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Predictive thermal inactivation model for Listeria monocytogenes with temperature, pH, NaCl, and sodium pyrophosphate as controlling factors.

The effects and interactions of heating temperature (55 to 65 degrees C), pH (4 to 8), salt (NaCl; 0 to 6%, wt/vol), and sodium pyrophosphate (SPP; 0 to 0.3%, wt/vol) on the heat inactivation of a four-strain mixture of Listeria monocytogenes in beef gravy were examined. A factorial experimental design comparing 48 combinations of heating temperature, salt concentration, pH value, and SPP content was used. Heating was carried out using a submerged-coil heating apparatus. The recovery medium was plate count agar supplemented with 0.6% yeast extract and 1% sodium pyruvate. Decimal reduction times (D-values) were calculated by fitting a survival model to the data with a curve-fitting program. The D-values were analyzed by second-order response surface regression for temperature, pH, NaCl, and SPP levels. Whereas increasing the NaCl concentration protected L. monocytogenes against the lethal effect of heat, high SPP concentrations increased heat sensitivity. Also, low pH values increased heat sensitivity of L. monocytogenes. The four variables interacted to affect the inactivation of the pathogen. Thermal resistance of L. monocytogenes can be lowered by combining these intrinsic factors. A predictive model that described the combined effect of temperature, pH, NaCl, and SPP levels on thermal resistance of L. monocytogenes was developed. The model can predict D-values for any combination of temperature, pH, NaCl, and SPP that are within the range of those tested. Using this predictive model, food processors should be able to design adequate thermal regimes to eliminate L. monocytogenes in thermally processed foods.

Animals↗

A predictive model to determine the effects of temperature, sodium pyrophosphate, and sodium chloride on thermal inactivation of starved Listeria monocytogenes in pork slurry.

The effects and interactions of 27 combinations of heating temperature (57.5 to 62.5 degrees C), sodium pyrophosphate (SPP) level (0 to 0.5%, wt/vol), and salt (NaCl) level (0 to 6%, wt/vol) on the thermal inactivation of starved Listeria monocytogenes ATCC 19116 in pork slurry were investigated. A split-split plot experimental design was used to compare all 27 combinations. L. monocytogenes survivors were enumerated on tryptic soy agar supplemented with 0.6% yeast extract. The natural logarithm (loge) of the means of decimal reduction times (D-values) were modeled as a function of temperature, SPP level, and NaCl level. Increasing concentrations of SPP or NaCl protected starved L. monocytogenes from the destructive effect of heat. For example, D-values for the pathogen at 57.5 degrees C in pork slurry with 0, 3, and 6% NaCl were 2.79, 7.75, and 14.59 min, respectively. All three variables interacted to affect the thermal inactivation of L. monocytogenes. A mathematical model describing the combined effect of temperature, SPP level, and NaCl level on the thermal inactivation of starved L. monocytogenes was developed. There was strong correlation (R2 = 0.97) between loge D-values predicted by the model and those observed experimentally. The model can predict D-values for any combination of variables that falls within the range of those tested. This predictive model can be used to assist food processors in designing thermal processes that include an adequate margin of safety for the control of L. monocytogenes in processed meats.

Animals↗

Acute sacroiliitis as a manifestation of calcium pyrophosphate dihydrate crystal deposition disease.

While radiographic lesions of the sacroiliac joint (SIJ) are common in patients with calcium pyrophosphate dihydrate (CPPD) crystal deposition disease, they are rarely accompanied by clinical symptoms. We report the case of a 69-year-old woman who presented with an acute sacoiliitis and a linear calcification in the right SIJ on CT scan. The patient recovered well after intra-articular steroid injections.

Acute Disease↗

Activation of C-C beta-chemokines in human peripheral blood gammadelta T cells by isopentenyl pyrophosphate and regulation by cytokines.

