Imaging inflammatory diseases with neutrophil-specific technetium-99m-labeled monoclonal antibody anti-SSEA-1.

UNLABELLED: Imaging inflammatory diseases with a 99mTc-labeled neutrophil-specific agent that can be injected directly intravenously continues to be a challenge. METHODS: The antibody, anti-SSEA-1, chosen from studies of 10 neutrophil-specific MAbs, recognizes CD-15 antigens (5.1 x 10(5)/human PMN) with a high association constant (kd = 10(-11) M). One hundred micrograms of MAb labeled with 10-20 mCi 99mTc either by a direct or DTPA conjugation method were injected intravenously into 12 patients (9 men, 3 women, aged 19-48 yr) with clinical evidence of ongoing inflammatory processes. Vital signs of all patients were recorded before and up to 3 hr following administration of the MAb. HAMA was determined in two patients. Anterior and posterior spot views and whole-body images were obtained. All patients except one underwent biopsy, US or CT examinations and/or surgical procedures. Blood samples collected from five patients were analyzed. In nine patients, quantitative organ distribution was determined and radiation dosimetry was calculated. RESULTS: Labeling yields were 94.8% +/- 1.4% and 95.8% +/- 3.5%, respectively. All patients had unequivocally positive images within 3 hr of the MAb injection. Eleven of these were confirmed by other modalities. One patient recovered on antibiotics and was sent home without surgery or other procedures. The lack of radioactivity in the thyroid or gastrointestinal tract indicated that the in vivo stability of the agent was excellent. At 3 hr postinjection, bladder activity in six patients was 1.3% +/- 0.4% of the administered dose. At this time, splenic uptake (7.7% +/- 1.0% ad. dose) and red marrow uptake (14 +/- 1.8%) were lower than those of 111In-WBC. At 49.0% +/- 3.2% administrated dose, liver uptake was at the upper limit with 111In-WBC uptake. Renal uptake was only 2.4% +/- 0.03% administered dose. At 2 hr postinjection, 14% to 51% of the radioactivity was associated with PMN. Radioactivity with lymphocytes was 0.7% to 10.9%, 1.2% to 4.3% with platelets and 1.1% to 2.4% with RBC. No HAMA were detectable in either patient, and no adverse reaction was detectable in any patient. CONCLUSION: Results are highly encouaging and have prompted us to prepare a kit for instant preparation and to initiate clinical trials.

99mTc-labeled vasoactive intestinal peptide analog for rapid localization of tumors in humans.

In recent years, imaging tumors with receptor-specific biomolecules has been the focus of increasing interest. Vasoactive intestinal peptide (VIP) has a high affinity for specific receptors that are expressed in high density on a large number of malignant tumors. VIP was modified (TP 3654) without compromising its biologic activity and labeled with 99mTc. Pharmacokinetics and feasibility studies were performed in 3 healthy volunteers and 11 patients with a history of cancer. Imaging was performed for up to 2 h after injection. Within 24 h after injection of 99mTc-TP 3654 (370-555 MBq/5 microg), approximately 70% of the tracer cleared through the kidneys and 20% through the liver. Blood clearance was rapid. No adverse reaction was noted in any subject. All known tumors were clearly delineated within 20 min. Findings were compared with the results of 99mTc-methoxyisobutyl isonitrile, CT, MRI, or histology. There was concordance in 9 patients. In the other 2 patients, only the VIP scan was positive for tumors known to express VIP receptors. The early results of imaging tumors with 99mTc-VIP are promising and warrant further study.

Do antioxidant strategies work against aging and age-associated disorders? Propargylamines: a possible antioxidant strategy.

The free radical theory of aging was initially proposed by Harman half a century ago primarily to explain biological aging processes. Although administration of so-called antioxidant chemicals, which have been tested in the past for several decades, turned out to be mostly ineffective in prolonging the life spans of animals, the same theory of age-associated diseases appears to be increasingly supported in the last two decades. Despite these difficulties, the success in extending life span of 4 different animal species (mice, rats, hamsters, and dogs) with (-)deprenyl (including a study of our group) indicates that there might exist another type of antioxidant strategy in addition to a simple administration of antioxidant chemicals. (-)Deprenyl has also been shown to increase superoxide dismutase (SOD) and catalase (CAT) activities selectively in brain dopaminergic tissues. Interestingly, we have recently shown that another propargylamine, rasagiline not only increases antioxidant enzyme activities (CAT and SOD) in brain dopaminergic regions as (-)deprenyl does, but also increases CAT and SOD activities in extrabrain catecholaminergic systems such as the heart and kidneys as well. These recent observations coupled with previous observations on the life span of animals with (-)deprenyl suggest that pharmacological modulation of endogenous antioxidant enzyme activities could be one potential antioxidant strategy against aging and age-associated disorders. If the causal relationship between the two effects of (-)deprenyl exists as we hypothesized, we might be able to advance the elucidation of mechanism(s) of aging based on the free radical theory of aging.

