Description and interpretation of interphalangeal lines in tetrapods

A previously unnoticed geometric pattern is present in the extremities of all tetrapods. Sets of straight and typically uninterrupted hinge lines pass through neighboring interphalangeal joints and across ungual tips. Four sets of these lines appear in basal polydactyl tetrapods, two medial sets, a transverse set and a lateral set. The two medial sets merge in primitive pentadactyl tetrapods. The resulting three line sets persist in later taxa, even when digits shrink and disappear. Primitively and typically the lines in each set are more or less parallel, but lines may converge, merge and shift as phalanges disappear or phalangeal proportions change. Confirming this geometric pattern, complex interphalangeal joint surfaces typically align with hinge lines and pad divisions parallel them. In addition, unguals rarely cross extensions of hinge lines and longer unguals may divert medially or laterally rather than cross them. Exceptions occur most commonly on ungual II. Line sets may exist because phalanges appear to flex and extend most efficiently in unison. Hinge line patterns appear to identify clades so they may, to a limited extent, be used taxonomically. Hinge lines also have predictive value in that missing phalanges, including unguals, can be reconstructed with confidence using hinge lines as size guides. Correct digit spread and metapodial configuration can also be determined in extinct taxa by seeking the appearance of continuous interphalangeal hinge lines in tested reconstructions.     

Interpreting the autopodia of tetrapods: interphalangeal lines hinge on too many assumptions

Recently Peters proposed the concept of ‘interphalangeal lines’, defined as sub-parallel lines that could supposedly be drawn across the joints of the digits of all tetrapods. The lines were viewed as potential axes of rotation, and it was suggested that they could be used to determine the resting position of the digits, reconstruct missing digital elements of fossil tetrapods, and provide information on systematic relationships. Evidence was adduced from the skeletons of recent and fossil vertebrates and from footprints. However, detailed analysis shows that these claims are largely unfounded. Linear alignments of joints on neighbouring digits are not consistently present in tetrapods, especially across locomotor cycles. Even if present, interphalangeal (IP) lines would rarely be in an appropriate orientation to facilitate joint movements during locomotion. There is no reason to believe that IP lines would be homologous across different taxa, so they cannot be used to infer systematic relationships. Finally, the alleged support from the ichnological record is undermined by the uncertain relationship between the joint structure of the skeleton and the form of the print. We conclude that IP lines cannot be consistently constructed on tetrapod extremities, and would have minimal functional relevance or predictive power in any case.     

Anti-predator defence drives parallel morphological evolution in flea beetles

Complex morphological or functional traits are frequently considered evolutionarily unique and hence useful for taxonomic classification. Flea beetles (Alticinae) are characterized by an extraordinary jumping apparatus in the usually greatly expanded femur of their hind legs that separates them from the related Galerucinae. Here, we examine the evolution of this trait using phylogenetic analysis and a time-calibrated tree from mitochondrial (rrnL and cox1) and nuclear (small subunits and large subunits) genes, as well as morphometrics of femora using elliptic Fourier analysis. The phylogeny strongly supports multiple independent origins of the metafemoral spring and therefore rejects the monophyly of Alticinae, as defined by this trait. Geometric outline analysis of femora shows the great plasticity of this structure and its correlation with the type and diversity of the metafemoral springs. The recognition of convergence in jumping apparatus now resolves the long-standing difficulties of Galerucinae-Alticinae classification, and cautions against the value of trait complexity as a measure of taxonomic significance. The lineage also shows accelerated species diversification rates relative to other leaf beetles, which may be promoted by the same ecological factors that also favour the repeated evolution of jumping as an anti-predation mechanism.     

