ONTOGENY OF CHICKEN HEMOGLOBIN. III. IMMUNOLOGICAL STUDY OF THE HETEROGENEITY OF HEMOGLOBIN IN DEVELOPMENT

By applying the double diffusion technique of Ouchterlony and the immunoelectrophoresis, sequence in the appearance of antigens reactive with antisera against HbCOs from 5-day embryos and adult chickens, and the major component of adult HbCO in the course of chicken development has been studied.
Antigenic components reacting to antiserum against HbCO from 5-day embryos have been detected throughout development. Three globin-like components specific to early embryos are detectable in embryos to 2 days of incubation. Two Hb components are detectable in embryos from 3 to 5 days of incubation; one is apparently specific to embryos, while the other seems to be somewhat different from adult Hb. After 6 days of incubation only one adult Hb component is detectable.
Antigenic components reacting to antiserum against HbCO from adults have also been detectable throughout embryonic life. One globin-like component specific to early embryos can be found in embryos to 2 days of incubation. One or two Hb components which are probably specific to embryos can be detected in embryos from 3 to 5 days of incubation. After 6 days of incubation one adult Hb component is detectable, while one globin-like component specific to adults can be found after 15 days of incubation. Further, the other globin-like component is detectable after 3 days of incubation.
Antigenic components reacting to antiserum against the major component of adult HbCO have been detected throughout development; one which seems to be somewhat different from either adult Hb components can be found in embryos from 2 to 5 days of incubation, while the other which is identical with the major component of adult Hb is detectable after 6 days of incubation.     

Fat Metabolism in Higher Plants. XXXVII. Characterization of the beta-Oxidation Systems From Maturing and Germinating Castor Bean Seeds

In the maturing castor bean seed (Ricinus communis), maximum β-oxidation appears at 28 days after flowering and in the germinating seed, 4 days after germination. Highest specific activities for both β-oxidation systems and their component enzymes are associated with cytosomal particles banding at a density of 1.25 g/ml in a sucrose gradient. Substrate specificity studies indicate that of several fatty acids, ricinoleate is oxidized most rapidly by the preparation from the maturing seed (28 days after flowering) while palmitate and linoleate are oxidized most rapidly by extracts obtained from tissue germinated for 4 days. The β-oxidation activities observed in both systems reflect the expression of activity of at least 3 of the component enzymes, crotonase, β-hydroxyacyl dehydrogenase and β-keto-thiolase, which rise and fall co-ordinately. Acyl thiokinase does not appear to play a limiting role in regulating β-oxidation per se under the conditions employed here.     

Accumulation of the four myelin basic proteins in mouse brain during development.

Myelin was isolated from the brains of mice 15, 20, 30, and 60 days after birth. The total amount of basic protein present in the isolated myelin was determined by radioimmunoassay. The 4 myelin basic proteins, with molecular weights of 21,500, 18,500, 17,000 and 14,000, were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and their relative amounts were determined densitometrically. The absolute amount of each of the basic proteins was calculated from its relative amount on the gel and from the total amount of myelin basic protein in the sample as determined by radioimmunoassay. The results show that between 10 and 30 days after birth each protein accumulates at a characteristic rate so that the molar ratios among the 4 basic proteins are (in descending order according to their molecular weights) 1:5:2:10 during this period. Between 30 and 60 days after birth the 14 K and 18.5 K proteins continue to accumulate at reduced rates while the 21.5 K and 17 K proteins begin to disappear from the myelin membrane; 60 days after birth the molar ratios among the 4 basic proteins are 1:10:3.5:35. These developmental patterns of accumulation are discussed in relation to the possible role of each of the 4 myelin basic proteins in myelination.     

Fleshy fungi in grass fields. II. Precipitation and fructification

An experimental field with grass had 192 parcels treated with combinations of NKP fertilizers and sown with 4 species of grass. Species of fleshy fungi in the parcels were counted over 4 years with 17–24 observations per year. Precipitation was measured at the field. The occurrence of the species varied with the distribution of the precipitation through the season. In early summer and after prolonged desiccation the fungus flora developed 25–30 days after heavy rain, while after mowing and after shorter desiccation periods, a new crop of fruit bodies had developed after 5 days. These intervals correspond to the maturation period of the mycelia, and to the period of the development of primordia already formed. Differences in the yearly periodicity of the individual species are discussed.     