Imaging tumors in humans with Tc-99m-VIP.

Vasoactive intestinal peptide (VIP) was modified at the C terminus with a spacer and four amino acids to serve as a chelating moiety. The modified peptide, TP 3654, was labeled with Tc-99m and evaluated in normal volunteers, as well as in patients with a history of cancer. Renal clearance (67%) was the primary route of excretion, with approximately 20% of the radioactivity clearing through the hepatobiliary system. No adverse reaction was noted in any of the subjects and all, except one small, of the known lesions as seen by CT, MRI, Tc-99m-MIBI, or mammography were correctly identified within a few minutes of an i.v. injection of approximately 10 mCi of Tc-99m-TP 3654 (specific activity 11.3 x 10(3) Ci/m mol). The scans were in concordance in nine patients. In the remaining two, one with a visible mass in the neck from high grade spindle cell sarcoma and the other with a palpable mass in a breast from ductal epithelial hyperplasia, were localized only with Tc-99m-TP 3654, but not with Tc-99m-MIBI. Both malignancies are known to express VIP receptors. The VIP analog promises to be a nontoxic and reliable agent for imaging cancers in humans that express VIP receptors.

Isolation and partial characterization of a compound with siderophore activity from Vibrio parahaemolyticus.

A compound with siderophore activity was purified by successive column and thin layer chromatographic procedures from Dowex 1 x 8 extracts of culture supernatants of Vibrio parahaemolyticus AQ 3354. The strain synthesized the compound in culture media containing less than 2 microM added FeCl3. Hydrolysis of the compound yielded alanine, ethanolamine, citric acid and 2-ketoglutaric acid. The 1H-NMR spectrum exhibited the presence of a residue from each of these components in the intact molecule. The fast-atom bombardment mass spectrum of the methyl ester derivative indicated a prominent ion at m/z 477, probably corresponding to [M + 1] ion. Other strains of V. parahaemolyticus were also found to produce this compound when grown in an iron-limited medium.

Enantio-selective occurrence of (S)-tetrahydropapaveroline in human brain.

Tetrahydropapaveroline is an endogenous complex alkaloid derived from dopamine through the oxidation by monoamine oxidase. This alkaloid is considered to be involved in the pathogenesis of alcoholism and to act as a false neurotransmitter. Recently the (S) enantiomer was proposed to be a precursor of morphine biosynthesis in the opium poppy. In this paper stereo-chemical characteristic of tetrahydropapaveroline in human brains was examined. In all four control human brains examined, only the (S)-tetrahydropapaveroline was detected. The concentrations were 0.12-0.22 pmol/g wet weight of brain tissue, and the presence of alcohol in blood did not affect the concentration. The results suggest that (S)-tetrahydropapaveroline may be enantio-selectively synthesized in human brain and it may be an intermediate of the de novo synthesis of morphine analogues.

Dopaminergic neurotoxins, 6,7-dihydroxy-1-(3', 4'-dihydroxybenzyl)-isoquinolines, cause different types of cell death in SH-SY5Y cells: apoptosis was induced by oxidized papaverolines and necrosis by reduced tetrahydropapaverolines.

Dopamine-derived 6,7-dihydroxy-1-(3', 4'-dihydroxybenzyl)-isoquinolines, papaverolines and tetrahydropapaverolines, have been proposed to be neurotoxin candidates related to the pathogenesis of Parkinson's disease. In this paper, the cytotoxicity of papaverolines and their N-methyl derivatives was examined using human dopaminergic neuroblastoma SH-SY5Y cells as a model of dopamine neurons. Apoptotic and necrotic cell death were assessed by morphological observation of cells after staining with propidium iodide and Hoechst 33342. Papaveroline and N-methyl-papaveroline induced apoptosis in almost all the cells with typical features of condensed and fragmented nuclei. On the other hand, (R)- and (S)-tetrahydropapaveroline caused necrosis in cells. Tetrahydropapaverolines markedly reduced adenosine triphosphate (ATP) level, whereas papaverolines did not, suggesting that the types of cell death induced by these isoquinolines, necrosis and apoptosis, depend on ATP concentrations in the cells.

