Anterior Pair Research Articles

Differential mesodermal expression of two amphioxus MyoD family members ( AmphiMRF1 and AmphiMRF2)

To explore the evolution of myogenic regulatory factors in chordates, we isolated two MyoD family genes ( AmphiMRF1 and AmphiMRF2) from amphioxus. AmphiMRF1 is first expressed at the late gastrula in the paraxial mesoderm. As the first somites form, expression is restricted to their myotomal region. In the early larva, expression is strongest in the most anterior and most posterior somites. AmphiMRF2 transcription begins at mid/late gastrula in the paraxial mesoderm, but never spreads into its most anterior region. Through much of the neurula stage, AmphiMRF2 expression is strong in the myotomal region of all somites except the most anterior pair; by late neurula expression is downregulated except in the most posterior somites forming just rostral to the tail bud. These two MRF genes of amphioxus have partly overlapping patterns of mesodermal expression and evidently duplicated independent of the diversification of the vertebrate MRF family.

Trilobite Tagmosis and Body Patterning from Morphological and Developmental Perspectives

The Trilobita were characterized by a cephalic region in which the biomineralized exoskeleton showed relatively high morphological differentiation among a taxonomically stable set of well defined segments, and an ontogenetically and taxonomically dynamic trunk region in which both exoskeletal segments and ventral appendages were similar in overall form. Ventral appendages were homonomous biramous limbs throughout both the cephalon and trunk, except for the most anterior appendage pair that was antenniform, preoral, and uniramous, and a posteriormost pair of antenniform cerci, known only in one species. In some clades trunk exoskeletal segments were divided into two batches. In some, but not all, of these clades the boundary between batches coincided with the boundary between the thorax and the adult pygidium. The repeated differentiation of the trunk into two batches of segments from the homonomous trunk condition indicates an evolutionary trend in aspects of body patterning regulation that was achieved independently in several trilobite clades. The phylogenetic placement of trilobites and congruence of broad patterns of tagmosis with those seen among extant arthropods suggest that the expression domains of trilobite cephalic Hox genes may have overlapped in a manner similar to that seen among extant arachnates. This, coupled with the fact that trilobites likely possessed ten Hox genes, presents one alternative to a recent model in which Hox gene distribution in trilobites was equated to eight putative divisions of the trilobite body plan.

Neural basis of a simple behavior: abdominal positioning in crayfish.

Crustaceans have been used extensively as models for studying the nervous system. Members of the Order Decapoda, particularly the larger species such as lobsters and crayfish, have large segmented abdomens that are positioned by tonic flexor and extensor muscles. Importantly, the innervation of these tonic muscles is known in some detail. Each abdominal segment in crayfish is innervated bilaterally by three sets of nerves. The anterior pair of nerves in each ganglion controls the swimmeret appendages and sensory supply. The middle pair of nerves innervates the tonic extensor muscles and the regional sensory supply. The superficial branch of the most posterior pair of nerves in each ganglion is exclusively motor and supplies the tonic flexor muscles of that segment. The extension and flexion motor nerves contain six motor neurons, each of which is different in axonal diameter and thus produces impulses of different amplitude. Motor programs controlling each muscle can be characterized by the identifiable motor neurons that are activated. Early work in this field discovered that specific central interneurons control the abdominal positioning motor neurons. These interneurons were first referred to as "command neurons" and later as "command elements." Stimulation of an appropriate command element causes a complex, widespread output involving dozens of motor neurons. The output can be patterned even though the stimulus to the command element is of constant interval. The command elements are identifiable cells. When a stimulus is repeated in a command element, from either the same individual or from different individuals, the output is substantially the same. This outcome depends upon several factors. First, the command elements are not only identifiable, but they make many synapses with other neurons, and the synapses are substantially invariant. There are separate flexion-producing and extension-producing command elements. Abdominal flexion-producing command elements excite other flexion elements and inhibit extensor command elements. The extension producing elements do the opposite. These interactions insure that interneurons of a particular class (flexion- or extension-producing) synaptically recruit perhaps twenty others of similar output, and that command elements promoting the opposing movements are inhibited. This strong reciprocity and the recruitment of similar command elements give a powerful motor program that appears to mimic behavior.