Effect of prolonged administration of insecticide (Cyhalothrin/Karate) on the blood and liver of rabbits.

A sublethal dose of Karate administered to rabbits produced a significant increase in the total erythrocyte count and packed cell volume after 15 days of administration, though no significant change was observed after 30 days. The transaminases (glutamate oxaloacetate transaminase, GOT; glutamate pyruvate transaminase, GPT) also increased after 15 days of treatment. The GPT activity increased 119% and 60% after 15 and 30 days, respectively. From amongst metabolites, glucose content increased 17% and 185%, while cholesterol decreased 40% and 66%, and bilirubin 84% and 61%, after 15 and 30 days, respectively. The hepatic AkP activity decreased 30%, while the GPT activity increased 44%. Other enzymes such as AcP, GOT and LDH remained unaffected. The concentration of other metabolites, except for FAA which increased 35%, remained unaffected. Histological changes were marked by atrophied hepatic cells and hypertrophied nuclei and nucleoli. A trend towards necrosis of hepatic cells was also observed. All these results indicate that Karate is moderately toxic to mammals.     

Histological changes in the orbital region of rats after orbital puncture.

To contribute to the assessment of the degree of discomfort in rats after orbital puncture, we have examined the histological changes in the intraorbital tissues caused by this technique of blood sampling. Orbits were studied from rats euthanized either within 1 min, 4 days, 28 days or 56 days after puncture while under diethyl-ether anaesthesia. The techniques of 2 animal technicians were compared, one using a broken haematocrit capillary and the other using an intact Pasteur's pipette. Non-punctured orbits served as controls. Microscopic slides containing the eye in situ at 2 horizontal levels in the orbital region were examined for 37 parameters; the slides were scored blind and in random order. Orbital puncture caused haemorrhages in the puncture track and, depending on the technique used, also in the periosteum. Four days after puncture, inflammatory reactions were present in the puncture track. Depending on the technique of puncture, these reactions were also seen in the eye muscles and periosteum or in the Harderian gland. Within 4 weeks after puncture, the lesions had healed without detectable scars. The different histological effects of the 2 techniques of orbital puncture are discussed in the light of the characteristics of these techniques.     

Biphasic hypothermia in mice infected with a parasitic nematode, Trichinella spiralis.

In BALB/c mice infected with Trichinella spiralis, changes in body temperature (Tb) were observed over 35 days after the infection. T. spiralis infection induced hypothermia two times at 7 and 28 days after infection. The initial decrease persisted for about one week with a peak (37.1 +/- 0.62 degrees C) around 10 days after the infection, while the later phase persisted for at least one week. Both 10 and 35 days after the infection, there were remarkable decreases in Tb. The serum glucose level of infected mice at 10 days was significantly (p < 0.01) decreased compared with that of control mice at the same number of days, while the level in infected mice at 35 days was not decreased. Moreover, the later phase of hypothermia was prevented by the cyclooxygenase inhibitor indomethacin (10 mg/kg i.p.), while the initial phase was not. We conclude that hypothermia was caused by two different mechanisms, involving the effects of hypoglycemia and prostaglandins.     

Timing of the water-to-land transition and metamorphosis in the land hermit crab Coenobita compressus H. Milne Edwards: evidence that settlement and metamorphosis are de-coupled

After metamorphosing from the last larval stage to the transitional megalopal stage in the marine plankton, the hermit crab Coenobita compressus moves ashore where it undergoes a second metamorphosis to the first juvenile instar on land. In two experiments using laboratory-reared crabs, I moved megalopae from water to land after different amounts of time at this stage and investigated the impact of this manipulation on the timing of and survival through the second metamorphosis. In the Involuntary Settlement experiment, megalopae were moved to land when they were 3, 6, 9, 12, or 15 days old. None of those moved between the ages of 3 and 6 days survived through metamorphosis, but the majority of 9-day-old megalopae survived, as did most 12- and 15-day-old megalopae. This suggests that developmental changes early in the megalopal stage prepare C. compressus for terrestrial life. Once on land, megalopae that had been moved to land at 9 days spent about nine additional days there before metamorphosing, while 12- and 15-day-old megalopae metamorphosed after spending about 5 and 4 days, respectively, on land. In the Voluntary Settlement experiment, megalopae were given access to land when they were 1, 5, 10, or 15 days old, but were not forced to make the transition. Those given access to land after 1 day voluntarily left their dishes for the first time after an average of 7 days in water. Those given access when they were 5 days old remained in water about 4 days longer, while those given access when they were 10 and 15 days old left after less than a day. In both experiments, the timing of metamorphosis relative to settlement (i.e., transition to land) showed that these events are dissociated to a degree and revealed the presence of a metamorphic clock. I discuss why the dissociation of settlement and metamorphosis may have been favored in the land hermit crab and in another anomuran crab.     

