Clinical trials of bestatin for leukemia and solid tumors

Dissociated cells from 13- and 17-day-old embryonic rat mesencephali have grown in primary cultures in order to compare the early and late influences of different agents - insulin, dexamethasone and nerve growth factor (NGF) - on the expression of cholinergic maturation process. We have studied cholin acetyltransferase (ChAT) activity, which is regarded as a specific marker for cholinergic function of the brain, and a widely used differentiation marker, the acetyl-cholinesterase (AchE) enzyme. Biochemical maturation of increasing specific activity of ChAT in both younger and older cells was taken into consideration. During cultivation the AchE activity was slightly increased in younger cells, but a dramatic decrease could be noted in older ones. Insulin in concentration from 10 to 27 µg mL^–1 causes a significant inhibition in ChAT activity in comparison with the enzyme activity measured in control cultures (insulin ranging from 1 to 100 ng), independently of embryos age. This polypeptide hormone is able to enhance AchE activity in the cultured cells, especially in older ones. With continuous treatment of the culture with dexamethasone, a synthetic glucocorticoid, the ChAT activity in younger cells reaches a maximum curve by day 9 (nine). At this time the AchE activity shows a slighter, no significant increase than at any other time during cultivation. In cell cultures taken from 17-day-old embryos however dexamethasone treatment evoked a significant decrease in ChAT activity with a concomitant increase of AchE activity which was compared to insulin treatment. In spite of the fact that the NGF is able to enhance the ChAT activity, no significant alteration in AchE activity can be measured in younger cell cultures. These results suggest an uneven expression of the enzymes in embryonic rat mesencephali in the presence of above agents depending on the age of cells.     

Pheromone Blend Does not Explain Old Male Mating Advantage in a Butterfly

In several insect species, male mating success is higher in older than in younger males, although condition diminishes dramatically with age. Two hypotheses are under debate to explain the counterintuitive pattern of old male mating advantage: first, an increased eagerness of older males to mate, driven by their low residual reproductive value, and second female preference for older males based on chemical cues such as sex pheromones (female choice hypothesis). In a series of experiments, we manipulated female olfaction, male pheromone blend and female age to test whether old male mating advantage prevails when the influence of male sex pheromones is controlled for, using the tropical butterfly Bicyclus anynana as model. We found that older males had a higher mating success than younger ones irrespective of female scent‐sensitivity and irrespective of male pheromone blend. Interestingly, older males were found to court more often and for longer time bouts than younger males. These results were independent of female age, although younger males courted younger females more often and for longer bouts than older females. Taken together, our results indicate that male courtship activity (1) is higher in older compared to younger males and (2) increases the mating success of older males. Olfaction and sensing pheromones, in contrast, were not a necessary prerequisite for old male mating advantage to occur and may use other cues than pheromones to assess male quality.     

Immobility reactions in socially behaving rabbits: Relation to hippocampal electrical activity

The presence of an intruder conspecific of the same sex caused a series of conflict reactions in rabbits living in a semi-natural environment. In some animals, defensive responses were predominant and two immobility reactions were observed. One consisted of freezing when the intruder was exploring at a distance; the other consisted of subdued posture in close contact with the intruder. Hippocampal EEG, recorded by telemetry, showed two different profiles. During freezing, rhythmic slow activity prevailed and during subdued posture an irregular pattern was predominant. Frequency increased when the intruder was in close contact with the resident. Aspects such as distance or behaviour of the intruder were found to affect both behavioural strategies and hippocampal electrical responses.     

Are extra‐pair males different from cuckolded males? A case study and a meta‐analytic examination

Traditional models for female extra‐pair matings assume that females benefit indirectly from extra‐pair mating behaviour. Under these so‐called adaptive models, extra‐pair males are hypothesized to have more compatible genotypes, larger body size, exaggerated ornaments or to be older than cuckolded males. Alternatively, (‘nonadaptive’) models that consider female extra‐pair matings to be a by‐product posit that female extra‐pair mating can be maintained even if there is no benefit to females. This could happen if, for example, males gained fitness benefits from extra‐pair mating, while female and male extra‐pair mating behaviours were genetically correlated. Extra‐pair males are also expected to be older and larger if this improves their ability to convince or coerce females to mate. We investigated whether a female's extra‐pair mates differed from her cuckolded mate in both genetic and phenotypic traits by analysing data from an insular house sparrow population. We found that extra‐pair males were older than cuckolded males, consistent with both models. However, in contrast to the expectations from from adaptive models, extra‐pair and cuckolded males were of similar genetic relatedness, and hence expected compatibility, with the female, and had comparable body size and secondary sexual traits. We also updated previous meta‐analyses examining differences between extra‐pair and cuckolded males. The meta‐analytic results matched results from our house sparrow case study. Although we cannot completely exclude indirect benefits for females, nonadaptive models may better explain female extra‐pair matings. These neglected alternative models deserve more research attention, and this should improve our understanding of the evolution of mating systems.     

