Growth characteristics of human melanoma xenografts

Abstract. The growth of twelve human malignant melanomas in athymic nude mice was studied. Gompertz curves were fitted to volumetric growth data. DNA histograms were obtained with flow cytometry. Each of the twelve melanomas exhibited a characteristic growth pattern, indicating that inherent properties of the tumours are important for the growth control. The theoretical maximum volumes (Vmax) ranged from 208 to 12,900 mm³, the volume doubling times ( T d) from 2.8 to 15.3 days (^V= 50 mm³) and from 3.8 to 64.6 days ( V = 200 mm³), and the fraction of cells in S from 5 to 21%. Tumours with short T d were characterized by a higher growth fraction and probably by a lower cell loss factor than those with long T d. The growth was also influenced by the nude mouse host, as indicated by the values for V max which were similar to those reported for mouse tumours (geometric mean = 8100 mm³), but considerably lower than the volumes of many tumours in man. Also the T d-values for the xenografts were generally lower than those reported for tumours in man, presumably due to a lower cell loss factor. During serial transplantation the growth rate of one of the melanomas increased abruptly, probably because of both an increased growth fraction and a reduced cell loss factor. The latter result demonstrates the necessity of keeping basic biological parameters of xenografts under observation during serial transplantation.     

Cytological ontogenesis and involution of the thymus and head‐kidney in juvenile and old domestic carp: Is ageing in fish a chronological or growth‐related phenomenon?

Samples of the thymus and head-kidney of domestic carp ( Cyprinus carpio ) were studied in order to evaluate the process of functional and cytological change during ageing. We examined larvae from 6 days post-fertilization, 1- to 10-year-old fish from fishponds, and experimentally stunted 15-year-old fish that had attained no more than 100–120 mm total length (TL) and 18–20 g weight. The primordium of the thymus appears 6 days after fertilization as a group of non-specialized cells 0.8 mm³ in volume. Differentiation of cell types, especially thymocytes, starts in fry of 6 mm TL, and the thymus continues to enlarge, attaining 200 mm³ (±3.4 mm³) in 8-year-old fish of 550 mm TL. Throughout its development the thymus remains active, containing various forms of reticulocytes and thymocytes. The head-kidney appears in pre-hatching embryos on either side of the pharynx as renal units, each with a single glomerulus and a short renal tubule. During development the protonephridia pass through three functional stages: (1) excretory, as the primordial kidney, (2) in a mixed renal and glandular function, producing blood cells and adrenergic cells, and (3) in young fish of 80 mm TL and above, they continue to function as blood producing and endocrine organs. At this last stage the renal elements become degraded. The largest head-kidney measured had a volume of 8 cm³ (±0.8 cm³) in an 8-year-old fish weighing 6.5 kg. Retention of thymal cytological integrity in reproductive older carp and other fish would indicate that ageing in fish, and possibly also in other exothermic vertebrates, is not simply a time-related phenomenon, thereby differing greatly from similar processes in endothermic vertebrate.     

Production and litter processing by crayfish in an Appalachian mountain stream

SUMMARY 1. Mean annual density and biomass () of Cambarus bartonii in an Appalachian mountain stream (U.S.A.) was 12 individuals m⁻² and 1669 mg (ash-free dry weight) m⁻².
2. Annual production ( P ) of C bartonii was 961 mg AFDW m⁻². Despite high biomass, low growth rates resulted in low production and a low P/ ratio of 0.58.
3. While C bartonii constituted 61% of the total macroinvertebrate biomass, it contributed only 13% of annual community secondary production.
4. Litter processing was positively related to temperature and crayfish size. Cambariis bartotnii was estimated to comminute 36 g m⁻² y⁻¹ of leaf litter (>1 mm²) to 24 g m−^>2 y⁻¹ fine particulate material (<1 mm²). The annual pattern of litter comminution by crayfish was regulated by temperature. As a result, >5()% of shredding activity by crayfish occurred from June to September which was also the period of lowest litter standing crops and activity of other shredding macroinvertebrates.
5. We speculate that during summer crayfish play an important role in temperate woodland streams by converting slowly processed leaf litter species (e.g. Rhododendron ) to fine particles which are then available to collector-gatherers (e.g. Chironomidae, Oligochaeta).     