Human gammadelta T lymphocytes respond to viral, bacterial, protozoal, and tumoral antigens, but their precise function remains unknown. In adults the major circulating gammadelta T-cell subset expresses the Vgamma9Vdelta2 T-cell receptor and responds to protease-resistant phosphorylated derivatives found in many pathogens. In this study we show that activation of Vdelta2(+) cells with the nonpeptidic antigen isopentenyl pyrophosphate (IPP) rapidly induces (within 4-12 hours) the C-C chemokines MIP-1alpha, MIP-1beta, and lymphotactin but not MCP-1. The most robust response was obtained for MIP-1beta. IPP induction of MIP-1alpha and MIP-1beta was not affected by costimulation with interleukin-4 (IL-4), IL-10, TGF-beta, or interferon-gamma (INF-gamma). However, IL-12 significantly enhanced IPP-induced expression and release of MIP-1alpha that was down-regulated by TGF-beta whereas the induction of MIP-1beta by IPP+IL-12 was refractory to cotreatment with TGFbeta indicating that these chemokines are differentially regulated by these cytokines. Vdelta2(+) T cells also expressed a wide range of C-C chemokine receptors including CCR1, CCR5, and CCR8, all of which were down-regulated following activation. We conclude that Vdelta2(+) cells can be rapidly induced by components of bacterial cell walls to express high levels of proinflammatory chemokines, supporting an important role for these cells in the early stages of the inflammatory responses to many common pathogens. (Blood. 2000, 95:39-47)

Adult↗

Calcium pyrophosphate dihydrate deposition disease causing thoracic cord compression: case report.

OBJECTIVE AND IMPORTANCE: Calcium pyrophosphate dihydrate (CPPD) deposition disease is being increasingly recognized. Spinal involvement in CPPD deposition disease is rare. When involved, the cervical and lumbar regions are commonly affected. We report a rare case of CPPD deposition disease that caused thoracic cord compression. CLINICAL PRESENTATION: A 45-year-old woman presented with clinical features suggestive of thoracic cord compression. Radiographic findings were consistent with calcification of the ligamenta flava in the lower thoracic levels causing cord compression. Calcification of the ligamentum flavum is commonly attributed to CPPD deposition disease. Evaluation for conditions that might be associated with CPPD deposition disease proved to be negative. INTERVENTION: Laminectomy with removal of the calcified ligamenta flava was performed. Histopathological examination of the excised ligaments revealed evidence of CPPD crystals. Postoperatively, the patient's spasticity decreased and sensations improved, with no significant improvement in motor power. CONCLUSION: Calcification of the ligamenta flava due to CPPD deposition disease is a rare cause of thoracic cord compression. CPPD deposition disease should be entertained in the differential diagnosis of thoracic cord compression.

Chondrocalcinosis↗

Calcium pyrophosphate dihydrate crystal deposition disease of the wrist: trapezioscaphoid joint abnormality.

OBJECTIVE: To determine whether trapezioscaphoid (TS) joint alterations are associated with calcium pyrophosphate dihydrate (CPPD) crystal deposition disease and, if so, to determine the nature of these alterations. METHODS: Radiographs of 160 wrists with evidence of chondrocalcinosis were evaluated with regard to TS joint abnormalities, and findings were compared with a similar number of radiographs in an age and sex matched control population in whom no evidence of chondrocalcinosis or other calcification in the wrist was seen. Two radiologists in consensus recorded radiographic findings in both groups, and a third radiologist blinded to the presence or absence of chondrocalcinosis reviewed wrist radiographs in both groups in a random order. Correlation of TS joint abnormalities with other changes in the wrist was also accomplished. RESULTS; TS arthropathy was found in 43.7% of CPPD wrists and in 14.4% of control wrists in the consensus evaluation. In the blind evaluation, 30% of CPPD wrists and 12.5% of control wrists had TS arthropathy. The degree of arthropathy was more extensive in the CPPD group than in the control group. Features associated with TS arthropathy in the patient population were first carpometacarpal arthropathy and subchondral cysts in the scaphoid or trapezium, or both bones. CONCLUSION: CPPD patients, compared to a control population, reveal frequent and significant radiographic abnormalities of the TS joint that may be suggestive of the diagnosis, even in patients in whom chondrocalcinosis is obscured or absent. The features associated with TS arthropathy are first carpometacarpal arthropathy and subchondral cysts in the scaphoid and trapezium.

