest radiography, a new diagnostic means must be found. We assessed whether population-based mass screening with a spiral computed tomography scanner could contribute substantially to detection of smaller cancers, and decrease mortality. METHODS: In 1996, we screened in a mobile unit 5483 individuals from the general population of Matsumoto, Japan, aged between 40 years and 74 years who had undergone annual chest radiography (miniature fluorophotography) and cytological assessment of sputum. All participants had a low-dose X-ray spiral computed tomography (CT) scan of the thorax; 3967 also underwent miniature fluorophotography. We compared smokers and non-smokers. Further assessments were done for probably benign but suspicious lesions; suspicion of cancer; and indeterminate small nodules by chest radiography and conventional CT, with additional transbronchial biopsy when possible. Thoracotomy was recommended when it was strongly suspected that the patients had lung cancer. FINDINGS: 19 patients were diagnosed as having lung cancer--14 with suspicion of lung cancer, three with benign but suspicious lesions, and two with indeterminate small nodules. 18 cases were surgically confirmed, and one was clinically diagnosed. The mean size of lesions was 17 mm (range 6-47). In four of 19 patients, lung abnormality was seen on CT and miniature fluorophotography. The lung-cancer detection rate with CT was 0.48%, significantly higher than the 0.03-0.05% for standard mass assessments done previously in the same area. CT missed one case that was found solely on a sputum cytology examination. INTERPRETATION: Our results show that miniature fluorophotography or conventional chest radiography, which have been the main diagnostic techniques for lung cancer, showed few small cancers. CT was more accurate in mass screening for lung cancer and led to early detection and an accurate diagnosis of lung cancer, and should be considered in future health plans.

Adult↗

Regional pharmacokinetics of 5-fluorouracil in dogs: role of the liver, gastrointestinal tract, and lungs.

The purpose of this study was to determine the presence and extent of pulmonary elimination for 5-fluorouracil (FUra). A secondary aim was to characterize the relative importance of the liver, gastrointestinal tract, splanchnic region, and lungs toward the overall elimination of FUra. A total of 10 mixed-breed male and female dogs were used in these acute studies in which FUra was administered through a cephalic vein. Six dogs were studied at sequentially escalated dose rates of 0.125, 0.250, 0.500, 0.750, and 1.00 micromol/min/kg (8-fold range); four dogs were studied at sequentially escalated dose rates of 0.0625, 0.250, 0.750, 1.50, and 2.00 micromol/min/kg (32-fold range). Each infusion lasted 2 h, at which time steady-state plasma concentrations were obtained (i.e., portal vein, carotid artery, hepatic vein, and pulmonary artery), perfusion rates were measured (hepatic artery, portal vein, and cardiac output), and pharmacokinetic parameters were directly assessed. Pulmonary elimination of FUra was conclusively demonstrated. Although only 17% of the drug was extracted by the lungs at the lowest dose rate, pulmonary clearance (16.0 ml/min/kg) was on the order of splanchnic clearance (13.5 ml/min/kg), or larger. As the dose rate increased, pulmonary clearance was more easily saturated than splanchnic clearance. Thus, it appears that at increasing dose rates, the splanchnic region becomes a more significant pathway, whereas the lungs have a reduced role in the overall elimination of FUra.

Animals↗

Total synthesis of brevetoxin A.

Brevetoxin A is the most potent neurotoxin secreted by Gymnodinium breve Davis, a marine organism often associated with harmful algal blooms known as 'red tides'. The compound, whose mechanism of action involves binding to and opening of sodium channels, is sufficiently toxic to kill fish at concentrations of nanograms per ml and, after accumulation in filter-feeding shellfish, to poison human consumers. The precise pathway by which nature constructs brevetoxin A is at present unknown, but strategies for its total synthesis have been contemplated for some time. The synthetic challenge posed by brevetoxin A reflects the high complexity of its molecular structure: 10 oxygen atoms and a chain of 44 carbon atoms are woven into a polycyclic macromolecule that includes 10 rings (containing between 5 and 9 atoms) and 22 stereogenic centres. Particularly challenging are the 7-, 8- and 9-membered rings which allow the molecule to undergo slow conformational changes and force a 90 degrees twist at one of its rings. Here we describe the successful incorporation of methods that were specifically developed for the construction of these rings into an overall strategy for the total synthesis of brevetoxin A in its naturally occurring form. The convergent synthesis reported here renders this scarce neurotoxin synthetically available and, more importantly, allows the design and synthesis of analogues for further biochemical studies.

