Bilateral simultaneous cataract surgery and intraocular lens implantation in an adult female chimpanzee (Pan troglodytes) inducing monovision.

BACKGROUND: Cataract is the major cause of visual impairment in humans. Phacoemulsification with intraocular lens (IOL) implantation is the standard technique for cataract treatment with a high success rate. In a few cases, the surgical cataract procedure and lens implantation have been applied in non-human primates. CASE DESCRIPTION: A +/- 40-year-old female chimpanzee (Pan troglodytes) in captivity was diagnosed with mature cataract optical density (OD) and posterior subcapsular cataract combined with cortical opacities OS after ophthalmic examination. To restore vision and facilitate far- and near sight, phacoemulsification OU with +22.5 diopter (D) IOL implantation OD and + 24 D OS were performed. Despite complicated surgery OD due to posterior capsular rupture, the outcome was successful during 1-year follow-up. The chimpanzee regained adequate vision, normal behavior, and was successfully re-introduced to her group of chimpanzees. CONCLUSION: This is the first case report of a simultaneous bilateral cataract surgery with IOL implantations in both eyes, targeting emmetropia OS and near vision OD resulting in monovision, in a chimpanzee. Vision was restored without postoperative complications.

The ecology of nitrogen fixation in cyanobacterial mats.

All cyanobacterial mats that have been investigated have been proven to be diazotrophic, i.e., use atmospheric dinitrogen (N(2)) as the source of nitrogen. Many cyanobacteria possess the capacity to fix N(2) and different species have evolved various ways to cope with the sensitivity of nitrogenase toward oxygen which is produced by these oxygenic phototrophs. These different strategies give rise to complex patterns of nitrogenase activity in microbial mats. Nitrogenase activity may exhibit complex variations over a day-night cycle but different types of microbial mats may also have their own characteristic patterns. Besides the cyanobacteria, numerous other members of the Bacteria as well as some Archaea are known to be diazotrophic. The complexity of the microbial community and of the observed patterns of nitrogenase activity makes it difficult to understand how the different groups of organisms contribute to N(2) fixation in microbial mats. Cyanobacteria have ample access to energy (sunlight) and reducing equivalents (water) and therefore easily satisfy the demands of nitrogenase. As well, since they also fix CO(2), they are able to synthesize the acceptor molecules for the fixed nitrogen. However, it is also feasible that other diazotrophs in a joint venture with cyanobacteria are responsible for the bulk of the fixed nitrogen. In this review we discuss the importance of cyanobacteria as diazotrophs in microbial mats, their interactions with other potential N(2)-fixing microorganisms, and the factors that control their activities.

Quantitative analysis of the 2002 phocine distemper epidemic in the Netherlands.

Phocine distemper virus (PDV) caused thousands of deaths among harbor seals (Phoca vitulina) from the North Sea in 1988 and 2002. To examine the effects of different factors on the pathology of phocine distemper, we performed necropsies and laboratory analyses on 369 harbor seals that stranded along the Dutch coast during the 2002 PDV epidemic. Diagnostic tests for morbillivirus infection indicated a differential temporal presence of morbillivirus in lung and brain. Seals of 3 years or older were significantly more often IgG positive than younger seals. The most frequent lesions in PDV cases were bronchopneumonia, broncho-interstitial pneumonia, and interstitial emphysema. Extra-thoracic emphysema was rare in <1-year-olds compared with older seals, even though severe pneumonia was more common. PDV cases generally had empty stomachs and less blubber than by-caught seals from before the epidemic. In PDV cases involving older animals, lung, kidney, and adrenal weights were significantly increased. Bordetella bronchiseptica was isolated from lungs in two thirds of the PDV cases examined. Our results indicate that brain should be included among the tissues tested for PDV by RT-PCR; that either phocine distemper has a longer duration in older seals or that there are age-related differences in immunity and organ development; that dehydration could play a role in the course and outcome of phocine distemper; and that bacterial coinfections in lungs are more frequent in PDV cases than gross lesions suggest. These results illustrate how quantitative analysis of pathology data from such epidemics can improve understanding of the causative disease.

Retroperitoneal haemorrhage caused by a renal angiomyolipoma.

