Bio Cell Research Articles

A Brief of Water and Soil Pollution Management (Recent Trend and Economic Perspective)

Soil and water pollution are the most crucial issues in the world. Various reports have informed that pollution has had long-term adverse effects on environmental sustainability and human health. Several methods have been reported to be effective in reducing pollutant parameters in water and soil. The methods often used in water and soil remediation are bioremediation (land-vetting, bio-cell. Bio cell, Phytoremediation, Land venting, Composite, Bio venting, Bio slurry), Adsorption, Pretreatment (Ultrasonic), Microwave, Electrokinetic disintegration, High-Pressure Homogenization/HPH, Thermal Hydrolysis, Acid hydrolysis (HCl, H2SO4, H3PO4, and HNO3), Ozonation, Fenton Oxidation, Fe (II) -activated persulfate, Protease, amylase, lipase. This paper will explain water and soil pollution and the methods used to overcome them. Information collection is carried out using secondary data; internationally reputable journals and accredited national journals are used to obtain information about the effectiveness and costs incurred. This paper will provide a holistic comparison of prices, effectiveness, and information on various remediating water and soil pollution methods around the world. Based on the NPV and IRR analysis, the water treatment method using the adsorption method is an economically feasible method with an IRR value of 51%.

A BRIEF OF WATER AND SOIL POLLUTION MANAGEMENT (RECENT TREND AND ECONOMIC PRESPECTIVE)

Soil and water pollution are the most crucial issues in the world. Various reports have informed that the pollution that has occurred has had long-term adverse effects on environmental sustainability and human health. Several methods have been reported to be effective in reducing pollutant parameters in water and soil. The methods often used in water and soil remediation are bioremediation (land-vetting, bio-cell. Bio cell, Phytoremediation, Land venting, Composite, Bio venting, Bio slurry), Adsorption, Pretreatment (Ultrasonic), Microwave, Electrokinetic disintegration. , High-Pressure Homogenization/HPH, Thermal Hydrolysis, Acid hydrolysis (HCl, H2SO4, H3PO4, and HNO3), Ozonation, Fenton Oxidation, Fe (II) -activated persulfate, Protease, amylase, lipase. This paper will explain water and soil pollution and the methods used to overcome them. Information collection is carried out using secondary data; internationally reputable journals and accredited national journals are used to obtain information about the effectiveness and costs that must be incurred. This paper will provide a holistic comparison of prices, effectiveness, and information on various methods of remediating water and soil pollution around the world. Based on the NPV and IRR analysis, the water treatment method using the adsorption method is an economically feasible method with NPV of 174,8742 and an IRR value of 39%.

Autoregressive parametric modeling combined ANOVA approach for label-free-based cancerous and normal cells discrimination

Label free based methods received huge interest in the field of bio cell characterizations because they do not cause any cell damage nor contribute any change in its compositions. This work takes a close outlook of cancerous cells discrimination from normal cells utilizing parametric modeling approach. Autoregressive (AR) modeling technique is used to fit the measured optical transmittance profiles of both cancer and normal cells. The transmitted light intensity, when passes through the cells, gets affected by their intercellular compositions and membrane properties. In this study, four types of cells: lung-cancerous and normal, liver-cancerous and normal, were suspended in their corresponding medium and their transmission characteristics were collected and processed. The AR coefficients of each type of the cell were analyzed with the statistical technique called Analysis of variance (ANOVA), which provided the significant coefficients. The poles extracted from the significant coefficients resulted in an improved demarcation for normal and cancer cells. These outcomes can be further utilized for cell classification using statistical tools.

