Meiotic drive by the Y-linked D gene in Aedes aegypti (L.) (Diptera: Culicidae) is associated with disruption of spermiogenesis, leading to premature senescence of spermatozoa.

Y chromosome meiotic drive in the mosquito Aedes aegypti, due to the gene D (Distorter) in coupling with M (male determination) [the MD haplotype], is associated with spermiogenic disruption, leading to senescence, at a rate proportionate to male excess. Spermiogenesis was compared between 'Enhanced Mutant' males with a strongly female-depleted sex ratio (8.9% females), 'Mutant' males showing a lesser degree of distortion (38.3% females), and two controls with normal sex ratios (51.2% and 49.2% females). Sections of testes dissected from mature pupae and adults aged 0, 4, 8, 12 and 16 days were examined by transmission electron microscopy. A difference between Mutant and control spermiogenesis was apparent as early as the pupal stage when some Mutant spermatids showed extra tail elements (axonemes and/or mitochondrial derivatives). The same was true of Enhanced Mutant males but to a more extreme degree. Sperm senescence was evident in Enhanced Mutant testes from day 0 of adult life but in Mutant testes not until day 4. Progressive disorganisation was associated with many loose organelles, and disturbance of the anterior-posterior axis of gamete differentiation within the testis. Degenerative changes of a similar kind in the controls did not become apparent until day 8. These findings are discussed with respect to other characteristics of this meiotic drive system, in terms of a theory of inhibition of reduction division in spermatogenesis associated with fragmentation of the X chromosome, leading to the formation of a restitution nucleus as early as metaphase 1.

Morphological evidence for a morphogenetic field in gastropod mollusc eggs.

Eggs of the marine gastropod Crepidula fornicata examined by confocal imaging of FITC-lectin binding to the surface, and cryoscopic-SEM both reveal a surface architecture of linear structures organized around the animal-vegetal axis, which is spatially related to the anterior-posterior (a-p) axis of the subsequent embryo. A series of structures is also orientated with reference to specific micromere quartets formed during spiral cleavage. Thus, the surface architecture may provide a visible marker for a morphogenetic field which generates the a-p axis and organizes the cleavage pattern. Moreover, this architecture is co-extensive with that found on the vegetal, polar lobe-bearing region of eggs, as described by others, and which varies between gastropod taxa with varied types of body form. Confocal imaging reveals a distinct localization of F-actin to the architecture of the lobe region. However, the integrity of this F-actin is not responsible for the maintenance of the surface architecture. The significance of these findings to our understanding of the generation of diversity within the Gastropoda and general ontogenic mechanisms is discussed.

Selected lines of Aedes aegypti with persistently distorted sex ratios.

A breeding scheme to isolate X chromosomes sensitive to drive by the T8 (Trinidad) Y chromosome of Aedes aegypti (the MD haplotype) is reported. Crosses with an Australian strain Th.I (Thursday Island) revealed not only sensitive and resistant X chromosomes but also some with the capacity to drive against the T8 Y chromosome. Four strains were created in which sex ratio was male-distorted (28-36 per cent Female) for 10 generations, with no regression towards sexual parity. The proportion of females varied significantly between generations in each of the four strains. Further selection produced strains with normal sex ratios, capable of generating fewer than 15 per cent Female on outcrossing to T8 males.

Seasonal changes in whole-cell metal levels in protozoa of activated sludge.

In this study the authors quantify, and correlate, whole-cell elemental levels of vorticellids in activated sludge, over a 1-year period, and Aspidisca cicada and Opercularia coarctata on one sampling date. The aim was to determine the extent, and seasonal variation, of metal uptake and/or accumulation, to give new information regarding the fate and dynamics of the cellular elements, and to determine how they are related to each other over time and between species, in an environment exposed to high levels of heavy metals. Samples of activated sludge were collected monthly, and whole-cell elemental levels of vorticellids determined by scanning electron microscopy electron probe X-ray microanalysis (XRMA). Positive intersample correlations were found between phosphorus and three elements, sulfur, potassium, and iron. Potassium and calcium levels correlated positively with sulfur levels. Iron levels were positively correlated with potassium levels. Levels in A. cicada sampled from activated sludge were also compared with the same species grown in vitro, previously isolated from the same sewage treatment works. The level of cellular elements varies considerably with time, and between species from the same environment. Vorticellids could tolerate varying levels of aluminum, copper, iron, and zinc. Decreased iron levels were associated with decreased phosphorus and potassium levels.