Ontogenetic allometry of the digital rays of the leopard gecko (Gekkota: Eublepharidae; Eublepharis macularius)

The role of allometry in producing the variation in autopodial morphology observed among the lizards is not well understood. Allometry of metapodial and digit lengths in the manus and pes of the primitively padless gekkotan (Eublepharis macularius) is explored using maximum‐likelihood repeated‐measures ANCOVAs with body length as the covariate. Estimated variance–covariance matrices differed significantly within and between autopodia, and integration was stronger among the metapodials than the digits. The first metapodial and the first digit of each autopodium exhibit the strongest covariances with each of the remaining components in each variance–covariance matrix, suggesting that the lengths of the first rays are important for allometric integration of both manus and pes. Metapodials scale isometrically and digits negatively allometrically; both display allometric heterogeneity among themselves in both autopodia. Both autopodia exhibit changes in proportion over the ontogenetic size range, attributable to variation in scaling among the components of the rays. Allometric coefficients do not vary among pedal digits, despite differences in phalanx number, although phalanx number is associated with differences in slope in the manual digits. This is suggestive of heterogeneity in allometry among the manual phalanges, which thus may be associated with variation in phalanx length within gekkotan digits.     

In defence of the calcichordates

The cornutes and mitrates, collectively known as stylophorans or calcichordates, are the most controversial members of the traditional group of the 'carpoid echinoderms'. In a recent review, Philip has rejected Ubaghs' interpretation of them as aberrant echinoderms with a single feeding arm, has also rejected my own interpretation of diem as primitive chordates, and has made some tentative proposals of his own. The present paper answers Philip's criticisms of the calcichordate interpretation and discusses his own proposals and his comments on Ubaghs' views.     

In vitro host range of the Hz-1 nonoccluded virus in insect cell lines

A total of 13 insect cell lines spanning 4 orders (Lepidoptera, Coleoptera, Diptera, and Homoptera) were tested for their ability to replicate the nonoccluded virus Hz-1. Only the Lepidopteran cell lines supported replication of the virus with TN-CL1 and BCIRL-HZ-AM1 producing the highest titers of 2.4 × 10⁸ tissue culture infective dose (TCID)₅₀/ml and 2.0 × 10⁸ TCID₅₀/ml, respectively. A codling moth cell line (CP-169) was the only Lepidopteran cell line that did not replicate the virus and transfection of this cell line with Hz-1 DNA failed to replicate the virus. Also, transfection with DNA from a recombinant baculovirus carrying the red fluorescent protein gene (AcMNPVhsp70 Red) was not expressed in CP-169 cells. The replication cycle of Hz-1 in BCIRL-HZ-AM1 cells showed that this virus replicated rapidly starting at 16 h postinoculation (p.i.) and reaching a peak titer of 1.0 × 10⁸ TCID₅₀/ml 56 h postinoculation. Hz-1 when compared with several other baculoviruses has the widest in vitro host spectrum.     

Characterization of invertebrate cell lines

Summary Cell extracts of five mosquito cell lines and a tick cell line were examined for four cellular isozymes using a cellulose-acetate electrophoretic technique. This method distinguished the cell lines that were derived from the different species. Intraspecies distinctions were not made using the cell lines tested; the significance of this finding is discussed. The usefulness of this technique in identifying a potentially mislabeled cell line was demonstrated. This research was supported by contracts, DADA 17-72C-2170 of the U.S. Army and N00014-78C-0104 of the U.S. Office of Naval Research and grants from the World Health Organization and the Rockefeller Foundation.     

