Cartilage calcification is associated with histological degeneration of the knee joint: a highly prevalent, age-independent systemic process.

OBJECTIVES: To investigate if cartilage calcification (CC) is a systemic process, the purpose of this study was to determine the prevalence and the amount of meniscal/hyaline CC of the knee joint in the general population by high-resolution imaging (DCR) and to evaluate the association between CC with cartilage degeneration and age. METHODS: Cross-sectional DCR-study of 180 knee joints of 90 donors (42 female/48 male, mean age 62.3y). Histological hyaline (OARSI) and meniscal (Krenn) cartilage degeneration was determined of all knees. RESULTS: CC was observed in 100% of the donors (bilaterally in 98%), hyaline cartilage calcification (HCC) in 92% and meniscal calcification (MC) in 100%. CC was detected in more than three out of six distinct cartilage areas in 84.4% of all knees. The mean amount of CC correlated between both sides of donors, the different analyzed areas of the knee joint and between the various types of cartilage structures. There was more calcification in meniscal than in hyaline cartilage (factor 5.3) and in the medial than the lateral compartment (factor 1.2). HCC/MC were already detectable with only mild cartilage lesions and the amount correlated with histological cartilage degeneration, but not with age. CONCLUSIONS: The present study provides evidence that meniscal and hyaline CC occurs in a pattern that is compatible with CC being a systemically driven process and that meniscal fibrocartilage is more prone to calcification than hyaline cartilage. Furthermore, the age-independent association between the amount of CC and the grade of degeneration in both hyaline and meniscal cartilage, suggests that CC is an obligatory early event in initiating cartilage degeneration.

[Refixation of osteochondral fragments with resorbable polylactid implants : Long-term clincal and MRI results]. / Refixation von osteochondralen Fragmenten mit resorbierbaren Polylactid-Implantaten : Klinische und MR-morphologische Langzeitergebnisse.

BACKGROUND: Refixation with resorbable implants is a common surgical treatment in patients who suffer an injury with shearing of an osteochondral flake due to trauma of the knee or the upper ankle joint. To date there are no studies which outline long-term outcomes for this procedure. The aim of this study was to evaluate long-term clinical and magnetic resonance imaging (MRI) results after refixation with resorbable polylactide (PLLA) implants. MATERIAL AND METHODS: In this retrospective study 12 patients with 13 injuries were examined 13.9 years (±1.2 years) after refixation of an osteochondral fragment of the knee (10 patients) and the upper ankle joint (2 patients) with a mean size of 3.33 cm2 (±2.33) by resorbable polylactide (PLLA) implants (nails, pins, screws, Bionx, Tampere, Finland). To objectify the clinical results eight established clinical scores (VASS, Tegner, Lysholm, McDermott, KSS, WOMAC, AOFAS, FADI+Sports) were used. Furthermore, the morphological integration of bone and cartilage was assessed by MRI (3 T) using proton-weighted and cartilage-sensitive 3D double-echo steady-state (DESS) sequences. The morphological results were objectified with a modified MRI score according to Henderson et al. RESULTS: After 13.9 years (±1.2) the patients with an injury of the knee as well as of the upper ankle joint showed good to excellent results (knee: VASS 1.2 (±1.7), Tegner 4.4 (±1.3), Lysholm 85.7 (±12.2), McDermott 90.7 (±8.6), KSS 189 (±14.2), WOMAC (6.16% (±8.45)) (upper ankle joint: VASS 2.5 (±2.5), Tegner 5.5 (±1.5), Lysholm 87 (±13), McDermott 88 (±12); WOMAC (8.54% (±8.54), AOFAS 75.5 (±24.5), FADI+Sports 118 (±18)). In all cases there was evidence of good integration of the osteochondral fragment in MRI. In five patients there was moderate subchondral cyst formation (∅ ≤1 mm); however, mild changes of the cartilage contour were found in all patients. The mean modified Henderson score achieved was 14.4 (±2.0, best 8, worst 32), which corresponds to a good morphological result. CONCLUSION: Because of good clinical and morphological results shown by MRI, refixation through resorbable implants (PLLA) can be recommended for treatment of traumatic osteochondral flakes.

[Promoting Young Talents in Trauma Surgery through Students-On-Call]. / Nachwuchsförderung in der Unfallchirurgie durch studentischen Rufdienst.