Intracellular calcium transients in suctorian protozoa (Trichophrya spp.): correlation with spontaneous tentacle contractions.

The luminescent photoprotein aequorin was used to measure intracellular free Ca2+ in three species of suctorian protozoon, Trichophrya riederi, Trichophrya collini and Trichophrya rotunda. Resting [Ca2+]i ranged from about 75-110 nM, and was unaffected by a change in temperature of the perfusate. Spontaneous Ca2+ transients were observed in all three species, with peak amplitudes ranging from 100-600 nM. In T. riederi and T. rotunda, three categories of transient (small, intermediate, large) were recorded; T. collini displayed only small transients. In both T. riederi and T. collini, raising the temperature from 5 degrees to 26 degrees C led to an increase in the frequency of transients. Furthermore, in T. riederi, large transients occurred only at the higher temperature. The frequency of spontaneous contractions of the tentacles of T. riederi was also temperature-dependent. Increasing the temperature over the range 5-26 degrees C led to a concomitant increase in contraction frequency and a decrease in mean tentacle length. The possible mechanisms of spontaneous Ca2+ transient generation and their role in the control of contraction are discussed.

Optimal therapy for small noncleaved cell lymphoma.

Small noncleaved cell lymphoma is now well recognized as a specific subtype of non-Hodgkin's lymphoma with distinctive clinicopathologic characteristics. Initial treatment results in children with endemic Burkitt's lymphoma also indicated a unique sensitivity to cytotoxic chemotherapy; to this day, Burkitt's lymphoma remains one of the few human tumors potentially curable with single-agent chemotherapy. However, the development of effective therapy has proved more difficult in nonendemic SNCL, where presentation with advanced stage and large tumor bulk occurs in most patients. The combination chemotherapy regimens currently considered standard for treatment of large-cell lymphoma have usually produced only transient responses in patients (both children and adult) with SNCL. Recently, several regimens of increased dose intensity have yielded encouraging results both in children and adults. High complete response rates and long-term disease-free survival rates in the 60% range have been reported from several institutions using such regimens. At present, we feel that adults without severe coexisting problems should be treated with high dose-intensity regimens, such as those developed at MD Anderson and Vanderbilt. Routine treatment of these patients with standard lymphoma regimens should be avoided, since the cure rate with this approach has been low. Curative therapy for these patients can be of brief duration, and maintenance therapy is not necessary. Although guidelines are unclear, it seems reasonable at present to include meningeal prophylaxis in the treatment of Stage III and IV patients. Since dose intensity has emerged as an important factor in the curative therapy of SNCL, further exploration of this concept in future clinical trials is critical. The role of growth factors is undefined; if the frequently espoused possibility that growth factors can increase curability by allowing intensification of therapy is to be realized in any human tumor, SNCL leads the list of candidates. The role of early high-dose therapy with bone marrow transplantation is also largely unexplored in SNCL. At the other end of the spectrum, the possibility of administering lesser therapy while maintaining a high cure rate in patients with clinical Stage I SNCL needs further investigation. It is likely that continued clinical investigation will continue to improve therapeutic results in this uncommon but highly distinctive lymphoma.

Hydrogenosomes in the rumen entodiniomorphid ciliate Polyplastron multivesiculatum.

The rumen entodiniomorphid ciliate protozoon Polyplastron multivesiculatum was shown, by biochemical and electron microscopic techniques, to possess hydrogenosomes. After differential centrifugation of whole cell homogenates the hydrogenosomal marker enzymes pyruvate:ferredoxin oxidoreductase and hydrogenase were recovered predominantly (61% and 70% of activity respectively) in the large granular fractions that were sedimented by centrifugation for 10(4) g-min (fraction P1) and 10(5) g-min (fraction P2). These subcellular fractions contained membrane-bound organelles that were approximately 0.4-0.6 microns in diameter and which had a mean equilibrium density of 1.22-1.24 g ml-1 after isopycnic centrifugation in sucrose gradients. Malate dehydrogenase (decarboxylating) activity, however, was predominantly non-sedimentable after centrifugation for 6 x 10(6) g-min. Numerous hydrogenosome-like organelles were present in the ectoplasm and endoplasm of the cell. Hydrogenase activity was demonstrated and localized in the protozoan cell using a novel staining procedure with distyryl nitroblue tetrazolium chloride (DSNBT).

