Incidental cardiac uptake of 99mTc-diphosphonates is predictive of poor outcome: data from 9616 bone scintigraphies.

BACKGROUND: Bone scintigraphy (BS) is highly diagnostic for amyloid transthyretin (ATTR) cardiomyopathy. Prevalence and prognostic value of BS cardiac uptake is not well established. Our aim was to assess the prevalence of subclinical cardiac ATTR amyloidosis in patients undergoing [99mTc]MDP/DPD scintigraphy and to define their phenotype and prognosis. METHODS AND RESULTS: BS scans performed for any clinical indications from 2009 to 2020 were reviewed. Patients were stratified according to Perugini visual score of cardiac uptake. Follow-up data were collected. Among 9616 BS scans, 0.7% (n = 67) showed cardiac uptake. In 47 (70%) patients, Perugini score was 1 and in 20 (30%) patients uptake was ≥ 2, suggesting cardiac ATTR amyloidosis. Forty subjects (61%) died during the follow-up (mean 47 ± 30 months). Compared with patients with Perugini score 1, those Perugini score ≥ 2 showed increased death rate (P = .018). Two (2/67) subjects were investigated for TTR gene mutations resulting negative. CONCLUSIONS: In patients undergoing BS for different clinical indications, cardiac uptake suggesting cardiac ATTR amyloidosis is a rare, but still neglected finding, thus preventing possible diagnosis of ATTR cardiomyopathy. Importantly, cardiac uptake negatively affects the survival. Physicians should be aware of this rare, but crucial finding for timely diagnosis and treatment.

Determination of travoprost and travoprost free acid in human plasma by electrospray HPLC/MS/MS.

A quantitative method for the analysis of AL-5848, the (+)-enantiomer of fluprostenol (FP), in human plasma is described. Plasma was spiked with a tetradeuterated analog of travoprost free acid (AL-5848X) as internal standard (IS) and acidified with 0.1 M formic acid. Sample clean up was performed using reversed phase solid-phase extraction. Following elution of the compounds of interest and evaporation to dryness, the residue was reconstituted in methanol:water (1:1) and chromatographed on an octadecylsilica (C18) column with negative ion electrospray ionization tandem mass spectrometry. The [M[bond]H](-) ions at m/z 457 and 461 for the analyte and IS, respectively, were subjected to collisional fragmentation with argon to yield the same intense 3-trifluoromethylphenolate (m/z 161) product ion. The validated concentration range was 0.010-3.00 ng/ml based on a 1.0 ml plasma aliquot. Fully adequate accuracy, precision, specificity, recovery and stability for routine use in clinical pharmacokinetic studies were demonstrated. Analysis of a second plasma aliquot following incubation with rabbit esterase allows the isopropyl ester pro-drug, travoprost (AL-6221), to be determined by difference.

Decentration of 3-piece versus plate-haptic silicone intraocular lenses.

PURPOSE: To compare the incidence of decentration with 2 types of silicone intraocular lenses (IOLs). SETTING: John A. Moran Eye Center, University of Utah Medical Center, Salt Lake City, Utah, USA. METHODS: Selection criteria for this retrospective study included patients who had uncomplicated cataract surgery with a clear cornea or scleral tunnel incision with a curvilinear capsulorhexis and capsular bag implantation of a silicone IOL. After a mean follow-up of 14 months (range 12 to 18 months), 54 eyes implanted with a 3-piece lens (AMO SI-30) and 58 eyes implanted with a plate-haptic lens (Staar AA403) were examined for posterior chamber IOL decentration. The decentration criterion was 0.5 mm or more from the center of the pupil. Detailed chart review of preoperative and postoperative measurements was performed for each patient. RESULTS: Eighteen of the 3-piece IOLs (33%) and 11 of the plate-haptic IOLs (20%) were decentered 0.5 mm or more (P = .129). Using photographic analysis, the mean IOL decentration with the 3-piece IOL (1.12 mm +/- 0.198 [SD]) was significantly greater than with the plate-haptic IOL (0.632 +/- 0.278 mm)(P < .001). No statistically significant correlation was found between the centered or decentered IOL groups' preoperative refraction, axial length, capsulorhexis size, type of incision, or rate of neodymium:YAG laser capsulotomy. CONCLUSIONS: No statistically significant difference was seen between the decentration rates of 3-piece and plate-hepatic IOLs; however, the amount of decentration with the 3-piece IOL was significantly greater than with the plate-hepatic IOL. Other factors did not contribute to IOL decentration.