特比萘芬短程口服加外用特比萘芬治疗中重度皮肤癣菌病的临床对照研究

目的为治疗中、重度皮肤癣菌病选择合适的方法。方法采用开放、随机对照的平行研究。将临床和真菌镜检确诊的中重度体、股癣和手、足癣患者随机分入A、B、C、D四组,A组口服特比萘芬片联合外用特比萘芬乳膏,疗程7d;B组常规口服特比萘芬片,疗程14d;C组常规外用特比萘芬乳膏,疗程14d;D组外用联苯苄唑乳膏,疗程4周。对各组的临床疗效、安全性、患者对治疗的满意度等进行分析。结果①真菌学疗效:停药后4周时A组手、足癣患者的真菌清除率最高为96.65%,与D组比较(65.00%)差异有显著性。②临床疗效:停药后2周和4周时A组手足癣患者的治愈率分别为93.33%和90.00%,与D组(68%、64%)比较差异有显著性。③患者满意度:在本研究的四组中,A组患者对疗效的满意度最高,与C组、D组比较差异有显著性;患者的综合满意度(包括对治疗费用、安全性等方面)A组最高,与C组、D组比较差异有显著性。D组患者对费用的接受程度最高,A组优于B组,有极显著的统计学差异。治疗结束后患者对安全性的接受程度四组间没有显著差异。结论特比萘芬口服加外用1周的“1 1”短程联合疗法治疗中、重度皮肤癣菌病的疗效高、疗程短、依从性好,患者满意度高。     

Fish cell lines: Establishment and characterization of nine cell lines from salmonids

Summary Nine permanent cell lines have been established from five species of salmonids native to America's Pacific Northwest. With the exception of a hepatoma from an adult trout, the lines were derived from normal tissues of embryonic or juvenile fish. Cells were routinely grown in Eagle's minimum essential medium with 10% fetal bovine serum. Optimum growth temperatures for these lines ranged from 21 to 24°C. All survived storage for at least 1 yr at −65°C and at least 5 yr in liquid nitrogen. Six of the lines were demonstrably free of any microbial contamination but mycoplasmas were found in three. Eight of the lines were heteroploid. The morphology of only one was fibroblastic. All the lines effectively replicated one or more of the common salmonid viruses. Isozyme patterns were consistent with those of the species of origin. These cell lines have significant application in fish virology. This work is a result of research sponsored in part by the Oregon State University Sea Grant College Program supported by NOAA Office of Sea Grant, U.S. Department of Commerce, under Grant NA79AA-D-0016 and by the Oregon Department of Fish and Wildlife under PL-89304 Anadromous Fish Act and is Oregon Agricultural Experiment Station Technical Paper 6857.     

In vitro proliferation and in vivo malignancy of cell lines simultaneously derived from a chemically-induced heterogeneous rat mammary tumor

Summary Identification of clones in primary tumors responsible for proliferation, invasion, and metastasis was carried out. Four different aneuploid established cell lines derived from a ductal infiltrating mammary rat tumor induced by 7,12-dimethylbenz[a]anthracene were studied for proliferative and growth features in vitro and for tumorigenic and metastatic potential in vivo in nude mice. Clones, named RM1, RM2, RM3, and RM4, were characterized by different proliferative activity. Clone RM1 showed the highest proliferative activity by both tritiated thymidine incorporation and S-phase flow cytometry, followed by clone RM4. Conversely, clones RM2 and RM3 showed a lower proliferation rate. Growth-promoting activity, tested on 3T3 Swiss cells, was high in all clones, although RM1 showed significantly lower growth factors—releasing activity. Nude mice tumorigenesis demonstrated a strong tumor induction of line RM1 (100% of the mice after 47±7 d) and a slightly lower tumor induction of line RM4 (70% of the mice after 69±9 d). Line RM3 showed tumor induction in 40% of the mice after 186±16 d. Lines RM2 showed no tumor induction. Metastasis occurred in mice treated with line RM1 only. Therefore, tumorigenesis and metastasis correlate with proliferation but not with the release of growth factors. In conclusion, flow cytometry monitoring of clones from heterogeneous primary tumors proved to be a suitable model for the study of in vivo malignancy and in vitro proliferation.     