The Chemoreceptive Lattice Organs in Cypris Larvae Develop from Naupliar Setae (Thecostraca: Cirripedia, Ascothoracida and Facetotecta)

Lattice organs are peculiar chemoreceptors found only in the Crustacea Thecostraca (Facetotecta, Ascothoracida, Cirripedia). In these taxa, five pairs occur in the head shield (carapace) of the terminal larval instar (y-cyprid, ascothoracid larva, cyprid), which is the settlement stage. Lattice organs represent an autapomorphy for the Thecostraca but their evolutionary origin and possible homologues in other Crustacea remain obscure. We have used scanning electron microscopy to describe the setation pattern of the head shield in late nauplii of one species of Ascothoracida, one species of Facetotecta and several species of the Cirripedia Thoracica, Acrothoracica, and Rhizocephala. The naupliar head shield always carries two pairs setae situated anteriorly near the midline. Each of these setae carry a single pore, and positional, structural and ontogenetic evidence show that these setae are homologous in all the examined species and that they represent precursors of the two anterior pairs of lattice organs of the succeeding larval stage, viz., the ascothoracid larva (Ascothoracida), y-cyprid (Facetotecta), and cyprid (Cirripedia). This leads us to infer that lattice organs are among the most highly modified sensilla in all Crustacea and they have in most cases lost all external resemblance to a seta. The nauplii of the Rhizocephala carry an additional three pairs of setae situated more posteriorly on the head shield and they could be precursors of the three posterior pairs of lattice organs. All other species examined lack these posterior setae, except the Facetotecta which have one posteriorly situated pair.

The Development of Three Identified Motor Neurons in the Larva of an Ascidian, Halocynthia roretzi

The generation of distinct classes of motor neurons underlies the development of complex motile behavior in all animals and is well characterized in chordates. Recent molecular studies indicate that the ascidian larval central nervous system (CNS) exhibits anteroposterior regionalization similar to that seen in the vertebrate CNS. To extend the understanding about the diversity of motor neurons in the ascidian larva, we have identified the number, position, and projection of individual motor neurons in Halocynthia roretzi, using a green fluorescent protein under the control of a neuron-specific promoter. Three pairs of motor neurons, each with a distinct shape and innervation pattern, were identified along the anteroposterior axis of the neural tube: the anterior and posterior pairs extend their axons toward dorsal muscle cells, whereas the middle pair project their axons toward ventral muscle. Overexpression of a dominant-negative form of a potassium channel in these cells resulted in paralysis on the injected side, thus these cells must constitute the major population of motor neurons responsible for swimming behavior. Lim class homeobox genes have been known as candidate genes that determine subtypes of motor neurons. Therefore, the expression pattern of Hrlim, which is a Lim class homeobox gene, was examined in the motor neuron precursors. All three motor neurons expressed Hrlim at the tailbud stage, although each down-regulated Hrlim at a different time. Misexpression of Hrlim in the epidermal lineage led to ectopic expression of TuNa2, a putative voltage-gated channel gene normally expressed predominantly in the three pairs of motor neurons. Hrlim may control membrane excitability of motor neurons by regulating ion channel gene expression.

Evidence for the association of FMRFamide-related peptides with the spermatheca of Locusta migratoria

The association of FMRFamide-related peptides (FaRPs) with the spermatheca of Locusta migratoria was demonstrated using radioimmunoassay and immunohistochemical techniques. The physiological effects of various FaRPs on the neurally evoked contractions of the spermatheca were also examined. FMRFamide-like immunoreactivity (FLI) was demonstrated in processes and cell bodies situated in the VIIIth (terminal) abdominal ganglion. These included an anterior, central and posterior pair of ventral cell bodies positioned near the midline of the ganglion, in addition to two bilaterally paired dorsal cell bodies in the posterior region of the VIIIth abdominal ganglion. Two axons displaying FLI proceed down the ventral ovipositor nerve (VON) and into the receptaculum seminis nerve which innervates the anterior regions of the spermatheca. FLI was also noted in processes on the spermathecal muscle with the highest density occurring on the spermathecal sac and coil duct. FaRPs applied to the spermathecal muscle included GQERNFLRFamide, NFIRFamide, ADDRNFIRFamide, YGGFMRFamide, FMRFamide, ADVGHVFLRFamide and SchistoFLRFamide (PDVDHVFLRFamide). Dose-dependent physiological effects were only noted for FMRFamide, ADVGHVFLRFamide and SchistoFLRFamide. FMRFamide led to a dose-dependent increase in the amplitude of neurally evoked contractions with a threshold of approximately 5 × 10 −7 M. SchistoFLRFamide, and ADVGHVFLRFamide, had an inhibitory effect, decreasing the amplitude of neurally evoked spermathecal contractions.