MESENCHYMAL STEM CELL THERAPY FOR ACUTE KIDNEY DISEASE IN A CAT – CASE REPORT

<h3>Background</h3> Mesenchymal stem cells (MSCs) are attracted to the inflammatory microenvironment, where they act by producing cytokines and growth factors, which provide the immunomodulatory properties and can assist in the recovery of damaged tissue in cases of acute and chronic kidney disease. In this case report, the response of an animal with acute kidney disease to treatment with adipose tissue derived MSC (ADSC) was evaluated. <h3>Methods</h3> A cat diagnosed with acute kidney disease – 14.29 mg/dL of creatinine and 402 mg/dL of urea – was conducted to BIO CELL Cell Therapy to receive the treatment with mesenchymal stem cells. Four applications of ADSC by intravenous route, with 21 days of interval, were performed. Quantification of creatinine and urea in the animal‘s blood was performed 7 days after the second application to verify the animal‘s response to treatment. The exams were repeated after 1 year. <h3>Results</h3> The animal showed an improvement in the clinical signs and in the evaluated exams. Clinically, the animal returned to eat, drink water and gained weight. In the initial exams, before cell therapy, the animal presented creatinine of 14.29 mg/dL and urea of 402 mg/dL, but between the applications, the parameters stabilized at 5.28 mg/dL of creatinine and 171.00 mg/dL of urea. One year after treatment, exams showed that the parameters improved to 2.74 mg/dL of creatinine and 127.7 mg/dL of urea. The values were still above the reference value (from 0.5 to 1.5 mg/dL of creatinine and 10 to 75 mg/dL of urea), but the animal showed clinical improvement, returning to eat, with no renal alteration observed in ultrasound even after 1 year of treatment. <h3>Conclusion</h3> Treatment with ADSC helped in the recovery of the animal‘s kidneys, as demonstrated in the creatinine and urea values, demonstrating to be a promising therapy for acute renal disease. However, more clinical studies should be performed to evaluate the best protocol for this disease.

A Creation Model from the Gell-Mann Standard Model to the Creation of Bio Cells: Based on the Assumption of Homogeneous 5D Space-Time Universe

In this paper, we briefly go over the homogeneous 5D model field theory: from the 5D space-time inception, to its quantum field solutions given in terms of Higgs vacuum, filled with magnetic monopole bose fields of all energies. Then through the space dimension reduction projections, the Gell-Mann standard model was obtained as well as a quantum to Classical connection was made via introducing Bose distribution to the monopoles to obtain the Perelman entropy and Ricci Flow mappings. This provided us a picture to the creation of Astronomical objects, from galaxies to stars and planets. This method of splitting the monopole energy into ranges is extended to show that below the basic rest mass range of the electron and Quark, it still can be applied to explaining for the creation of the chemical elements periodic table. But perhaps the most interesting is in the lowest hundreds of Hz energy range, obtained from yet another 3 fold space symmetry breaking, into 2D × 1D, producing bio nitrogenous bases composed of 3 Carbon 12 in hexagon structures, due to preservation of the 1D monopole standing waves of this low frequencies. From that by imposing gauge changes the monopole states into DNA spectra. Since such spectra states retain the DLRO, it induces formation of charge carriers periodicity in a spherical bio cell.. It was then argued that due to cell’s surface proteins, the structure must contain partial filled VB, with “p” state hole density, and empty CB, separated from VB by a positive band gap. Such band structures resemble known HTC Cuprate ceramics. Since the HTC goes through a Superconductivity transition via the simultaneous bose exciton condensation, providing a Coulomb pressure, which reduces the band gap substantially, and induces the ODLRO transition of the hole density. The same obviously applies to the bio cells. Because of the near continuous exciton levels generated, a matching to the DNA spectra then can always occur by selective choices of proteins on the cell surface. Judging from a numerical study, we did years ago on YBCO, with doping. We found with a large enough VB hole density, the exciton induced superconducting gap can easily lead to Tc in the room temperature range. In fact by EMF excitation can increase the exciton pressure and trigger the ODLRO transition Tc upward. In fact, numerical results then suggest there do exist coherent EMF spectra from three key elements: Water, Carbon and Hydrogen, together with Oxygen, as studied over the years by numerous people, starting from Schrödinger to most recently Geesink.

Energy harvesting from a bio cell

This work shows experimentally how electrical energy can be harvested directly from cell membrane potential and used to power a wireless communication. The experiment is performed by exploiting the membrane potential of Xenopus oocytes taken from female frogs. Electrical potential energy of the membrane is transferred to a capacitor connected to the cell via a proper electrical circuit. Once the capacitor has reached the planned amount of energy, the circuit is disconnected from the cell and the stored energy is used to power a radio frequency communication that carries bio-sensed information to a distanced receiving circuit. Our result shows that electrical energy can be harvested directly from biological cells and used for a number of purposes, including wireless communication of sensed biological quantities to a remote receiving hub.

