Interspecific Competitions between Frankliniella intonsa and Frankliniella occidentalis on Fresh Lentil Bean Pods and Pepper Plants.

BACKGROUND: Flower thrips (Frankliniella intonsa, IFT) and west flower thrips (Frankliniella occidentalis, WFT) are often found together on the host plant in China. WFT is an important invasive species that seems to outcompete the native IFT. In order to clarify the interspecific competitions between the two thrips, this study measured the population development of IFT and WFT under sexual and parthenogenetic reproductive modes on two hosts (fresh lentil bean pods with/without honey and pepper plants at seedling/flowering stages) in the laboratory. RESULTS: When reared on fresh lentil bean pods (with/without honey), WFT population size was lower in mixed species populations compared to single species populations but the presence of WFT had nor negative effect on IFT population size. These results were dependent of the reproductive mode. When honey was supplied, the ratio of female-to-male in the progeny of WFT produced under sexual reproductive mode increased significantly in the presence of IFT. On pepper seedlings, mixed populations were more favorable to the population development of IFT at the seedling stage, but more favorable to WFT at the flowering stage. CONCLUSIONS: In the early stage of WFT invasion and colonization, the emergence of flowering and honey (nectar) sources may have a positive effect on the population development of WFT.

Establishment and characterization of the Bactrocera dorsalis (Diptera: Tephritidae) embryonic cell line QAU-Bd-E-2.

In this study, we successfully established a Bactrocera dorsalis (Diptera: Tephritidae) embryonic cell line, i.e., QAU-Bd-E-2, from the insect eggs. The cells have been stably passaged for more than 60 times in TNM-FH medium with 10% fetal bovine serum (FBS). QAU-Bd-E-2 cells are adherent cells. Most of the cells were round, spindle-shaped, and rod-shaped. Round cells accounted for 82.3%, with a diameter of 13.9 ± 2.6 µm; spindle-shaped cells accounted for 9.8%, with the size of 51.2 ± 11.2 µm × 10.3 ± 3.1 µm; the rod-shaped cells accounted for 7.9%, with the size of 35.2 ± 9.4 µm × 12.0 ± 2.5 µm. The mitochondrial cytochrome oxidase I subunit (CoI) gene from QAU-Bd-E-2 cells was amplified, and the 657 bp fragment had a 100% similarity with the CoI gene of B. dorsalis, suggesting that the cell line was derived from B. dorsalis. The chromosome number of QAU-Bd-E-2 cells was mostly 12, which is the same as the B. dorsalis chromosome number. The cell density of QAU-Bd-E-2 cells reached the maximum (3.4 × 106 cells/mL) at 192 h, and the population doubling time was 31.9 h. Bactrocera dorsalis cripavirus (BdCV) could replicate in QAU-Bd-E-2 cells, suggesting that this cell line could be used for in-depth study of the relationship between virus and host.

A retrospective study of treatment of genu valgum/varum with guided growth: Risk factors for a lower rate of angular correction.

The rate of angular correction (ROAC) is very unpredictable and may be affected by various factors in the treatment of genu valgum and varum by means of guided growth. The purpose of this study was to assess the ROAC in cases from our institution and to identify risk factors associated with the occurrence of lower ROAC.We retrospectively reviewed the chart records of 68 patients undergoing guided growth with figure-eight plate for the correction of genu valgum and varum. Based on the data from these patients, the annual increment of physeal growth was calculated and compared with data from the Anderson chart. The associations between patient characteristics and ROAC were evaluated with the use of univariate logistic regression.The mean rate of femoral angular correction was 10.29 degrees/year, while the mean rate of tibial angular correction was 7.92 degrees/year. In a univariate logistic regression analysis, the variables associated with a higher risk of lower ROAC included non-idiopathic coronal deformity of the knee (odds ratio = 13.58, p < 0.001) and body weight at or above the 95th percentile for children (odds ratio = 2.69, p = 0.020).Obesity and non-idiopathic coronal deformity of the knee are risk factors for lower ROAC. It is still uncertain whether severity of deformity, race, and operative procedure have a substantial effect on the rate of correction.Level III evidence.

A cell clone strain from Mythimna separata (Lepidoptera: Noctuidae) highly susceptible to Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and M. separata NPV (MsNPV).