Animals↗

Blockade of long chain fatty acid oxidation by non-steroidal anti-inflammatory drugs may contribute to inhibition of proliferation of human colorectal carcinoma cell lines.

Non-steroidal anti-inflammatory drugs (NSAIDs) reduce the growth of colorectal carcinoma (CRC) cell lines. Although the mechanism appears to be independent of cyclooxygenases, the inhibitory target has not previously been defined. We now report for the first time that NSAIDs inhibit oxidation of the long chain fatty acid palmitate in human CRC cell lines with potencies which are in good agreement with the potencies of NSAIDs as inhibitors of cell proliferation. The absence of inhibition of acetate oxidation rules out an effect on mitochondrial functions. We conclude that the long chain fatty acid oxidation pathway may be a novel target for some of the inhibitory effects of NSAIDs on the growth of CRC cell lines.

Anti-Inflammatory Agents, Non-Steroidal↗

Distinct cellular interactions of secreted and transmembrane Ebola virus glycoproteins.

The mechanisms by which Ebola virus evades detection and infects cells to cause hemorrhagic fever have not been defined, though its glycoprotein, synthesized in either a secreted or transmembrane form, is likely involved. Here the secreted glycoprotein was found to interact with neutrophils through CD16b, the neutrophil-specific form of the Fc gamma receptor III, whereas the transmembrane glycoprotein was found to interact with endothelial cells but not neutrophils. A murine retroviral vector pseudotyped with the transmembrane glycoprotein preferentially infected endothelial cells. Thus, the secreted glycoprotein inhibits early neutrophil activation, which likely affects the host response to infection, whereas binding of the transmembrane glycoprotein to endothelial cells may contribute to the hemorrhagic symptoms of this disease.

Cell Line↗

Mice that lack thrombospondin 2 display connective tissue abnormalities that are associated with disordered collagen fibrillogenesis, an increased vascular density, and a bleeding diathesis.

Thrombospondin (TSP) 2, and its close relative TSP1, are extracellular proteins whose functions are complex, poorly understood, and controversial. In an attempt to determine the function of TSP2, we disrupted the Thbs2 gene by homologous recombination in embryonic stem cells, and generated TSP2-null mice by blastocyst injection and appropriate breeding of mutant animals. Thbs2-/- mice were produced with the expected Mendelian frequency, appeared overtly normal, and were fertile. However, on closer examination, these mice displayed a wide variety of abnormalities. Collagen fiber patterns in skin were disordered, and abnormally large fibrils with irregular contours were observed by electron microscopy in both skin and tendon. As a functional correlate of these findings, the skin was fragile and had reduced tensile strength, and the tail was unusually flexible. Mutant skin fibroblasts were defective in attachment to a substratum. An increase in total density and in cortical thickness of long bones was documented by histology and quantitative computer tomography. Mutant mice also manifested an abnormal bleeding time, and histologic surveys of mouse tissues, stained with an antibody to von Willebrand factor, showed a significant increase in blood vessels. The basis for the unusual phenotype of the TSP2-null mouse could derive from the structural role that TSP2 might play in collagen fibrillogenesis in skin and tendon. However, it seems likely that some of the diverse manifestations of this genetic disorder result from the ability of TSP2 to modulate the cell surface properties of mesenchymal cells, and thus, to affect cell functions such as adhesion and migration.

Animals↗

Different proliferative properties of smooth muscle cells of human arterial and venous bypass vessels: role of PDGF receptors, mitogen-activated protein kinase, and cyclin-dependent kinase inhibitors.