In-line mammalian cell concentration measurement with a Bio Cell Vitality Analyzer

Background Cell cultivations in a 30L bioreactor were in situ and on-line monitored using the Bio Cell Vitality Analyzer (BCVA, Sequip S+E, Dusseldorf, Germany). Based on the technique of optical reflectance measurement the BCVA is able to detect mammalian cells by combining reflectance measurement with a three dimensional moving focus point and a time of flight measurement. The aim of the experiments was to verify the process usability of the BCVA in bioreactor cultivation. Furthermore the BCVA shall be compared with the two already established Casy and Cedex devices (both Roche, Basel Switzerland). In contrast to the Cedex and Casy where a sample preparation is necessary the BCVA is able to measure the cells in situ. Moreover the particle distribution over time can be plotted.

Robust Hybrid Control for Two-Dimensional Handheld Micromanipulator

&lt;div class=""abs_img""&gt;&lt;img src=""[disp_template_path]/JRM/abst-image/00260003/06.jpg"" width=""300"" /&gt;A handheld micromanipulator&lt;/span&gt;&lt;/div&gt; This paper proposes a two-dimensional handheld micromanipulator oriented toward bio cell handling. The micromanipulator consists of two flexible links, each of which is constructed with a parallel beam structure. Electric coils and permanent magnets are used to produce an actuator to form double drivers. An explicit model predictive control combined with a PID controller called a robust hybrid control is proposed not only to achieve robust tracking performance, but also to dampen the vibration of the mechanism. The experimental results are compared to results from a standard PID controller to investigate the effectiveness of the micromanipulator. &lt;/span&gt;

The Role of MXD3 in Human Precursor B Cell Acute Lymphoblastic Leukemia

AbstractAbstract 3523Disregulation of the Hedgehog (Hh) signaling pathway is known to drive proliferation in several cancers, including hematopoietic malignancies. The role of the Hh pathway in precursor B cell (preB) acute lymphoblastic leukemia (ALL), the most common leukemia in children, is unclear; however, recent reports suggest that Hh signaling is involved in the development of B cell precursors as well as the self-renewal and survival of B cell malignancies.MXD3 is a transcription factor previously shown in our lab to be a novel member of the Hh pathway (Yun, Rust, Ishimaru, Diaz, Mol Bio Cell, 2007). We have previously shown that MXD3 is expressed in medulloblastoma (the most common brain tumor in children) and that its knockdown reduces proliferation of human medulloblastoma cell lines (Barisone et al., PLoS, 2012). In the current study, we investigated a possible role for MXD3 in preB ALL cell proliferation. Using quantitative real-time reverse-transcription PCR (qRT-PCR) we observed high levels of MXD3 mRNA expression in 8 primary preB ALL samples as well as in the preB ALL cell lines Reh and JM1. However, MXD3 levels were very low in mobilized peripheral blood mononuclear cells from healthy donors, mouse bone marrow and spleen. Indeed, MXD3 levels in the primary ALL cells and cell lines ranged between 13 and 35 fold increase when compared to the normal cells. Immunoblot analysis with anti-MXD3 monoclonal antibodies confirmed that the protein was present in the ALL samples but not in normal cells. We investigated the role of MXD3 in cell proliferation and survival, by silencing MXD3 in the Reh cell line. We used lentiviral delivery to knockdown MXD3 using an RNA interference approach. Briefly, lentiviruses were produced carrying a short hairpin RNA sequence specific for MXD3 (shMXD3) or a negative control sequence (shCTRL). Upon transduction by the viral vector, MXD3 knockdown was confirmed at both the RNA and protein level. Within 48 hours after transduction, MXD3 protein levels were reduced >90% in cells infected with the shMXD3 virus but not with the control virus. Interestingly, MXD3 knockdown resulted in decreased proliferation in Reh cells, supporting our hypothesis that it may be involved in the maintenance of this malignancy. As a first step toward understanding the mechanisms by which MXD3 exerts its function in leukemia cells, we analyzed cell cycle progression and apoptosis levels after knockdown using flow cytometry. We observed no significant differences in the G0/G1, S or G2/M populations between experimental and control samples. However, samples where MXD3 had been knocked down showed higher levels of apoptosis when compared to controls.Taken together, our results suggest that MXD3 is important for preB ALL cell proliferation, possibly by acting as an anti-apoptotic factor. Therefore, MXD3 may represent a suitable candidate for future efforts in developing targeted therapies against preB ALL. Disclosures:No relevant conflicts of interest to declare.