Ultrastructure of the rumen ciliate Dasytricha ruminantium.

The ciliate Dasytricha ruminantium possesses cilia over the entire surface. Both the oral and somatic kineties are composed of monokinetids comprising a kinetosome, a tangential ribbon of 5 transverse microtubules, a weakly convergent bundle of 5 postciliary microtubules, a short kinetodesmal fibre, and a single microtubule homologous to T(2) of other litostomate ciliates [15]. The reversed orientation of the kineties within and around the vestibulum may be a consequence of the evolutionary migration of the vestibulum from the anterior to the posterior of the cell. The vestibulum leads to the cytostome and a cytopharynx of the rhabdos type [3]. Microtubules lining the exit canals of the posterior contractile vacuole and cytoproct are believed to originate from somatic kinetids. The ecto-endoplasmic boundary layer (eeb) is composed of two microfibrillar layers. A large extension of the eeb connects the vestibulum to the cell cortex but does not form a karyophore as seen in the closely related genus Isotricha [12].

Tentacle contraction in Heliophrya erhardi (Suctoria): the role of inositol phospholipid metabolites and cyclic nucleotides in stimulus-response coupling.

Extracellular stimulation of Heliophrya erhardi with 15 V, 100 ms induces tentacle contraction. The effects of a number of pharmacological agents on response latency (time from stimulus to start of contraction) and contraction time (time for tentacles to reach 20% of their original length) were recorded. Contraction was enhanced in the presence of phorbol ester TPA and R59022, both of which increase activation of diacylglycerol-dependent protein kinase C. Lithium ions, which are known to inhibit dephosphorylation of inositol 1-phosphate, also affected the response, causing a decrease in latency and increase in contraction time. These results strongly implicate products of inositol phospholipid metabolism in stimulus-contraction coupling. The latency of contraction was reduced in the presence of IBMX, a nonspecific inhibitor of cyclic nucleotide phosphodiesterases. Application of membrane-permeable cyclic nucleotide analogs led to an increase in latency and contraction time for cAMP analog, but a decrease in both parameters for cGMP. This suggests that the two cyclic nucleotides have opposing roles in modulating the response. The possible roles and interactions of membrane phospholipid metabolites and cyclic nucleotides in the control of tentacle contraction of H. erhardi are discussed.

Spermatozoon ultrastructure of Platichthys flesus.

The ultrastructure of the spermatozoon of Platichthys flesus (flounder) has been determined. The cell is of the primitive type and lacks an acrosome. The bullet-shaped, homogeneously electron-dense nucleus has a deep, caudal articular fossa containing proximal and distal centrioles. Both centrioles are embedded in pericentriolar material which is intimately associated with the nuclear envelope. Around the proximal centriole the material is organised into a dense ring which bears nine fibres associated with the triplets, and a large fibrous body, all connected to the nuclear envelope. Connections exist also between the pericentriolar material of the proximal and distal centrioles. The material around the distal centriole is organised into a complex collar. The anterior surfaces of eight spherical mitochondria lie within shallow depressions of the nuclear caudal surface. They form a ring surrounding the posterior opening of the articular fossa and the proximal region of the 9 + 2 flagellar axoneme.

Spermiogenesis in Platichthys flesus.

Early spermatids of Platichthys flesus (flounder) have a central spherical nucleus and cytoplasm with numerous dispersed mitochondria and a pair of peripheral centrioles. One of the centrioles acts as a kinetosome for axoneme formation. After the start of chromatin condensation the centrioles and developing axoneme migrate to adopt a tangential orientation adjacent to a flattened, lateral margin of the nucleus. During migration, pericentriolar material becomes reorganised. The proximal centriole becomes surrounded by nine fibres, and the distal centriole by a complex collar. As the nucleus condenses it undergoes a rotation which shifts it to the anterior of the centrioles and deepens the articular fossa which houses, and is connected to, the two centrioles and their associated structures. This rotation also results in the mitochondria becoming relocated to form a ring around the proximal region of the axoneme to give the cell its mature ultrastructure.