BACKGROUND: Due to restrictions on admission to medical school, changing claims to an optimized work-life balance and occupational perspectives, surgical professions in particular are struggling with strategies to motivate young academics. Surgical disziplines aim towards a profound transfer of knowledge and pique student's interest by ensuring a sustainable education at university. OBJECTIVES: The goal of this study was to evaluate a Students-On-Call System (SOCS) and to identify a financial benefit. MATERIALS AND METHODS: In this study the SOCS was compared pre-/postevaluation using questionnaires and the supporting X­rays within a curricular teaching module of orthopedic trauma surgery, with students in the fourth semester of specialism and those in the practical semester at medical school. RESULTS: The students of SOCS showed significantly better results prior to the course and afterwards than the two other groups. By establishing SOCS medical students get involved into the treatment of emergency patients in the trauma resuscitation unit (TRU) and operating room (OR). Students get the chance to enhance their comprehension of diagnostics, therapy and decision making in surgical context. This highly valuable traineeship combines a minimized teaching effort with an effective motivation of young academcis for the surgical profession. A SOCS has reduced the workload of medical colleagues. Establishing SOCS spare the residents being on call and results in reduced costs of 23,659.86 Euro per year. CONCLUSION: The results presented show that the SOCS leads to an excellent cost-benefit balance, which has been established in multiple surgical departments at the medical school of the University of Göttingen. Apart from practice-oriented surgical teaching, the SOCS is a way of promoting successful young talent saving resources in the medical on-call services.

[Mid-term clinical and MRI results after refixation of osteochondral fractures with resorbable implants]. / Mittelfristige klinische und kernspintomografische Ergebnisse nach Refixation osteochondraler Fragmente mit resorbierbaren Implantaten.

AIM: Refixation of osteochondral fractures with resorbable implants is a common surgical treatment. There are almost no studies that prove good clinical outcomes. Hence, the aim of the study was to evaluate the mid-term results after refixation of osteochondral fractures. METHODS: The results of 12 patients were recorded 6.5 (±1) years after refixation of osteochondral fractures measuring 3.4 cm (2) (±2.5) of the knee (8 ×) or the ankle joint (4 ×) with resorbable inplants. Clinical scores and a modified MRI score based on that of Henderson et al. were used. RESULTS: The clinical scores showed good to excellent results after 6.5 (±1) years (VAS pain: 1.9 [±2.4], Tegner: 5.0 [±1.7], Lysholm: 84.8 [±14.3], McDermott: 91.3 [±7.9], Knee Society: 189.4 [±12.1]). MRI showed with one exception good integration of the fractures. In 3 cases subchondral cysts could be found. In 7 cases changes in the chondral outline occurred. The effect of this was a modified Henderson score of 12.6 (±3.7). The MRI results did not correlate with the clinical outcome. CONCLUSION: Because of its good clinical results the refixation with resorbable implants can be recommended to treat osteochondral fractures.

[The treatment of patellar dislocation: a systematic review]. / Die Therapie der Patellaluxation: eine systematische Literaturanalyse.