BACKGROUND: Internal mammary artery (IMA) bypass grafts have a higher patency than saphenous vein (SV) grafts. Intimal hyperplasia of SV grafts is due to smooth muscle cell (SMC) proliferation and migration. We hypothesized that different SMC growth activity exists in IMA and SV, which may explain the different patencies of arterial and venous grafts. METHODS AND RESULTS: SMCs were isolated from IMA and SV by explant culture and stimulated with serum or platelet-derived growth factor-BB (PDGF-BB). Cell growth was analyzed by explant outgrowth rate, 3H-thymidine incorporation, or cell counting. PDGF receptor expression and autophosphorylation, regulation of mitogen-activated protein kinases (MAPKs), and cyclin-dependent kinase inhibitors (p27Kip1 and p21Cip1) were analyzed by molecular techniques. SMC outgrowth from explants by serum (20%) over a 20-day period was more pronounced in SV (37+/-5%) than in IMA (4+/-3%; P<.001) of the same patients. Serum (10%) increased cell number more rapidly in SV (2 x 10(4)/well to 18+/-4 x 10(4)/well; P<.05) than in IMA (2 x 10(4)/well to 9+/-4 x 10(4)/well; P<.05) over an 8-day period. PDGF-BB (0.01 to 10 ng/mL) stimulated 3H-thymidine incorporation (1347+/-470% above control levels) and increased cell number in SV (2 x 10(4)/well to 5+/-1 x 10(4)/well; P<.05) but not in IMA. PDGF alpha- and beta-receptors were similarly expressed and were activated in both SV and IMA. PDGF-BB induced a similar MAPK activation (kinetics and maximal activity) in both SV and IMA cells but increased MAPK protein level only in SV. Furthermore, PDGF-BB markedly downregulated the cell cycle inhibitor p27Kip1 in SV, but this was much less pronounced in IMA. CONCLUSIONS: SMCs from SVs exhibit enhanced proliferation compared with IMA in spite of functional growth factor receptor expression and MAPK activation. However, PDGF increased MAPK protein level only in SV and downregulated cell cycle inhibitor (p27Kip1) more potently in SV than in IMA. This may explain the resistance to growth stimuli of IMA SMCs and may contribute to the longer patency of arterial versus venous grafts.

Blotting, Northern↗

Cyclooxygenase-2-selective antagonists do not inhibit growth of colorectal carcinoma cell lines.

Non-steroidal anti-inflammatory drugs (NSAIDs) reduce the incidence of colorectal carcinoma. We now report that the potent cyclooxygenase-1 inhibitor indomethacin had no effect on the growth of human colorectal carcinoma cell lines in vitro at concentrations up to 30 microM. The selective cyclooxygenase-2 inhibitors L-745337 and NS-398 reduced cyclooxygenase activity, but had no effect on cell growth at concentrations as high as 100 microM. Our results provide direct evidence that inhibition of cyclooxygenase activity does not necessarily inhibit the growth of colorectal carcinoma cell lines and imply that the growth-inhibitory effects of NSAIDs in vitro are not mediated by inhibition of cyclooxygenases.

Anti-Inflammatory Agents, Non-Steroidal↗

Apolipoprotein(a) gene enhancer resides within a LINE element.

Apolipoprotein(a), (apo(a)), is the distinguishing protein portion of the lipoprotein(a) particle, elevated plasma levels of which are a major risk factor for cardiovascular disease. A search for enhancer elements that control the transcription of the apo(a) gene led to the identification of an upstream element that contains target binding sites for members of the Ets and Sp1 nuclear protein families. The enhancer element functions in either orientation to confer a greater than 10-fold increase in the activity of the apo(a) minimal promoter in cultured hepatocyte cells. Unexpectedly, the enhancer element is located within a LINE retrotransposon element, suggesting that LINE elements may function as mobile regulatory elements to control the expression of nearby genes.

Apolipoproteins↗

Changes in interfilament spacing mimic the effects of myosin regulatory light chain phosphorylation in rabbit psoas fibers.