Lattice organs in y‐cyprids of the Facetotecta and their significance in the phylogeny of the Crustacea Thecostraca

AbstractScanning and transmission electron microscopy (SEM and TEM) were used to study lattice organs in facetotectan y‐cyprids from the White Sea and from Norwegian and Bahamian waters. The larvae represent at least four and possibly five different species of Facetotecta. Y‐cyprids have five pairs of lattice organs in the head shield (carapace) organized into two anterior pairs and three posterior pairs. Both groups of lattice organs are arranged around a large central pore. The facetotectan lattice organs are elongate areas with a longitudinal keel, just as in the Ascothoracida and some Cirripedia Acrothoracica. The terminal pore of the organs is situated posteriorly in all five pairs. TEM confirms that the organs have the same general morphology as in the Cirripedia and Ascothoracida, namely, a cuticular chamber into which project ciliary segments from the chemosensory cells. Unlike Cirripedia the cuticular roof of the chamber lacks any pores. We conclude that five pairs of lattice organs represent an autapomorphy for the Thecostraca, which supports the monophyly of this taxon. In the ground pattern the terminal pore is posterior in all five pairs. The anterior position of the pore in lattice organ pair 2 is apomorphic for the Cirripedia, while within this taxon an anterior position also in pair 1 is apomorphic for a monophylum comprising the Thoracica and the Rhizocephala. Minute pores in the roof of the organs is another apomorphy of the Cirripedia, but its elaboration into pores visible with SEM may have been subject to some homoplasy. Since lattice organs are omnipresent in the settling instar of the Thecostraca they probably serve a critical role for the function of these cypris or cypris‐like larvae.

Transepithelial potential differences and Na+ flux in isolated perfused gills of the crabChasmagnathus granulatus(Grapsidae) acclimated to hyper- and hypo-salinity

SUMMARYWe studied the transepithelial potential difference (TEPD) and 22Na flux across isolated perfused gills (anterior pair 5 and posterior pairs 6–8) of the crab Chasmagnathus granulatus acclimated to either hypo- or hyper-osmotic conditions.The gills of crabs acclimated to low salinity, perfused and bathed with 10 ‰ saline solutions, produced the following TEPDs (hemolymph side with respect to bath side): 0.4±0.7, –10.2±1.6, –10.8±1.3 and –6.7±1.3 mV for gills 5, 6, 7 and 8, respectively. Gills 6, 7 and 8 did not differ significantly. Reducing the saline concentration of bath and perfusate from 30 ‰ to 20 ‰ or 10 ‰ increased significantly the TEPDs of these gills. TEPDs of gill 6 (representative of posterior gills) were reduced by 69±5 % and 60±5 % after perfusion with ouabain or BaCl2 (5 mmol l–1 each), respectively. The same gill showed a net ouabain-sensitive Na+ influx of 1150±290 μequiv g–1 h–1.Gill 6 of crabs acclimated to high salinity produced TEPDs of –1.5±0.1 and –1.3±0.09 mV after perfusion with 30 ‰ or 40 ‰ salines, respectively. Perfusion with ouabain or BaCl2 reduced TEPDs by 76±7 % and 86±4 %, respectively. A net ouabain-sensitive Na+ efflux of 2282±337 μequiv g–1 h–1 was recorded in gill 6 perfused with 38 ‰ saline.

The dorsal fin engine of the seahorse (Hippocampus sp.).