Renal angiomyolipoma (AML) is a benign renal tumour and is nowadays considered a relatively common lesion. When an AML increases in size or becomes symptomatic, embolisation via the renal artery should then be considered, because rupture is an important complication and interventional therapies are required to stop bleeding. We present a 21 year old woman who was seen at the emergency department following a low velocity trauma. After a period of 9 weeks, clinical examination and radiological examination revealed a haemorrhage from a renal AML, which was treated by selective embolisation. A discussion of the relevant literature is also presented.

[Endodontically treated teeth: use of adhesive core build-up procedures]. / Endodontisch behandelde gebitselementen: weefselbehoud en adhesieve opbouw beste optie.

Endodontically treated teeth can be restored in several ways. The prognosis of these teeth does not only depend on an adequate endodontic treatment, but also on a solid restoration that protects the weakened tooth against fracture. Adhesive core build-up procedures, possibly with the application of new generation glass fibre posts, become more and more in favour. This article is dealing with the considerations that should be made when adhesive core build-up procedures are used and more specific with core build-up procedures for endodontically treated premolars.

Influence of fatigue loading on the performance of adhesive and nonadhesive luting cements for cast post-and-core buildups in maxillary premolars.

PURPOSE: This study evaluated the influence of fatigue loading on the performance of an adhesive and a nonadhesive cement for cast post-and-core restorations in maxillary premolars. MATERIALS AND METHODS: The adhesive cement used was Panavia 21, a resin-based composite cement, and the nonadhesive cement was PhosphaCem/C, a zinc-oxy-phosphate cement. The coronal sections of single-rooted human maxillary premolars were removed at the level of the proximal CEJ. After endodontic treatment, a cast post and core was prepared for each tooth and cemented into the root canal with either Panavia 21 (n = 8) or PhosphaCem/C (n = 8). Half of the specimens from each cement group were exposed to fatigue loading almost perpendicular to the axial axis; the other half were used as controls. Three parallel transverse root sections were cut from each specimen and used for evaluation of the influence of fatigue loading. For each section, cement integrity was studied by SEM, and retention strength of the cemented post section was determined with a push-out test. RESULTS: For SEM evaluation and the push-out test, Panavia 21 proved significantly better than PhosphaCem/C. However, fatigue loading did not show any effect. CONCLUSION: Under the conditions of this study, fatiguing of cemented cast post-and-core restorations was not decisive as a single test to evaluate the quality of the cement.

The gene for a halophilic beta-galactosidase (bgaH) of Haloferax alicantei as a reporter gene for promoter analyses in Halobacterium salinarum.

Investigations of transcriptional regulation and the characterization of promoters in homologous expression systems are most easily performed using suitable reporter genes. Presumably because of the high internal salt concentration in halophilic Archaea, the successful application of the commonly used reporter genes has not been reported so far. Recently, the gene for an extremely halophilic beta-galactosidase (bgaH) from Haloferax alicantei has become available. After transformation of Halobacterium salinarum with a vector-carrying bgaH, the enzyme activity in cell lysates could be readily determined by a simple colorimetric assay and colonies could be screened for activity on plates containing Xgal substrate. Expression of bgaH under the control of various halobacterial promoters of known strength led to different specific beta-galactosidase activities in the lysates. Using Northern blot hybridization and semiquantitative RT-PCR, it was shown that the bgaH transcript level corresponded to the specific enzyme activity. Therefore, the bgaH gene of Haloferax alicantei appears to be a useful tool for in vivo studies of gene expression in Halobacterium salinarum and possibly other halophilic Archaea.

Mechanisms of multidrug transporters.

Drug resistance, mediated by various mechanisms, plays a crucial role in the failure of the drug-based treatment of various infectious diseases. As a result, these infectious diseases re-emerge rapidly and cause many victims every year. Another serious threat is imposed by the development of multidrug resistance (MDR) in eukaryotic (tumor) cells, where many different drugs fail to perform their therapeutic function. One of the causes of the occurrence of MDR in these cells is the action of transmembrane transport proteins that catalyze the active extrusion of a large number of structurally and functionally unrelated compounds out of the cell. The mode of action of these MDR transporters and their apparent lack of substrate specificity is poorly understood and has been subject to many speculations. In this review we will summarize our current knowledge about the occurrence, mechanism and molecular basis of (multi-)drug resistance especially as found in bacteria.