Phase I and pharmacokinetic trial of aminopterin in patients with refractory malignancies.

PURPOSE: Aminopterin (AMT) is a potent folate analog that is no longer in routine clinical use. Because of laboratory data that suggests improved metabolism of AMT versus methotrexate (MTX) in lymphoblasts, we developed a phase I trial to determine the maximum-tolerated dose (MTD), dose-limiting toxicity (DLT), and pharmacokinetic profile of AMT. PATIENTS AND METHODS: Twenty patients with refractory malignancies were treated. The starting dose of AMT was 2.5 mg/m2 every 12 hours for two doses weekly: the dose of AMT was decreased and leucovorin (LV) rescue was added after the DLT was observed. Pharmacokinetics were performed after both intravenous (i.v.) and oral AMT administration. RESULTS: Mucosal toxicity was dose-limiting and resulted in the need for a dose reduction (dose level 2: AMT 2 mg/m2 every 12 hours for two doses weekly) and, subsequently, the addition of scheduled LV rescue (dose level 3: AMT 2 mg/m2 every 12 hours for two doses followed by LV 5 mg/m2 orally every 12 hours for two doses, starting 24 hours after the second dose of AMT). The mean areas under the curve (AUC) for the i.v. (n = 14) and oral (n = 13) doses were 1.20 +/- 0.09 (SE) and 1.05 +/- 0.14 micromol x h/L respectively. The half-life was 3.64 +/- 0.28 hours and the oral bioavailability in 12 matched subjects was 83.5% +/- 8.3%. One patient with endometrial adenocarcinoma achieved a complete response (CR) and remains on therapy at 11+ months. Seven patients had stable disease (SD) for 8 weeks or greater, which included one patient with a metastatic nerve sheath tumor who was stable for 9 months. CONCLUSION: We conclude that AMT has good oral bioavailability and that, when given on a q12 hour x two weekly schedule, the MTD is 2 mg/m2 with delayed LV rescue.

[Geographic distribution of digestive system tumor mortality in Italy, 1970-87]. / Distribuzione geografica della mortalità per tumori dell'apparato digerente in Italia, 1970-1987.

The geographical distribution of mortality rates from tumours of digestive tract in Italy is analyzed in this paper. The analysis is based on official mortality data collected by the National Institute of Statistics (ISTAT). Age-adjusted mortality rates for stomach cancer presented the highest values in some provinces of the North and the Center, and the lowest values in the South of Italy. A same geographical pattern was observed for men and women, and during the whole considered period. Colorectal cancer presented the highest rates in the North-West of the country, and in some provinces of Liguria and Tuscany. The lowes rates were observed in the South, particularly in Calabria and Sicily. Mortality for this cancer was positively associated with degree of urbanization. The geographical pattern remained fairly constant in time, but the North-South differences narrowed during the years 1980s. A similar geographical distribution, characterized by the highest mortality levels in the north-eastern regions of Italy, was observed for cancers of the oral cavity and pharynx, of the oesophagus, of the pancreas, and for male tumours of the liver. Female liver cancer presented, on the contrary, the highest mortality levels in the southern regions.

Accumulation of 5-methyltetrahydrofolic acid and folylpolyglutamate synthetase expression by mitogen stimulated human lymphocytes.

The accumulation of 5-methyl[3H]tetrahydrofolic acid (5CH3[3H]FH4) by phytohaemagglutinin stimulated lymphocytes (PHA-L) cultured in folate free media was investigated to determine the mechanism of uptake of 5CH3FH4 and the requirement of the cells for this vitamin as assessed by monitoring de novo thymidine synthesis. When grown in 20 nM 5CH3[3H]FH4 PHA-L accumulate radiolabel at a rate of 0.04 pmol/h/10(6) cells. This doubles the endogenous folate pool of unstimulated cells (0.6 +/- 0.16 pmol/10(6) cells) in about 15 h. Uptake proceeded via a saturable process, independent of a high affinity folate receptor as assessed by ligand binding and by Northern and Western blot analysis. However, transport was blocked by probenecid, which is consistent with an anion carrier mechanism. Unstimulated cells lacked folypolyglutamate synthetase (FPGS) activity and did not express significant amounts of FPGS mRNA. After 48 h of mitogen stimulation there was a 4-10-fold increase in FPGS mRNA and folypolyglutamate formation (Glu > or = 5) was essentially simultaneous with 5CH3[3H]FH4 transport. Increasing extracellular folate to 2 microM only increased intracellular folate 8-fold, but the length of the folylpolyglutamates decreased. The increased folate did not increase de novo thymidine synthesis compared to cells grown in physiological folate. We conclude that mitogen stimulation activates the process(es) for folate accumulation, especially FPGS, and that physiological uptake (0.04 pmol/h/10(6) cells) is adequate for meeting the cells' need for the vitamin.