Adult↗

[Interaction of HIV-1 reverse transcriptase and bacteriophage T7 RNA polymerase with NTP phosphonate analogs and inorganic pyrophosphate].

We have examined the interaction of human immunodeficiency virus reverse transcriptase (HIV RT) and T7 RNA polymerase (T7 RNAP) with modified nucleoside triphosphates and inorganic pyrophosphate (PPi) analogs containing nonhydrolyzable bisphosphonate groups. We have synthesized a number of derivatives of bisphosphonic acid having different aromatic and nonaromatic side substituents, as well as the NTP derivatives whose incorporation into the growing nucleotide chain during the polymerization reaction results in formation of bisphosphonates as leaving groups. The competitive character of inhibition of both enzymes has been revealed for all the compounds under study, and the inhibition constants have been estimated. One of PPi analogs containing a bulky aromatic substituent is characterized by similar inhibition constants for both T7 RNAP and RT. The universal character of this inhibitor can serve as evidence for a similar structure of the NPT-binding sites in the two polymerases. It has been shown that nonsubstituted methylenebisphosphate is a better leaving group than that containing additional methyl and hydroxyl groups. The NTP analogs are very weak inhibitors of T7 RNAP, whereas HIV-1 RT is more sensitive to this type of compounds. On the basis of the X-ray crystallographic data on the T7 RNAP complex with a template and NTP, we have modeled the binding of some derivatives of bisphosphonic acid in the active center of the enzyme. The peculiarities observed in the model correlate well with the experimental data on inhibition.

DNA-Directed RNA Polymerases↗

[Joint scintigraphy using 99mTc pyrophosphate].

Joint scintigraphy was performed in 85 patients suffering from a variety of rheumatic diseases, using a gamma camera and line scanner. Tc-pyrophosphate was the radio nuclide employed; it accumulates selectively in the juxta-articular parts of the bone. The method provides an objective demonstration of inflammatory and degenerative rheumatic joint changes. More over scintigraphic changes can be demonstrated in joint disease which is too early to be clinically apparant or before there are any corresponding changes in serological parameters. The method is useful both in the localisation and staging of disease, in the evaluation of treatment and as an objective control of clinical skills.

Arthritis, Infectious↗

Pseudotumor of temporomandibular joint: destructive calcium pyrophosphate dihydrate arthropathy.

The clinical and pathological features of a tumor of the temporamandibular joint occurring in a 55 year old man, and subsequently identified as a calcium pyrophosphate dihydrate (CPPD) arthropathy, are reported. Crystalline deposits were identified by compensated light microscopy and confirmed with X-ray diffraction, transmission, and scanning electron microscopy. The relationship of this unique case to other clinical presentations of CPPD deposition disease and the implications of the histological features to the pathogenesis of pseudogout are discussed. This case demonstrates that CPPD arthropathy should be included in the differential diagnosis of an arthrosis or of a tumor involving the temporomandibular joint.

Arthritis↗

Mitochondrial F-type ATPases: the glycine-rich loop of the beta-subunit is a pyrophosphate binding domain.

The beta-subunit of the mitochondrial ATP synthase complex comprises the bulk, if not all, of the catalytic nucleotide binding site on the enzyme. A region of homologous sequence rich in glycines (G) and containing a basic lysine (K) and a threonine (T) is found in the beta-subunit as well as many other purine nucleotide binding proteins. The consensus sequence of this region is Gx4GKT, where x represents any amino acid, and is called the A region or glycine-rich loop. The related function of these proteins implies that the glycine-rich loop is directly involved in nucleotide binding. Here we directly test the involvement of the beta-subunit's glycine-rich region in adenine nucleotide binding using two independent approaches. A synthetic fifty amino acid peptide, PP-50, containing the glycine-rich region and the surrounding sequence was assessed for secondary structure and interaction with potential ligands. Circular dichroism spectropolarimetry indicates that PP-50 assumes a predominantly beta-sheet conformation in solution. Significantly, the peptide precipitates from solution when ATP, ADP, GTP, ITP, and pyrophosphate are added, but not when AMP or phosphate are included. Magnesium is not required for the interaction with the purine nucleotides. Complimentary to these studies, the sequence around the Gx4GKT motif was deleted from a recombinant rat liver beta-subunit overexpressed in E. coli. While the wild type beta-subunit showed specificity for the tri- and diphosphonucleotides, the deletion mutant bound tri-, di-, and monophosphate nucleotides with equal affinity.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenine Nucleotides↗