Water-relation parameters of giant and normal cells of Capsicum annuum pericarp

Abstract. Measurements of the water-relation parameters of the giant subepidermal cells (volume, V = 0.119 to 1.658 mm³; = 0.53±0.35 mm³, SD, n = 23) and the smaller mesocarp parenchyma cells ( V = 0.10 to 0.79×10⁻³ mm³; = 0.36±0.27×10⁻³ mm³, SD, n = 6) of the inner pericarp surface of Capsicum annuum L. were made using the Jülich pressure probe. The volumetric elastic modulus ɛ for the large cells was between 1.5 and 27 MPa for a pressure range of 0.09 to 0.41 MPa. For the small cells ɛ was 0.1 to 0.6 MPa for a pressure range of 0.22 to 0.39 MPa. The turgor pressure P , the half-time of water exchange T _1/2, and the hydraulic conductivity L _p were as follows, with SD and number of replicates: large cells, P = 0.27±0.06 MPa (23), T _1/2=2.7±2.2 s (46), L _p=5.8±3.7 pm s⁻¹ Pa⁻ (46); small cells, P = 0.33±0.07 MPa (6), T _1/2= 33±10s (12), L _p=0.21±0.07 pm s⁻¹ Pa⁻¹ (12). The determination of these basic water-relation parameters is considered as a prerequisite for future ecotoxicological and phytopathological studies. The differences between the large and the small cells are discussed in relation to a desirable biophysical definition of succulence. Further, for the large cells a pressure and volume dependence of ɛ was demonstrated.     

Heterotrophic bacterioplankton production and grazing mortality rates in an Ethiopian rift-valley lake (Awassa)

SUMMARY. 1. Heterotrophic bacterioplankton growth and production rates were estimated in a tropical lake by various methods. Mean growth rates, determined by tritiated thymidine incoporation into DNA, frequency of dividing cells and increase in cell density varied between 0.013 and 0.014 (with a range of 0.006–0.026) h⁻¹ corresponding to bacterial production of 1.16–1.22 (0.34–3.63) mg C m⁻³ h⁻¹.
2. Heterotrophic bacterial production estimated from oxygen and inorganic carbon consumption in the dark were compared with these values. The oxygen method gave similar results, while values from dark carbon uptake were as much as 2.5 times higher.
3. Although the different estimates of rates of bacterial production showed different patterns, the existence of spatial (vertical) and temporal (diel and seasonal) variation was demonstrated. Bacterial production was 13–41% of the net primary production and 10–30% of gross primary production.
4. Bacterial grazing mortality rate was estimated from size-fractionation and metablic inhibitor experiments. Average grazing rates were between 0.34 and 3.77 mg C m⁻³ h⁻¹ corresponding to 76–120% of the mean bacterial production rate. Organisms 1–12 μm in size, possibly mainly ciliates. were implicated as important bacterial grazers.     

Long-term study of the ecology of wild Atlantic salmon smolts in a small Norwegian river

Backcalculated lengths at the end of the first growth season in wild Atlantic salmon Salmo salar differed significantly between parr smolting at age 1, 2 and 3 years over a period of 11 years (i.e. 1983–1993). Mean body lengths of the respective age groups at the end of the first growth period were 11·1, 6·2 and 4·7 cm, respectively. The mean percentage distribution of fish smolting at age 1, 2 and 3 was 14, 78 and 7%, and the mean smolt age was 1·95 years. Mean lengths at smolting of age groups 1, 2 and 3 were 13·6, 15·8 and 17·5 cm, respectively. Females outnumbered males among the downstream migrating smolts with a mean sex ratio (females/ males) estimated at 1·61, with a significant female surplus in 7 of the 11 years sampled. Of the smolts sampled, 14% exhibited enlarged gonads indicative of parr maturation, and all were males (37% of the parr males sampled). Mean annual smolt density from 1975 to 1996 was 13·4 individuals 100 m⁻² ranging between 0·3–31 smolts 100 m⁻². Mean densities (100 m⁻²) of the smolts aged 1, 2 and 3 years were 1·5, 9·3 and 0·9 fish, respectively. Mean annual biomass for the 22-year period (1975–1996) was estimated at 437 g 100 m⁻², with a range of variation from 136 to 683 g 100 m⁻². Smolt age 2 made up 81% of the mean annual biomass (355 g 100 m⁻²) and smolt age 1 and 3, 8% (35 g 100 m⁻²) and 11% (47 g 100 m⁻²), respectively.     

