[Stress in the dental practice: 'Suddenly I wasn't able to work anymore']. / Stress in de tandartspraktijk: 'Ineens lukte werken niet meer'.

Recent research indicates 20% of dentists to experience a high degree of stress, physical pain and distress, and insomnia, which is associated with loss of productivity, both in quantity and quality. Physical distress and pain, but also difficulties with colleagues, demanding patients or loss of productivity are clearly associated with the prevalence of stress. Stress complaints are often recognised too late. 2 fictitious cases from the daily practice of occupational health experts/ergonomists are discussed. The first case concerns a 45-year-old dentist who was referred to an occupational health expert regarding severe fatigue, listlessness and a sense of loss of control. After the occupational health expert had inventoried his full workload, it was shown not to have increased. With the help of relaxation therapy, a professional organiser and an organisational and business psychologist, the dentist was able to return to work full time after 9 months. In the second case, a 55-year-old endodontist presented with neck pain. The occupational health expert/ergonomist observed postural problems and stress. The client reported she suffered from fatigue. The endodontist did not acknowledge these stress issues and wanted to address improvement of posture in particular, which was effective. The occupational health expert was nevertheless able to introduce measures to reduce stress and fatigue; after some time, the endodontist enjoyed her return to work.

[Spotlight on light]. / Zicht op licht.

An adequate perception is essential for a dentist because details are often just or just not visible. A proper use of light in the oral cavity (mouth of the patient), around the mouth and in the rest of the oral surgery, together with the use of light colours ought to make possible an optimal perception and colour matching. This can also prevent tiring adaptation of the eyes to too strong varying levels of light. Principles of a proper use of light and colours are described.

[Profession-related disorders of the locomotor apparatus. A multi-causal problem?]. / Beroepsgebonden aandoeningen van het bewegingsapparaat. Een multicausaal probleem?

This article provides information on the prevention of disorders among dentists and dental hygienists, for instance neck, shoulders, upper extremity, wrist and hand disorders. Many daily (strained) operating postures are described, and research data from the Sonde prevention project is mentioned. This data is compared with the newly implemented NEN-ISO-standard 11226. The standard lists the limits of static working postures. It becomes clear from this data that many dentists exceed these limits daily and work in overstrained postures. The article will further go into the complicated interrelationship of and connection between various interactive factors which intentionally and often unintentionally lead to static use of muscles. It may be a matter of cumulative effects that are multi-causally related, with the possible consequences. A further awakening within the dental profession, conditioning of young students into adopting a correct operating posture and purchasing ergonomically sound equipment may contribute to a healthier situation.

Cold shock proteins of Lactococcus lactis MG1363 are involved in cryoprotection and in the production of cold-induced proteins.

Members of the group of 7-kDa cold-shock proteins (CSPs) are the proteins with the highest level of induction upon cold shock in the lactic acid bacterium Lactococcus lactis MG1363. By using double-crossover recombination, two L. lactis strains were generated in which genes encoding CSPs are disrupted: L. lactis NZ9000 Delta AB lacks the tandemly orientated cspA and cspB genes, and NZ9000 Delta ABE lacks cspA, cspB, and cspE. Both strains showed no differences in growth at normal and at low temperatures compared to that of the wild-type strain, L. lactis NZ9000. Two-dimensional gel electrophoresis showed that upon disruption of the cspAB genes, the production of remaining CspE at low temperature increased, and upon disruption of cspA, cspB, and cspE, the production of CspD at normal growth temperatures increased. Northern blot analysis showed that control is most likely at the transcriptional level. Furthermore, it was established by a proteomics approach that some (non-7-kDa) cold-induced proteins (CIPs) are not cold induced in the csp-lacking strains, among others the histon-like protein HslA and the signal transduction protein LlrC. This supports earlier observations (J. A. Wouters, M. Mailhes, F. M. Rombouts, W. M. De Vos, O. P. Kuipers, and T. Abee, Appl. Environ. Microbiol. 66:3756-3763, 2000). that the CSPs of L. lactis might be directly involved in the production of some CIPs upon low-temperature exposure. Remarkably, the adaptive response to freezing by prior exposure to 10 degrees C was significantly reduced in strain NZ9000 Delta ABE but not in strain NZ9000 Delta AB compared to results with wild-type strain NZ9000, indicating a notable involvement of CspE in cryoprotection.

The role of cold-shock proteins in low-temperature adaptation of food-related bacteria.