AIM: The diagnosis and treatment of patellar dislocation is very complex. The aim of this study is to give an overview of the biomechanics of the patellofemoral joint and to point out the latest developments in diagnosis and treatment of patellar dislocation. METHOD: The authors electronically searched Medline, Cochrane and Embase for studies on the biomechanics of the patellofemoral joint and for conservative and surgical treatments after patellar dislocation. We extracted baseline demographics, biomechanical, conservation and surgical details. RESULTS: Understanding the biomechanics of the patellofemoral joint is necessary to understand the pathology of patellar dislocation. The patellofemoral joint consists of a complex system of static, active and passive stabilising factors. Patellar instability can result from osseous and soft-tissue abnormalities, such as trochlear dysplasia, patella alta, a high tibial tuberosity trochlear groove (TTTG) distance, weaknesses of the vastus medialis obliquus or a lesion of the medial retinaculum. Recent studies have focused on the medial patellofemoral ligament (MPFL) and have shown that the MPFL is the most significant passive stabiliser of the patella. Following patellar dislocation, an MRI should be standard practice to detect an MPFL rupture, osteochondral lesions or other risk factors for redislocation. An acute first-time patellar dislocation without osteochondral lesions and without severe risk factors for a redislocation should follow a conservative treatment plan. If surgical treatment is required, the best postoperative results occur when the MPFL is reconstructed, leading to a redislocation rate of 5%, this includes cases that have a dysplastic trochlea. Duplication of the medial retinaculum show very inconsistent results in the literature, possibly due to the fact that the essential pathomorphology of patellar dislocation is not addressed. Addressing the exact location of the rupture of the MPFL with a suture is possibly more convenient, especially after first-time dislocation with associated risk factors for a redislocation. Recent literature does not encourage the use of lateral release, since this can increase patellar instability. Indications for lateral release include persistent patellar instability or pain reduction in an older arthritic subject. For correcting a patellofemoral malalignment, the TTTG distance should be measured and a medial transposition of the anterior tibial tubercle hinged on a distal periosteal attachment should be considered. Cartilage lesions on the medial facet of the patella are a contra-indication for medial tubercle transposition. For cartilage lesions of the lateral facet, antero-medialization of the tibial tubercle can be successful. A tubercle osteotomy can be efficiently combined with MPFL reconstruction. We believe that patients with open epiphyseal plates should be treated with duplication of the medial retinaculum. In the presence of patellar maltracking, an additional subperiostal soft tissue release with medialisation of the distal part of the patellar tendon can be performed. CONCLUSION: It seems that the predominating factors for patellar dislocation are heterogenic morphology in combination with individual predisposition. Non-surgical treatment is typically recommended for primary patellar dislocation without any osteochondral lesions and in the absence of significant risk factors for redislocation. If surgical treatment is deemed necessary, addressing the essential pathomorphology has become the primary focus.

Analysis of light-controlled accumulation of carotenoids in mustard (Sinapis alba L.) seedlings.

Carotenoid accumulation in the cotyledons of the mustard seedling (Sinapis alba L.) is controlled by light. Besides the stimulatory function of phytochrome in carotenogenesis the experiments reveal the significance of chlorophyll accumulation for the accumulation of larger amounts of acrotenoids. A specific blue light effect was not found. The data suggest that light exerts its control over carotenoid biogenesis through two separate mechanisms: A phytochrome regulation of enzyme levels before a postulated pool of free carotenoids, and a regulation by chlorophyll draining the pool by complex-formation.

Trypanosoma cruzi is a potent inducer of interleukin-12 production in macrophages.

Cytokines produced after infection with Trypanosoma cruzi have been shown to be crucial in the determination of resistance or susceptibility. Interferon-gamma (IFN-gamma) is the predominant cytokine produced after infection and has been shown to protect susceptible mice from infection. IFN-gamma production by natural killer cells and T cells is induced by interleukin-12 (IL-12). Therefore, the aim of our study was to analyze the ability of T. cruzi to induce IL-12 production. Spleen cells and bone marrow-derived macrophages incubated with T. cruzi trypomastigotes induced high amounts of IL-12p40 mRNA as shown by reverse transcriptase-polymerase chain reaction. Lipopolysaccharide (LPS) was less efficient in inducing IL-12p40-specific mRNA. Furthermore, biologically active IL-12, detected by the capacity of the supernatant of infected macrophages to induce IFN-gamma production in spleen cells, was produced at very high levels. In comparison, macrophages stimulated with LPS secreted drastically less IL-12. Interestingly, only live, UV- or gamma-irradiated trypanosomes, but not heat-killed parasites or lysates, were functional in this respect. In a kinetic study, in the supernatant obtained from cultures of infected macrophages, IL-12 was already detectable at 2 h after infection, peaked at 32 h and declined after 45 h.

Infection with Trypanosoma cruzi selectively upregulates B7-2 molecules on macrophages and enhances their costimulatory activity.