In this study, we describe a cell line, Ms-10C, cloned from the line QAU-Ms-E-10 (simplified Ms-10), an embryonic line from Mythimna separata. The cloned cell line was significantly more sensitive to nucleopolyhedrovirus (NPV). Ms-10C cells were mainly spherical with a diameter of 14.42 ± 2.23 µm. DNA amplification fingerprinting (DAF) confirmed the profile of PCR-amplified bands of the cloned cell line was consistent with those of the parental cell line, Ms-10. The sequencing result of the mitochondrial cytochrome c oxidase I (mtCO I) fragment confirmed that the amplified 636-bps mtCOI fragment was 100% identical to that of M. separata. Its chromosomes exhibited the typical characters of lepidopteran cell lines. Its population doubling time was 42.2 h at 27°C. Ms-10C was more sensitive than Ms-10 to both Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and M. separata nucleopolyhedrovirus (MsNPV). At 4 d post infection, the infection rates of two viruses reached 94.2 and 92.3%, respectively. The availability of this cell clone strain will provide a useful tool for the basic research on nucleopolyhedrovirus and for potential application in expression of recombinant proteins with baculovirus expression vector system.

Establishment of insect cell lines expressing green fluorescent protein on cell surface based on AcMNPV GP64 membrane fusion characteristic.

Displaying a protein on the surface of cells has been provided a very successful strategy to function research of exogenous proteins. Based on the membrane fusion characteristic of Autographa californica multiple nucleopolyhedrovirus envelope protein GP64, we amplified and cloned N-terminal signal peptide and C-terminal transmembrane domain as well as cytoplasmic tail domain of gp64 gene into vector pIZ/V5-His with multi-cloning sites to construct the cell surface expression vector pIZ/V5-gp64. To verify that the vector can be used to express proteins on the membrane of insect cells, a recombinant plasmid pIZ/V5-gp64-GFP was constructed by introducing the PCR amplified green fluorescent protein (GFP) gene and transfected into insect cell lines Sf9 and H5. The transected cells were screened with zeocin and cell cloning. PCR verification results showed that the GFP gene was successfully integrated into these cells. Green fluorescence in Sf9-GFP and H5-GFP cells was observed by using confocal laser scanning microscopy and immunofluorescence detection indicated that GFP protein was located on the cell membrane. Western blot results showed that a fusion protein GP64-GFP of about 40 kDa was expressed on the membrane of Sf9-GFP and H5-GFP cells. The expression system constructed in this paper can be used for localization and continuous expression of exogenous proteins on insect cell membrane.

A new cell line derived from embryonic tissues of Holotrichia parallela (Coleoptera:Scarabaeidae).

Holotrichia parallela is an important agricultural underground insect pest and also an edible and medicinal insect. Establishing a new cell line of H. parallela will provide a rapid and convenient tool for the studies on its physiology, pathology, and gene functions. In this study, by using the embryonic tissue of H. parallela as the material, we established a new cell line named Hp-E-1. The microscopic observation of its morphological characteristics revealed that its cellular morphology was mainly in the spherical morphology with a mean cellular diameter of 17.71 ± 2.34 µm, accounting for 67% of the total cells. The spindle-shaped cells accounted for 33% of the total cells with a mean size of 23.51 ± 4.37 × 13.98 ± 2.05 µm. The chromosomal number varied from 7 to 40, with about 50% of the cells having a diploid chromosome number of 2n = 20. Random amplified polymorphic DNA (RAPD) analysis indicated that the profiles of PCR-amplified fragments of this cell line were basically similar to those of the embryonic tissues of H. parallela but were obviously different from those of cell line BTI-Tn5B1-4 of Trichoplusia ni and cell line Sf-9 of Spodoptera frugiperda. The DNA fragment encoding mitochondrial cytochrome C oxidase subunit I (COI) gene of this cell line shared 99.7% homology with that of the embryonic tissue of H. parallela, confirming that this cell line is indeed derived from H. parallela. The results of growth curve measurement indicated that the population doubling time of this cell line was 136.7 h. Cell line Hp-E-1 could not be infected by three viruses Autographa californica multiple nucleopolyhedrovirus (AcMNPV), Bombyx mori nucleopolyhedrovirus (BmNPV), and Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV).