The occurrence and ecological importance of dissolved ATP in fresh water

SUMMARY. Dissolved ATP, defined as ATP which passes through 0.2 μm filters, was found in fresh water. During the spring diatom bloom in two eutrophic Danish lakes, concentrations of dissolved ATP varied between 0.1 and 3.8 μgl⁻¹, constituting 14–76% of the total ATP (particulate plus dissolved ATP). The kinetics of the light emission obtained from mixing firefly enzyme with dissolved ATP demonstrated that the major proportion of the dissolved ATP was in fact ATP. Despite some variations, the seasonal changes in dissolved ATP paralleled the changes in the increasing phytoplankton population during the rise of the diatom blooms. The dissolved ATP increased after the diatom peak, indicating that release of ATP from the phytoplankton due to mortality may be a major source of dissolved ATP.
Consumption of dissolved ATP was evaluated in uptake experiments using ³H-ATP. Rates of uptake of ³H-ATP by micro-organisms (diameter 0.2–0.6 μm) proved to be close to the rates for ³H-D-glucose uptake. The variations in ³H-ATP uptake during the diatom blooms showed non-systematic changes and ranged between 1.0 and 15.8% h⁻¹ (mean = 4.9% h⁻¹) of the quantity added. Turnover rates for dissolved ATP varied between 12 and 730 ng l⁻¹ h⁻¹ (mean = 175 ng l⁻¹). These rather high rates of turnover suggest that dissolved ATP is an important compound in the metabolism of freshwater bacteria.     

Annual changes in the nutritive state of North Sea dab

The nutritive state of dab Limanda limanda was investigated over a 2-year period at a fixed sampling site northwest of Helgoland (German Bight, North Sea), with respect to feeding habits and the accumulation of biochemical storage products. Ophiuroids formed the main weight of food organisms (50%) while polychaetes (10%), molluscs and crustaceans (<5% each) were less frequent. Feeding activity in males varied between summer and winter, while females fed more constantly. The condition factor and the hepatosomatic index showed characteristic seasonal cycles in both sexes. The glycogen content in the liver reached 40–60 mg g⁻¹ FW in summer and fell to about 10–20 mg g⁻¹ FW in late winter. Total lipids of the liver showed a distinct seasonal cycle with 400 mg g⁻¹ FW in summer and a minimum of 50–100 mg g⁻¹ FW in spring. The lipid content of the muscle ranged from 5 to 6 mg g⁻¹ FW and did not vary significantly between seasons.     

Single-cell vs. bulk activity properties of coastal bacterioplankton over an annual cycle in a temperate ecosystem

The connections between single-cell activity properties of heterotrophic planktonic bacteria and whole community metabolism are still poorly understood. Here, we show flow cytometry single-cell analysis of membrane-intact (live), high nucleic acid (HNA) content and actively respiring (CTC+) bacteria with samples collected monthly during 2006 in northern Spain coastal waters. Bulk activity was assessed by measuring ³H-Leucine incorporation and specific growth rates. Consistently, different single-cell relative abundances were found, with 60–100% for live, 30–84% for HNA and 0.2–12% for CTC+ cells. Leucine incorporation rates (2–153 pmol L⁻¹ h⁻¹), specific growth rates (0.01–0.29 day⁻¹) and the total and relative abundances of the three single-cell groups showed marked seasonal patterns. Distinct depth distributions during summer stratification and different relations with temperature, chlorophyll and bacterial biovolume suggest the existence of different controlling factors on each single-cell property. Pooled leucine incorporation rates were similarly correlated with the abundance of all physiological groups, while specific growth rates were only substantially explained by the percentage of CTC+ cells. However, the ability to reduce CTC proved notably better than the other two single-cell properties at predicting bacterial bulk rates within seasons, suggesting a tight linkage between bacterial individual respiration and biomass production at the community level.     