T-cell-mediated immune responses are essential for protection against infection with the protozoan Trypanosoma cruzi. In this study, we investigated the influence of infection of murine macrophages with T. cruzi on costimulatory signals for T lymphocytes provided by these cells. We demonstrate that bone marrow-derived macrophages (BMMph) selectively and strongly upregulate expression of B7-2 molecules after infection, while the expression of other costimulatory molecules such as B7-1, intercellular adhesion molecule 1, lymphocyte function-associated antigen 3, and heat-stable antigen is not significantly affected. Infection by live trypanosomes was required. As a consequence of the strong B7-2 upregulation, the infected macrophages are able to induce proliferation of splenic CD4+ T cells in the presence of anti-CD3 antibodies with much higher efficiency than uninfected macrophages. Costimulation could be inhibited by an antibody to B7-2. Furthermore, costimulatory activity for established T-cell clones of Th1 and Th2 phenotype was also strongly enhanced in BMMph by infection with T. cruzi. Th1 cells stimulated either via anti-CD3 antibodies or via specific antigen proliferated with higher efficiency in the presence of infected macrophages than in the presence of uninfected cells. BMMph stimulated with gamma interferon (IFN-gamma), expressing elevated levels of B7-2 molecules, are also able to enhance Th1 cell proliferation, which was highest, using macrophages which were infected and in parallel were stimulated with IFN-gamma. Noteworthy, for cloned Th2 cells, the mechanism of costimulation differed, because costimulation of Th2 cells was not dependent on B7-2 upregulation but was due to secretion of interleukin-1alpha. These findings demonstrate that infection of macrophages with T. cruzi transforms the macrophage into a potent costimulatory cell based on the induction of two different costimulatory activities.

Regulation of enzyme levels by phytochrome in mustard cotyledons: Multiple mechanisms?

Phytochrome controls the appearance of many enzymes in the mustard (Sinapis alba L.) cotyledons. The problem has been whether the effect of phytochrome on the appearance of enzymes in this organ is due to a common initial action of Pfr, e.g. due to the liberation of a "second messenger". We have compared the modulation by light (phytochrome) of the appearance of phenylalanine ammonia lyase (PAL)(+) and ribulosebisphosphate carboxylase (Carboxylase)(+). PAL becomes detectable in the mustard cotyledons at 27 h after sowing while Carboxylase starts to appear only at 42 h after sowing (starting points, 25° C). The starting points cannot be shifted by light. As a major result, in the case of PAL the inductive effect of continuous red light (given from the time of sowing) remains fully reversible by 756 nm-light up to the starting point (27 h after sowing) while with Carboxylase full reversibility in continuous red light is lost at approximately 15 h after sowing. While the induction of Carboxylase is already saturated at a very low level of Pfr (e.g. continuous 756 nm-light saturates the response) and does not depend on irradiance (e.g. continuous 675 mW m(-2) red light and 67.5 mW m(-2) red light lead to the same time course), PAL induction is a graded response over a wide range of Pfr doses and depends strongly on the fluence rate (high irradiance response, HIR). It is concluded that PAL induction and Carboxylase induction are not only separated in time but differ in every regard except that both responses are mediated by phytochrome.The present data support the previous conclusion that the specification of the temporal and spatial pattern of development is independent of phytochrome even though the realization of the pattern of development can only occur in the presence of phytochrome (Pfr). It seems that there is no feedback from pattern realization to pattern specification.

Light control of plastogenesis and ribulosebisphosphate carboxylase levels in mustard seedling cotyledons.

We have studied the problem whether the phytochrome-mediated accumulation of ribulosebisphosphate carboxylase ("carboxylase"; EC 4.1.1.39) in the cotyledons of sinapis alba L. is related to size, ultrastructure, or organization of the plastid compartment. We have shown that under different light conditions (e.g. continuous far-red light, continuous white light) which lead to conspicuously different plastids the time course of the enzyme levels remains precisely the same. It is concluded that the onset and the rate of carboxylase accumulation is not related to the organizational state of the plastid compartment as discernible under the electron microscope.

In vitro effects of monoterpenes on chloroplast membranes.

A chloroplast preparation was extracted from squash (Cucurbita pepo (L.) var. Senator). Enrichment of intact chloroplasts was achieved by continuous free-flow electrophoresis. The addition of monoterpenes, detergent and free fatty acids changed the elecrophoretic separation pattern characteristically. Monoterpene-dependent degradation of envelope membranes could be prevented by addition of α-tocopherol prior to monoterpene incubation.Photosynthetic electron transport of photosystem II was completely inhibited by ß-pinene, Triton X-100 and linolenic acid. Inhibition could be modulated by addition of α-tocopherol or lecithin (phosphatidylcholine) either before or after inhibition by monoterpenes and detergent.Percentage reconstitution of photosynthetic electron transport inhibited by ß-pinene depended on light conditions and incubation time.

Oxalate accumulation from citrate by Aspergillus niger. II. Involvement of the tricarboxylic acid cyclase.