The cervical spine in calcium pyrophosphate dihydrate deposition disease. A prevalent case-control study.

OBJECTIVE: To test the hypothesis that calcium pyrophosphate dihydrate (CPPD) deposition disease is a risk factor for neck pain. METHODS: A prevalent case-control study was conducted to assess cervical calcifications and neck pain between patients with and without known peripheral CPPD deposition disease. CPPD cases were included if diagnosed with CPPD deposition disease of peripheral joints, and excluded if their chief complaint was neck pain. Controls were randomly selected among consecutive patients, hospitalized for conditions unrelated to CPPD deposition disease or neck pain, and matched to CPPD cases by age and sex. Cervical calcifications were assessed by lateral cervical radiographs and computed tomography scans of the upper cervical spine; neck pain and cervical function were appraised by a validated questionnaire. RESULTS: Cervical calcifications were found in 24 out of 35 patients (69%) in the CPPD group compared to 4 out of 35 patients (11%) in the control group (p < 0.001). Patients with CPPD deposition disease reported significantly more neck pain and discomfort than controls (p < 0.001), and were 5 times more likely to report any neck pain (odds ratio 5.5; 95% confidence interval: 1.9, 21.9). Among male patients, more extensive cervical calcified deposits correlated with more severe neck pain (rs = 0.58, p = 0.03). CONCLUSION: These results suggest that CPPD deposition disease frequently involves the cervical spine and may be associated with the development of neck pain.

Aged↗

Massive calcium pyrophosphate dihydrate crystal deposition disease: a cause of pain of the temporomandibular joint.

Calcium pyrophosphate dihydrate deposition (CPDD) disease is a disorder that occasionally affects the temporomandibular joint (TMJ) and temporal bone, causing pain (66.6% of cases), swelling (50%), trismus (36.8%), and hearing loss (22.2%). Diagnosis of CPDD is challenging because clinical symptoms and imaging features are not characteristic and may mimic a chondrosarcoma. When the diagnosis of CPDD of the TMJ is under consideration, conventional radiographs of the wrist or the knee may contribute to the final diagnosis. Imaging features of CPDD are discussed with a review of the literature.

Aged↗

Liver mevalonate 5-pyrophosphate decarboxylase is responsible for reduced serum cholesterol in stroke-prone spontaneously hypertensive rat.

Spontaneously hypertensive rat (stroke-prone) (SHRSP) has an interestingly low serum cholesterol level due to a reduced biosynthesis of cholesterol in the liver (Iritani, N., Fukuda, E., Nara, Y., and Yamori, Y. (1977) Atherosclerosis 28, 217-222). In this study, we examined the mechanism underlying the reduction of hepatic cholesterol biosynthesis in the rat. Our initial findings in SHRSP, as compared with normotensive Wistar Kyoto rat (WKY), showed that 1) the incorporation of [14C]acetate into cholesterol in the liver slices was markedly less, 2) 3-hydroxyl-3-methylglutaryl (HMG) CoA reductase activity was not reduced, and 3) the incorporation of [3H]mevalonic acid into both cholesterol and squalene was significantly less. The above initial findings suggested that the reduction in the hepatic cholesterol biosynthesis took place in one or more enzymatic processes starting with mevalonic acid and continuing to squalene. When the incorporation of [3H]mevalonic acid into phosphomevalonate derivatives was studied using an ion exchange column, only the radioactivity incorporated into isopentenyl-pyrophosphate (isopentenyl-PP) was less in SHRSP. Furthermore, the specific activity of diphosphomevalonate (mevalonate-PP) decarboxylase in the liver-soluble fractions was reduced 50% in SHRSP as compared with WKY. Kinetic studies using liver crude extracts indicated a lower Vmax value in SHRSP (SHRSP, 0.47; WKY, 2.05 nmol/min/mg), and an unchanged Km value (SHRSP, 18.2; WKY, 19.6 microM). The activity of mevalonate-PP decarboxylase was also found to be reduced in other tissues, including the brain, testis, small intestine, and cultured vascular smooth muscle cells. From the above observations, we concluded that the lower activity of mevalonate-PP decarboxylase was responsible for the reduced cholesterol biosynthesis in the liver of SHRSP.