The muscles, fin ray joints, and supporting structures underlying the dorsal fin are described for two seahorse species: Hippocampus zosterae and Hippocampus erectus. A fan-shaped array of cartilaginous bones, the pterigiophores, form the internal supporting structure of the dorsal fin. Each pterigiophore is composed of a proximal radial that extends from a vertebra to the dorsal side of the animal, where it fuses to a middle radial. The middle radials fuse with each other to form a dorsal ridge upon which sit the spheroidal distal radials. Each distal radial articulates with a fin ray on its dorsal side and is attached to the dorsal ridge on its ventral side by a material that has been histologically identified as elastic cartilage. Together these connections form a two-axis joint that permits elevation, depression, and inclination of the ray. Each fin ray is actuated by two bilateral pairs of muscles, an anterior pair of inclinators, and a posterior pair of depressors. The anteriormost fin ray is actuated by three bilateral pair of muscles, the inclinators, the depressors, and a pair of elevator muscles that are positioned anterior to the inclinators. Preliminary examinations of the ray joints of the pectoral and anal fins of adult H. zostera and the pectoral fins of newborn H. erectus revealed structures similar to that seen in the dorsal fins. To further explore the structure and function of the dorsal fin gross dissections and simple functional tests were performed on H. erectus and H. barbouri and behavioral observations were made of all three species plus Hippocampus kuda.

Possible Roles of Protein Kinase A in Cell Motility and Excystation of the Early Diverging Eukaryote Giardia lamblia

Since little is known of how the primitive protozoan parasite, Giardia lamblia, senses and responds to its changing environment, we characterized a giardial protein kinase A (gPKA) catalytic subunit with unusual subcellular localization. Sequence analysis of the 1080-base pair open reading frame shows 48% amino acid identity with the cyclic AMP-dependent kinase from Euglena gracilis. Northern analysis indicated a 1.28- kilobase pair transcript at relatively constant concentrations during growth and encystation. gPKA is autophosphorylated, although amino acid residues corresponding to Thr-197 and Ser-338 of human protein kinase A (PKA) that are important for autophosphorylation are absent. Kinetic analysis of the recombinant PKA showed that ATP and magnesium are preferred over GTP and manganese. Kinase activity of the native PKA has also been detected in crude extracts using kemptide as a substrate. A myristoylated PKA inhibitor, amide 14-22, inhibited excystation with an IC(50) of 3 microm, suggesting an important role of gPKA during differentiation from the dormant cyst form into the active trophozoite. gPKA localizes independently of cell density to the eight flagellar basal bodies between the two nuclei together with centrin, a basal body/centrosome-specific protein. However, localization of gPKA to marginal plates along the intracellular portions of the anterior and caudal pairs of flagella was evident only at low cell density and higher endogenous cAMP concentrations or after refeeding with fresh medium. These data suggest an important role of PKA in trophozoite motility during vegetative growth and the cellular activation of excystation.

SUCKLING BEHAVIOR OF PINE VOLES (MICROTUS PINETORUM)

Abstract The suckling behavior of some mammals is characterized by preferences for anterior or posterior nipples and consistent return by individual young to particular nipples or nipple pairs. Some murid species also display tenacious nipple attachment. Pine voles (= woodland voles, Microtus pinetorum) have tenaciously clinging young and 2 pairs of abdominal mammae. We examined whether young pine voles preferred particular nipple pairs and whether young on the 2 pairs were differentially groomed or dislodged by their mothers. We also examined whether young showed fidelity to suckling location. Young pine voles preferred the hindmost nipples and were dislodged less frequently from those nipples than from the more anterior pair. We found no evidence that mothers differentially groomed young on the 2 pairs. Fidelity to nipple and nipple pair was greater in small than in large litters, which may reflect less competition for hind nipples in small litters and the need to consistently stimulate a nipple to ensu...

Myth: idiopathic wounds are often due to brown recluse or other spider bites throughout the United States.

Spiders, especially brown recluses (Loxosceles reclusa) (figure 1), are frequently incriminated as causative agents in idiopathic wounds, even though diagnoses are usually made solely on the basis of dermatologic symptoms. Rarely is a spider seen inflicting the wound or captured while biting, and it is difficult to either prove or disprove spider participation in the event. The biologic distribution of the brown recluse and related recluse species indicates that many diagnoses made on cases occurring in the western United States are incorrect.1 Figure 1 Dorsal view of the brown recluse spider, Loxosceles reclusa. The most consistent diagnostic feature for identification is the pattern of 6 eyes, grouped in pairs, with 1 anterior pair and 2 lateral pairs (most spiders have 8 eyes in 2 rows of 4). Although ... SOURCES OF INFORMATION Because of my research interests in spider identification and medically important spiders, I have been avidly studying recluse spiders and their occurrence in the western United States. My experience is greatest in California, but the information presented here is pertinent to northern and western states as well. In almost a decade of research, I have found verification of fewer than 10 brown recluse specimens in California in more than 40 years of records. Sources include medical and arachnologic literature and correspondence with scores of arachnologists, county entomologists, vector control personnel, and the California Department of Food and Agriculture, which handles all exotic pest identifications in the state. Most verified specimens were individuals intercepted from facilities receiving goods from out of state. There are no populations of brown recluses in California.