5,10-Dideazatetrahydrofolic acid (DDATHF) transport in CCRF-CEM and MA104 cell lines.

5,10-Dideazatetrahydrolic acid (DDATHF) is representative of a new class of antifolates acting through inhibition of de novo purine synthesis. We report here the transport characteristics of the diastereomers of DDATHF, which differ in configuration at C6, and comparison studies with other folate and antifolate analogs. (6R)-DDATHF showed high affinity for the influx system of CCRF-CEM cells with a Km of 1.07 microM and an influx Vmax of 4.04 pmol/min/10(7) cells. Comparative studies with methotrexate yielded an influx Km of 4.98 microM and a Vmax of 6.64 pmol/min/10(7) cells, and with 5-formyltetrahydrofolate an influx Km of 2.18 microM and a Vmax of 6.84 pmol/min/10(7) cells. Uptake of (6R)-DDATHF was competitively inhibited by (6S)-DDATHF, methotrexate (MTX), and 5-formyltetrahydrofolate, all with Ki values similar to their influx Km. The (6S)-DDATHF diastereomer had an influx Km of 1.04 microM, similar to that of (6R)-DDATHF; however, the Vmax of 1.72 pmol/min/10(7) cells was 2.3-fold lower than for (6R)-DDATHF. The transport properties of DDATHF were also studied in a mutant cell line (CEM/MTX), resistant to MTX based on impaired drug transport. In this system (6R)-DDATHF showed an influx Km of 1.49 microM and a decreased influx Vmax of 0.60 pmol/min/10(7) cells. A similar effect was shown for MTX (Km of 7.48 microM, Vmax of 1.02 pmol/min/10(7) cells). The number of binding sites in CCRF-CEM cells was similar for (6R)-DDATHF, (6S)-DDATHF, and MTX, 0.74, 0.71, and 0.76 pmol/10(7) cells, respectively. These values were slightly higher in the CEM/MTX cell line (1.07 and 1.09 pmol/10(7) cells for (6R)-DDATHF and MTX, respectively). Treatment of CCRF-CEM cells with either the N-hydroxysuccinimide ester of MTX or the corresponding N-hydroxysuccinimide ester of (6R)-DDATHF caused substantial inhibition (> 90%) of the influx of (6R)-[3H]DDATHF and [3H]MTX, respectively. These results suggest strongly that DDATHF and MTX share a common influx mechanism through the reduced folate transport system. The internalization of DDATHF by monkey kidney epithelial MA104 cells, which express a high affinity folate receptor, was also studied. Competitive binding studies using purified folate receptor and radiolabeled 5-methyltetrahydrofolate showed that (6S)- and (6R)-DDATHF both had I50 values lower than 5-methyltetrahydrofolate (12 nM). Further studies indicate that both DDATHF isomers are actively intracellularly concentrated through this route and are also rapidly converted to high chain length polyglutamates. Transport via this system was inhibited in folate-depleted cells by 10 nM folic acid.(ABSTRACT TRUNCATED AT 400 WORDS)

Effect of human sex hormones on cytomegalovirus growth and Fc receptor expression.