The role of transport processes in survival of lactic acid bacteria. Energy transduction and multidrug resistance.

Lactic acid bacteria play an essential role in many food fermentation processes. They are anaerobic organisms which obtain their metabolic energy by substrate phosphorylation. In addition three secondary energy transducing processes can contribute to the generation of a proton motive force: proton/substrate symport as in lactic acid excretion, electrogenic precursor/product exchange as in malolactic and citrolactic fermentation and histidine/histamine exchange, and electrogenic uniport as in malate and citrate uptake in Leuconostoc oenos. In several of these processes additional H+ consumption occurs during metabolism leading to the generation of a pH gradient, internally alkaline. Lactic acid bacteria have also developed multidrug resistance systems. In Lactococcus lactis three toxin excretion systems have been characterized: cationic toxins can be excreted by a toxin/proton antiport system and by an ABC-transporter. This cationic ABC-transporter has surprisingly high structural and functional analogy with the human MDR1-(P-glycoprotein). For anions an ATP-driven ABC-like excretion systems exist.

Multidrug resistance mediated by a bacterial homolog of the human multidrug transporter MDR1.

Resistance of Lactococcus lactis to cytotoxic compounds shares features with the multidrug resistance phenotype of mammalian tumor cells. Here, we report the gene cloning and functional characterization in Escherichia coli of LmrA, a lactococcal structural and functional homolog of the human multidrug resistance P-glycoprotein MDR1. LmrA is a 590-aa polypeptide that has a putative topology of six alpha-helical transmembrane segments in the N-terminal hydrophobic domain, followed by a hydrophilic domain containing the ATP-binding site. LmrA is similar to each of the two halves of MDR1 and may function as a homodimer. The sequence conservation between LmrA and MDR1 includes particular regions in the transmembrane domains and connecting loops, which, in MDR1 and the MDR1 homologs in other mammalian species, have been implicated as determinants of drug recognition and binding. LmrA and MDR1 extrude a similar spectrum of amphiphilic cationic compounds, and the activity of both systems is reversed by reserpine and verapamil. As LmrA can be functionally expressed in E. coli, it offers a useful prokaryotic model for future studies on the molecular mechanism of MDR1-like multidrug transporters.

Energetics and mechanism of drug transport mediated by the lactococcal multidrug transporter LmrP.

The gene encoding the secondary multidrug transporter LmrP of Lactococcus lactis was heterologously expressed in Escherichia coli. The energetics and mechanism of drug extrusion mediated by LmrP were studied in membrane vesicles of E. coli. LmrP-mediated extrusion of tetraphenyl phosphonium (TPP+) from right-side-out membrane vesicles and uptake of the fluorescent membrane probe 1-[4-(trimethylamino)phenyl]-6-phenylhexa-1,3,5-triene (TMA-DPH) into inside-out membrane vesicles are driven by the membrane potential (Deltapsi) and the transmembrane proton gradient (DeltapH), pointing to an electrogenic drug/proton antiport mechanism. Ethidium bromide, a substrate for LmrP, inhibited the LmrP-mediated TPP+ extrusion from right-side-out membrane vesicles, showing that LmrP is capable of transporting structurally unrelated drugs. Kinetic analysis of LmrP-mediated TMA-DPH transport revealed a direct relation between the transport rate and the amount of TMA-DPH associated with the cytoplasmic leaflet of the lipid bilayer. This observation indicates that drugs are extruded from the inner leaflet of the cytoplasmic membrane into the external medium. This is the first report that shows that drug extrusion by a secondary multidrug resistance (MDR) transporter occurs by a "hydrophobic vacuum cleaner" mechanism in a similar way as was proposed for the primary lactococcal MDR transporter, LmrA.

Multidrug resistance in Lactococcus lactis: evidence for ATP-dependent drug extrusion from the inner leaflet of the cytoplasmic membrane.