Acetates↗

Tonoplast H+-ATPase activity in barley roots is regulated by ATP and pyrophosphate contents under NaCl stress.

The changes in ATP and PPi contents in roots under NaCl stress, and the effect of PPi on tonoplast H(+)-ATPase activity were studied with two barley cultivars differing in salt tolerance. The results showed that tonoplast H(+)-ATPase activity in salt-tolerant barley cultivar "Tanyin 2" roots increased obviously under NaCl 200 mmol/L (containing half-strength Hoagland solution) for 2 d, and decreased gradually. Tonoplast H(+)-PPase activity always decreased under NaCl stress. For salt-sensitive barley cultivar "Kepin 7", both tonoplast H(+)-ATPase and H(+)-PPase activities decreased during NaCl stress, as compared with control(grown in half-strength Hoagland solution), ATP content of "Tanyin 2" roots increased under NaCl stress for 2 d, and decreased on 4 d after NaCl stress, as compared with control. In "Kepin 7" roots, ATP accumulation was inhibited during NaCl stress. Pyrophosphate (PPi) content increased slightly both in "Tanyin 2" and "Kepin 7" roots under NaCl stress. Tonoplast H(+)-ATPase activity was inhibited competitively by PPi. These results indicated that the increase in rate of ATP accumulation might be essential for the stimulation of tonoplast H(+)-ATPase activity induced by NaCl. The decrease in ATP content and the inhibition of PPi might result in the decrease of tonoplast H(+)-ATPase activity in barley roots under NaCl stress.

Adenosine Triphosphate↗

[Properties of native and ortho- and pyrophosphate-modified Fe-humic complexes of the soil and their effect on solubility and toxicity of the products of exogenous Zn binding].

The properties of soil Fe-humic sorbent complexes proved to determine the level of exogenous Zn binding and the toxicity of Zn-compounds for soil microbiota. The properties of Fe-humic complexes were profoundly altered under the action of phosphate anions as potential ligands. Distinct properties of Fe-humic complexes modified by ortho- and pyrophosphates were revealed.

Diphosphates↗

Complete nucleotide sequence of human phosphoribosyl pyrophosphate synthetase subunit I (PRS I) cDNA and a comparison with human and rat PRPS gene families.

cDNA clones for human phosphoribosyl pyrophosphate synthetase subunit I (PRS I) were isolated from a glioblastoma cell line MGC 1 cDNA library. The longest clone contained 2,075 base pairs (bp) almost covering the 2.3-kb mRNA and the base sequence of the coding region (954 bp) had a 92.0% sequence homology with that of rat PRS I cDNA. The deduced amino acid sequences were identical between human and rat PRS I. This perfect conservation has heretofore not been reported for other enzymes involved in nucleotide metabolism and glycolysis. A comparison with other isoforms of this enzyme, PRS II and PRS III, showed that the human PRS I was 79.9 and 92.2% homologous in the coding sequence and 95.3 and 94.0% in the deduced amino acid sequence to human PRS II and PRS III, respectively. The high value of the synonymous difference between PRS I and PRS II cDNAs places their time of divergence long before that of the radiation of mammals. Based on the evolutionary rate of amino acid substitution, the PRS I and II genes probably diverged about 760 million years ago.

Amino Acid Sequence↗