Ipsilateral retinal projections into the tectum during regeneration of the optic nerve in the cichlid fish Haplochromis burtoni: a Dil study in fixed tissue.

Retinal projections were experimentally manipulated in a bony fish to reveal conditions under which considerably enlarged ipsilateral projections developed and persisted. Three experimental groups were studied: animals after unilateral enucleation, after unilateral nerve crush, and after enucleation and crush of the remaining optic nerve. At 29 days after unilateral enucleation alone, no enhanced ipsilateral projection had developed. After nerve crush, however, large numbers of retinal fibers regenerated into the ipsilateral tectum. Retrogradely filled, ipsilaterally projecting ganglion cells were distributed throughout the entire retina. After 15 days regenerating retinal fibers covered the entire ipsilateral tectum. At later stages the ipsilateral projection showed progressive reduction in coverage of the tectum. Combining enucleation with nerve crush led to an ipsilateral projection that covered the tectum at 28 days and later. In this experimental situation the development of an ipsilateral projection appears to be a two-step process: (1) Fibers are rerouted to the ipsilateral side at the diencephalon, and (2) ipsilateral fibers persist in the tectum only in the absence of a contralateral projection while they appear to be eliminated in the other cases.     

Effects of fluoroacetate on the testis of the rat.

Rats recieving 20, 6.6 or 2.2 p.p.m. sodium fluoroacetate in the drinking water were killed daily during the 7 days of treatment and at more widely spaced intervals in the succeeding 21 days. Testicular weight and ATP concentrations decreased in rats receiving 20 or 6 p.p.m. fluoroacetate, while citrate concentrations were elevated and morphological damage was seen in the testes of all the treated rats. Initial cellular changes common to the three treatment groups included altered appearance and decreased numbers of spermatids, and formation of spermatid and spermatocyte giant cells. At the two higher concentrations damage progressed to marked seminiferous tubule atrophy. Regeneration of the seminiferous tubules was complete by 7 days after treatment, in the rats given 2 p.p.m. but regeneration was not complete by Day 21 after treatment in those receiving the higher doses. Spermatogenesis was abnormal in some instances during the regneration period in these groups. The findings are consistent with impaired energy production via blockage of the Krebs cycle, and subsequent impairment of carbohydrate metabolism through the Embden-Meyerhof pathway.     

Induction and floral determination in the terminal bud of Nicotiana tabacum L. cv. Maryland Mammoth,a short-day plant

Floral determination in the terminal bud of the short-day plant Nicotiana tabacum cv. Maryland Mammoth has been investigated. Plants grown continuously in short days flowered after producing 31.4±1.6 (SD) nodes while plants grown continuously in long days did not flower and produced 172.5±9.5 nodes after one year. At various ages, expressed as number of leaves that were at least 1.0 cm in length above the most basal 10-cm leaf, one of three treatments was performed on plants grown from seed in short days: 1) whole plants were shifted from short days to long days, 2) the terminal bud was removed and then rooted and grown in long days, and 3) the terminal bud was removed and then rooted and grown in short days. Whole plants flowered only when shifted from short days to long days at age 15 or later. Only rooted terminal buds from plants at age 15 or older produced plants that flowered when grown in long days. Only terminal buds from plants at age 15 or older that were rooted and grown in short days produced the same number of nodes as they would have produced in their original locations while buds from younger plants produced more nodes than they would have in their original locations. Thus, determination for floral development in the terminal bud, as assayed by rooting, is simultaneous with the commitment to flowering as assayed by shifting whole plants to non-inductive conditions.Abbreviations LD long day(s) - SD short day(s) - DN dayneutral     