Calcium homeostasis in larval and adult Drosophila melanogaster.

Calcium homeostasis in Drosophila melanogaster was examined in response to the challenges imposed by growth, reproduction and variations in dietary calcium content. Turnover time for calcium, calculated as the time for (45)Ca(2+)to accumulate to half the steady state value of 3.46 nmol/fly, was 3.3 days. Although larvae weighed 2x as much as adults, they contained 3-4x as much calcium. Anterior Malpighian tubules (Mts) contain much more calcium than posterior Mts, accounting for 25-30% of the calcium content of the whole fly. In response to a 6.2-fold increase in dietary calcium level, calcium content of whole flies increased only 10%. Hemolymph calcium concentration ( approximately 0.5 mM) was similar in males and females and in animals raised on diets differing in calcium content. Fluid secretion rate, secreted fluid calcium concentration, and transepithelial calcium flux in tubules isolated from flies raised on high and low calcium diets did not differ significantly. Malpighian tubules secrete calcium at rates sufficient to eliminate whole body calcium content in 0.5 and 3 days for tubules secreting fluid at basal and maximal rates, respectively. It is suggested that flies absorb high quantities of calcium from the diet and maintain homeostasis through the combined effects of elimination of calcium in fluid secreted by the Malpighian tubules and the sequestration of calcium in granules, especially within the distal segment of the anterior pair of Malpighian tubules.

Investigating the function of follicular subpopulations during Drosophila oogenesis through hormone-dependent enhancer-targeted cell ablation.

Although it is known that the establishment of polarity during Drosophila oogenesis is initiated by signalling from the oocyte to the overlying follicle cells, much less is understood about the role of specific follicular subpopulations. One powerful approach for addressing this question, toxigenic cell ablation of specific subpopulations, has not previously been applicable to studying follicular subpopulations because many of the genes and Gal4 enhancer trap insertions that are expressed in the ovary are also expressed at earlier times in development. To overcome this problem, we have utilized a fusion protein between Gal4 and the human estrogen receptor to achieve hormone-dependent, tissue-specific gene expression of UAS-linked transgenes in flies. We used this system to study the role of the polar subpopulations of follicle cells during oogenesis by expressing within them a modified form of diphtheria toxin that causes cell death. Our results confirmed previous functions ascribed to these cells, and also demonstrated a previously undescribed role for the border cells in facilitating the migration of the anterior Fasciclin III-expressing polar pair cells to the edge of the oocyte.

Canal and otolith afferent activity underlying eye velocity responses to pitching while rotating.

Pitching the head while rotating (PWR) combines periodic activation of the semicircular canals and the otoliths to generate pitch and roll eye deviations and continuous horizontal nystagmus. Monkeys were tested after individual pairs of semicircular canals were plugged and single units were recorded in the vestibular nerve while the animals were sinusoidally pitched 20-40 deg about a spatial horizontal axis with 5- and 16-s periods and simultaneously rotated about a spatial vertical axis at 30-120 deg/s. As previously shown, the steady-state horizontal response disappeared after plugging the vertical semicircular canals, but was maintained when the lateral canals were plugged. When the left anterior and right posterior canal (LARP) pair was left intact, the steady-state response depended on the axis about which the pitching took place. When the axis was normal to the LARP plane, there was no steady-state response. When the pitching axis was perpendicular to the LARP normal, the response was maximal. Firing rates of otolith units were approximately in phase with pitch position, and the addition of rotation about a vertical axis did not change the response. Lateral canal units did not have a steady-state modulation during pitch or constant velocity rotation. During PWR, they oscillated at twice the pitch frequency. This corresponded to the frequency at which the canal was maximally activated as it aligned with the plane of rotation. The amplitude of modulation increased proportionally to rotational velocity, but the phase remained the same. These characteristics were unchanged during roll while rotating (RWR), which induces little continuous nystagmus. Anterior and posterior canal units were maximally excited near pitch-velocity maxima and minima, respectively, during pure pitching. During PWR, however, the phases of both components simultaneously shifted toward each other and toward being in phase with otolith units. The peak excitation tended toward a forward-pitch position when the rotation was to the ipsilateral side, and toward a backward pitch position when the rotation was to the contralateral side. With 120-deg/s rotation during a 16-s pitch period, the phase difference between anterior and posterior canal units was as small as 17 deg. These data support the postulate that the correlation between vertical canal and otolith units is the critical factor in generating continuous unidirectional horizontal nystagmus during PWR.