There is a considerable interest in the cold adaptation of food-related bacteria, including starter cultures for industrial food fermentations, food spoilage bacteria and food-borne pathogens. Mechanisms that permit low-temperature growth involve cellular modifications for maintaining membrane fluidity, the uptake or synthesis of compatible solutes, the maintenance of the structural integrity of macromolecules and macromolecule assemblies, such as ribosomes and other components that affect gene expression. A specific cold response that is shared by nearly all food-related bacteria is the induction of the synthesis so-called cold-shock proteins (CSPs), which are small (7 kDa) proteins that are involved in mRNA folding, protein synthesis and/or freeze protection. In addition, CSPs are able to bind RNA and it is believed that these proteins act as RNA chaperones, thereby reducing the increased secondary folding of RNA at low temperatures. In this review established and novel aspects concerning the structure, function and control of these CSPs are discussed. A model for bacterial cold adaptation, with a central role for ribosomal functioning, and possible mechanisms for low-temperature sensing are discussed.

Changes in glycolytic activity of Lactococcus lactis induced by low temperature.

The effects of low-temperature stress on the glycolytic activity of the lactic acid bacterium Lactococcus lactis were studied. The maximal glycolytic activity measured at 30 degrees C increased approximately 2.5-fold following a shift from 30 to 10 degrees C for 4 h in a process that required protein synthesis. Analysis of cold adaptation of strains with genes involved in sugar metabolism disrupted showed that both the phosphoenolpyruvate-dependent sugar phosphotransferase system (PTS) subunit HPr and catabolite control protein A (CcpA) are involved in the increased acidification at low temperatures. In contrast, a strain with the PTS subunit enzyme I disrupted showed increased acidification similar to that in the wild-type strain. This indicates that the PTS is not involved in this response whereas the regulatory function of 46-seryl phosphorylated HPr [HPr(Ser-P)] probably is involved. Protein analysis showed that the production of both HPr and CcpA was induced severalfold (up to two- to threefold) upon exposure to low temperatures. The las operon, which is subject to catabolite activation by the CcpA-HPr(Ser-P) complex, was not induced upon cold shock, and no increased lactate dehydrogenase (LDH) activity was observed. Similarly, the rate-limiting enzyme of the glycolytic pathway under starvation conditions, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), was not induced upon cold shock. This indicates that a factor other than LDH or GAPDH is rate determining for the increased glycolytic activity upon exposure to low temperatures. Based on their cold induction and involvement in cold adaptation of glycolysis, it is proposed that the CcpA-HPr(Ser-P) control circuit regulates this factor(s) and hence couples catabolite repression and cold shock response in a functional and mechanistic way.

Physiological and regulatory effects of controlled overproduction of five cold shock proteins of Lactococcus lactis MG1363.

The physiological and regulatory effects of overproduction of five cold shock proteins (CSPs) of Lactococcus lactis were studied. CspB, CspD, and CspE could be overproduced at high levels (up to 19% of the total protein), whereas for CspA and CspC limited overproduction (0.3 to 0.5% of the total protein) was obtained. Northern blot analysis revealed low abundance of the cspC transcript, indicating that the stability of cspC mRNA is low. The limited overproduction of CspA is likely to be caused by low stability of CspA since when there was an Arg-Pro mutation at position 58, the level of CspA production increased. Using two-dimensional gel electrophoresis, it was found that upon overproduction of the CSPs several proteins, including a number of cold-induced proteins of L. lactis, were induced. Strikingly, upon overproduction of CspC induction of CspB, putative CspF, and putative CspG was also observed. Overproduction of CspB and overproduction of CspE result in increased survival when L. lactis is frozen (maximum increases, 10- and 5-fold, respectively, after 4 freeze-thaw cycles). It is concluded that in L. lactis CSPs play a regulatory role in the cascade of events that are initiated by cold shock treatment and that they either have a direct protective effect during freezing (e.g., RNA stabilization) or induce other factors involved in the freeze-adaptive response or both.

Cold shock proteins and low-temperature response of Streptococcus thermophilus CNRZ302.