Recovery from monocular deprivation in the monkey. III. Reversal of anatomical effects in the visual cortex

Transneuronal autoradiography was used to study the effects of visual deprivation on the ocular dominance stripes in layer IVc of the striated cortex of Erythrocebus patas (Old World) monkeys. The animals were studied after: (a) 21-28 days of monocular deprivation starting at, or within, a few days of birth; (b) the same treatment followed by a further 3, 6, 15 or 126 days of monocular vision through both eyes (reopening). One other monkey was monocularly deprived from birth to 1890 days. In most cases the behaviour of the ocular dominance stripes formed by the initially closed eye was studied. After 24 days of monocular deprivation from birth, the input from the normal eye was distributed uniformly within layer IVc, with no periodicity evident. After 21 days of deprivation, the deprived eye's input formed narrow stripes occupying about 38% of layer IVc in the operculum. Seven months of monocular deprivation reduced this to about 29%. Opening the closed eye after the deprivation produced no change in the area innervated: when periods of 15 or 96 days of binocular vision followed the deprivation, the areas innervated by the initially deprived eye were 26 and 30% respectively. However, in both cases the deprived eye's input formed blobs and spots, rather than uniformly narrow stripes. In contrast to reopening, reverse suturing increased the fraction of layer IVc occupied by input form the initially deprived eye. In the operculum, the effects of reverse suturing appeared to be fully developed after only 6 days of reversal: the initially deprived eye's stripes having expanded to occupy about 50% of layer IVc. A further 9 days' reversal produced little change in this. In the visual cortex in the calcarine fissure, the effect of the initial deprivation ws more severe, and the expansion induced by reverse suturing more pronounced. The initial deprivation caused the stripes to shrink to occupy 24% of layer IVc; after 6 days of reverse sulture the proportion increased to 52%, while after 15 days of reverse suture about 88% of IVc was occupied. These results show that reverse suturing can cause fresh growth of afferent axons in regions of layer IVc from which they had been at least partially removed, either by the normal process of segregation, or as a consequence of monocular deprivation. Taken in conjuction with the findings of the accompanying two papers (Blackemore et al...     

Quantitative changes and ultrastructural alterations of the cornea in response to ultraviolet light. II. Effects of amphibia; elucidation of desmosomal structure and basement membrane synthesis.

The effects of far ultraviolet light irradiation upon an amphibian cornea were studied to compare the effects observed both quantitatively and ultrastructurally with data obtained after UV irradiation of mammalian corneas. The ultimate goal of this series of investigations is the elucidation of the alterations and the regeneration mechanisms, which might reflect existing morphological diversities among the species, observed in vertebrate corneas following exposure to UV light. It was found that while the epithelial cells undergo oedema after low dose exposures and are gradually damaged after high doses of UV light, 2-4 days leter a new epithelium has been formed. Intercellular permeability is increased by low dose exposure as was detected by the penetration of Ruthenium Red into the intercellular clefts. Under these conditions desmosomal structure revealed a 21-laminar configuration. The basement membrane of the amphibian, unlike that of the mammal, does not dissolve away upon exposure but shows localized disruptions which are thought to accommodate the passage of leucocytes from stroma to epithelium. That a new basement membrane is subsequently formed is evident by the existence of extracellular and intracellular secretion granules. In comparison to irradiated rabbit corneas, this stroma remains remarkably at the same thickness following a high dose exposure although a noticeable disorganization of collagen arrangement is apparent. Finally, as in the case of the rabbit corneas, a secondary degeneration of endothelium was observed 4 days after a moderate dose exposure.     

Experimental infections with Cooperia oncophora (Railliet, 1918) in calves. Results of single infections with two graded dose levels of larvae.