Leg autotomy in a spider has minimal costs in competitive ability and development

A number of species have the ability to autotomize limbs voluntarily, but animals that have lost limbs often face substantial costs. We examined the frequency of leg loss and its effects on competitive ability and development in the spiderHolocnemus pluchei(Araneae: Pholcidae), a family of spiders known for its readiness to autotomize legs. Leg loss was common in field populations, with 7.5% of all surveyed spiders missing at least one leg, most commonly one of the anterior pair. More spiders were missing multiple legs than expected by chance, suggesting that leg loss events are not independent. Large adult spiders were missing legs more frequently than were small spiders. The competitive ability of injured males was tested in three contexts. In the field, no effect of leg loss was found on the ability of spiders to remain in webs into which they were introduced. In the laboratory, no effect of leg loss was found on the ability to fight with a single opponent over a prey, except that injured spiders were more likely to lose high-intensity fights. There was no difference between intact and injured males in their ability to compete with three females for limited prey. Leg loss significantly affected development time. The moult interval during the instar in which the injury occurred increased by approximately 15%. However, the growth rate for injured spiders was slightly but not significantly faster in the instar following leg loss, and total development time of the two instars together did not differ significantly between treatments. No spider showed any signs of regeneration. We conclude that, although there were some statistically significant differences between intact and injured males, these are unlikely to have major impacts on fitness, in contrast to findings in other species.

Projections and firing properties of down eye-movement neurons in the interstitial nucleus of Cajal in the cat.

To clarify the role of the interstitial nucleus of Cajal (INC) in the control of vertical eye movements, projections of burst-tonic and tonic neurons in and around the INC were studied. This paper describes neurons with downward ON directions. We examined, by antidromic activation, whether these down INC (d-INC) neurons contribute to two pathways: a commissural pathway to the contralateral (c-) INC and a descending pathway to the ipsilateral vestibular nucleus (i-VN). Stimulation of the two pathways showed that as many as 74% of neurons were activated antidromically from one of the pathways. Of 113 d-INC neurons tested, 44 were activated from the commissural pathway and 40 from the descending pathway. No neurons were activated from both pathways. We concluded that commissural and descending pathways from the INC originate from two separate groups of neurons. Tracking of antidromic microstimulation in the two nuclei revealed multiple low-threshold sites and varied latencies; this was interpreted as a sign of existence of axonal arborization. Neurons with commissural projections tended to be located more dorsally than those with descending projections. Neurons with descending projections had significantly greater eye-position sensitivity and smaller saccadic sensitivity than neurons with commissural projections. The two groups of INC neurons increased their firing rate in nose-up head rotations and responded best to the rotation in the plane of contralateral posterior/ipsilateral anterior canal pair. Neurons with commissural projections showed a larger phase lag of response to sinusoidal rotation (54.6 +/- 7.6 degrees ) than neurons with descending projections (45.0 +/- 5.5 degrees ). Most neurons with descending projections received disynaptic excitation from the contralateral vestibular nerve. Neurons with commissural projections rarely received such disynaptic input. We suggest that downward-position-vestibular (DPV) neurons in the VN and VN-projecting d-INC neurons form a loop, together with possible commissural loops linking the bilateral VNs and the bilateral INCs. By comparing the quantitative measures of d-INC neurons with those of DPV neurons, we further suggest that integration of head velocity signals proceeds from DPV neurons to d-INC neurons with descending projections and then to d-INC neurons with commissural projections, whereas saccadic velocity signals are processed in the reverse order.