Pretreatment of human embryonic lung cells with estradiol, progesterone, and testosterone was found to have little effect on cytomegalovirus (CMV) replication in vitro. The three hormones in combination delayed the expression of CMV-induced Fc receptors in this cell line. However, similar effects on Fc receptor expression were observed at both first trimester and third trimester concentrations of the hormones. CMV grew well in a primary cell line derived from endometrial stromal cells. However, in this female genital line, as in the (male somatic) human embryonic lung cells, estradiol, progesterone, and testosterone had little effect on CMV replication. CMV-induced Fc receptors could not be demonstrated in endometrial stromal cells. These results suggest that the variation in cervical CMV excretion during gestation is mediated by some mechanism other than a direct effect of sex hormones on viral replication or Fc receptor expression.

Immunoreactivity of cytomegalovirus-induced Fc receptors.

The present studies were undertaken to characterize the affinity of CMV-induced Fc receptors for each of the subclasses of human IgG and to define the specific region of the IgG Fc fragment interacting with such receptors. To do this, we infected confluent human embryonic lung (HEL) cell monolayers with CMV (strain AD169) and then used a double radiolabel assay to measure adherence of antibody-coated E. coli 06 to such monolayers. Preincubating monolayers with each of the 4 subclasses of human IgG (but not IgA, IgM, or human or bovine albumin) abrogated the enhancing effect of CMV infection on adherence of antibody-coated E. coli 06 to HEL monolayers. Pepsin-derived, purified Fc fragments of human IgG had a similar abrogative effect. Preincubating these with staphylococcal protein A did not reduce their capacity to interfere with binding of antibody-coated E. coli to CMV-induced Fc receptors. These observations establish a broad range of immunoreactivity for CMV-induced Fc receptors, that encompasses all 4 subclasses of human IgG. They also provide indirect evidence that the reaction site of CMV-induced Fc receptors is in the CH2 domain of the Fc fragment.

Effects of cancer chemotherapy on the human aerobic oropharyngeal flora.

Since various agents used in cancer chemotherapy exhibit antimicrobial activity in vitro, we performed sequential quantitative cultures of saline gargles obtained from patients receiving cancer chemotherapy to determine if such chemotherapy alters the composition of the aerobic oropharyngeal flora. When we compared results of cultures obtained from 12 patients just before and at various times after receiving courses of cancer chemotherapy, we observed small, though significant reductions in the numbers of total bacteria, alpha-hemolytic streptococci and inhibitory streptococci two to seven days after courses of chemotherapy. A concomitant increase in the percentage of patients colonized by gram-negative bacilli occurred. Of the chemotherapeutic agents used to treat our subjects, only doxorubicin exhibited antimicrobial activity in vitro. All four alpha-hemolytic streptococci, but none of the seven strains of gram-negative bacilli examined, were inhibited by doxorubicin at concentrations of less than or equal to 12.5 mg/l. Doxorubicin had a modest enhancing effect on in vitro adherence of gram-negative bacilli to human embryonic lung cells. These data suggest that cancer chemotherapy might play a role in colonization of cancer patients by gram-negative bacilli by creating a microbiologic vacuum conducive to such colonization. In this way, cancer chemotherapy might contribute to the high incidence of gram-negative bacillary pneumonia among patients with malignant neoplasms.

Bactericidal activity of antiseptics against methicillin-resistant Staphylococcus aureus.

Various commonly used antiseptics were tested against three strains of methicillin-resistant Staphylococcus aureus (MRSA) at stock strength and in serial 10-fold dilutions. The stock solutions of 4% chlorhexidine gluconate-alcohol (Hibiclens), 1% p-chloro-m-xylenol (Acute-Kare), and 3% hexachlorophene (Phisohex) produced 2-log reductions of MRSA after a 15-s exposure, but even after 240 s, these solutions failed to kill all the MRSA. Povidone-iodine (Betadine) solution was maximally effective at the 1:100 dilution, killing all the MRSA within 15 s; other dilutions were less effective, though each killed the MRSA within 120 s. Similar results were obtained with three different strains of methicillin-susceptible S. aureus. Thus, of the four most commonly used antiseptics, povidone-iodine, when diluted 1:100, was the most rapidly bactericidal against both MRSA and methicillin-susceptible S. aureus.

Antibody-mediated bacterial adhesion to cytomegalovirus-induced Fc receptors. Potential relationship to secondary infections complicating herpesvirus infections.