Carbon-14 was incorporated into oxalate and CO2 from either citrate-1,5-14C, succinate-1,4-14C, or fumarate-1,4-14C by cultures of Aspergillus niger pregrown on a medium which contained glucose as the sole carbon source and which did not allow citrate accumulation. In cell-free extracts of mycelium forming oxalate and CO2 from added citrate the following enzymes of the tricarboxylic acid (TCA) cycle were identified: citrate synthase CE 4.1.3.7), aconitate hydratase (EC4.2.1.3), NAD and NADP-dependent isocitrate dehydrogenase (EC 1.1.1.41, 1.1.1.42), (alpha-oxoglutarate dehydrogenase (EC 1.2.4.2), succinate dehydrogenase (EC 1.3.99.1), fumarate hydratase (EC 4.2.1.2), and malate dehydrogenase (EC 1.1.1.37). The in vitro activity of aconitate hydratase and of NADP-dependent isocitrate dehydrogenase was shown to be almost identical to the rate of in vivo degradation of citrate or to exceed this rate. The degradation of citrate to oxalate was inhibited completely by 9 mM fluoroacetate. It is concluded that the TCA cycle is involved in the formation of oxalate from citrate.

The capacity of bone marrow-derived macrophages to process bovine insulin is regulated by lymphokines.

The antigen presentation capacity of bone marrow-derived macrophages (BMMph) was shown previously to be increased after stimulation with the lymphokines IFN-gamma or granulocyte macrophage colony stimulating factor (GM-CSF) respectively. Using bovine insulin (BI) as antigen, activation of BMMph with GM-CSF resulted in the generation of highly effective presenting cells. In contrast, IFN-gamma-treated macrophages, although better presenters than untreated BMMph, stimulated BI-specific T hybridoma cells only weakly to IL-2 production despite the fact that they expressed drastically more MHC class II molecules than GM-CSF-activated BMMph. Therefore we analyzed whether the observed differences in the presentation function of GM-CSF- and IFN-gamma-pulsed BMMph might be a consequence of differences in their capability to process BI. By blocking thiol and serine proteases with specific inhibitors or by raising the intracellular pH with chloroquine during BI pulse, the presentation capacity of IFN-gamma-activated BMMph was significantly enhanced, while the presentation function of GM-CSF-pulsed macrophages was not positively influenced. These findings suggest that the activity of thiol/serine proteases in BMMph is differently influenced by the two cytokines. A regulatory influence of the cytokines on the activity of metallo and acidic proteases was not observed. Thus, the weaker BI presentation capacity of IFN-gamma-treated macrophages as compared with GM-CSF-pulsed cells seems to be the consequence of a more excessive degradation of BI and destruction of the antigenic epitope.

Granulocyte-macrophage colony-stimulating factor activates macrophages derived from bone marrow cultures to synthesis of MHC class II molecules and to augmented antigen presentation function.

The effects of granulocyte-macrophage (GM)-CSF on the synthesis of MHC class II molecules and on the Ag presentation capacity by bone marrow derived macrophages (BMM phi) was investigated. BMM phi obtained by in vitro culture in the presence of macrophage-CSF were negative for synthesis of I-A molecules and induced the Ag-mediated proliferation of insulin-specific T clone cells with lower efficiency than splenic accessory cells. After pulse treatment with GM-CSF for 24 to 48 h, day 12 BMM phi exhibited highly efficient Ag presentation function which was superior to that induced by IFN-gamma. Expression of membrane-bound IL-1 was augmented significantly by GM-CSF, but not by IFN-gamma. However, the T cell clone used to probe for accessory cell function of BMM phi was not dependent on IL-1 for optimal proliferation. Concomitantly, GM-CSF induced the de novo synthesis of I-A molecules, although to a lesser extent than optimal doses of IFN-gamma. Thus GM-CSF appears to elicit properties in addition to Ia molecule synthesis and membrane IL-1 expression in BMM phi being essential for efficient accessory cell function to the T clone cells. The activation of BMM phi by GM-CSF was reversible and could be repeated. These data show that GM-CSF exerts a modulatory influence on preformed BMM phi, reversibly activating cells to Ia biosynthetic potential and pronounced accessory cell capacity, thus rendering the explanation unlikely that differentiation of precursor cells into a constitutively functional state had occurred.

Trypanosoma cruzi induces strong IL-12 and IL-18 gene expression in vivo: correlation with interferon-gamma (IFN-gamma) production.