Development and characterization of new immortalized human breast cell lines

New human breast cell lines were developed from metastatic breast cancer tissues and normal breast tissues. Primary cultures were initiated from cellular outgrowths of explanted tissues or from mechanically isolated cells in two serum-free media. Cell cultures derived from both cancer and normal tissues were immortalized with pRSV-T plasmid to generate permanent breast cell lines that exhibited an epithelial morphology. Cell lines generated in this study were characterized with respect to morphology, growth rate, karyotype, presence of specific genes, and the expression of epithelial and breast markers. The cell lines expressed the epithelial cell markers, cytokeratins 8 and 18, and retained the capacity to produce human milk fat globulin. They also express the BRCA-1, erbB2, and EGF receptor genes and possess the H-ras, K-ras, and p53 genes. Preliminary data showed that one of the new cancer cell lines was highly sensitive to the cytotoxic action of taxol. It is envisioned that the new breast cell lines will be useful as targets for identification of therapeutic agents against breast cancer and as models for carcinogenesis studies. This revised version was published online in June 2006 with corrections to the Cover Date.     

Characterization of invertebrate cell lines

Summary A reliable, simple method for the separation of four cellular isozymes utilizing celluloseacetate electrophoresis is described. Cell extracts of 16 cell lines were examined previously, utilizing starch gel electrophoretic techniques. These same cell extracts were retested for their isozyme phenotype using the cellulose-acetate electrophoretic system. These data indicate that the results of the two electrophoretic systems are comparable. Isozyme analyses of freshly prepared cell extracts of theAedes albopictus (ATC-15) andSpodoptera frugiperda (IPLB-SF-21AE) cell lines resulted in identical isozyme mobilities when compared with their respective stored counterparts. Since the cellulose-acetate electrophoretic system was able to separate cellular isozymes into characteristic patterns, distinctions between cell lines were made. The application of this technique for identification and characterization of invertebrate cell lines is discussed. This research was supported in part by the Rockefeller Foundation and U.S. Public Health Service Grant AI-13727.     

Characterization of invertebrate cell lines

Summary The usefulness of four serologic techniques for distinguishing five selected lepidopteran cell lines was evaluated; the techniques included complement fixation, hemagglutination. immunodiffusion, and immunoelectrophoresis. The five selected lepidopteran cell lines represent three taxonomic families of Lepidoptera with one family, Noctuidae, containing two cell lines derived from insects within the same genus. The five cell lines were crossreactive in complement-fixation tests, but the lines were distinguishable at a familic level when two units of antigens were used in the test. Agglutination of goose erythrocytes was not observed with the antigens over a pH range of 5.8 to 7.2 at 4°C or ambient temperature. Immunodiffusion tests demonstrated a common cross-reactive antigen(s), but spurs of partial identity and the presence of extra precipitin bands were indicative that differentiation at a familic level was possible. Immunoelectrophoresis of the cellular antigens also revealed common cross-reactive precipitin arcs, but the number and clarity of arcs in homologous systems was increased such that four of the five cell lines were distinguishable. A basic protein was consistently seen in the homologous system, but it was absent in the heterologous systems. Although these data suggest that immunoelectrophoresis was the best serologic technique for distinguishing the five lepidopteran cell lines, the shortcomings of this approach are also discussed. This research was supported in part by the World Health Organization, The Rockefeller Foundation, and U.S. Public Health Service Grant AI-13727.     

Harris lines: A study of age-associated bias in countingand interpretation

Harris lines are regularly used in paleopathology as indicators of episodic nonspecific stress, but the methodology for their use has not been clearly established. We studied radiographs of the distal shaft of the tibia in 82 immature and 49 mature subjects from a medieval burial site and compared the number of Harris lines and observer error according to age categories. We found statistically significant differences in both line counts and in observer error by age groups. In conclusion, studies of Harris lines must take into account age variation in order to be validated. Am J Phys Anthropol 103: 209–217, 1997. © 1997 Wiley-Liss, Inc.     

Measurements of housing density along transmission lines

One of the fundamental inputs to models of population exposure to the power-frequency fields of high-voltage transmission lines is the density of population about the lines. We examined aerial photographs of over 1,700 km of transmission corridor to characterize the density of nearby housing units. The data show that the density of houses within 200 m of transmission lines 1) is typically smaller than the average density of houses in the local utility's service area, 2) tends to increase with distance from the line, and 3) is inversely correlated with line voltage.     