Lactococcus lactis possesses an ATP-dependent drug extrusion system which shares functional properties with the mammalian multidrug resistance (MDR) transporter P-glycoprotein. One of the intriguing aspects of both transporters is their ability to interact with a broad range of structurally unrelated amphiphilic compounds. It has been suggested that P-glycoprotein removes drugs directly from the membrane. Evidence is presented that this model is correct for the lactococcal multidrug transporter through studies of the extrusion mechanism of BCECF-AM and cationic diphenylhexatriene (DPH) derivatives from the membrane. The non-fluorescent probe BCECF-AM can be converted intracellularly into its fluorescent derivative, BCECF, by non-specific esterase activities. The development of fluorescence was decreased upon energization of the cells. These and kinetic studies showed that BCECF-AM is actively extruded from the membrane before it can be hydrolysed intracellularly. The increase in fluorescence intensity due to the distribution of TMA-DPH into the phospholipid bilayer is a biphasic process. This behaviour reflects the fast entry of TMA-DPH into the outer leaflet followed by a slower transbilayer movement to the inner leaflet of the membrane. The initial rate of TMA-DPH extrusion correlates with the amount of probe associated with the inner leaflet. Taken together, these results demonstrate that the lactococcal MDR transporter functions as a 'hydrophobic vacuum cleaner', expelling drugs from the inner leaflet of the lipid bilayer. Thus, the ability of amphiphilic substrates to partition in the inner leaflet of the membrane is a prerequisite for recognition by multidrug transporters.

The Lactococcal lmrP gene encodes a proton motive force-dependent drug transporter.

To genetically dissect the drug extrusion systems of Lactococcus lactis, a chromosomal DNA library was made in Escherichia coli and recombinant strains were selected for resistance to high concentrations of ethidium bromide. Recombinant strains were found to be resistant not only to ethidium bromide but also to daunomycin and tetraphenylphosphonium. The drug resistance is conferred by the lmrP gene, which encodes a hydrophobic polypeptide of 408 amino acid residues with 12 putative membrane-spanning segments. Some sequence elements in this novel membrane protein share similarity to regions in the transposon Tn10-encoded tetracycline resistance determinant TetA, the multidrug transporter Bmr from Bacillus subtilis, and the bicyclomycin resistance determinant Bcr from E. coli. Drug resistance associated with lmrP expression correlated with energy-dependent extrusion of the molecules. Drug extrusion was inhibited by ionophores that dissipate the proton motive force but not by the ATPase inhibitor ortho-vanadate. These observations are indicative for a drug-proton antiport system. A lmrP deletion mutant was constructed via homologous recombinant using DNA fragments of the flanking region of the gene. The L. lactis (delta lmrP) strain exhibited residual ethidium extrusion activity, which in contrast to the parent strain was inhibited by ortho-vanadate. The results indicate that in the absence of the functional drug-proton anti-porter LmrP, L. lactis is able to overexpress another, ATP-dependent, drug extrusion system. These findings substantiate earlier studies on the isolation and characterization of drug-resistant mutants of L. lactis (Bolhuis, H., Molenaar, D., Poelarends, G., van Veen, H. W., Poolman, B., Driessen, A. J. M., and Konings, W. N. (1994) J. Bacteriol. 176, 6957-6964).

Proton motive force-driven and ATP-dependent drug extrusion systems in multidrug-resistant Lactococcus lactis.

Three mutants of Lactococcus lactis subsp. lactis MG1363, termed EthR, DauR, and RhoR, were selected for resistance to high concentrations of ethidium bromide, daunomycin, and rhodamine 6G, respectively. These mutants were found to be cross resistant to a number of structurally and functionally unrelated drugs, among which were typical substrates of the mammalian multidrug transporter (P-glycoprotein) such as daunomycin, quinine, actinomycin D, gramicidin D, and rhodamine 6G. The three multidrug-resistant strains showed an increased rate of energy-dependent ethidium and daunomycin efflux compared with that of the wild-type strain. This suggests that resistance to these toxic compounds is at least partly due to active efflux. Efflux of ethidium from the EthR strain could occur against a 37-fold inwardly directed concentration gradient. In all strains, ethidium efflux was inhibited by reserpine, a well-known inhibitor of P-glycoprotein. Ionophores which selectively dissipate the membrane potential or the pH gradient across the membrane inhibited ethidium and daunomycin efflux in the wild-type strain, corresponding with a proton motive force-driven efflux system. The ethidium efflux system in the EthR strain, on the other hand, was inhibited by ortho-vanadate and not upon dissipation of the proton motive force, which suggests the involvement of ATP in the energization of transport. The partial inhibition of ethidium efflux by ortho-vanadate and nigericin in the DauR and RhoR strains suggest that a proton motive force-dependent and an ATP-dependent system are expressed simultaneously. This is the first report of an ATP-dependent transport system in prokaryotes which confers multidrug resistance to the organism.