Changes in serum immunoreactive inhibin and follicle-stimulating hormone during gonadal development in male and female rats.

We have measured serum and ovarian immunoreactive inhibin alpha (irI alpha) and serum FSH in fetal and neonatal rats from 20 days of gestation until 40 days of age. For animals aged 10 days or older, serum measurements were made on intact and gonadectomized animals. Serum irI alpha was detectable in intact male and female rats at all ages studied. In females, irI alpha levels were low until Day 5 and then increased steadily to peak at Day 25. Thereafter they declined until Day 35 to reach levels typical of adult females. There was a significant decrease in irI alpha levels 24 h after ovariectomy at all ages. Serum FSH levels in females were low until Day 7, then increased rapidly to plateau from Days 10-15. The levels then declined until Day 25 and were generally unchanged after that time. There was a significant increase in FSH 24 h after ovariectomy in rats aged 20 days and older, and in younger rats by 48 h after ovariectomy. In male rats, serum irI alpha levels were significantly higher than females until Day 7. The levels increased at Day 7 and then remained relatively constant until Day 20, after which they declined to reach typical adult male levels. Serum irI alpha levels were significantly lower in males than females from Days 25-40. There was a significant decrease in serum irI alpha 24 h after castration at all ages studied. Serum FSH levels in males were low until Day 20, increased at Day 25, and thereafter remained relatively unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)     

Do males evaluate female age for precopulatory mate guarding in the two-spotted spider mite?

In the two-spotted spider mite Tetranychus urticae (Acari: Tetranychidae), because only the first mating results in fertilisation, adult males guard quiescent deutonymph females, the stage immediately before adult emergence. Previous studies showed that T. urticae males prefer to guard older rather than younger females. However, as previous experimental designs have included uncertain factors, findings have not always been sufficiently rigorous. Therefore, we reexamined whether T. urticae males discriminate between the females closest to becoming receptive and younger females. One male was introduced onto leaf squares with two differently aged quiescent deutonymph females, with time lags between the females of 3, 6, 12 or 23 h, and whether the older or younger female was guarded was recorded. When the time lags between the females were 3, 6 or 12 h, males showed no preferences. Males preferred older females over younger ones only when the time lag was 23 h. Under non-choice conditions, fewer males guarded the 23-h younger females than other older females, suggesting that the attractiveness of the younger females is weaker than older females. Our results are not fully consistent with previous studies; T. urticae males do not obviously discriminate between females closest to becoming receptive and younger females.     

Behavioral characteristics of female Wag/Rij rats

Behavior of male and female WAG/Rij and Wistar rats was compared in the tests assessing the level of anxiety (light-dark choice, open field) and depression-like state (sucrose intake and preference, forced swimming). Females of WAG/Rij rats like males of the same strain exhibited symptoms of depression-like behavior: increased immobility in the forced swimming test and decreased sucrose intake and preference (anhedonia). In contrast to males, females of WAG/Rij rats displayed more distinct signs of increased anxiety as compared to Wistar rats. Both WAG/Rij and Wistar females exhibited increased locomotor and exploratory activity in the open field as compared to males.     

Female Age Determines Remating Behavior in Wild Mexican Fruit Flies

Male and female age are important factors that can influence mating and remating behavior. Females can discriminate against or prefer older males, but there have been relatively fewer studies on how female and male age influence female remating. Here we showed in wild flies of the Mexican fruit fly Anastrepha ludens (Loew), that when females were given a choice between males of different ages, younger females preferred to mate with younger males over older males, while older females were less selective. Also, when given a choice between males of different ages, older females had longer copulation durations than younger females. On the other hand, older males and females had lower mating success, compared with young and middle-aged flies under no choice conditions. However, middle-aged females mated faster compared to young females and young males mated faster compared to middle-aged males. Male age did not influence female remating, while female age strongly determined female remating, with no females remating when they were old. It is unclear if female receptivity mechanisms are switched off at older ages, or if females are reluctant to remate due to possible costs of mating. We discuss our results in terms of how male and female age can influence mating decisions.     