Establishment and characterization of three embryonic cell lines of beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae).

Three cell lines (QAU-Se-E-1, -2 and -3, or Se-1, -2 and -3 for short) were established from eggs of beet armyworm (Spodoptera exigua) that have been passaged stably for more than 60 times in TNM-FH medium supplemented with 10 % fetal bovine serum. The cell lines consisted of round and spindle-shaped cells. The round cells accounted for 96.82, 84.34 and 83.16 % of the cells in the three cell lines, respectively, with cell diameters of 16.21 ± 0.72, 15.63 ± 0.58 and 13.06 ± 0.44 µm. Random amplified polymorphic DNA and analysis of the CO I gene showed that the three cell lines were all derived from S. exigua. Growth curves at passage 30 were determined and the results showed that the cell population doubling times were 59.03, 49.08 and 49.91 h, respectively. The three cell lines can be infected by S. exigua multiple nucleopolyhedrovirus (SeMNPV). Se-3 was extremely susceptible to the virus with an infection rate of 97.52 % 4 days after the inoculation and produced 2.02 × 10(6) OBs per mL of culture. Flow cytometry analysis showed that some of Se-1 and Se-2 cells had apoptosis after infection, whereas Se-3 cells did not. Bioassays showed that the virulence of the SeMNPV proliferated from Se-3 was similar to that from the insect with LC50 of 5.55 × 10(5) and 2.64 × 10(5) OBs/mL. Therefore, the cell lines can be used to study the SeMNPV-host interactions and mechanisms underlying the interactions.

Characteristics and indications of hydrogen and oxygen isotopes distribution in lake ice body.

Stable isotopes have been used to identify the characteristics of precipitation, evaporation, basin hydrology, and residence times. However, lakes in the cold regions are usually covered by ice for 5-6 months. To get a better understanding of stable isotopes characteristics and indications in lake ice bodies, ice and water were sampled during the icebound season in both the ice and water bodies in Dali Lake, and deuterium, oxygen-18 total nitrogen (TN), and the major ions were analyzed. The results showed that deuterium and oxygen-18 compositions (δD-δ¹8O) compositions in the ice body were greater than in the water body beneath, scattered on a straight line, and deviating downward from the global meteoric water line in the top right. The ice profile showed that the δD-δ¹8O compositions increased from the ice surface downward and decreased near to the bottom. In contrast, the TN and the major ions in the ice decreased from the ice surface downward and increased near to the bottom, meaning that the concentrations of δ¹8O had a negative correlation with the concentrations of TN and major ions. These indicated that stable isotopes can be used for tracing the nutriment and ion transport processes in the ice body.

Serumfree culture of the suspension cell line QB-Tn9-4s of the cabbage looper, Trichoplusia ni, is highly productive for virus replication and recombinant protein expression.

Serumfree cultures of insect cells play an important role in the fields of protein engineering, medicine, and biology. In this paper, the suspension cell line QB-Tn9-4s of Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) was successfully adapted to serumfree Sf-900 III medium and passaged for 52 generations. The adapted QB-Tn9-4s cells grew faster. Their population doubling time shortened from 27.4 hr in serum-containing medium to 24.1 hr, and their maximal density increased by 1.83-fold, reaching 3.50 ×10(6) cells/mL in serumfree culture in T-flasks. The cells readily adapted to spinner culture, with maximum cell density of 4.40 × 10(6) cells/mL in a spinner flask. Although the infection rate of Autographa californica multiple nucleopolyhedrovirus and production of occlusion bodies (OBs) of the adapted QB-Tn9-4s cells were 91.0% and 85.4 OBs/cell, respectively, similar to those of QB-Tn9-4s cells cultured in serum-containing medium and control BTI-Tn5B1-4 cells, their budded virus titer was 4.97 ×10(7) TCID50/mL, significantly higher than those of the latter two. In addition, the expression levels of ß-galactosidase at six days postinfection and secreted alkaline phosphatase at seven days postinfection in the adapted QB-Tn9-4s cells reached 2.98 ± 0.15×10(4) IU/mL and 3.34 ± 0.13 IU/mL, respectively, significantly higher than those of QB-Tn9-4s and control BTI-Tn5B1-4 cultured in serum-containing media. The above findings establish a foundation for industrial production of virus and recombinant proteins in QB-Tn9-4s serumfree culture.