Heart rate and heart rate variability as indexes of sympathovagal balance.

According to the Rosenblueth-Simeone model, the heart rate (HR) is proportional to the sympathovagal balance. The individual proportionality constant is the intrinsic heart rate, which can only be determined invasively. The normalized low-frequency heart rate variability power (LF) has been raised as a calibrated noninvasive alternative. To concrete this assumption, we studied the individual LF-HR relation during incremental head-up tilt (0, 10, 20, 30, 40, 45, 50, 55, 60, 65, 70, 75, and 80 degrees) in 21 young, healthy males. HR (means +/- SD) increased from 61.0 +/- 9.1 beats/min at 0 degree to 85.9 +/- 18.3 beats/min at 80 degrees. LF increased from 45.8 +/- 16.7 nu at 0 degrees to 79.8 +/- 13.8 nu at 80 degrees (nu meaning normalized units). Individual regressions of LF on HR yielded correlation coefficients of 0.80 +/- 0.13 (means +/- SD). The demonstrated linear relation between LF and HR confirms the potential significance of heart rate variability as a noninvasive means of assessing the sympathovagal balance.

The efflux of a fluorescent probe is catalyzed by an ATP-driven extrusion system in Lactococcus lactis.

Many bacteria, both gram positive and gram negative, extrude in an energy-dependent manner the fluorescent pH indicator 2',7'-bis-(2-carboxyethyl)-5[and -6]-carboxyfluorescein (BCECF) (D. Molenaar, T. Abee, and W. N. Konings, Biochim. Biophys. Acta 1115:75-83, 1991). This efflux was studied in detail in Lactococcus lactis, and several indications that a transport system is involved were found. This transport system is most likely driven by ATP or a related compound. The evidence is that BCECF extrusion (i) occurs against a BCECF gradient, (ii) is strictly correlated with ATP concentration and not with the proton motive force, and (iii) is inhibited by vanadate and to a lesser extent by N,N'-dicyclohexylcarbodiimide. Most convincingly, a UV mutant with a strongly reduced efflux rate was isolated. Such a mutant was isolated from a BCECF-loaded and lactose-energized population by selection of highly fluorescent cells in a flow cytometer-cell sorter. The physiological function of this extrusion system is unknown, but its characteristics classify it among the traffic ATPases.

Giant synovial cyst of the hip joint presenting with femoral vein compression.

A patient is presented with signs mimicking deep venous thrombosis of the leg, which proved to be compression of the femoral vein, without thrombosis, caused by a large cyst from the adjacent hip joint. Clinical presentation, diagnostic imaging procedures and therapy are presented with a review of the literature.

Influence of pharmacological doses of calcitonin on serum beta 2 microglobulin concentration.

The effect has been studied of continuous infusion of calcitonin in 14 hypercalcemic patients and 5 patients with Paget's disease of the bones. In all hypercalcemic patients but one, a good serum calcium lowering effect was obtained. In all subjects there was a significant decrease of serum beta 2 microglobulin concentration during calcitonin infusion (4.1 +/- 3.4 vs 2.9 +/- 2.5 mg . l-1; P less than 0.01). Especially in patients with an initial increased serum beta 2 microglobulin, a pronounced decrement of this serum beta 2 microglobulin was achieved. Moreover, a positive correlation was found between the drop in serum calcium concentration and the serum beta 2 microglobulin concentration before calcitonin infusion (r = 0.83; P less than or equal to 0.01). Urinary beta 2 microglobulin excretion did not change significantly during calcitonin infusion. These results led to the speculation that the serum calcium lowering effect of calcitonin is not only the result of the direct antiosteoclastic effect of this hormone but that some immunologic modulating effect of calcitonin on the monocyte macrophage system of the bones is contributory to this hypocalcemic effect of calcitonin.