Two experiments were carried out in which calves reared parasite-free were infected with a single dose of 3rd-stage larvae of Cooperia oncophora. In the first experiment the calves received 20 000 or 200 000 infective larvae and they were autopsied 28 or 56 days after the infection. In the second experiment the doses were the same but the animals were killed 14, 84 or 140 days after infection. If a dose of 20 000 was given, clinical signs were never observed, while at a dose level of 200 000 the weight gain was less on 56 and 84 days after the infection compared with the low-infected groups or the control animals. After 170 days the differences in weight gain were compensated. Faecal egg output was higher in the 200 000 groups only in the first period of patency, thereafter the calves in the 20 000 groups produced more. No obvious differences between the two infection levels were observed with regard to the haematological data. In the low-infected groups worm counts were only slightly lower when the results after 28 days were compared with those after 56 days. Also, the worm numbers after 14 days were almost equal to those after 84 days, while at 140 days 1 animal still had the same number, the other one had lost its worm burden. In the high-infected groups the worm loss was much quicker. After 28 days a great part of the population had already been lost. Obviously, at the 200 000 level the reaction of the host against the parasite was much stronger. Adult worms were expelled at a higher rate than early 4th-larval stages. In the first experiment worm measurements revealed differences between the length of females, males and spicules of males, these being significantly longer in the low-infected groups. Analysis of the distribution of worms over the small intestine showed that in the low-infected groups worms were mainly restricted to the first 6 metres. In the high-infected groups the worm population was distributed more evenly over the whole small intestine.     

Cytodifferentiation in the accessory glands of Tenebrio molitor. III. Fine structure of the spermathecal accessory gland in the pupa

To establish indices for studying the hormonal control of differentiation of the accessory reproductive glands of insects, the ultrastructural development of the spermathecal accessory gland (SAG) of female mealworm beetles has been analyzed. Over the 9 days between adult and pupal ecdysis, the SAG transforms from a stubby sac of columnar epithelium into an elongate cylindrical gland, lined with cuticle, and containing several distinct types of differentiated cells. The first phase of pupal differentiation is one of cell division and overall gland morphogenesis which lasts 3--4 days; at its close, two populations of cells can be distinguished. One of these populations will produce the cuticular ductules while the other will yield the three-cell secretory units or organules. In the second phase which lasts 2 days, the three cells of each organule become wrapped around one another and then the innermost puts out a pseudocilium and retracts within the next ensheathing cell. In the third phase which lasts 4 days, the cuticles of the axial duct, of the efferent ductule, of the vestibule upon which the ductules converge, and of the end apparatus, are deposited. The ciliary process degenerates, and after ecdysis, the secretory cells undergo peak differentiation.     

Dynamics of pancreatic tissue cells in the rat exposed to long-term caerulein treatment. 2. Comparative analysis of the various cell types and their growth

This study was undertaken to evaluate the effects of caerulein, a CCK analog, on the different cell populations of the pancreatic tissue and their respective turnover. Rats received saline or caerulein subcutaneously and 3H-thymidine intraperitoneally three times a day for 4 days. They were sacrificed immediately after termination of treatment and 2, 15 and 50 days later. With age, the proportion of acinar cells decreased significantly whereas those of the ductal and interstitial cells increased. Although caerulein induced preferential acinar cell growth, it did not modify the proportion of this cell population with regard to the other cells. However, at specific times after termination of treatment, caerulein induced modifications in the ductal, endothelial and interstitial cell populations. The growth promoting effect of caerulein was evident from the specific increases in total DNA content and DNA synthesis. The labeling indices indicate that all cell populations except the endocrine system were stimulated to grow in response to caerulein. Furthermore, all new cells remained for at least 50 days after termination of treatment. These data indicate that caerulein induced uniform growth of the pancreatic tissue during intensive treatment. The normal growth rate of these stimulated cells was, however, arrested for the following 50 days while that of the control group cell population proceeded normally.     

Moderate red wine consumption protects the rat against oxidation in vivo.

The effect of the moderate consumption of red wine on the antioxidant system in rat liver, kidney and plasma has been evaluated. Wistar rats were treated in separate groups as follows: control; red wine for 45 days or 6 months; and 13.5% ethanol for 45 days or 6 months. The consumption of alcoholic beverages was free because the rat could always choose between the alcoholic beverage and the water. In liver, red wine ingestion resulted in higher hepatic superoxide dismutase and glutathione peroxidase activities after 45 days of treatment. The data indicate that wine and ethanol ingestion resulted in lower hepatic malondialdehyde and enhanced hepatic catalase activity in both of the periods studied. In kidney, the reduced glutathione/oxidized glutathione ratio was higher after 45 days of wine consumption, and the malondialdehyde was lower after 6 months of wine consumption. In plasma, malondialdehyde was lower after 6 months of both treatments, but plasmatic vitamin E was higher after red wine consumption while it was lower after ethanol consumption for this period of time. The present study shows that the moderate and prolonged consumption of red wine is consistent with higher protection against oxidation in vivo.     