Potentially pathogenic vibrios associated with mussels from a tropical region on the Atlantic coast of Brazil

Mussels ( Perna perna ) harvested on the coast of Ubatuba, in three different stations in the State of São Paulo, Brazil, were examined for Vibrio spp. over a 1 year period. The ranges of most probable number (MPN 100 g^-1) were: Vibrio alginolyticus (<3–24000), V. parahaemolyticus (<3–24000), V. fluvialis (<3–1100), V. cholerae non-O1 (<3–23), V. furnissi (< 3–30), V. mimicus (< 3–9) and V. vulnificus (< 3–3). The highest incidence was observed for V. alginolyticus (92–100%), followed by V. parahaemolyticus (67–92%), V. fluvialis (34–67%), V. vulnificus (8–17%), V. furnissii (0–17%), V. mimicus (0–17%) and V. cholerae non-O1 (0–8%). Tests for virulence factors were positive in 34.1% of the vibrios in the rabbit ileal loop and 31.7% in the Dean test. Positive results in the Kanagawa test were obtained with 0.51% of V. parahaemolyticus strains. The mean values (MPN 100 g^-1) of faecal coliforms in mussels from the three regions varied from 1100 to 44000, and seawater collected at the same stations gave average values for faecal coliforms in the range 18–3300 MPN 100 ml^-1. These results highlight the potential risks of food poisoning associated with raw or undercooked seafood.     

Quantitative observations on the pulmonary anatomy of the domestic Muscovy duck (Cairina moschata)

The lungs of five female domestic Muscovy ducks, mean body weight 1.627 kg, total lung volume 48.07 cm³, were analysed by standard morphometric methods. Principal results obtained are: lung volume per unit body weight, 30.17 cm³/g; volume densities of exchange tissue relative to lung volume, 49.24%, blood capillaries relative to exchange tissue, 29.63%, tissue of the blood gas (tissue) barrier relative to exchange tissue, 5.88%; surface area of the blood-gas (tissue) barrier per unit body weight, 30.04 cm²/g; ratios of the surface area of the blood-gas (tissue) barrier per unit volume of the lung and per unit volume of exchange area, 979 cm²/cm³ and 200.06 mm²/mm³, respectively; harmonic and arithmetic mean thicknesses of the tissue barrier, 0.199 μm and 0.303 μm, respectively. The anatomical diffusing capacity of the tissue barrier for oxygen ( DtO₂ ) and the total pulmonary diffusing capacity ( DLO₂ ), 49.58 ml O₂/min/mmHg/kg and 4.55 ml O₂/min/mm Hg/kg, respectively. The lungs of the domestic Muscovy duck appear to be about as well adapted anatomically for gas exchange as the lungs of wild anatid species, and there is no clear evidence that domestication has been associated with any deterioration in the anatomical capacity for oxygen uptake. The weight-specific anatomical diffusing capacity of the lung for oxygen ( DLO₂/W ) was about 3.6 times greater than the weight-specific physiological value, a factor which falls within the expected range.     

Influence of different rations and water temperatures on the growth rates of shortfinned eels and longfinned eels

Juvenile (12–152 g) shortfinned eels Anguilla australis and longfinned eels A. dieffenbachia caught in New Zealand streams were fed squid mantle Nototodarus spp. 4 days per week in laboratory experiments. A linear multiple regression equation showed the amount of food eaten (0–2·7% w day⁻¹) explained 77·7% of the variation in specific growth rates (–0·60 to +1·07% w day⁻¹) among individual eels, while previous growth rates, water temperature (10·0–20·6°C), and eel weight (12–152 g) explained a further 5·6, 1·4 and 0·8%, respectively. Growth in length ranged from –0·3 to +0·9 mm day⁻¹. Eels which were starved and then given high rations grew substantially faster than expected. Once growth rates were adjusted for differences in ration and other factors, there were no significant differences in growth rates between species or individual fish. Growth of shortfinned eels fed maximum rations of commercial eel food depended on fish size and water temperatures and ceased below 9·0°C. Growth rates in the wild were substantially less than the maximum possible, after seasonal changes in water temperatures were taken into account, indicating that food supplies and not low water temperatures were controlling growth rates in the wild.     