IFN-gamma, produced after infection with Trypanosoma cruzi, has been shown to be crucial in the determination of resistance or susceptibility. We have performed a detailed study on the expression of IFN-gamma and of the IFN-gamma-inducing cytokines IL-12 and IFN-gamma-inducing factor (IGIF)/IL-18 with regard to time course and tissue localization. IFN-gamma was present in high amounts in the serum and in the supernatants of unseparated spleen cells and isolated CD4+ and CD8+ T cells from the spleens of infected mice which were stimulated ex vivo with T. cruzi. Using the in situ hybridization technique we demonstrate that IL-12 p40 messages were expressed in the spleen and increased during infection, correlating with the expression of IFN-gamma transcripts. Furthermore, we show for the first time that the mRNA for the cytokine IL-18 was induced by a parasitic infection and that this expression increased during infection with T. cruzi. Interestingly, the message for IL-18 was produced earlier during infection and already had declined until day 38, when IFN-gamma and IL-12 p40 transcripts were optimally expressed. Surprisingly, the changes in IL-12 and IL-18 mRNA production were clearly seen only by in situ hybridization, but less clearly by quantitative reverse-transcriptase polymerase chain reaction (RT-PCR). This is possibly due to the extensive activation and proliferation of spleen cells observed during infection leading to a dilution of these specific mRNAs.

The long term effects of traumatic brain injury on the roles of caregivers.

This study was conducted to identify the long term effects of traumatic brain injury (TBI) on the roles of caregivers. The subjects consisted of 155 caregivers of survivors with TBI who were randomly selected from 15 midwestern state brain injury association databases. A questionnaire was developed by the researchers to determine factors affecting role changes of caregivers. The Role Checklist, by Barris, Oakley and Kielhofner, was also included with the questionnaire. Both were mailed to each selected caregiver and used for data gathering. The data obtained were analysed to determine existing trends in the data. Graphs were utilized to depict the trends that was identified. The following trends and conclusions established by this research include: (a) behavioural effects of the survivor with a TBI are associated with the number of role changes experienced by caregivers; (b) participation in support systems is associated with the number of role changes experienced by caregivers; and (c) caregivers who care for a person with a TBI in the home will show a larger number of role changes than those who do not provide direct care for a person with a TBI.

Ribulose bisphosphate carboxylase capacity and chlorophyll content in developing seedlings of Chenopodium rubrum L. growing under light of different qualities and fluence rates.

In order to evaluate the aclimation of Chenopodium seedlings to different quantum fluence rates of R and BL, kinetics of Rubisco capacity, Chl content and chloroplast structure were studied. Under monochromatic light photoreceptors are stimulated selectively and their influence on biosynthetis capacities during chloroplast development can be studied.R irradiations saturate Rubisco capacity even at the lowest quantum fluence rates applied, whereas Chl a+b synthesis depends strongly upon fluence rate of R. Under BL irradiations, both Rubisco capacity and Chl content are fluence rate dependent. R irradiations favour Chl b synthesis relative to Chl a, whereas under BL Chl a content is high relative to Chl b. Under R irradiation Pfr is the main photoreceptor involved in regulation of Rubisco capacity whereas under BL a specific BL absorbing photoreceptor may control the response. From the fluence rate dependency under BL irradiations it is concluded that the blue region of the day light spectrum may be the sensor for monitoring fluence rate and causing the characteristic changes in shade and high/low WL adaptation with respect to Rubisco levels in Chenopodium.

Modulation of proliferation and lymphokine secretion of murine CD4+ T cells and cloned Th1 cells by proteins of the extracellular matrix.