Establishment and characterization of three new cell lines from the embryonic tissue of Holotrichia oblita Faldermann (Coleoptera: Scarabaeidae).

The establishment of new insect cell lines plays important roles in the researches of insect pathology, insect toxicology, insecticide screening and activity assay, etc. Using embryos of Holotrichia oblita Faldermann (Coleoptera: Scarabaeidae) as materials, this study describes the establishment of three cell lines designated as QAU-Ho-E-3 (Ho-3), QAU-Ho-E-4 (Ho-4), and QAU-Ho-E-6 (Ho-6), respectively. Currently, the three cell lines have been passaged more than 50 times in the TNM-FH insect cell medium containing 10% fetal bovine serum (FBS). All of them showed adherent growth. The majority of Ho-3 cells are spindle-shaped, with a size of 24.35 ± 5.29 × 11.56 ± 1.67 µm. The Ho-4 cells were either spindle-shaped or oblong, with a size of 38.07 ± 8.57 × 17.62 ± 2.48 µm. The Ho-6 cells were primarily round in shape with a diameter of 14.54 ± 1.96 µm. The Ho-3 and Ho-4 cell lines contained 20 chromosomes (i.e., diploid, 2n = 20) at passages 14 and 45. The Ho-6 cell line contained 20 chromosomes (i.e., diploid, 2n = 20) at passage 14 but 40 chromosomes (i.e., polyploidy, 4n = 40) at passage 45. The results of random amplified polymorphic DNA (RAPD) analysis showed that the RAPD fingerprint of the three cell lines was consistent with that of H. oblita eggs, but clearly different from that of BTI-Tn5B1-4 and Sf-9 cells, demonstrating that the three cell lines Ho-3, Ho-4, and Ho-6 are H. oblita cell lines. The results of the growth curve test showed that the population doubling times of Ho-3, Ho-4, and Ho-6 were 101.1, 105.2, and 83.6 h, respectively. The viral infection assay indicated that these H. oblita cell lines were not permissive to infection by Autographa californica multiple nucleopolyhedrovirus (AcMNPV) or Bombyx mori nucleopolyhedrovirus (BmNPV).

[Bioavailability of dissolved organic nitrogen components in the lake sediment to alage].

Samples in the sediments of Wuliangsuhai and Erhai Lake were selected, and the technologies of XAD-8 resins separation and three dimensional fluorescence excitation-emission matrix (3DEEM) spectra were applied, in order to study the bioavailability of dissolved organic nitrogen components to alage under the room cultivation. The obtained results showed that: (1) Average loss of DON and DOC from sediments was below 5% after dividing DON into different groups, which means the technology of XAD- 8 resins separation could be used in the study of DON components in the lake sediment. (2) Through 3DEEM analysis, hydrophilic and hydrophobic DON was dominated by protein-like and humic-like materials in the lake sediment, respectively. (3) Under the hydrophilic component condition, growth curves of algae tended to show an "S" shape with a straight upward trend in the sediments of Wuliangsuhai and Erhai lake,with the maximum algal density reaching 535.5 x 10(4) and 709.5 x 10(4) per milliliter, respectively. Meanwhile, the DON concentrations were significantly reduced after cultivation to 2.46 and 2.98 mg x L(-1), respectively, which suggests that hydrophilic DON in the lake sediment was the bioavailable organic nitrogen for alage. (4) Under the hydrophbic component conditions, growth curves of algae tended to show a "unimodal" shape in the sediments of Wuliangsuhai and Erhai lake,with the maximum algal density reaching 113.5 x 10(4) and 275.5 x 10(4) per milliliter,respectively. The DON concentrations were significantly reduced during the early cultivation period, and then kept stable in the late period, which suggests that the hydrophobic DON component bioavailable to alage was low in short-term, and the hydrophobic DON component had hardly any positive effect on the growth of algae.

[Influence on emerged plant in the process of numerical simulation about lake water quality].

