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Therapeutic drug monitoring (TDM) for tacrolimus (Tac) is universally applied. However, the concentration–effect relationship for Tac is poorly defined. This study investigated whether Tac concentrations are associated with acute rejection in kidney transplant recipients. Data from three large trials were pooled. We used univariate and multivariate analysis to investigate the relationship between biopsy‐proven acute rejection (BPAR) and Tac predose concentration at five time points (day 3, 10 and 14, and month 1 and 6 after transplantation). A total of 136/1304 patients experienced BPAR, giving an overall incidence of 10.4%. We did not find any significant correlations between Tac predose concentrations and the incidence of BPAR at the different time points. In the multivariate analysis, only delayed graft function (DGF) and the use of induction therapy were independently correlated with BPAR, with an odds ratio of 2.7 [95% CI: 1.8–4.0; p < 0.001] for DGF and 0.66 [95% CI: 0.44–0.99; p = 0.049] for induction therapy. The other variables, including the Tac predose concentrations, were not statistically significantly associated with BPAR. We did not find an association between the Tac predose concentrations measured at five time points after kidney transplantation and the incidence of acute rejection occurring thereafter. Based on this study it is not possible to define the optimal target concentrations for Tac.

The Symphony study showed that at 1 year posttransplant, a regimen based on daclizumab induction, 2 g mycophenolate mofetil (MMF), low-dose tacrolimus and steroids resulted in better renal function and lower acute rejection and graft loss rates compared with three other regimens: two with low-doses of cyclosporine or sirolimus instead of tacrolimus and one with no induction and standard cyclosporine dosage. This is an observational follow-up for 2 additional years with the same endpoints as the core study. Overall, 958 patients participated in the follow-up. During the study, many patients changed their immunosuppressive regimen (e.g. switched from sirolimus to tacrolimus), but the vast majority (95) remained on MMF. During the follow-up, renal function remained stable (mean change: ?0.6 mlmin), and rates of death, graft loss and acute rejection were low (all about 1 per year). The MMF and low-dose tacrolimus arm continued to have the highest GFR (68.6 ± 23.8 mlmin vs. 65.9 ± 26.2 mlmin in the standard-dose cyclosporine, 64.0 ± 23.1 mlmin in the low-dose cyclosporine and 65.3 ± 26.2 mlmin in the low-dose sirolimus arm), but the difference with the other arms was not significant (p 0.17 in an overall test and 0.077, 0.039 and 0.11, respectively, in pair-wise tests). The MMF and low-dose tacrolimus arm also had the highest graft survival rate, but with reduced differences between groups over time, and the least acute rejection rate. In the Symphony study, the largest ever prospective study in de novokidney transplantation, over 3 years, daclizumab induction, MMF, steroids and low-dose tacrolimus proved highly efficacious, without the negative effects on renal function commonly reported for standard CNI regimens.

Renal transplantation faces challenges: the organ shortage resulting in extended waiting times and an aging population resulting in death with a functioning graft. The Eurotransplant Senior Program (ESP) allocates kidneys within a narrow geographic area from donors aged ?65 years to recipients ?65 years regardless of HLA. This analysis investigates the impact of the ESP on waiting time, graft and patient survival. The ESP group (n 1406, old to old) was compared to two groups allocated via the Eurotransplant Kidney Allocation System (ETKAS) with either similar donor age (old to any OA, donor age ?65, n 446) or recipient age (any to old, AO, recipient age 60-64, n 1687). All patients were transplanted between 1999 and 2004. Since initiation of the ESP (1999), availability of elderly donors doubled and waiting time for ESP patients decreased. Local allocation led to shorter cold ischemia time (11.9 vs. >17.0 h, p < 0.001) and less delayed graft function (DGF, ESP 29.7 vs. OA 36.2, p 0.047) but 5-10 higher rejection rates. Graft and patient survival were not negatively affected by the ESP allocation when compared to the standard allocation. The ESP age matching of elderly donors and recipients is an effective allocation system for organs from elderly donors.

Rate constants for the attachment of CH(CN)2^- to Fischer carbene complexes of the type (CO)5M&dbd;C(XR)C6H4Z with M = Cr, W and XR = OMe, OEt, SMe in 50% MeCN−50% water (v/v) at 25 °C are reported. The malononitrile anion shows a much higher reactivity toward these carbene complexes than OH^-, primary aliphatic amines (e.g. n-butylamine), or secondary alicyclic amines (e.g. piperidine) but is slightly less reactive than thiolate ions (e.g. HOCH2CH2S^-). The alkoxycarbene complexes react more quickly than the thiomethyl derivatives, consistent with previous findings for alkoxide ion, OH^-, amine, and thiolate ion nucleophiles. Hammett ρ values are 0.69 for the reaction with (CO)5Cr&dbd;C(SMe)C6H4Z and 2.88 for the reaction with (CO)5Cr&dbd;C(OMe)C6H4Z. The much larger ρ value for the methoxycarbene complexes fits a pattern observed previously with other nucleophiles and can be explained by the π-donor effect of the methoxy group, which is much stronger than that of the thiomethyl group.