Giardia agilis: Ultrastructure of the Trophozoites in the Frog Intestine

Intestine samples of Bufo sp. tadpoles with parasitism confirmed for Giardia agilis were studied by transmission electron microscopy. The G. agilis trophozoites were long and thin. The plasma membrane was sometimes undulated and the cytoplasm, adjacent to the dorsal and ventral regions, showed numerous vacuoles. The two nuclei presented prominent nucleoli. The cytoplasm was electron-dense with free ribosomes, glycogen and rough endoplasmic reticulum-like structures. Polyhedral inclusions were observed in the cytoplasm and outside the protozoan; some of these inclusions exhibited membrane disruption. The flagella ultrastructure is typical, with the caudal pair accompanied by the funis. Next to the anterior pair, osmiophilic material was noticed. The ventro-lateral flange was short and thick, supported by the marginal plates that penetrated into its distal extremity; only its distal portion had adjacent osmiophilic filament. The G. agilis trophozoites showed the general subcellular feature of the genus. However, the ventro-lateral flange ultrastructure was an intermediate type between G. muris and G. duodenalis.

Activity of the Extrinsic and Intrinsic Muscle Networks of the Malpighian Tubules and the Ureter in the Female Stable Fly, Stomoxys calcitrans (L.)

The four Malpighian tubules observed in the female stable fly, Stomoxys calcitrans (L.), are divided into an anterior dorsal pair and a posterior ventral pair. Intrinsic muscles were found only in the short proximal section (ureter) of each pair of tubules that emptied into the midgut. Isolated preparations of the ureter demonstrated four types of organized rhythmic activity: compression, peristalsis, reverse peristalsis, and segmentation. Compression caused a momentary (0.4 to 0.6 sec) shortening of the longitudinal muscle fibers in a localized region of the ureter. Peristalsis was the dominant type of activity. Each wave progressed along the duct with a duration that ranged from 0.9 to 1.7 sec. Segmentation consisted of a localized annular constriction of the ureter without progression that lasted for a duration of 0.3 to 0.6 sec. The large distal ends of the dorsal pair of tubules showed spontaneous movement in situ. Close examination revealed a network of extrinsic muscle cells attached to the distal tubular ends. The motile properties of the extrinsic muscles were slower and not as complex as those of the ureter. The sequence from the resting state to contraction and return was much longer with durations that ranged from 10.8 to 27.1 sec. Also, the time for peak tension was 5 to 25.9 sec in duration. The extrinsic muscles did not show the organized and rhythmic activity observed in the ureter.

Carcinoma of the nasopharynx treated by radiotherapy alone: determinants of distant metastasis and survival

Purpose: This retrospective study was conducted to identify the prognostic factors for distant metastasis and survival in a population of 378 patients with nasopharyngeal carcinomas treated by radiation therapy alone. Materials and methods: All patients were treated at the University of Texas M.D. Anderson Cancer Center between 1954 and 1992, following a consistent dose and volume prescription policy. There were 286 males and 92 females. The median age was 52 years (range: 16–86 years). The majority of the patients were white Caucasians (282 patients,75%). Tumors were classified as squamous cell carcinomas (193; 51%), lymphoepitheliomas (154; 41%), or unclassified carcinomas (31, 8%). Three fourths of the patients presented with AJCC Stage IV disease (T4, N0–3, 118 patients; T1–3, N2–3 164 patients). The treatment techniques included opposed lateral fields with or without an anteroposterior or an anterior oblique pairs for dose supplementation to the primary site. Average total doses per T-stage ranged between 60.2 and 72.0 Gy. Median follow-up time was 10 years (range 0.3 to 28.6 years). Results: A total of 103 patients (27%) developed distant metastases at a median time of 8 months (range: 1–90 months). Actuarial rates for distant metastasis were 30%, 32%, 32% at 5, 10, and 20 years, respectively. Actuarial rates for disease specific survival at the same time points were 53%, 45%, and 39% with 184 patients (49%) dying of their nasopharyngeal cancer. Advanced T-stage, N-stage, and non-lymphoepithelioma histology were independent adverse prognostic factors for disease specific survival. Advanced N-stage and low neck disease were independent adverse prognostic factors for distant metastasis with a very high rate of distant metastases for those patients who presented with both adverse factors (relative risk 7.86). On average, patients with distant metastasis lived 5 months after they were diagnosed with metastatic disease (range: 0–172 months), although four patients (4%) survived more than 5 years after diagnosis. Conclusions: This study demonstrates good long term survival rates after definitive radiotherapy for patients with nasopharyngeal carcinomas. Patients with advanced and lower neck disease have the highest risk of developing distant failures. Such patients can be considered the reference risk group to test the value of adjunctive chemotherapy.