Cytomegalovirus (CMV) and other viruses within the herpes group have recently been shown to induce Fc receptors in infected monolayers. We have examined the possibility that such receptors might facilitate the adherence of antibody-coated bacteria to CMV-infected cells. To do this, we infected confluent human embryonic lung (HEL) cell monolayers with CMV (strain AD 169) and then used a double radiolabel assay to measure adherence of Escherichia coli 06 to both infected and control monolayers. We examined infected monolayers 48 h after viral seeding, at which time 30-60% of the cells exhibited characteristic cytopathic changes. We compared the adherence of untreated E. coli 06 with the adherence of E. coli 06 that had been preincubated for 1 h at 37 degrees C with either nonimmune or anti-E. coli 06 antiserum. Pretreatment of the E. coli 06 with specific antiserum significantly enhanced its adherence to CMV-infected, but not to control, monolayers (P less than 0.01 by the Mann-Whitney U test). We did not see such enhancement when we used anti-E. coli 06 antiserum to treat a nontypable E. coli. The augmented adherence of antibody-coated E. coli 06 to CMV-infected monolayers was abrogated by pretreating the monolayers with nonimmune serum or purified Fc fragments, but not by pretreating with IgA, IgM, or 1 mM trypan blue. Preincubating HEL cell monolayers with 100 U/ml human leukocyte interferon for 72 h at 37 degrees C did not affect the adherence of antibody-coated E. coli 06 to the monolayers. To determine if antibody-coated bacteria that adhered to the surface of CMV-infected monolayers might themselves act as receptors for microorganisms with Fc binding potential, we compared the adherence of Cowan strain Staphylococcus aureus to CMV-infected and control monolayers that had been preincubated with antibody-coated E. coli 06. The S. aureus adhered significantly better to the former monolayers (P less than 0.001). These results illustrate a previously unrecognized mechanism by which certain herpesviruses might enhance the adherence of secondary pathogens to nonphagocytic cell populations. Such a mechanism, if active in vivo, might facilitate the colonization of mucosal surfaces by these pathogenic microorganisms, and in this way might contribute to both the reported predisposition of CMV-infected patients to secondary infections and to the high prevalence of S. aureus in the vaginal flora of women with histories of genital herpes.

Hyperthermic enhancement of serum antimicrobial activity: mechanism by which fever might exert a beneficial effect on the outcome of gram-negative sepsis.

We have previously demonstrated heightened antibiotic activity at temperatures at the upper end of the physiological range. In the present studies we examined the effect of physiological variations in temperature on the antibacterial activity of antibiotic-free pooled human serum by comparing serum minimal inhibitory and bactericidal titers for gram-positive and gram-negative bacterial strains at 33, 37, and 41 degrees C. We observed a progressive rise in both minimal inhibitory and minimal bactericidal titers with temperature for all classes of gram-negative bacilli studied. However, gram-positive cocci were generally resistant to serum, even at the highest experimental temperature. Bacterial strains adapted to growth at temperatures normally encountered on body surfaces were more susceptible to the enhancing effect of hyperthermia on serum inhibition than were strains adapted to 37 degrees C. In addition, in vitro adaptation of one bacterial strain to different temperatures within the physiological range resulted in readily apparent variations in colonial morphology. These in vitro observations indicate that serum antibacterial activity and bacterial morphology may vary in response to minor changes in either the temperature to which bacteria are adapted before examination or the temperature of the assay system. If similar principles operate in vivo, hyperthermically augmented serum antimicrobial activity might represent one mechanism by which fever exerts a beneficial effect on the outcome of gram-negative sepsis.

Effects of temperature on antimicrobial susceptibility of bacteria.

The effects of variations in temperature within the physiologic range on minimal inhibitory concentrations (MICs) and on the serum bactericidal activity of 17 different antimicrobial agents for 432 strains of bacteria were studied. Comparison of 3,053 experimental MICs performed at 41.5, 40, 38.5, 37, or 35 C with duplicate standard MICs performed at 35 C showed a progressive increase in antimicrobial activity as the temperature was raised within the experimental range. At the highest temperature (41.5 C), 17.1% of MICs were four or more times lower, 7% were eight or more times lower, and 2% were greater than or equal to 16 times lower than the standard MICs performed at 35 C. Binding to proteins in serum neither accentuated nor diminished the augmenting effect of temperature on antimicrobial activity. Comparison of serum bactericidal activity determined at 35 C and 40 C revealed similar hyperthermic augmentation of antimicrobial activity.