According to the Principle of Nonperfect Synchronization (PNS), a product‐stabilizing factor, the development of which at the transition state lags behind bond changes, increases the intrinsic barrier of a reaction, but lowers it if it develops ahead of bond changes. Because of the generality of this principle, details of transition state structures can be deduced from comparisons of intrinsic barriers within classes of reactions. A major conclusion that has emerged from such comparisons is that reactions that lead to charge delocalization/resonance stabilized products have transition states in which the development of charge delocalization/resonance stabilization lags behind bond changes and, hence, have relatively high intrinsic barriers. This paper deals with reactions that lead to aromatic molecules or ions. The question we are asking is whether or not aromaticity follows the pattern of common resonance effects and lags behind bond changes at the transition state. Evidence based on experimentally determined intrinsic barriers of proton transfers from rhenium carbene complexes in solution, as well as that based on ab initiocalculations of proton transfers in the gas phase, suggests that aromaticity development at the transition state may actually be aheadof bond changes. Copyright © 2004 John Wiley & Sons, Ltd.

Objectives To compare the antiviral efficacy of a salvage therapy combining lopinavir and amprenavir with 200 mg/d or 400 mg/d ritonavir, together with nucleoside reverse transcriptase inhibitors, over a 26-week period in HIV-infected patients in whom multiple anti-retroviral regimens had failed.Design Phase IIb, randomized, open-label, multicentre trial. Patients were eligible if they had <500 CD4+ cells/mm3and >4 log10copies/ml HIV-RNA after treatment with at least two protease inhibitors (PIs) and one non-nucleoside reverse transcriptase inhibitor.Results At baseline (n=37), the median CD4+ cell count was 207/mm3and the median plasma HIV-1 RNA level was 4.7 log10copies/ml; the median number of PI mutations was seven and the median decrease in phenotypic susceptibility to lopinavir and amprenavir was 9.7 and 2.6, respectively. The mean number of antiretrovirals received prior to randomization was 7.7. The fall in the median HIV-1 RNA level at week 26 was -1.4 log10copies/ml in the 200 mg/d ritonavir group and -2.5 log10copies/ml in the 400 mg/d group (P=0.02). Viral load fell below 50 copies/ml in 32% and 61% of patients, respectively (P=0.07). After adjustment for the ritonavir dose, a smaller number of PI mutations was the only baseline characteristic associated with a better virological response at week 26. Amprenavir concentrations were significantly lower in presence of lopinavir. The lopinavir inhibitory quotient at week 6 correlated weakly with the change in the HIV-RNA level at week 26.Conclusion Combination of amprenavir, lopinavir and 400 mg/d ritonavir shows significant virological efficacy without increased toxicity in HIV-infected patients in whom multiple antiretroviral regimens have failed.

A kinetic study of the aminolysis of (CO)5Cr&dbd;C(OEt)C6H5 (7-Cr) and (CO)5Cr&dbd;C(OEt)C6H5(Cr(CO)3) (8-Cr) by five primary aliphatic amines and of the hydrolysis of 8-Cr in 50% MeCN−50% water (v/v) at 25 °C is reported. The introduction of the Cr(CO)3 group has only a minor effect on the electrophilic reactivity of the carbene carbon; that is, the nucleophilic attack by OH^- and the amines on 7-Cr and 8-Cr occurs at similar rates. However, the Cr(CO)3 group exerts a strong influence on the mechanisms of both types of reactions, which results in changes of what steps are rate limiting. In the hydrolysis reaction these changes lead to a more complex rate versus pH profile for 8-Cr compared to the previously reported one for 7-Cr. In the aminolysis the effect is to change the mechanism of general base catalysis from rate-limiting leaving group departure for the reactions of 7-Cr to rate-limiting proton transfer for the reactions of 8-Cr. We provide a detailed analysis of the reasons for these changes.

We evaluated the impact of genetic changes within p6Gaggene on the virological response (VR, mean decrease in plasma viral load at week 12) to unboosted amprenavir (APV). Gag-protease fragments, including gag p2, p7, p1, p6 regions and whole protease (PR) were sequenced from baseline plasma specimens of 84 highly pre-treated but APV-naive patients included in the NARVAL (ANRS 088) trial. The correlation between baseline p6Gagpolymorphism, PR mutations, baseline characteristics and VR to APV was analysed in univariate analysis. Insertions (P459Ins) within p6 protein, leading to partial or complete duplication of the PTAPP motif, were significantly associated with a decreased VR (P459Ins versus wild-type; –0.3 ±0.8 vs –1.1 ±1.2 log copies/ml, P=0.007) and were more frequent when the V82A/F/T/S PR mutation was present (P=0.020). In multivariate analysis, after adjustment on the predictive factors of the VR in the NARVAL trial and on the PR mutations linked with response, there was a strong trend to an association (P=0.058) between the presence of P459Ins and an altered VR. In conclusion, these results suggest that insertions in the p6 region of HIV-1 gag gene may affect the VR, in highly pre-treated patients receiving an unboosted APV-containing regimen.