99mTc labeled VIP analog: evaluation for imaging colorectal cancer.

Early and reliable diagnosis of colorectal cancer continues to be demanding and challenging. Colorectal cancer cells express Vasoactive Intestinal Peptide (VIP) receptors in high density. We have prepared a VIP analog (TP3654), labeled it with (99m)Tc, and evaluated it in experimental animals as an agent for imaging colorectal cancer. The tissue distribution of (99m)Tc-TP3654 has been compared with that of (111)In-DTPA-Octreotide and (99m)Tc-anti-CEA scan in nude mice bearing human colorectal cancer LS174T. Finally, pharmacokinetic and tissue distribution studies of (99m)Tc-TP3654 have been performed in four normal human volunteers. Data suggest that (99m)Tc-TP3654 can be prepared efficiently without loss of its receptor specificity and biological activity. Although the 24 hr tumor uptake of (99m)Tc-TP3654 in the animal model used was modest (0.21 +/- 0.07% I.D./g), the tissue distribution profile was more favorable than that of (111)In-DTPA-Octreotide or (99m)Tc-anti-CEA scan. Human studies indicated that (99m)Tc-TP3654 had no adverse effect in any subject. Within 24 hours, approximately 70% of the injected dose cleared through the kidneys, and approximately 20% through the hepatobiliary system. In these non-fasting volunteers hepatobiliary clearance was slow and in cancer patients tumor uptake was rapid. Data suggest that (99m)Tc-TP3654 is a promising agent for imaging colorectal cancer.

Common properties for propargylamines of enhancing superoxide dismutase and catalase activities in the dopaminergic system in the rat: implications for the life prolonging effect of (-)deprenyl.

(-)Deprenyl has been reported to prolong the life span of different animal species. Further, the drug effectively increases antioxidant enzyme activities such as superoxide dismutase (SOD) and catalase (CAT) in brain dopaminergic regions. We have found that the effect of the drug on antioxidant enzyme activities is highly dose dependent, increasing with an increasing dose, however, a higher dose becomes less effective and an excessive dose becomes adversely effective. Most importantly, an optimal dose for the effect varies widely depending on animal species, strain, sex, age and duration of the treatment, which may at least partly explain discrepancies reported among different studies in the past. From the parallelism of the dose-effect relationship of the drug between life span extension and increasing endogenous antioxidant enzyme activity, we have suggested that the above two effects of (-)deprenyl may be causally related. This review summarizes our past series of studies and also reports our very recent observation that other propargylamines such as rasagiline and (R)-N-(2-heptyl)-N-methylpropagylamine (R-2HMP) also share the property of enhancing antioxidant enzyme activities. Further, our most recent study has found that these propargylamines increase antioxidant enzyme activities not only in brain dopaminergic regions but in extra-brain dopaminergic tissues such as the heart and kidneys. These observations are discussed in relation to the life prolonging effect of (-)deprenyl reported in the past.

Enhancing effect of rasagiline on superoxide dismutase and catalase activities in the dopaminergic system in the rat.

Rasagiline [N-propargyl-l(R)-aminoindan] is a selective irreversible MAO-B inhibitor as is (-)deprenyl. The effect of the drug on antioxidant enzyme activities on dopaminergic tissue was examined in male F-344 rats (8.5-months-old). Two experimental groups were infused subcutaneously with rasagiline saline solutions by means of osmotic minipumps implanted subcutaneously in the back of the rats. Control animals were also similarly implanted with saline filled mini-pumps. Three-and-one-half weeks later, animals were sacrificed and selected tissue samples removed from brain, kidney and heart. Two doses of rasagiline (0.5 mg/kg/day, 1.0 mg/kg/day, both for 3.5 weeks) significantly increased catalase activities about 2-fold in substantia nigra and striatum but not in hippocampus. Interestingly, in both renal cortex and medulla. catalase (CAT) activities were significantly increased. Both Mn- and Cu,Zn-superoxide dismutase (SOD) activities were increased 2 to 4 fold in substantia nigra, striatum and renal cortex and heart. Several groups, including our own have reported an extension of survival of deprenyl-treated animals of different species. Although the mechanism(s) of the life extension by deprenyl remains unresolved, it would be interesting to investigate the effect of rasagiline on the survival of animals, since deprenyl also was shown to increase antioxidant enzyme activities in brain dopaminergic regions.