Sumoylation in neurodegenerative diseases.

The yeast SUMO (small ubiquitin-like modifier) orthologue SMT3 was initially discovered in a genetic suppressors screen for the centromeric protein Mif2 (Meluh and Koshland in Mol Bio Cell 6:793-807, 1). Later, it turned out that the homologous mammalian proteins SUMO1 to SUMO4 are reversible protein modifiers that can form isopeptide bonds with lysine residues of respective target proteins (Mahajan et al. in Cell 88:97-107, 2). This was the discovery of a post-translational modification called sumoylation, which enzymatically resembles ubiquitination. However, very soon it became clear that SUMO attachments served a far more diverse role than ubiquitination. Meanwhile, numerous cellular processes are known to be subject to the impact of SUMO modification, including transcription, protein targeting, protein solubility, apoptosis or activity of various enzymes. In many instances, SUMO proteins create new protein interaction surfaces or block existing interaction domains (Geiss-Friedlander and Melchior in Nat Rev in Mol Cell Biol 8:947-956, 3). For the past few years, sumoylation attracted increasing attention as a versatile regulator of toxic protein properties in neurodegenerative diseases. In this review, we summarize the growing knowledge about the involvement of sumoylation in neurodegeneration, and discuss the underlying molecular principles affected by this multifaceted and intriguing post-translational modification.

Bimorph Cantilevers Actuated by Focused Laser from the Side

The thin and long bimorph cantilever is mechanically responded in motion depending on the position of the heating laser which is focused from the side. The bimorph cantilever was designed in order to detect the heat from a bio cell in liquid. In the calibration process of the cantilever using a focused laser in vacuum and air, we found the quick and interesting responses of bimorph cantilever, which is vibrated and statically deflected due to the position of the local heat source on the cantilever. The deflections of cantilever have been investigated with a developed model. The responses are expressed by the modeled system with heat loss condition into ambient. To define the temperature distributions on the cantilever using a model, the temperature increments at the end of cantilever are successfully estimated to be +20 K in air and +110 K in vacuum with a laser power of 0.17 mW.

Development of Piezo Driven Inchworm Micro X-Y Stage and Hemispherical Tilting Positioner with Microscope Head

This paper deals with small piezo driven inchworm X-Y stage for a cell-manipulation in SEM chamber. Generally the electric noise from the moto causes the image destorsion in SEM operation. Thus the piezo driven inchworm small X-Y stage has been designed and fabricated as well as another tilting mechanism has been developed so that a micro pipettee can be implemented onto this tilting stage and manipulate such bio cell samples. Here we also developed a manipulation tool with micro force indicating system by using acoustic modulation technique since the microscopic operation in SEM without tactile information is difficult for the operator to control the tool precisely. On the other hand, a self-walking wafer inspection system is also described. The unique system is composed of the hemispherical manipulator and a self-walking piezo driven stage. This small walking mechanism can move on the glass plate that covers on the micro patterned plate. The hemispherical ball with microscope lens that can be rotated by the manner of stick-slip with three point contact piezo actuators is mounted on this self-walking stage so that the microscope lens can be focused onto the target sample to investigate the defect of micro patterns on the printed plate.

Abstract LB-227: Receptor tyrosine kinase phosphorylation: Simultaneous detection of 10 kinases upon inhibitor treatment

Abstract Receptor tyrosine kinases have been shown to be major pharmaceutical targets involved in disease states such as diabetes and cancer. Target driven therapeutics like small molecules or monoclonal antibodies are already in clinical use and have tremendous influence on new cancer therapies. In most cases either ligand binding to the extracellular domain followed by receptor dimerization or ATP binding to the intracellular kinase domain is inhibited. However, it is still challenging to gain specificity of those inhibitors for certain receptor tyrosine kinase targets. Bead-based assay technologies, such as the xMAP® platform, are ideal tools for the multiplex analysis of 5-20 proteins. Low sample volumes, simple assay protocols, flexibility and robustness are characteristics of bead-based assays that make them well suited for the immunological detection of bio marker and cell signaling proteins. In this presentation we will show results for the simultaneous detection of the ten most relevant receptor tyrosine kinases. The expression profile of more than 50 cell lines as well as breast tumor samples were evaluated with this powerful multiplex tool in a very robust and convenient way. Inhibitor profiling of small molecules under physiological conditions on ten receptor tyrosine kinases will be demonstrated by the detection of their phospho-Tyrosine level. These specificity profiles will help to improve the drug development process and to obtain potentially better drugs in future. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-227.