Low-temperature adaptation and cryoprotection were studied in the thermophilic lactic acid bacterium Streptococcus thermophilus CNRZ302. S. thermophilus actively adapts to freezing during a pretreatment at 20 degrees C, resulting in an approximately 1, 000-fold increased survival after four freeze-thaw cycles compared to mid-exponential-phase cells grown at an optimal temperature of 42 degrees C. No adaptation is observed when cells are exposed to a temperature (10 degrees C) below the minimal growth temperature of the strain (just below 15 degrees C). By two-dimensional gel electrophoresis several 7-kDa cold-induced proteins were identified, which are the major induced proteins after a shift to 20 degrees C. These cold shock proteins were maximally expressed at 20 degrees C, while the induction level was low after cold shock to 10 degrees C. To confirm the presence of csp genes in S. thermophilus, a PCR strategy was used which yielded products of different sizes. Sequence analysis revealed csp-like sequences that were up to 95% identical to those of csp genes of S. thermophilus ST1-1, Streptococcus dysgalactiae, Streptococcus pyogenes, and Lactococcus lactis. Northern blot analysis revealed a seven- to ninefold induction of csp mRNA after a temperature shift to 20 degrees C, showing that this thermophilic bacterium indeed contains at least one cold-inducible csp gene and that its regulation takes place at the transcriptional level.

Microbial stress response in minimal processing.

"Bacteria have evolved adaptive networks to face the challenges of changing environments and to survive under conditions of stress. Therefore, the efficiencies of inactivation and preservation methods need to be assessed, especially with regard to the enormous potential of food pathogens to adapt to a wide variety of stress conditions. All adaptive responses, whether to changing nutrients or to various stresses encountered in minimal processing, involve a series of genetic switches that control the metabolic changes taking place. A common regulatory mechanism involves the modification of sigma (sigma) factors whose primary role is to bind to core RNA polymerase conferring promoter specificity directing expression of specialty regulons involved in heat-shock response, the chemotactic response, sporulation, and general stress response. Examples of the latter regulon in Gram-positive bacteria (the sigmaB regulon) and in Gram-negative bacteria (the RpoS regulon) will be discussed in more detail. Cellular adaptive mechanisms to starvation, cold shock, heat shock, (weak) acids, high osmolarity and high hydrostatic pressure will be described and their significance in food preservation and safety will be discussed."

Physiological activity of Campylobacter jejuni far below the minimal growth temperature.

The behavior of Campylobacter jejuni at environmental temperatures was examined by determining the physiological activities of this human pathogen. The minimal growth temperatures were found to be 32 and 31 degrees C for strains 104 and ATCC 33560, respectively. Both strains exhibited a sudden decrease in growth rate from the maximum to zero within a few degrees not only near the maximal growth temperature but also near the minimal growth temperature. This could be an indication that a temperature-dependent transition in the structure of a key enzyme(s) or regulatory compound(s) determines the minimal growth temperature. Oxygen consumption, catalase activity, ATP generation, and protein synthesis were observed at temperatures as low as 4 degrees C, indicating that vital cellular processes were still functioning. PCR analysis showed that cold shock protein genes, which play a role in low-temperature adaptation in many bacteria, are not present in C. jejuni. The fact that chemotaxis and aerotaxis could be observed at all temperatures shows that the pathogen is able to move to favorable places at environmental temperatures, which may have significant implications for the survival of C. jejuni in the environment.

A possible role of ProP, ProU and CaiT in osmoprotection of Escherichia coli by carnitine.

Exogenously provided carnitine (beta-hydroxy-L-tau-N-trimethyl aminobutyrate) was found to stimulate aerobic growth of enterohaemorrhagic Escherichia coli O157:H7 in a medium of inhibitory osmotic strength. Its osmoprotective ability is comparable with that of betaine. As carnitine is an important compound in mammalian tissues, it is suggested that it might play a role in the growth of the pathogen on low water activity (aw) meat products. Using specific uptake mutants of E. coli K-12, it was established that, under osmotic stress, carnitine accumulates in the cytoplasm following import through the ProP and ProU transport systems. Betaine and carnitine also protect E. coli cells while growing anaerobically at inhibitory osmolarity. Under these conditions, an E. coli K-12 strain with lesions in both proP and proU accumulates low levels of L-carnitine but fails to accumulate betaine when these compounds are supplied in the external medium. This is probably a result of uptake of L-carnitine by the secondary transporter CaiT. The caiT gene forms part of the caiTABCDE operon which encodes the carnitine pathway, and is transcribed during anaerobic growth in the presence of carnitine. However, further experiments revealed that the carnitine pathway, including CaiT, does not play a significant role in osmoregulation of E. coli during anaerobiosis. Together, the results indicate that ProP and ProU are the sole transport systems involved in carnitine influx, both in aerobically and anaerobically osmotically stressed E. coli cells.