Super-CHO - A cell line capable of autocrine growth under fully defined protein-free conditions

Chinese Hamster Ovary (CHO) cells are widely used for the large scale production of recombinant biopharmaceuticals. Growth of the CHO-K1 cell line has been demonstrated in serum-free medium containing insulin, transferrin and selenium. In an attempt to get autocrine growth in protein-free medium, DNA coding for insulin and transferrin production was transfected into CHO-K1 cells. Transferrin was expressed well, with clones secreting approximately 1000 ng/10(6)cells/24h. Insulin was poorly expressed, with rates peaking at 5 ng/10(6)cells/24h. Characterisation of the secreted insulin indicated that the CHO cells were incompletely processing the insulin molecule. Site-directed mutagenesis was used to introduce a furin (prohormone converting enzyme) recognition sequence into the insulin molecule, allowing the production of active insulin. However, the levels were still too low to support autocrine growth. Further investigations revealed insulin degrading activity (presumably due to the presence of insulin degrading enzymes) in the cytoplasm of CHO cells. To overcome these problems insulin-like growth factor I (instead of insulin) was transfected into the cells. IGF-1 was completely processed and expressed at rates greater than 500 ng/10(6)cells/24h. In this paper we report autonomous growth of the transfected CHO-K1 cell line expressing transferrin and IGF-1 in protein-free medium without the addition of exogenous growth factors. Growth rates and final cell densities of these cells were identical to that of the parent cell line CHO-K1 growing in insulin, transferrin, and selenium supplemented serum-free media.

Role of galectin-1 in the olfactory nervous system

The olfactory nervous system is responsible for the detection of odors. Primary sensory olfactory neurons are located in a neuroepithelial sheet lining the nasal cavity. The axons from these neurons converge on to discrete loci or glomeruli in the olfactory bulb. Each glomerulus consists of the termination of thousands of primary axons on the dendrites of second-order olfactory neurons. What are the molecular mechanisms which guide growing olfactory axons to select sites in the olfactory bulb? We have shown that subpopulations of these axons differentially express cell surface carbohydrates and that these different subpopulations target and terminate in particular regions of the olfactory bulb. Interestingly, the olfactory neurons and glial components in the olfactory pathway between the nose and brain express galectin-1. By using in vitro assays of neurite outgrowth we found that both galectin-1 and it's ligands were capable of specifically stimulating neurite elongation. Examination of the olfactory system in galectin-1 null mutants revealed that a subpopulation of axons failed to navigate to their target site in the olfactory bulb. This is the first phenotypic effect observed in galectin-1 null mutants and indicates that galectin-1 has a role in the growth and/or guidance of a subpopulation of axons in the olfactory system during development.

Effect of body composition at selection on reproductive development in large white gilts

Fifty-four Large White gilts were used to determine the effect of body composition at selection (145 d of age) on the onset of puberty and subsequent reproductive development until 202 d of age. Gilts were assigned to one of three groups based on their backfat depth at selection: 10 to 12 mm (L), 13 to 15 mm (M), and 16 to 18 mm (F). All of the F gilts, 92% of the M gilts, and 67% of the L gilts reached puberty by slaughter at 202 d of age. Data from a subgroup (first 67% to reach puberty in each group; L = Lp, M = Mp, and F = Fp) was also used. The M (Mp) and F (Fp) gilts reached puberty at 172 d (166 d) and 170 d (166 d) of age, respectively, but the L (Lp) gilts at 184.5 d were 12 d (18 d) older than M(P < .05), Mp(P < .001), and F(P < .01), Fp (P < .001) gilts. The Lp (97.68 kg) and Mp (98.33 kg) gilts were lighter (P < .01) than Fp (108.72 kg) gilts at puberty. There were no differences (P < .05) among the L, M, and F gilts in terms of backfat depth or weight at puberty. The L (Lp) gilts had a mean of 1.16 (1.75) estrous cycles, which was lower (P < .01) than for M (Mp) and (P < .01) F (Fp) gilts, with 1.96 (2.29) and 2.25 (2.33) cycles, respectively. L (Lp) gilts had fewer (P < .05) follicles, 13.14 (12.63), than either M (Mp), 19.08 (18.71), or F (Fp), 18.25 (17.42) gilts. The number of corpora lutea was not influenced (P > .05) by grouping at selection, but Fp gilts had fewer (P < .05) corpora lutea than Mp or Fp gilts. Live weight at slaughter was not influenced (P > .10) by grouping at selection or subgrouping at puberty. The L gilts with a mean of 18.05 mm of backfat at slaughter were leaner (P < .05) than the F (21.66 mm) but not (P > .10) the M gilts (19.41 mm). Subgrouping had no effect. Fat deposition and protein deposition were higher (P < .05) in those animals that attained puberty. We conclude that the rate of fat and protein deposition seems to be one of the determinants of puberty attainment.