The Role of the Calcium Sensing Receptor in Regulating Intracellular Calcium Handling in Human Epidermal Keratinocytes

Calcium is critical for controlling the balance of proliferation and differentiation in epidermal keratinocytes. We previously reported that the calcium sensing receptor (CaR) is required for mediating Ca2+ signaling and extracellular Ca2+ (Ca2+(o))-induced differentiation. In this study, we investigated the mechanism by which CaR regulates intracellular Ca2+ (Ca2+(i)) and its role in differentiation. Membrane fractionation, fluorescence immunolocalization, and co-immunoprecipitation studies were performed to assess potential interactions between CaR and other regulators of Ca2+ stores and channels. We found that the glycosylated form of CaR forms a complex with phospholipase C gamma1, IP3 receptor (IP3R), and the Golgi Ca2+-ATPase, secretory pathway Ca2+-ATPase 1, in the trans-Golgi. Inactivation of the endogenous CaR gene by adenoviral expression of a CaR antisense cDNA inhibited Ca2+(i) response to Ca2+(o), decreased Ca2+(i) stores, decreased Ca2+(o)-induced differentiation, but augmented store-operated channel activity and Ca2+ uptake by intracellular organelles. Our results indicate that CaR regulates keratinocyte differentiation in part by modulating Ca2+(i) stores via interactions with Ca2+ pumps and channels that regulate those stores.

The Effect of GSK3β Inhibitor on Murine Multipotent Adult Progenitor Cells (mMAPCs).

The Wnt/β-catenin pathway is important for maintenance of the undifferentiated state of human embryonic stem cells (ESCs), hematopoietic stem cells (HSCs) and other stem cells. We here tested if β-catenin activation, by inhibiting GSK3 β with BIO (6-Bromoindirubin-3′-oxime), affects the differentiation status of murine multipotent adult progenitor cells (mMAPCs). mMAPCs were cultured with or without 0.1–2 μ M BIO, LIF, EGF and PDGF-BB, at low density (plating density of 100/cm2 and cells passed every 2 days) or at high confluency (like ESCs). Quantitative RT-PCR (Q-RT-PCR) was done weekly for the ESC-specific pluripotency transcripts, Oct4, Rex1, Nanog, UTF1 and E-Ras, and lineage commitment genes, vWF, VE-cadherin, Sox1, Nestin, CK19, Albumin, HNF3β and AFP. After 2–4 weeks, cells were analyzed by FACS for ESC/MAPC specific antigens, and by immunohistochemsitry for Oct4, β-catenin and E-cadherin protein. Furthermore, cells were differentiated into endothelial cells and hepatocytes by using VEGF-A and HGF/FGF4, respectively for 2 weeks. Q-RT-PCR was used to demonstrate lineage differentiation.BIO caused a dose-dependent clustering of mMAPCs from 4 days after treatment at both cell densities. FACS phenotype of low- and high-density mMAPCs was not affected by BIO. Nanog mRNA was not detected in any of the cell populations. However, Oct4, Rex1, vWF, and CK19 mRNA levels decreased in a BIO dose-dependent manner in mMAPCs maintained at low density, while they acquired higher levels of Sox1 mRNA (P=NS). At high-density, Oct4 and Rex1 mRNA level decreased in a BIO dose-dependent manner for the first week. On the other hand, 0.1 μ M BIO-treatment induced higher levels of Oct4 and Rex1 in cells maintained at high density. β-catenin and E-cadherin protein was highly expressed in clusters of low-density mMAPCs induced by BIO as well as BIO-treated mMAPCs maintained at ESC densities. In both cell densities, only mMAPCs treated with 1 or 2 μM BIO contained large dense mMAPCs clusters that expressed very high levels of β-catenin and E-cadherin. There was no statistical difference of differentiation potential between BIO conditions and cell densities.BIO may not allow culture of mMAPCs without loss of Oct4 as it did for ESCs, but induces clusters like ESCs in a BIO-dose dependent manner. High concentrations of BIO enhances β-catenin and E-cadherin protein expression. BIO may induce lineage commitment of mMAPCs. Despite loss of Oct4 mRNA and protein, MAPCs treated with BIO continue to have the ability to differentiate. Ongoing studies are testing whether BIO treated cells maintained at high density also maintain in vivo engraftment and differentiation potential.