Differential survival of juvenile sockeye and coho salmon exposed to low dissolved oxygen during winter

Juvenile sockeye salmon (43–78 mm) survived 100% for 24 h in cages in ice–covered Black Lake, Alaska at oxygen saturations >65% (9 mg l^–1), but only 45% at 24% saturation (3·0–3·3 mg l^–1) and none at <17% saturation (2·3 mg l^–1). All juvenile coho (50–120 mm) survived 100% for 24 h down to 21% oxygen saturation (3·1 mg l^–1), and all 50 coho survived 4–5 days at 23–24% saturation (3·2–3·3 mg l^–1).     

Indispensability of Iron for the Growth of Cultured Chick Cells

In order to clarify the role of iron in the growth promoting effect of transferrin (Tf), the effects of the following substances were examined in cultured chick skeletal myogenic cells: transition metal ions (Fe²⁺, Fe³⁺, Cr³⁺, Cu²⁺, Mn²⁺, Co²⁺, Cd²⁺, Zn²⁺ and Ni²⁺), Tf complexes with these metals and metal-free apoTf.
The cells did not grow well when incubated in a culture medium composed of Eagle's minimum essential medium and horse serum. But they grew well in the presence of Fe²⁺ or Fe³⁺ (10–100 μM) or iron-bound Tf (10–500 nM) in the medium. None of the transition metal ions other than iron was effective. Neither apoTf nor Tf complexes with these metals showed the growth promoting effect. The generality of the requirement of iron for cell growth was ascertained in the primary culture of other types of chick embryonic cells: fibroblasts, cardiac myocytes, retinal pigment cells and spinal nerve cells.
The results show that iron is one of the indispensable substances for cell growth and suggest that Tf protein plays a role in facilitating the transport of iron into the cells.     

δ13C of wood in growth-rings indicates cambial activity of drought-stressed trees of Eucalyptus globulus

1. Growth, density and δ¹³C of wood and leaf area were measured in two adjacent stands of 6 year-old Eucalyptus globulus growing in the 600–700 mm year^–1 rainfall region of south-western Australia. Study sites were identical except for differences in the availability of water owing to physical properties of soil profiles and location of sites within the landscape.
2. Abundance of ¹³C (expressed as δ¹³C) in wood of trees growing on the drought-prone site (– 24·8‰±1·4) was greater than in other trees (– 25·8‰±1·2, P <0·001) throughout the 6 years and, with further development, the δ¹³C signatures of wood may become useful indices of drought-susceptibility in plantations within a few years of establishment. The seasonal pattern of δ¹³C of wood appeared to reflect seasonal variation in water availability and duration of cambial activity.
3. Basic density of wood of trees growing on the more drought-prone site (496±14·0 kg m^–3) was reduced compared to other trees (554±5·3 kg m^–3, P <0·001). δ¹³C of wood across boundaries of growth-rings suggested that drought stopped cambial activity resulting in less production of late wood and less dense wood.
4. The stand growing on the drought-prone site had reduced growth, wood yield and leaf area but identical specific leaf area. Annual growth was correlated with the previous season's rainfall. Together, these results suggested that within the same evaporative climate, drought reduces growth primarily by reducing leaf area and that there is a lag between onset of drought and reduced productivity.     

Normal ranges of some blood chemistry parameters in adult farmed Atlantic salmon, Salmo salar

Blood samples from healthy adult Atlantic salmon fed an optimal diet in net sea pens were collected at intervals from October to May. Haematological determinations and biochemical serum analyses were carried out on 20 fish in each of seven samples. The ranges of haemato-logical values for sample means were: haematocrit 44–49%, haemoglobin 8.9–10.4 g 100ml⁻¹, red blood cell count 0.85–1.10 × 10¹² l⁻¹, MCV 441–553 × 10⁻¹⁵ 1, MCH 94–106 × 10⁻⁶ g, MCHC 19.4–21.7 g 100ml⁻¹ and leucocrit 0.43–0.96%. The ranges of enzyme activities in serum, for sample means, were: alkaline phosphatase 647–988Ul⁻¹, aspartate aminotrans-ferase 202–351 Ul⁻¹ and alanine aminotransferase 4–8 Ul⁻¹. The ranges of the other parameters analyzed in serum were: total protein 41.6–56.6 gl⁻¹, albumin 18.3–24.3 gl⁻¹, albumin/total protein ratio 39.3–44.0%, creatinine 26–46 μmol, triglycerides 2.53–4.98 μmol and cholesterol 9.3–12.8 μmol. These values are considered to be the normal ranges in healthy fish. Variations due to seasonal changes, and the clinical significance of the selected parameters, are discussed. Data showing the reproducibility of the biochemical analyses in serum are presented.     