The modulatory effect of myosin regulatory light chain phosphorylation in mammalian skeletal muscle, first documented as posttetanic potentiation of twitch tension, was subsequently shown to enhance the expression and development of tension at submaximal levels of activating calcium. Structural analyses demonstrated that thick filaments with phosphorylated myosin regulatory light chains appeared disordered: they lost the near-helical, periodic arrangement of myosin head characteristic of the relaxed state. We suggested that disordered heads may be more mobile than ordered heads and are likely to spend more time close to their binding sites on thin filaments. In this study we determined that the physiological effects of phosphorylation could be mimicked by decreasing the lattice spacing between the thick and the thin filaments, either by osmotic compression with dextran or by increasing the sarcomere length of permeabilized rabbit psoas fibers. Phosphorylation of regulatory light chains by incubation of permeabilized fibers with myosin light chain kinase and calmodulin, followed by low levels of activating calcium, potentiated tension development at resting or lower sarcomere lengths in the absence of dextran but had no additional effect on tension potentiation or development in fibers with decreased lattice spacing due to either osmotic compression or increased sarcomere length.

Animals↗

Structural and functional responses of mammalian thick filaments to alterations in myosin regulatory light chains.

The ordered array of myosin heads, characteristic of relaxed striated muscle thick filaments, is reversibly disordered by phosphorylating myosin regulatory light chains, decreasing temperature and/or ionic strength, increasing pH, and depleting nucleotide. In the case of light chain phosphorylation, disorder, most likely due to a change in charge affecting the light chain amino-terminus, reflects increased myosin head mobility, thus increased accessibility to actin, and results in increased calcium sensitivity of tension development. Thus, interactions between the unphosphorylated regulatory light chain and the filament backbone may help maintain the overall order of the relaxed filament. To define this relationship, we have examined the structural and functional effects of such manipulations as exchanging wild-type smooth and skeletal myosin light chains into permeabilized rabbit psoas fibers and removing regulatory light chains (without exchange) from such fibers. We have also compared the structural and functional parameters of biopsied fibers from patients with severe familial hypertrophic cardiomyopathy due to a single amino acid substitution in the regulatory light chains to those exhibited by fibers from normal relatives. Our results support a role for regulatory light chains in reversible ordering of myosin heads and suggest that economy of energy utilization may provide for evolutionary preservation of this function in vertebrate striated muscle.

Actin Cytoskeleton↗

Synonymous and nonsynonymous rate variation in nuclear genes of mammals.

A maximum likelihood approach was used to estimate the synonymous and nonsynonymous substitution rates in 48 nuclear genes from primates, artiodactyls, and rodents. A codon-substitution model was assumed, which accounts for the genetic code structure, transition/transversion bias, and base frequency biases at codon positions. Likelihood ratio tests were applied to test the constancy of nonsynonymous to synonymous rate ratios among branches (evolutionary lineages). It is found that at 22 of the 48 nuclear loci examined, the nonsynonymous/synonymous rate ratio varies significantly across branches of the tree. The result provides strong evidence against a strictly neutral model of molecular evolution. Our likelihood estimates of synonymous and nonsynonymous rates differ considerably from previous results obtained from approximate pairwise sequence comparisons. The differences between the methods are explored by detailed analyses of data from several genes. Transition/transversion rate bias and codon frequency biases are found to have significant effects on the estimation of synonymous and nonsynonymous rates, and approximate methods do not adequately account for those factors. The likelihood approach is preferable, even for pairwise sequence comparison, because more realistic models about the mutation and substitution processes can be incorporated in the analysis.

Animals↗

The effect of antibody against vascular endothelial growth factor on tumor growth and metastasis.

Vascular endothelial growth factor (VEGF), a very important in the process of tumor angiogenesis, was chosen as a target in a study to determine whether manipulation of angiogenesis with antibody against VEGF may interrupt tumor growth and metastasis. Anti-VEGF antibody was obtained from immunized rabbits, purified on an affinity column, and identified as neutralized antibody by Mile's assay. IVTA2MA891, a murine spontaneous breast cancer with a high rate of metastasis in lung in TA2 x 615 F1 mice, was chosen as an animal model in this study, because of the high expression of VEGF in the primary tumor as well as in the lung metastatic tumor. The anti-VEGF antibody could inhibit growth of S180 sarcoma in a dose-dependent manner, and the inhibition rate could reach 41.0% with a dose of 200 microg mouse(-1) day(-1). Anti-VEGF antibody could inhibit tumor growth by 76.2% in nude mice bearing human gastric cancer (MGC 803). When anti-VEGF antibody was combined with 131I-3H11, a murine monoclonal antibody conjugated with 131I, only one of five nude mice developed tumor and 84.0% more inhibition of tumor growth was obtained in comparison with treatment by 131I-3H11 alone. The growth of the primary tumor was inhibited by 44.0% and the number and size of the metastatic foci in the lungs were reduced by 73.0% and 83.7% respectively in the animal model, with a high rate of metastasis in lung. The anti-VEGF antibody may be potentially useful for clinical treatment of cancer and metastasis.