In this study we investigated the co-stimulatory signaling capacity of diverse proteins of the extracellular matrix (ECM) for murine resting CD4+ T cells and Th1 clone cells, activated by immobilized anti-CD3 mAb. ECM proteins used in various concentrations had no effect on IL-2 production or proliferation of highly purified CD4+ T cell populations. When the preparation of CD4+ T cells contained contaminating accessory cells, IL-2 secretion and proliferation was enhanced in the presence of co-immobilized collagens or fibronectin. However, the level of proliferation attainable by added irradiated splenocytes was not reached. Using Th1 cell clone M4, enhanced production of IL-2 in the presence of immobilized ECM proteins was observed. At a submitogenic anti-CD3 mAb dose, proliferation of M4 T cells was augmented by the ECM proteins in a concentration range that optimally induced IL-2 production. IL-2R p55 was up-regulated on M4 T cells by collagen type IV and fibronectin to the same level that was induced by exogenously added IL-2, whereas added accessory cells induced a higher level of IL-2R p55 expression. Likewise, in dot-blot analysis a comparable quantity of IL-2R p55- and p75-specific transcripts was induced by collagen type IV or fibronectin and by IL-2, which was lower than that induced by antigen-presenting cells. Our data suggest that the enhanced proliferation of M4 T cells induced by ECM proteins is not the consequence of direct up-regulation of IL-2R, but appears to be due indirectly to elevated secretion of IL-2. At an optimal anti-CD3 mAb dose the collagens inhibited M4 T cell proliferation. Diminished cell surface expression of IL-2R p55 following stimulation with anti-CD3 mAb plus collagen type IV compared with anti-CD3 mAb alone was observed and may be responsible for growth inhibition.

Inhibition of carotenoid biosynthesis by the herbicide SAN 9789 and its consequences for the action of phytochrome on plastogenesis.

Treatment of the mustard (Sinapis alba L.) seedling with the herbicide SAN 9789 inhibits synthesis of colored carotenoids and interferes with the formation of plastid membrane lipids without affecting growth and morphogenesis significantly. In farred light, which is hardly absorbed by chlorophyll, development of plastid ultrastructure, synthesis of ribulosebisphosphate carboxylase and synthesis of chlorophyll are not affected by SAN 9789. It is concluded that normal phytochrome actions on plastid structural development, protein and chlorophyll syntheses are not affected by the absence of carotenoids provided that there is no significant light absorption in chlorophyll. The findings show that the inhibition of synthesis of one set of plastid membrane components (the carotenoids) does not stop synthesis of other components such as chlorophyll and does not halt membrane assembly. Supplementary experiments with the closely related compound SAN 9785, which affects the amount and composition of plastid lipids but not carotenoid and chlorophyll syntheses, suggest that the effect of the herbicide SAN 9789 is due exclusively to its inhibition of synthesis of colored carotenoids. In the presence of SAN 9789 white or red light at high fluence rate causes photodestruction of chlorophyll and ribulosebisphosphate carboxylase and photodecomposition of thylakoids. These effects are interpreted as resulting exclusively from the self-photooxidation and photosensitizing action of chlorophyll once the protection by carotenoids of chlorophyll against self- and sensitized photooxidation is lost.

Expression of macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and RANTES genes in lymph nodes from HIV+ individuals: correlation with a Th1-type cytokine response.

The in vivo response of the immune system after HIV infection in regard to cytokine production and C-C chemokine synthesis is not well known. Here we have analysed cytokine and chemokine mRNA production in lymph nodes with follicular hyperplasia (FHLN) of HIV-infected patients by in situ hybridization using anti-sense mRNA probes. The synthesis of mRNAs for interferon-gamma (IFN-gamma), IL-12p35, IL-12p40, IL-4, and for the C-C chemokines RANTES, MIP-1alpha, and MIP-1beta was compared with that of lymph nodes from non-infected individuals to define HIV-specific events. Only few cells expressing IFN-gamma, RANTES, MIP-1alpha, and MIP-1beta mRNAs were detectable in the T-dependent area of lymph nodes from HIV-negatives. In contrast, in FHLN from HIV+ patients a high number of IFN-gamma, RANTES, MIP-1alpha, and MIP-1beta mRNA-containing cells were detectable. Remarkably, only single individual IL-12p35 mRNA-producing cells were present in the T-dependent area from both HIV+ and HIV lymph nodes. Furthermore, the low number of IL-12p40 mRNA-expressing cells did not differ between HIV+ and HIV- lymph nodes. This indicates that IFN-gamma is expressed independently of IL-12, possibly by a direct T cell-mediated reaction. IL-4 mRNA-producing cells were hardly detectable in infected and control lymph nodes. The same findings were made in a limited number of samples from patients with advanced disease. Thus, these results demonstrate that a high IFN-gamma production is accompanied by a strong expression of MIP-1alpha, MIP-1beta, and RANTES in the lymph node after HIV infection. This favours the idea that a Th1-type immune response correlates with a preferential production of C-C chemokines in FHLN of HIV+ patients.