Functional asymmetries in the movement kinematics of patients with Tourette's syndrome

Objectives-This study adopted a concurrent task design and aimed to quantify the efficiency and smoothness of voluntary movement in Tourette's syndrome via the use of a graphics tablet which permits analysis of movement profiles. In particular, the aim was to ascertain whether a concurrent task (digit span) would affect the kinematics of goal directed movements, and whether patients with Tourette's syndrome would exhibit abnormal functional asymmetries compared with their matched controls. Methods-Twelve patients with Tourette's syndrome and their matched controls performed 12 vertical zig zag movements, with both left and right hands (with and without the concurrent task), to large or small targets over long or short extents. Results-With short strokes, controls showed the predicted right hand superiority in movement time more strongly than patients with Tourette's syndrome, who instead showed greater hand symmetry with short strokes. The right hand of controls was less force efficient with long strokes and more force efficient with short strokes, whereas either hand of patients with Tourette's syndrome was equally force efficient, irrespective of stroke length, with an overall performance profile similar to but better than that of the controls' left hand. The concurrent task, however, increased the force efficiency of the right hand in patients with Tourette's syndrome and the left hand in controls. Conclusions-Patients with Tourette's syndrome, compared with controls, were not impaired in the performance of fast, goal directed movements such as aiming at targets; they performed in certain respects better than controls. The findings clearly add to the growing literature on anomalous lateralisation in Tourette's syndrome, which may be explained by the recently reported loss of normal basal ganglia asymmetries in that disorder.

Altered loading patterns and femoral bone mineral density in children with unilateral Legg-Calve-Perthes disease

Maximization of bone accrual during the growing years is thought to be an important factor in minimizing fracture risk in old age. Mechanical loading through physical activity has been recommended as a modality for the conservation of bone mineral in adults; however, few studies have evaluated the impact of different loading regimes in growing children. The purpose of this study was to compare bone mineral density (BMD) in weight-bearing and non-weight-bearing limbs in 17 children with unilateral Legg Calve Perthes Disease (LCPD). Children with this condition have an altered weight-bearing pattern whereby there is increased mechanical loading on the noninvolved normal hip and reduced loading on the involved painful hip. Thus, these children provide a unique opportunity to study the impact of differential mechanical loading on BMD during the growing years while controlling for genetic disposition. BMD at four regions of the proximal femur (trochanter, intertrochanter, femoral neck, total of the regions) was measured using dual energy x-ray absorptiometry (DXA), and the values were compared between the involved and noninvolved sides of the children with LCPD. The BMD of both sides also were compared with normative values based on both chronological and skeletal age data. A significantly higher BMD was found on the noninvolved side over the involved side for all regions (P

Customer emotions in service failure and recovery encounters

Emotions play a significant role in the workplace, and considerable attention has been given to the study of employee emotions. Customers also play a central function in organizations, but much less is known about customer emotions. This chapter reviews the growing literature on customer emotions in employee–customer interfaces with a focus on service failure and recovery encounters, where emotions are heightened. It highlights emerging themes and key findings, addresses the measurement, modeling, and management of customer emotions, and identifies future research streams. Attention is given to emotional contagion, relationships between affective and cognitive processes, customer anger, customer rage, and individual differences.

Gender, marital status, farm size and other factors influencing the extent of cash cropping in Kenya: A case study

This article examines the effects of commercialisation of agriculture on land use and work patterns by means of a case study in the Nyeri district in Kenya. The study uses cross sectional data collected from small-scale farmers in this district. We find that good quality land is allocated to non-food cash crops, which may lead to a reduction in non-cash food crops and expose some households to greater risks of possible famine. Also the proportion of land allocated to food crops declines as the farm size increases while the proportion of land allocated to non-food cash crops rises as the size of farm increases. Cash crops are also not bringing in as much revenue commensurate with the amount of land allocated to them. With growing commercialisation, women still work more hours than men. They not only work on non-cash food crops but also on cash crops including non-food cash crops. Evidence indicates that women living with husbands work longer hours than those married but living alone, and also longer than the unmarried women. Married women seem to lose their decision-making ability with growth of commercialisation, as husbands make most decisions to do with cash crops. Furthermore husbands appropriate family cash income. Husbands are less likely to use such income for the welfare of the family compared to wives due to different expenditure patterns. Married women in Kenya also have little or no power to change the way land is allocated between food and non-food cash crops. Due to deteriorating terms of trade for non-food cash crops, men have started cultivation of food cash crops with the potential of crowding out women. It is found that both the area of non-cash crops tends to rise with farm size but also the proportion of the farm area cash cropped rises in Central Kenya.