Animals↗

Pretreatment and biodegradability enhancement of DSD acid manufacturing wastewater.

Two advanced oxidation processes, Fenton's reagent oxidation and ozonation, were used for pretreatment of a 4,4'-diaminostilbene-2,2'-disulfonic acid (DSD-acid) manufacturing wastewater on a laboratory scale. The results showed that both methods are suitable for partial removal of chemical oxygen demand (COD) and color as well as the enhancement of biodegradability of DSD-acid manufacturing wastewater before the conventional biological treatment. Fenton's reagent oxidation by 9.0 g/L H2O2 and 150 mg/L Fe2+ and ozonation by 7.5 g/L O3 led to an improved biodegradability (BOD5/COD = 0.3). Fenton's reagent oxidation in combination with Fe3+ coagulation has shown to effectively remove COD and color. About 90% of COD and 95% of color removals were obtained at the optimum condition, oxidation by 150 mg/L Fe2+ and 2 g/L H2O2, followed by two-stage coagulation with 5 g/L and 2 g/L FeCl3, respectively.

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfo↗

DNA cytometry and chromosome 9 aberrations by fluorescence in situ hybridization of irrigation specimens from bladder cancer patients.

OBJECTIVES: To determine the sensitivity and specificity of combining fluorescence in situ hybridization (FISH) measurement of chromosome 9 and DNA cytometry of bladder irrigation specimens in the detection of bladder cancer. METHODS: Bladder irrigation specimens were obtained from 37 normal control patients and 317 bladder cancer patients during cystoscopic examinations. Bladder cancer patients were sampled in the absence of observable tumor (256 specimens) and concurrently with tumor (204 specimens). Chromosome 9 copy number was determined on a cellular basis by FISH, and cellular DNA content was determined by Feulgen DNA staining and image cytometry. RESULTS: Sensitivity of chromosome 9 FISH was 42% for all tumors and was not correlated to transitional cell carcinoma tumor grade, while the sensitivity of DNA cytometry was 55% and improved with increasing grade from 38% for grade 1 to 90% for grade 3 tumors. The results of FISH and DNA cytometry were combined, resulting in specificity of 92% and sensitivity of 69% for grade 1, 76% for grade 2, and 97% for grade 3 tumors. CONCLUSIONS: The lack of increase with grade in the percentage of positive specimens by FISH supports the hypothesis that chromosome 9 aberrations are critical events in bladder tumorigenesis for many patients. These data demonstrate the presence of cells in irrigation specimens with specific genomic lesions of chromosome 9 and DNA content. Combining FISH on chromosome 9 and DNA cytometry provides an increase in sensitivity to transitional cell carcinoma over either test alone.

Carcinoma, Transitional Cell↗

Interaction of Et2SnCl2 with 5'-IMP and 5'-GMP.

The interactions of Et2SnCl2 with 5'-IMP and 5'-GMP have been studied in aqueous solutions by 1H- and 31P-NMR spectroscopy as a function of pH. At low pH values (< 4.0) Sn(IV) interacts with the pyrophosphate oxygens of these nucleotides. At intermediate pH values (4-9.5) no interaction of the metal with the nucleotides take place, while at pH > 9.5 the sugar O'2 and O'3 atoms are the preferred coordination sites. In addition, the solid adducts obtained from aqueous solutions at pH = 3-4 of the above interactions correspond to formulae; (Et2Sn)2(5'-IMP)2(H2O) and (Et2Sn)3(5'-GMP)2(OH)2(H2O)2 as their elemental analysis show. IR spectra and solid state 13C, 31P-NMR spectra 119Sn Mössbauer and solution 119Sn-NMR spectra once more confirm the pyrophosphate involvement in bonding with Sn(IV) in oligomeric or polymeric structures and trigonal bipyramidal or octahedral geometries.

Guanosine Monophosphate↗