An eutrophication model on Wuliangsuhai Lake in Inner Mongolia is established through combining the environmental fluids dynamics code model with CE-QUAL-ICM model. In this model, the distribution of emergent plants, density, height, diameter and other morphological indicators to be considered. At the same time, nitrogen and phosphorus are calculated, compared and analyzed by whether to consider the impact of emergent plants to nitrogen and phosphorus in the simulation process. The result shows that the trend of simulated and measured values on nitrogen and phosphorus by considering the impact of emergent plants is consistent; however simulated values can't reflect the measured values if not to considering the impact of emergent plants. It can clearly be seen that emergent plants plays a key role in migration and transformation of nitrogen and phosphorus, which improve the efficiency and simulation accuracy. Overall eutrophication model considering the impact of emergent plants can better reflect dynamic simulation results on the concentration of nitrogen and phosphorus in the region of a lake; also can provide some reference to lake management.

[pH and ion balance in wheat-wheatgrass under salt- or alkali stress].

Different intensities of salt- or alkali stress were established by mixing different concentrations of NaCl and Na2SO4 or NaHCO3 and Na2CO3, respectively, and wheat-wheatgrass (Triticum aestivum L. -Agropyron intermedium) seedlings were grown under the stresses for 12 days. The pH value and the Na(+), K(+), free Ca(2+), Cl(-), SO4(2-), NO3(-), H2PO4(-), and organic acid concentrations in the fresh shoots of stressed wheat-wheatgrass seedlings were determined, aimed to approach the characteristics of pH and ion balance in wheat-wheatgrass under salt- or alkali stress. The results showed that intracellular pH was relatively stable under both stresses. Cl(-) concentration increased sharply and organic acid concentration changed less under increasing intensity of salt stress, while it was, in adverse under increasing intensity of alkali stress. Under both stresses, the cations in the fresh shoots were mainly Na(+) and K(+), but the anions were different. Under salt stress, inorganic anions were the dominant components and contributed 61.3 %-66.7% to the total negative charge, while under alkali stress, the contribution of organic acid to total negative charge increased from 38.35% to 61.6% with increasing stress intensity, and became the dominant component. It was concluded that organic acid accumulation might be a key physiological response of wheat-wheatgrass for its keeping pH and ion balance under alkali stress.

[Expression and insecticidal activity of a novel gene cry2ab4 from Bacillus thuringiensis strain B-Pr-88].

The full length cry2Ab gene was cloned by PCR-RFLP method from Bt strain B-Pr-88, which was isolated in China with high toxicity to the Lepidopteran insect pests. Nucleic acid sequence analysis showed that this gene was 1902 base pairs encoding 633 amino acids. This cry gene was named cry2Ab4 as a novel gene by Bacillus thuringiensis Delta Endotoxin Nomenclature Committee. The full open reading frame sequence of the cry2Ab4 gene was amplified with a pair of PCR primers L2ab5/L2ab3 designed according to its DNA sequence,and inserted into the BamH I /EcoR I sites of E. coli expression vector pET21b to obtain the recombinant plasmid pET-2Ab4. The result of SDS-PAGE proved that Cry2Ab4 could be expressed as a 60 kD protein in E. coli BL21 (DE3)strain induced by IPTG. Bioassay of the expressed product of the cry2Ab4 gene showed that Cry2Ab4 was highly toxic to the larvae of Helicoverpa armigera and Leguminivora glycinivorella, moderately active to the larvae of Plutella xylostella and Chilo suppressalis, but not insecticidal to the larvae of Spodotera exigua and Ostrinia furnacalis. Our result indicated that cry2Ab4 gene could be used as a novel gene for generation of transgenic plants and engineered microorganism.

[The expression of vegf in rat fracture reparative callus].

The present study has determined the histological changes, VEGF (Vascular endothelial growth factor) mRNA alternative expressions and VEGF mRNA levels in chondrogenic-callus, osteogenic-callus and total callus by histology, RT-PCR and Northern blot during rat fracture repair. The results showed that the cellular events of intramembranous ossification, chodrogenesis and endochondral ossification appeared in the callus simultaneously and sequentially. The VEGF mRNAs were expressed in the osteogenic-callus and the chondrogenic-callus, and the strongest expressions were VEGF120 and VEGF164 mRNA. The VEGF mRNA levels in the total callus increased with time after fracture. Take together, these findings suggest that osteogenesis-derived VEGF may play a potential role to modulate the differentiation process of osteogenic and chondrogenic cells during fracture repair.