Automatic Insemination and Incubation by Male and Female Micro Robots

The micro robot technology has the great potential benefit because they can provide very precise operation with the resolution of sub micro to nano meter easily. Recently it is increased that the demand for fine manipulation in the bio cell and sometime it should be done automatically. This paper describes the piezo based micro robots system which can provide the automatic insemination and incubation processes. These small robots with several range sensors can communicate and navigate themselves with the simple control sequences. And the experimental results show that the male and the female robots succeeded in make accurate navigation, insemination and incubation by using fish egg.

マイクロロボットファクトリの現状と将来

This paper describes the current status and future works of micro robots factory. For the latest demand of micro production, the miniaturized mechanical system such micro-mechatronics and instruments is the one of key technologies for providing several micro parts with a higher accuracy. Although it is known that the new technology of molecular machine has the great benefit to change the industries as well as human lives, the micro mechanical system ranging a few milimeters is still important to realize the reliable productivity. For these years, an unique micro robots system which is organized many tiny robots with tools and sensors has been developed. In our group, the small robots that are composed of the piezo elements and the electromagnets for providing the accurate inch worm locomotion, are employed as the base motion mechanism and several unique micro tools have been implemented on them. In the experiments, several actual applications such micro machining, bio cell operating as well as surface defect detecting are demonstrated.

Inhibitory effect of essential oils on apical growth of Aspergillus fumigatus by vapour contact.

The inhibitory effect of seven essential oils on the apical growth of hyphae of Aspergillus fumigatus was studied using a bio cell tracer by vapour contact in a sealed vessel. Based on the inhibitory pattern, these essential oils were classified into three groups. The first group, composed of citron, lavender and tea tree oils, stopped the apical growth in a loading dose of 63 micrograms ml-1 air, but allowed the regrowth of the hyphae after removal of the vapour, indicating fungistatic action. The second group, consisting of perilla and lemon-grass oils, stopped the apical growth in a loading dose of 6.3 micrograms ml-1 air, and did not allow the regrowth after gaseous contact at 63 micrograms ml-1 air, indicative of fungicidal action. The third group, consisting of cinnamon bark and thyme oils, retarded the growth in a dose of 6.3 micrograms ml-1 air, stopped it in a dose of 63 micrograms ml-1 air, and incompletely suppressed regrowth of the hyphae. Gas chromatographic analysis revealed that vapours of essential oils were absorbed on fungal mycelia and agar medium most abundantly by the first group, followed by the second and third groups, reflecting the volatility of the respective groups. Suppression of the apical growth by vapour contact was ascribed to the direct deposition of essential oils on fungal mycelia, together with an indirect effect via the agar medium absorbed.

A Novel Alternatively Spliced Form of Murine Vascular Endothelial Growth Factor, VEGF 115

Murine immortal fibroblasts express a form of vascular endothelial growth factor (VEGF) that was cloned, characterized and named VEGF 115. It differs from VEGF 120 by 37 amino acids at the carboxyl terminus. VEGF 115-specific sequence reacted to a single transcript in mouse tissues. Reverse transcription-polymerase chain reaction was performed in mouse tissues and in fibroblasts of normal and immortal divisional phenotypes. The data from mouse tissues suggested that VEGF 115 is not a tissue-specific isoform of VEGF 120, whereas a functional relevance with immortalization is indicated from the latter. The novel cDNA was expressed in Escherichia coli, and the His-tagged VEGF 115 (17.2 kDa) thus obtained was recognized by anti-VEGF antibody. A mammalian expression plasmid, pCMVneo+, encoding for VEGF 115 was transfected to NIH 3T3 cells, and the conditioned medium of stable transfectants was found to have fibroblast growth factor-replacing activity for human umbilical vein endothelial cells. Two independent genomic P1 clonings with primers specific for VEGF 164 and VEGF 115, respectively, resulted in isolation of identical P1 clones. We analyzed these three P1 clones on Southern blots with common and specific probes for VEGF 164 and VEGF 115. The results support the hypothesis that VEGF 115 is a new alternatively spliced form of mouse VEGF.