The ultrastructure of the renal corpuscle of a lizard.

The few and small renal corpuscles of the lizard Podarcis (= Lacerta) taurica are composed of a tuft of three to four capillaries (glomerulus), Bowman's capsule and mesangium. The thin interdigitated capillary endothelial cells are, in most regions, in contact with the mesangium. In some regions, however, they rest on a bilaminate basement membrane with an electron-dense lamina densa and a less dense lamina rara. Bowman's capsule is composed of visceral and parietal layers. The epithelial cells (podocytes) of the visceral layer bear trabeculae connected to pedicels with microvilli. The pedicels rest on a bilaminate basement membrane which in some regions has a double-layered densa with connecting bands. Generally, this basement membrane is thicker than that of the capillary endothelial cells. The mesangium is composed mostly of irregular satellite cells with large nuclei and cytoplasmic processes, but also has smaller cells with kidney-shaped nuclei and cytoplasmic processes containing microfilaments. The mesangium cells are embedded in a collagenous matrix which extends to invade the area between the epithelial basement membrane and the capillary endothelium. These observations are discussed in relation to the structure and function of vertebrate renal corpuscles with special reference to the mesangium.

Tentacle contraction in glycerinated Discophrya collini and the localization of HMM-binding filaments.

The contractile tentacles of the suctorian Discophrya collini contain a central microtubular axoneme as well as filamentous structures in the cortical epiplasm and in a fibrous collar around the axoneme at the tentacle base. The nature and possible roles of these components has been investigated by the use of reactivatable glycerinated cells. In these a mean tentacle contraction to 70% of the original length could be achieved by a 5-min treatment with a reaction mixture containing ATP, calcium and magnesium ions, the same treatment giving retraction to 30% in living cells. Both the microtubules of the axoneme and the filaments of the fibrous collar and epiplasm were present in the glycerinated cells, suggesting that these components consist of large water-insoluble molecules. The addition of heavy meromyosin to whole glycerinated cells resulted in the appearance of 36-50-nm spaced "tails" or filaments attached to the epiplasmic fibres and the aggregation of 3-6-nm filaments and electron-dense material in the region of the fibrous collar. Neither of these 2 features was apparent after treatment with ATP. It is suggested that actin-like filaments are localized in the region of the fibrous collar and in the epiplasm, and that these are involved in tentacle retraction; whilst the microtubules of the axoneme are concerned with feeding, and play only a cytoskeletal role in the contractile mechanism.

Distribution of calcium in the suctorian Discophrya collini: an x-ray microanalytical study.

Discophrya collini is a free-living suctorian with tentacles which can be induced to contract by means of a range of experimental stimuli, including the application of CaCl2 and MgCl2 but not BaCl2. X-ray microanalysis of glutaraldehyde-only fixed cells shows Ca to be present in the cytoplasmic ground substance and elongate dense bodies (EDB). In 10(-1) M CaCl2-treated cells, calcium levels remain unchanged except for a three-fold increase in the EDB. Treatment of cells with 10(-1) M MgCl2 and 10(-1) M BaCl2 does not result in their detection in the cell. It is suggested that EDB may act as reservoirs controlling levels of calcium.

Ribosomes and Polysomes in Cucumber Leaves during Growth and Senescence.

The quantity of RNA in the ribosomal fraction of the first leaf of cucumber (Cucumis sativus) increases during growth, reaches a maximum before the final fresh weight is attained, and then decreases. The main changes are in the free ribosome fraction, the quantity of membrane-bound ribosomes remaining about constant. Few 65.5S chloroplast ribosomes are present in small leaves; however, they increase in quantity rapidly during growth and form about half of the ribosomes present in the mature fully green leaf. The cytoplasmic ribosomes have a sedimentation coefficient of 77.6S. Ribonuclease-sensitive polysomes were present in leaves of all ages except possibly the very oldest. The proportion of ribosomes in polysome form decreases during growth and then remains roughly constant during senescence. Following maturation of the leaf, the rate of incorporation of (32)P into ribosomal-fraction RNA begins to decline. This decline could account for the loss of ribosomes during the early stages of senescence. The possibility that leaf ribonuclease might be responsible for the final, more rapid loss of RNA, is discussed.