Mounting frequency and number of cloacal contacts increase with age in common terns Sterna hirundo

We investigated courtship feeding and copulatory behaviour of common terns Sterna hirundo in relation to age. We found a significant positive correlation between the number of courtship feedings delivered to females and the number of mountings. Moreover, both courtship feeding and mounting rates were positively correlated with female and male age. Also, the number of cloacal contacts per mounting increased significantly with male or female age, which may indicate an increase in copulatory ability with age. The number of mountings for older birds was up to three times that for younger birds. Presuming that transfer of sperm is proportional to the number of cloacal contacts, older birds may have transferred up to six times more sperm than younger birds during the pre-laying and laying period. These results may account for the preference of young females for older mates as well as for older extra-pair mates.     

Interhemispheric asymmetry of neocortex and hippocampus in orienting exploratory behavior of rabbits

Correlation of discharges of cortical neurons in symmetrical points of the visual and parietal cortices and left and right hippocampal CA1 neurons was studied in freely moving rabbits during exposure to emotional stimuli. Crosscorrelation histograms were plotted. As compared to the initial state, during an active orienting exploratory reaction to stimuli, the left-side influence on right-hemispheric cortical neurons with a delay about 100 ms increased, which led to asymmetry in interhemispheric interaction with the left-side dominance. During freezing, the left-side influence became weaker, and the effects of the right hemisphere prevailed. Hippocampal asymmetry in neuronal activity was in reciprocal relationship with neocortical asymmetry. In the hippocampus, the right-side influence with a delay about 200 ms increased during the active exploratory reactions resulting in the right-side dominance. Freezing was accompanied by strengthening of the left-side influence (the left-side dominance). During the active locomotion, neuronal interaction in the hippocampus was predominantly realized in the theta-range frequency, whereas freezing was characterized by the delta-range correlation. It was concluded that the active or passive nature of a behavioral reaction to emotional stimuli was correlated with changes in asymmetry in the interhemispheric neuronal interactions at the cortical and hippocampal levels.     

Effect of male and female age on the mating success of the obliquebanded leafrollerChoristoneura rosaceana (Lepidoptera: Tortricidae) under different ecological conditions

The present study was undertaken to test the hypothesis that male and female age, as well as temperature, can affect the mating success ofChoristoneura rosaceana, given the role that these two factors play in the pheromone biology of this species. In the laboratory, the mating success of females generally declined linearly with age, whereas in males, it increased during the first 3 days and then decreased. The decline in female mating success was more pronounced under warm than cool thermocycles, while the changes observed in males were greater under fluctuating than constant temperature regimes. The onset time of mating was unaffected by male age, however, older females always mated earlier than younger ones, with the advance being more pronounced at cool than warm temperatures. Similar results were also obtained under field conditions. When a single 3-day-old male was provided with 0-, 3-, and 5-day-old females simultaneously, older females obtained mates significantly more often than younger individuals at all temperatures, indicating that calling earlier may afford a reproductive advantage to older females. At both constant temperatures, the time spent mating was longer in older than in younger females, but not under fluctuating thermocycles. Very young and very old males generally spent more time in copula than middle-aged individuals both at constant temperatures and under the warm thermocycle. Under the cool thermocycle, the duration of mating was considerably prolonged at all ages, which could increase the risk of predation.     

Relative roles of oestradiol and of the uterus in the maintenance of the corpus luteum in the pseudopregnant brown hare (Lepus europaeus)

Brown hares were made pseudopregnant by sterile matings or PMSG-hCG treatment (day of mating or hCG injection = Day 0 of pseudopregnancy). Progesterone secretion by the CL began 3-4 days after the ovulatory stimuli, reached maximum on Days 8 to 11 and decreased thereafter to reach low levels from Day 9 to 18, depending on the female. Cauterization of all large ovarian follicles on Day 7 resulted in an immediate luteolysis in young females, but had no effect in older ones. Oestradiol capsules implanted from Day 7 to Day 46 were able to maintain progesterone secretion until at least Day 30, in intact females as well as in females with all large follicles cauterized. Hysterectomy on Day 7 or 8 was followed by an immediate drop in progesterone concentrations; oestradiol capsules implanted at the time of hysterectomy prevented the drop in progesterone values, which remained elevated until Day 38. The induction of ovulation in females hysterectomized 2 months before resulted in CL of slightly shortened life-span. The injection of PGF-2 alpha on Day 7 of pseudopregnancy was followed by an immediate luteolysis. These results suggest that oestradiol secreted by the large ovarian follicles is the main luteotrophic factor in the brown hare. In old hares, the large amount of interstitial tissue could secrete oestrogens, and thus maintain pseudopregnancy. On Day 7 of pseudopregnancy, the uterus secretes a luteotrophic substance acting either directly on the ovary, or via the pituitary, to maintain oestradiol secretion by the follicles. In long-term hysterectomized females, the CL would be able to develop independently of any trophic substance, but for a reduced duration.     