Imaging rheumatic joint diseases with anti-T lymphocyte antibody OKT-3.

T lymphocytes play an important role in the pathogenesis of rheumatoid arthritis (RA). Murine monoclonal antibody OKT-3 (IgG2a), known to be specific for T lymphocyte 20 kD glycoprotein CD3 receptor was labelled with 5 mCi 99Tcm and given intravenously (i.v.) to seven RA and two psoriatic arthritis patients following informed consent to identify inflamed synovium. Anterior and posterior whole body scans and specific regional imaging was commenced 20 min later. At 1 h, approximately 20% of 99Tcm was associated with the lymphocytes. In these patients, all 41 asymptomatic joints and 43 joints with mild pain or minimal tenderness had normal scans. All 34 joints with moderate to severe pain had moderate to marked uptake of radioactivity. Two patients experienced shaking chills for 20-30 min within an hour of 99Tcm-OKT-3 infusion. These results suggest that 99Tcm-OKT-3 imaging serves as an objective surrogate for joint inflammation and could be useful as a measurement of therapeutic effectiveness in RA and other diseases with inflamed synovium. The side effect profile may limit the utility of 99Tcm-OKT-3 but other forms of antibodies directed toward lymphocyte subsets may be useful.

Imaging infection with LeuTech.

LeuTech is a 99Tcm labelled, anti-CD15, IgM, murine monoclonal antibody shown to have high affinity (Kd = 10(-11) M) for CD15 receptors (5.1 x 10(5)/cell) expressed on human neutrophils. LeuTech was injected directly, intravenously, and its efficacy in imaging infection in 46 consecutive patients was determined. Human anti-mouse antibody (HAMA) response was examined in 30 normal volunteers using a standard LeuTech dose reconstituted with decayed 99Tcm solution. There were 38 true positive, six true negative, and two false negative scans. Of the 38 positive images, 33 (92%) were positive within 10 min after injection of LeuTech. LeuTech accuracy in this group of patients was 96%, sensitivity 95%, specificity 100%, positive predictive value (PPV) 100%, and negative predictive value (NPV) 75%. No elevation of the HAMA titre was observed in any of the 30 normal volunteers and no adverse reaction was noted in any patient. LeuTech is a highly promising agent for rapid imaging of infectious foci.

Laboratory study of biological retention for urban stormwater management.

Urban stormwater runoff contains a broad range of pollutants that are transported to natural water systems. A practice known as biological retention (bioretention) has been suggested to manage stormwater runoff from small, developed areas. Bioretention facilities consist of porous soil, a topping layer of hardwood mulch, and a variety of different plant species. A detailed study of the characteristics and performance of bioretention systems for the removal of several heavy metals (copper, lead, and zinc) and nutrients (phosphorus, total Kjeldahl nitrogen [TKN], ammonium, and nitrate) from a synthetic urban stormwater runoff was completed using batch and column adsorption studies along with pilot-scale laboratory systems. The roles of the soil, mulch, and plants in the removal of heavy metals and nutrients were evaluated to estimate the treatment capacity of laboratory bioretention systems. Reductions in concentrations of all metals were excellent (> 90%) with specific metal removals of 15 to 145 mg/m2 per event. Moderate reductions of TKN, ammonium, and phosphorus levels were found (60 to 80%). Little nitrate was removed, and nitrate production was noted in several cases. The importance of the mulch layer in metal removal was identified. Overall results support the use of bioretention as a stormwater best management practice and indicate the need for further research and development.

[A study on the psychological status of perioperative patients (II)].