The production of brown trout, Salmo trutta in tributaries of the Upper Wye, Wales

The population density, age structure, biomass, growth and production of brown trout were investigated in four tributaries of the upper River Wye. The populations at each site were largely maintained by immigration from nursery areas. Abundance of separate year classes at sites on the three largest tributaries reached a peak at age 2+. On the smallest stream numbers reached a peak at 1+. Recruitment occurred throughout the year but decreased with age of year classes. Maximum O+ densities ranged from 0.04 to 0.89 m⁻², and >0+ densities from 0.13 to 0.59 m⁻². Average total biomass in 1975 ranged from 2.6 to 14.2 g m⁻². Within the study sites annual trout production in 1975 ranged from 2.9–19.7 g m⁻². Production values were dependent on age structure and population mobility at the study sites. In the three largest streams 2+ and 3+ fish contributed 66.3–88.3% of total production whilst 1+ and 2+ fish contributed 74.5–84.5 % of the total in the smallest stream. The mobile (non-resident) component of the population accounted for up to 60–70% of production at certain times of the year, but over the year (1976) accounted for =30 % of total production. The resident component of the highest annual production value (19.7 g m⁻²) was estimated to be between 15.0 and 18.2 g m⁻².     

The acquisition and accumulation of inorganic carbon by the unicellular green alga Chlorella ellipsoidea

Abstract. The uptake and accumulation of inorganic carbon has been investigated in Chlorella ellipsoidea cells grown at acid or alkaline pH. Carbonic anhydrase (CA) was detected in ceil extracts but not in intact cells and CA activity in acid-grown cells was considerably less than that in alkali-grown cells. Both cell types demonstrates low K_1/2 (CO₂) values in the range pH 7.0–8.0 and these were unaffected by O₂ concentration. The CO₂ compensation concentrations of acid- and alkali-grown cells suspended in aqueous media were not significantly different in the range of pH 6.0–8.0, but at pH 5.0, the CO₂ compensation concentrations of acid-grown cells (57.4cm³ m⁻³) were lower than those of alkali-grown cells (79.2cm³ m⁻³). The rate of photo-synthetic O₂ evolution in the range pH 7.5–8.0 exceeded the calculated rate of CO₂ supply two- to three-fold, in both acid- and alkali-grown cells, indicating that HCO₃⁻ was taken up by the cells. Accumulation of inorganic carbon was measured at pH 7.5 by silicone-oil centri-fugation, and the concentration of unfixed inorganic carbon was found to be 5.1 mol m⁻³ in acid-grown and 6.4mol m⁻³ in alkali-grown cells. These concentrations were 4.6- and 5.9-fold greater than in the external medium. These results indicate that photorespiration is suppressed in both acid- and alkali-grown cells by an intracellular accumulation of inorganic carbon due, in part, to an active uptake of bicarbonate.     

Energy and ration requirements of juvenile Pacific halibut (Hippoglossus stenolepis) based on energy consumption and growth rates

Growth of captive juvenile Pacific halibut was linearly related to energy consumption (J g⁻¹ day⁻¹) at 4°C by the following equation: growth (% body weight (b.w.) day⁻¹)=0–007 (consumption J g⁻¹ day⁻¹)– 0.192; r² =0.81. Weight gain was independent of size for fish between 9 and 7000 g when growth was expressed as a function of consumption in J g⁻¹ day⁻¹. Maintenance ration determined in feeding–growth experiments averaged 27.4 J g⁻¹ day⁻¹ at 4–0°C. Small halibut ate significantly more food than large fish. Single meals following 2 day fasts averaged 4.1% b.w. for halibut under 100 g, 1.72% b.w. for 1.2 kg fish and 1.1% B.W. for 6.8 kg fish. Both large and small size categories of halibut tended to evacuate their meal in about 3 days even though small fish ate relatively larger meals. Minimum estimates for daily ration to achieve growth rates observed in the Gulf of Alaska were approximately 0.5 to 2.4% b.w. day⁻¹ depending on fish size and whether northern shrimp or yellowfin sole were their prey.