The effect of age on encounters between male crab spiders

In males that compete aggressively for females, size and agemay determine which males obtain access to these females. Inthe present study, we use the crab spider, Misumena vatia, aspecies with males that do not grow after becoming sexuallymature adults, to test the hypothesis that age affects the successof males competing for access to females. M. vatia is an excellentspecies to test this hypothesis because it is possible to disentangleage from size, characters that typically vary together in thespecies usually tested. We staged encounters between similar-sizedolder and younger adult male M. vatia in the presence of a femaleto determine the role of age in male access to females. Encountersbetween the males occurred during 63.3% of these pairings. Youngermales won significantly more (70.2%) of the encounters thandid older ones, but did not initiate significantly more encountersthan did older ones (62.5%). Although older males won only 29.8%of these encounters, they initiated significantly more (76.5%)of them than predicted by chance. This design also allowed usto test Parker's hypothesis that older individuals should exhibita higher level of aggression than younger ones. However, attacksby younger males were most likely to include extensive bodilycontact, whereas attacks by older males involved significantlyless contact. These results counter the frequent assertion thatolder individuals usually prevail over younger ones in contestsfor access to females, and that older males are more likelyto engage in highly overt aggression than are younger ones.Aging may decrease reproductive opportunities and success ratesof male M. vatia, affecting as many as nearly one-fourth oftheir encounters.     

Tonic immobility in the squirrel monkey (Saimiri sciureus)

Tonic immobility and several accompanying behavioral changes were examined in the squirrel monkey (Saimiri sciureus). It was found that either the occurrence of a loud noise immediately before induction or the presence of a stuffed Cooper's hawk increased the duration of immobility in some subjects. Subspecies differences in durations of immobility were also found. The number of inductions required to produce immobility in squirrel monkeys were unusually large, and were negatively correlated with duration of the response. Behavioral variables accompanying tonic immobility showed qualitative differences when compared to what is known concerning their relation to immobility in other species. The findings support the belief that tonic immobility evolved as a predator defense in squirrel monkeys or one of their ancestors, but suggest that it no longer plays an active role in their survival.     

ChAT::Cre transgenic rats show sex-dependent altered fear behaviors,ultrasonic vocalizations and cholinergic marker expression

The cholinergic system is a critical regulator of Pavlovian fear learning and extinction. As such, we have begun investigating the cholinergic system's involvement in individual differences in cued fear extinction using a transgenic ChAT::Cre rat model. The current study extends behavioral phenotyping of a transgenic ChAT::Cre rat line by examining both freezing behavior and ultrasonic vocalizations (USVs) during a Pavlovian cued fear learning and extinction paradigm. Freezing, 22 kHz USVs, and 50 kHz USVs were compared between male and female transgenic ChAT::Cre+ rats and their wildtype (Cre-) littermates during fear learning, contextual and cue-conditioned fear recall, cued fear extinction, and generalization to a novel tone. During contextual and cued fear recall ChAT::Cre+ rats froze slightly more than their Cre- littermates, and displayed significant sex differences in contextual and cue-conditioned freezing, 22 kHz USVs, and 50 kHz USVs. Females showed more freezing than males in fear recall trials, but fewer 22 kHz distress calls during fear learning and recall. Females also produced more 50 kHz USVs during exposure to the testing chambers prior to tone (or shock) presentation compared with males, but this effect was blunted in ChAT::Cre+ females. Corroborating previous studies, ChAT::Cre+ transgenic rats overexpressed vesicular acetylcholine transporter immunolabeling in basal forebrain, striatum, basolateral amygdala, and hippocampus, but had similar levels of acetylcholinesterase and numbers of ChAT+ neurons as Cre- rats. This study suggests that variance in behavior between ChAT::Cre+ and wildtype rats is sex dependent and advances theories that distinct neural circuits and processes regulate sexually divergent fear responses.     