In order to study the changes of the psychological status during perioperative period, we performed four different psychological tests, Spielberger's rating scale "The State-Trait Anxiety Inventory (STAI)", Self-rating Depression Scale (SDS), Maudsley Personality Inventory (MPI) and Baum test, in 63 patients who received simple total hysterectomy for myoma of uterus. The anxiety ratings showed a significant decrease postoperatively. On the contrary, the SDS index that expresses the level of depressive mood showed no significant change during perioperative period. Moreover the patients who had shown a tendency of nervousness in MPI kept the high level of the SDS index during perioperative period than the other patients who had not shown a tendency of nervousness in MPI. In Baum test, more than 75% of patients manifested psychologic fragility or lability. These results suggest that depressive mood affects strictly the psychological status in postoperative period more than anxiety. We presume that the depressive mood during perioperative period is responsible for the postoperative psychological disturbance.

[Significance of the measurement of arterial ketone bodies during abdominal surgery].

Ketone body (3-hydroxy butyric acid) present in the blood was successively examined during surgical operations to ascertain the condition of energy metabolism and its clinical significance. In 10 patients (R group) to whom only the lactated Ringer's solution was infused during the operation, values of 3-hydroxy butyric acid increased remarkably in the 30 minutes after the operation began. In 7 patients (GI group), to whom the lactated Ringer's solution in 5% of glucose was administered, the values also increased in the 30 minutes after the operation was started. However, after the administration of glucose the values decreased, and returned to the normal value with administration of insulin. Furthermore, blood sugar readings which were examined at the same time increased significantly immediately after the beginning of the operation in both groups. Insulin readings showed considerably low values during the operation in R group. The readings of free fatty acid tended to show high values in R group, while they tended to decrease after the dose of glucose in GI group. With regard to epinephrine and norepinephrine readings, both groups showed significantly high values 30 minutes after the start of operation.

[Perioperative psychological status of patients undergoing radical mastectomy and total hysterectomy].

Using four different psychological tests, namely Spielberger's rating scale [The State-Trait Anxiety Inventory (STAI)], Self-Rating Depression Scale (SDS), Maudsley Personality Inventory (MPI) and Baum test, we investigated the perioperative psychological status of patients undergoing radical mastectomy for breast cancer and compared data with those of patients undergoing total hysterectomy for uterine myoma. Parameters reflecting the psychological status of hysterectomy patients disclosed lower postoperative anxiety, and slight nervousness and persistent depression perioperatively. In contrast, in patients with breast cancer, those with nervousness had persistent anxiety and depression, while those without nervousness showed the same persistent depression as in hysterectomy patients, as well as a high anxiety level postoperatively. Therefore, we speculate that the persistence of depressive mood during the perioperative period is a factor in the psychological disturbance of mastectomy patients with breast cancer, and moreover, they appear markedly apprehensive because of cancer. These results suggest that the persistence of anxiety and depressive mood associated with cancer is a prime factor in the etiology of postoperative psychological disturbance.

Factors affecting postoperative ventilatory support in patients with cardiac cachexia.

Factors affecting early v late weaning from ventilatory support after cardiac surgery were retrospectively compared in 15 patients with cardiac cachexia. These patients were divided into two groups based on the duration of postoperative ventilatory support, group I (n = 7) less than two days (22 +/- 8 hours) and group II (n = 8) over three days (140 +/- 125 hours). Findings in 14 randomly selected patients without cardiac cachexia who underwent cardiac valve replacement were also examined. Comparisons were made between the groups in regards to the following parameters: (1) preoperatively: anthropometric, biochemical, and immunologic parameters of nutritional status, hepatic function, cardiothoracic ratio (CTR) on chest x-ray, and data from cardiac catheterization; (2) intraoperatively and postoperatively: anesthetic drugs, duration of surgery, cardiopulmonary bypass (CPB), aortic cross-clamp, and postoperative use of inotropes. The anthropometric measurements, cell-mediated immunity, and hepatic function were significantly decreased in all patients with cachexia, but with no significant differences between the groups. Serum transferrin levels, which increased in patients with cachexia, showed no significant difference between the groups. The levels of serum albumin were significantly lower in cachectic patients than in patients without cachexia; they were significantly higher in group II than in group I. The durations of surgery and CPB were significantly longer in group II, but there was no significant difference in the duration of aortic clamping between the groups. The postoperative use of inotropes was higher in group II. These results indicate that group II patients were nutritionally as well sustained as group I. However, they had less satisfactory cardiac function, and therefore required a longer duration of postoperative ventilatory support.