Regulation of alternative splicing of Bcl-x by IL-6, GM-CSF and TPA.

The splicing of many alternative exons in the precursor messenger RNA (pre-mRNA) is regulated by extracellular factors but the underlying molecular bases remain unclear. Here we report the differential regulation of Bcl-x pre-mRNA splicing by extracellular factors and their distinct requirements for pre-mRNA elements. In K562 leukemia cells, treatment with interleukin-6 (IL-6) or granulocyte-macrophage colony stimulating factor (GM-CSF) reduced the proportion of the Bcl-xL variant mRNA while treatment with 12-O-tetradecanoylphorbol 13-acetate (TPA) had no effect. In U251 glioma cells, however, TPA efficiently increased the Bcl-xL level. These regulations were also seen for a transfected splicing reporter mini-gene. Further analyses of deletion mutants indicate that nucleotides 1-176 on the downstream intron are required for the IL-6 effect, whereas additional nucleotides 177-284 are essential for the GM-CSF effects. As for the TPA effect, only nucleotides 1-76 are required in the downstream intron, Thus UK-6, GM-CSF and TPA differentially regulate Bcl-x splicing and require specific intronic pre-mRNA sequences for their respective effects.

[Regulation of pre-mRNA alternative spicing in U251 cells by TPA].

By using the mini-gene construct containing partial sequence of Bcl-X gene as model, we examined the function of TPA on Bcl-X pre-mRNA alternative splicing in vivo and vitro with RT-PCR and site-directed mutagesis assay. The results show that PKA signaling system can regulate Bcl-X pre-mRNA alternative splicing, the possible mechanism is that the responsible sequence affect the choice of the 5'-downstream or upstream splice site of Bcl-X pre-mRNA.

[Immunogenecity of combined hepatitis A and B vaccine].

OBJECTIVE: To observe the immunogenicity of combined hepatitis A and B vaccine (HAB). METHODS: The combined HAB vaccine was prepared and different concentrations of HAB were administered on mice in week 0, 4 and 24, and then we tested the antibodies to both hepatitis A virus and B virus. After the first injection, we tested the hepatitis A antigen-induced and hepatitis B surface antigen-induced stimulation indices in spleen monocyte as well as changes of CD4+ and CD8+ cell numbers. RESULTS: The serum antibody positive rates were 100% in all three groups, and the antibody induced by HAB vaccine were earlier than by monovalent vaccine. The hepatitis A antibody and hepatitis B surface antibody titers after the combined vaccine inoculation were not significantly higher than those after the monovalent vaccine inoculation. On the other hand, after the first injection of the combined vaccine, the hepatitis A antigen-induced and hepatitis B surface antigen-induced stimulation indices in spleen monocyte were detected. The numbers of CD4+ and CD8+ cells increased. CONCLUSIONS: HAB vaccine has reliable immunogenicity.

Effects of tachyplesin on the morphology and ultrastructure of human gastric carcinoma cell line BGC-823.

AIM:To investigate the morphological and ultrastructural changes in the human gastric carcinoma cell line BGC-823 after being treated with tachyplesin.METHODS:Tachyplesin was isolated from acid extracts of Chinese horseshoe crab (Tachypleus tridentatus) hemocytes. BGC-823 cells and the cells treated with 2.0mg/L tachyplesin were examined respectively under light microscope, scanning and transmission electron microscope.RESULTS: BGC-823 cells had undergone the restorational alteration in morphology and ultrastructure after tachyplesin treatment. The changes were as follows: the shape of cells was unanimous, the volume enlarged and cells turned to be flat and spread, the nucleo-cytoplasmic ratio lessened and nuclear shape became rather regular, the number of nucleolus reduced and its volume lessened,heter-chromatin decreased while euchromatin increased in nucleus. In the cytoplasm, mitochondria grew in number with consistent structure relatively, Golgi complex turned to be typical and well-developed,rough endoplasmic reticulum increased and polyribosome decreased. The microvilli at cellular surface were rare and the filopodia reduced while lamellipodia increased at the cell edge.CONCLUSION:Tachyplesin could alter the malignant morphological and ultrastructural charact-eristics of human gastric carcinoma cells effectively and have a certain inducing differen-tiation effect on human gastric carcinoma cells.