Old‐male paternity advantage is a function of accumulating sperm and last‐male precedence in a butterfly

Old‐male mating advantage has been convincingly demonstrated in Bicyclus anynana butterflies. This intriguing pattern may be explained by two alternative hypotheses: (i) an increased aggressiveness and persistence of older males during courtship, being caused by the older males' low residual reproductive value; and (ii) an active preference of females towards older males what reflects a good genes hypothesis. Against this background, we here investigate postcopulatory sexual selection by double‐mating Bicyclus anynana females to older and younger males, thus allowing for sperm competition and cryptic mate choice, and by genotyping the resulting offspring. Virgin females were mated with a younger virgin (2–3 days old) and afterwards an older virgin male (12–13 days old) or vice versa. Older males had a higher paternity success than younger ones, but only when being the second (=last) mating partner, while paternity success was equal among older and younger males when older males were the first mating partner. Older males produced larger spermatophores with much higher numbers of fertile sperm than younger males. Thus, we found no evidence for cryptic female mate choice. Rather, the findings reported here seem to result from a combination of last‐male precedence and the number of sperm transferred upon mating, both increasing paternity success.     

Effects of ovariectomy and estrogen treatment on learning and hippocampal neurotransmitters in mice

This study examined the effects of long-term estrogen treatment (sc 17 beta-estradiol minipellets) on learning in C57BL/6J female and male mice using a position discrimination task in the T-maze and a win-stay task (1/8 arms baited) in the radial arm maze (RAM). In addition, hippocampal monoamines and ChAT activity were measured at the end of the study and correlated to task performance. Female sham-operated (gonadally intact) and ovariectomized (OVX) mice were treated with estrogen either for 7 or 40 days before the behavioral tests and intact male mice for 7 days before the behavioral tests. In sham-operated mice the 40-day estrogen treatment improved RAM performance and in OVX mice both the 7- and 40-day estrogen treatments improved the performance in both maze tasks. The estrogen treatment also improved RAM performance in males. The hippocampal ChAT, NA, 5-HIAA, and DOPAC levels were decreased in OVX mice. Furthermore, the effects of estrogen treatment on the levels of hippocampal 5-HT and its metabolite 5-HIAA were different in sham-operated than in OVX mice. We could find no correlation between cognitive measures and neurochemical variables. This study gives new information about the effects of estrogen on learning and hippocampal neurotransmitters in mice.     

Distinct Clinical and Experimental Characteristics in the Patients Younger than 60 Years Old with Myelodysplastic Syndromes

Myelodysplastic syndromes (MDS) mainly occur in elderly individuals in Western countries. However, MDS is commonly found in young individuals (<60 years) in Asia. The reason for the high incidence in younger individuals is still unclear, and the differences in disease features between young and elderly patients with MDS have been not well recognized. To explore these issues, in this study, we analyzed the clinical and experimental characteristics of MDS in the patients younger and older than 60 years old and characterized the potential age-associated differences. The results showed that over half of the patients with MDS (61.9%) were younger than 60 years old upon the first diagnosis. The younger patients were more likely to be female, who have lower risk and less advanced MDS. The occurrence of trisomy 8 and bone marrow failure were more frequent in the younger patients than the older ones. The marrow CD34+ cells in the younger patients showed lower proliferation and higher apoptosis in comparison with that in the older ones. Obvious amplification of T cells and low CFU formation could be found in the younger patients. CFU formation was significantly increased in the younger patients after the removal of activated T cells. In addition, the younger patients had a lower frequency of p15^INK4B methylation, longer survival expectancy and less AML transformation. In summary, the younger patients with MDS in China may show more benign disease features than the older ones. Enhanced immunological response may be involved in the pathogenesis of MDS in the patients younger than 60 years.     

Nocturnal behaviour of female Mythimna convecta

Nocturnal behaviour of Mythimna convecta (Walker) (Lepidoptera: Noctuidae) virgin females was studied in the laboratory under 20 °C and 16:8 LD conditions. The periodicity of activity, feeding, calling, pre-oviposition extrusion and oviposition varied with female age and hour of the scotophase. Females called for the first time between the 2nd and 11th scotophases with the peak in the 4th scotophase. Maximum calling occurred on the 7th hour of the scotophase. Young moths called more frequently with shorter bouts while old moths called less often but with longer bouts. In the presence of older females, moths spent significantly more time in pre-oviposition extrusion and resting and less in activity and feeding than they did when only females of the same age or younger were present. There were no significant differences for calling suggesting that pheromones of older calling females did not affect calling of younger females.     

Male size and reproductive success in the communal courtship system of the fiddler crab Uca rapax

Reproductive success of male fiddler crabs Uca rapax depends on the numbers of females a male can attract, and the fecundity of these females. The latter correlates directly with fenale size. The size of a female in a mated pair is directly correlated with the size of the male. Therefore, older larger males experience greater reproductive success than smaller younger males. Females appear to be highly selective in mating, while males are not. The question of why females should select a male whose size is correlated with hers cannot be answered from the present study.