Differences in red cell shape of two inbred chicken lines

Summary Comparison of two inbred chicken lines (F_x > 99.9%) revealed significant differences in shape of the red blood cells (RBC). The length-width index was lower for both sexes in IC-line (1.46) when compared to CB-line chickens (1.88). Phenotypic expression of this character in F₁ hybrids and both backcross groups corresponded to the common manifestations of the metric parameters. The index in F₁ hybrid chickens deviated from intermediate values with the dominant tendency to oval RBC. An analysis of the segregating first backcross generation chickens did not show any association between RBC shape and the genotype in the blood group systems B, C, I, and D and the IgG allotypes. The differences in RBC shape were probably not associated with the survival of RBC in the blood circulation.     

Development of pre-isogenic lines for rice blast-resistance by marker-aided selection from a recombinant inbred population

To increase the available set of near-isogenic lines (NILs) for blast-resistance in rice, we have developed a general method for establishing NILs from populations of fixed recombinants that have been used for gene mapping. We demonstrated the application of this method by the selection of lines carrying genes from the rice cultivar Moroberekan. Moroberekan is a West African japonica cultivar that is considered to have durable resistance to rice blast. Multiple genes from Moroberekan conferring complete and partial resistance to blast have previously been mapped using a recombinant inbred (RI) population derived from a cross between Moroberekan and the highly and broadly susceptible indica cultivar CO39. To analyze individual blast-resistance genes, it is desirable to transfer them individually into a susceptible genetic background. This RI population, and the associated data sets on blast reaction and restriction fragment length polymorphism (RFLP) genotypes, were used for selection of lines likely to carry individual blast-resistance genes and a minimum number of chromosomal segments from Moroberekan. Because skewed segregation in the RI population favored CO39 (indica) alleles, resistant lines carrying 8.7–17.5% of Moroberekan alleles (the proportion expected after two or three backcrosses) could be selected. We chose three RI lines carrying different complete resistance genes to blast and two RI lines carrying partial resistance genes to blast as potential parents for the development of NILs. These lines were subjected to genetic analysis, which allowed clarification of some issues that could not be resolved during the initial gene-mapping study.     

Aedes novalbopictus: establishment and characterization of larval cell lines

Two diploid cell lines were established from larval tissues of the mosquito Aedes novalbopictus. Many morphologically different types of cells were detected in primary cultures. However, only three types of cells became established in the cell lines. They were: epithelial-like cells (80–90%), fibroblast-like cells (5–10%), and giant cells (5–10%). Cell number increased about 15-fold during the first 6 days of culture and the population doubling time during the period of active growth was 18 hr. Seventy to 80% of the cells were diploid (2n = 6) and the rest were tetra-or polyploids. Cells from these two cultures so far (December, (1971)) have been subcultured 36 and 34 times, respectively. The growth pattern and general morphologic features of the cells of A. novalbopictus cultures closely resembled those of A. albopictus cultures.     

Three new continuous cell lines from marine fishes of asia

Summary Three new continuous cell lines were established from two species of marine fishes economically important in Asia. Perch (Lateolabrax japonicus) heart (PH), perch liver (PL), and grouper (Epinephelus amblycephalus) kidney (GK) lines were established in Eagle’s minimum essential medium with 10% fetal bovine serum and have been subcultured over 120 times. The optimum growth temperature was 25°C for the PK and GK lines and 30° C for the PH line. The modal chromosome numbers for each cell line are: PH (49), PK (50), and GK (65). None of the lines was susceptible to the rhabdovirus infectious hematopoietic necrosis virus (IHNV) or channel catfish herpesvirus (CCV); however, all three cell lines were susceptible to a variety of fish birnaviruses, including infectious pancreatic necrosis virus (IPNV), the EVE eel virus, and newly isolated birnaviruses from a variety of fish and shellfish in Taiwan. This research was funded by National Science Foundation grant INT-810447, the University of Main/University of New Hampshire Sea Grant College Program, and the Maine Agriculture Experiment Station publication no. 1165.