Recreational fishing: Its expansion, its economic value and aquaculture's role in sustaining it

Economic growth usually leads to a substantial increase in the demand for recreational fishing, and China is likely to follow this trend. Factors influencing this expansion in demand are identified. Recreational fishing is of major economic importance in higher income countries and indicators of its economic significance are given. Growing demand for recreational fishing results in intensified involvement of recreational fishers in conflicts about resource use. With increasing demand for recreational fishing, recreational fishers face growing competition with one another for limited fish stocks and with commercial fishers. Their concerns for environmental threats to fish stocks also intensify. Furthermore, some strategies of recreational fishers are increasingly criticised by conservationists. Governments, therefore, are put under pressure to adopt policies to address these conflicts. Some of the policy measures adopted to help sustain the fisheries and reduce conflict are outlined. These include limits on the catch and exclusive zones for recreational fishing. However, wild stocks of fish are likely to remain under mounting harvesting and other pressures. Therefore, we need to consider the role that aquaculture can play in overcoming these problems. The possible ways in which aquaculture can do this are outlined and discussed.

Marital status, farm size and other influences on the extent of cash cropping in Kenya: A household case study

This article examines the effects of marital status, farm size and other factors on the extent of cash cropping (and allocation of land use) by means of a case study in the Nyeri district in Kenya. It was found that married women are involved in the production of a relatively greater amount of output of cash crops than unmarried women since husbands prefer to have more land under cash crops than food crops. Farmers with better quality land allocate a high proportion of it to non-food cash crops, which may expose some households to greater risks of possible famine. The proportion of land allocated to food crops declines as the farm size increases while the proportion of land allocated to non-food cash crops rises as the size of farm increases. Age is also inversely associated with subsistence. Education, though inversely associated with subsistence farming does not appear to be statistically very significant as an influence on the composition of land use and composition of farm output. With growing commercialisation, married women work more hours than unmarried ones, working not only on non-cash food crops but also on non-food cash crops. Married women seem to lose their decision-making ability with growth of agricultural commercialisation, as husbands make most decisions to do with cash crops. Married women in Kenya also have little or no power to change the way land is allocated between food and non-food cash crops.

Closing the gender gap in Bangladesh: Inequality in education, employment and earnings

Assesses the status of women in Bangladesh by analysing the dynamics of female participation in labour force and education as well as gender earnings differentials at the macro level. The study finds evidence of growing commercialisation of women’s work in Bangladesh. Although the bulk of the female labour force is engaged in self-employment activities in the rural area or in low-skilled textile and readymade garment industries in the urban area, women’s participation in high-skill and entrepreneurial jobs as well as various decision-making bodies is also on the rise. While the gender wage differentials have been considerably reduced in many industries, in general women tend to be paid less than men. There have been remarkable improvements in women’s educational attainments compared to men. Further, female access to education is found to be highly correlated with overall female labour force participation, and relative to male participation. The overall results are suggestive of an improvement in the status of women in Bangladesh.

Economic incentives for global conservation of wildlife: New international policy directions

Growing economic globalisation by extending the operation of markets is a two-edged sword as far as nature conservation is concerned. In some circumstances, it threatens the conservation of nature and in other cases, it provides economic incentives that foster the conservation of biodiversity. This article shows how global policy directions have altered in that regard. Initially the World Conservation Union (IUCN) favoured bans on trade in endangered species. This view was enshrined in the Convention on International Trade in Endangered Species (CITES). Subsequently, with the upsurge of support for market-based economic liberalism, IUCN recognised that economic and market incentives, if linked to appropriate property rights, could foster biodiversity conservation. This is reflected in the International Convention on Biological Diversity. While there is conflict between this convention and CITES, its extent has been exaggerated. As explained, in certain cases, trade restrictions of the type adopted in CITES are appropriate for nature conservation whereas the market-oriented policy of the Convention on Biological Diversity can be effective in some different situations. Whether or not the extension of markets in wildlife and wildlife products and growing economic globalisation favours nature conservation varies according to the circumstances.

Consequences of traumatic injury in fossil and recent dipnoan dentitions

Traumatic injury to the dentition of dipnoans, indirectly as a result of jaw fracture, or directly from damage to the tooth tissues, is present throughout the history of this group, in fossil and in Recent material. Bones heal, but traces of the injury are retained in the tooth tissues, permanently if the proliferative regions of the tooth plate are injured, or until the damaged dentines are removed by wear if the growing regions are left intact. Lack of resorption and repair of damaged dental hard tissues in dipnoans has implications for some models of tooth plate growth in lungfish with a permanent dentition, because this indicates that lungfish tooth plates may not have the capacity to form reparative dentine as part of the normal growth processes.