Effect of thyroid hormones on microsomal fatty acid chain elongation synthesis in rat liver.

Evidence is presented that rat liver microsomal fatty acid chain elongation synthesis and desaturation, as well as acetyl-CoA carboxylase and fatty acid synthetase, are strongly influenced by thyroid hormone level. Conversely, the fatty acid chain elongation system in mitochondria, unlike the oxidative capacity of palmitate, NADH, succinate and malate, does not seem significantly affected by the thyrotoxic state. In triiodothyronine-induced or thyroxine-induced hyperthyroidism, rat liver acetyl-CoA carboxylase, fatty acid synthetase and microsomal chain elongation and desaturation reactions are not greatly affected after the first 10 days of treatment, while after longer intervals a respective increase in these activities is shown of up to 87, 116 and 65% after 22 days. In propylthiouracil-induced hypothyroidism, all the above synthetic activities are strongly reduced immediately after three days of drug administration and diminished no further following longer periods. Although the pattern of synthesized fatty acids in the thyrotoxic state is similar to that obtained from normal subcellular rat fractions, the esterification process of fatty acids in microsomal lipids appears to be slightly inhibited in hypothyroid rats and increased following triiodothyronine or thyroxine administration. Finally, a reduction in the hepatic cyclic AMP level of about 41% is reported after 19 days of triiodothyronine-administration to rats. On the basis of the observed insensitivity of the mitochondrial fatty acid chain elongation system to the thyrotoxic state, a tentative interpretation of its role in the hepatic cell is postulated.     

Reproductive timetable for the tropical Vireya rhododendron,R. Macgregoriae

Summary The reproductive development of Rhododendron macgregoriae F.v.M., Section Vireya (Ericaceae) has been followed from 10 days before anthesis to the production of mature germinable seeds about 145 days after anthesis. The species is self-compatible but shows both protandry and physical separation of pollen from the receptive stigma. Pollen is mature and viable from shortly before anthesis until the corolla and attached anthers abscise some 9–12 days after the flowers open. Spontaneous dehiscence, however, occurs mostly in the first few days after opening. The stigma becomes receptive 6–7 days after anthesis, and nectar is produced at this time. Female gametophytes are not mature until about 10 days after anthesis. Fertilization occurs about 6–7 days after pollination, and although the endosperm commences development immediately, development of the embryo proper does not begin until some 40–45 days later. Pollinations made with fresh pollen between 5 and 14 days after anthesis were successful, but those made on dry stigmas in the first 4 days after anthesis, or on senescing pistils 21 days after anthesis, gave little or no seed set. In nature, autogamy is probably uncommon, the majority of pollinations are likely to be geitonogamous, but there is considerable potential for outcrossing.     

Fructan metabolising enzymes in rhizophores of Vernonia herbacea upon excision of aerial organs.

The activities of fructan metabolising enzymes and fructan contents are reported for rhizophores of Vernonia herbacea (Vell.) Rusby induced to sprouting by shoot excision. The activities of fructan exohydrolase (1-FEH), sucrose: sucrose fructosyltransferase (1-SST), fructan: fructan fructosyltransferase (1-FFT) and invertase (INV) and the fructan contents were analysed every 3-4 days for 1 month by colorimetric and chromatographic methods. Sprouting of new shoots started on day 9. 1-FEH activity increased after day 13 and reached its maximum value 20 days after shoot excision. A gradual decrease in 1-SST activity was detected between days 3 and 9. 1-FFT activity exhibited fluctuations throughout the experimental period and a peak of activity for invertase was detected 9 days after shoot excision. Variation in fructan contents in vivo included a decrease until day 13 after which, levels remained practically unchanged. Fructan depolymerization and sprouting are concomitant processes in V. herbacea and can be induced by shoot excision at any phenological phase. 1-FEH and 1-FFT seemed to act in a concerted way to catalyse fructan depolymerization, while 1-SST was inhibited, possibly due to interruption of sucrose supply to rhizophores from the aerial organs.