Guadalupian algae-sponge reefs in siliciclastic environments - the reefs at Lengwu (South China) compared with the reef at Iwaizaki (Japan)

Guadalupian reefs occur locally in Guangxi, Guizhou, Yunnan and Western Zhejiang, South China. Two types of Guadalupian reefs can be recognized, one is developed in carbonate platforms, e.g. those in the juncture areas of Guangxi, Yunnan and Guizhou; the other occurs in a littoral clastic shelf. The Lengwu reef in Western Zhejiang is a representative of the latter type, which is a major topic of this paper. Lengwu algae-sponge reef, more than one hundred meters in thickness, are composed mainly of sponges, hydrozoans, algae, bryozoans, microbes and lime mud. Reef limestones sit on the mudstone interbedded with fine sandstone of the proximal prodelta facies and are overlain by coarse clasts of the delta front sediments. Lengwu reef displays a lens-shaped relief, dipping and thinning from the reef core, which is remarkably different from the surrounding sediments, showing a protruding relief. Sponges and microbe/algae form bafflestone, bindstone and framestone of the reef core facies. Fore-reef facies is characterized by lithoclastic rudstone and bioclastic packstone. Reef limestone sequence is composed of three cycles and controlled by sea level changes and sediment influx. Such reef is unique among the Guadalupian reefs in South China, but seems similar in some aspects to lwaizaki reef limestones of south Kitakami in Japan. Algae and microbes growing around sponges to form rigid structure in Lengwu reef are a typical feature, which is distinctly different to Guadalupian reefs in a stable platform facies of Guizhou, Yunnan and Guangxi, South China.

Phenotypic and genetic analyses of a short measure of psychosis-proneness in a large-scale Australian twin study

Previous genetic analyses of psychosis proneness have been limited by their small sample size. For the purposes of large-scale screening, a 12-item questionnaire was developed through a two-stage process of reduction from the full Chapman and Chapman scales. 3685 individuals (including 1438 complete twin pairs) aged 18–25 years and enrolled in the volunteer Australian Twin Registry returned a mail questionnaire which included this psychosis proneness scale and the Eysenck Personality Questionnaire. Despite the brevity of the questionnaire, item and factor analysis identified four unambiguous and essentially uncorrelated scales. There were (1) Perceptual Aberration – Magical Ideation; (2) Hypomania – Impulsivity/Nonconformity; (3) Social Anhedonia and (4) Physical Anhedonia. Model-fitting analyses showed additive genetic and specific environmental factors were sufficient for three of the four scales, with the Social Anhedonia scale requiring also a parameter for genetic dominance. There was no evidence for the previously hypothesised sex differences in the genetic determination of psychosis-proneness. The potential value of multivariate genetic analysis to examine the relationship between these four scales and dimensions of personality is discussed. The growing body of longitudinal evidence on psychosis-proneness suggests the value of incorporating this brief measure into developmental twin studies.

Veterinary drug delivery: Potential for skin penetration enhancement

A range of topical products are used in veterinary medicine. The efficacy of many of these products has been enhanced by the addition of penetration enhancers. Evolution has led to not only a highly specialized skin in animals and humans, but also one whose anatomical structure and skin permeability differ between the various species. The skin provides an excellent barrier against the ingress of environmental contaminants, toxins, and microorganisms while performing a homeostatic role to permit terrestrial life. Over the past few years, major advances have been made in the field of transdermal drug delivery. An increasing number of drugs are being added to the list of therapeutic agents that can be delivered via the skin to the systemic circulation where clinically effective concentrations are reached. The therapeutic benefits of topically applied veterinary products is achieved in spite of the inherent protective functions of the stratum corneum (SQ, one of which is to exclude foreign substances from entering the body. Much of the recent success in this field is attributable to the rapidly expanding knowledge of the SC barrier structure and function. The bilayer domains of the intercellular lipid matrices within the SC form an excellent penetration barrier, which must be breached if poorly penetrating drugs are to be administered at an appropriate rate. One generalized approach to overcoming the barrier properties of the skin for drugs and biomolecules is the incorporation of suitable vehicles or other chemical compounds into a transdermal delivery system. Indeed, the incorporation of such compounds has become more prevalent and is a growing trend in transdermal drug delivery. Substances that help promote drug diffusion through the SC and epidermis are referred to as penetration enhancers, accelerants, adjuvants, or sorption promoters. It is interesting to note that many pour-on and spot-on formulations used in veterinary medicine contain inert ingredients (e.g., alcohols, amides, ethers, glycols, and hydrocarbon oils) that will act as penetration enhancers. These substances have the potential to reduce the capacity for drug binding and interact with some components of the skin, thereby improving drug transport. However, their inclusion in veterinary products with a high-absorbed dose may result in adverse dermatological reactions (e.g., toxicological irritations) and concerns about tissue residues. These a-re important considerations when formulating a veterinary transdermal product when such compounds ate added, either intentionally or otherwise, for their penetration enhancement ability. (C) 2001 Elsevier Science B.V. All rights reserved.