A kinetic study of the reactions of (CO)5Cr&dbd;C(SMe)C6H4Z (Z = 4-CF3, 4-Cl, 4-F, 4-Me, 4-MeO, and 4-Me2N) with n-butylamine, 2-chloroethylamine, piperidine, and 1-(2-hydroxyethyl)piperazine in 50% MeCN−50% water (v/v) at 25 °C is reported. In all cases the nucleophilic attachment of the amine to the carbene complex is rate limiting, which contrasts with the previously reported aminolysis of the methoxy analogues ((CO)5Cr&dbd;C(OMe)C6H4Z), where leaving group departure is rate limiting in most cases. The substituent effect on the nucleophilic attachment step is much weaker (average ρ = 0.55) than for the methoxy analogue (ρ = 2.03). It is shown that the small ρ value for (CO)5Cr&dbd;C(SMe)C6H4Z is “normal” and reflects the near cancellation of the substituent effects on the partial positive and partial negative charges at the transition state. These findings provide evidence that the high ρ value for (CO)5Cr&dbd;C(OMe)C6H4Z is “abnormal” and the result of the strong π-donor effect of the methoxy group on the reactant carbene complex.

The kinetics of the reactions of (CO)5Cr&dbd;C(SMe)Ph (4-Cr) with five primary aliphatic and four secondary alicyclic amines have been investigated in 50% MeCN−50% water (v/v) at 25 °C. With n-butylamine, 2-methoxyethylamine, 2-chloroethylamine, glycinamide, piperidine, piperazine, and 1-(2-hydroxyethyl)piperazine, the reaction is strictly second order, i.e., first order in 4-Cr and first order in the amine. With morpholine, base catalysis by the amine and by OH^- was observed while with aminoacetonitrile, OH^- catalysis but no amine catalysis was detected. These results are interpreted in terms of a three-step reaction mechanism (Scheme 1):  the first step is nucleophilic attachment of the amine to the carbene complex, to form a zwitterionic intermediate, %@mt;sys@%T%@sx@%A%@be@%%@ital@%±%@rsf@%%@sxx@%%@mx@% ; the second step is a fast equilibrium deprotonation of %@mt;sys@%T%@sx@%A%@be@%%@ital@%±%@rsf@%%@sxx@%%@mx@% to generate the anionic intermediate %@mt;sys@%T%@sx@%A%@be@%-%@sxx@%%@mx@% ; the third step is MeS^- expulsion by two concurrent pathways, one being spontaneous (solvent catalyzed) expulsion of MeS^-, the other being RR‘NH2⁺ catalyzed loss of MeS^- from %@mt;sys@%T%@sx@%A%@be@%-%@sxx@%%@mx@% . For the reactions that follow the second-order rate law, the first step is rate limiting, while for the reactions that show amine and/or OH^- catalysis, the third step is rate limiting. The factors that determine which steps are rate limiting are discussed in detail.

A kinetic study of the reversible cyclization of (((2-hydroxyethyl)thio)arylcarbene)pentacarbonyltungsten(0) complexes, 2-SR(Z), in 50% MeCN−50% water (v/v) at 25 °C is reported. The results are consistent with Scheme 1, which shows a pathway in basic solution that involves rapid deprotonation of the OH group followed by rate-limiting cyclization and a pathway in acidic solution that involves direct nucleophilic attack by the OH group. Rate and equilibrium constants for the various elementary steps were determined as a function of the phenyl substituent. They were found to correlate well with the Hammett equation. The ρ values provide insights into (a) the reasons that the equilibrium constants are highly sensitive to substituent effects despite the fact that the negative charge of the adducts is strongly dispersed, (b) the mechanistic details of the cyclization pathway in acidic solution, and (c) the extra stabilizing effect of soft−soft interactions in previously studied adducts between a soft thiolate ion and the extra-soft carbene complex (2-SR(Z)).

Abstract:  Renal failure (RF) in multiple myeloma (MM) is considered an ominous complication even though, when timely therapy is started in patients with minimal damage, a high percentage of cases can achieve a regression. The evaluation of renal involvement usually relies on serum creatinine or its clearance, but these parameters have proved to be inadequate to identify initial damage. The aim of this study was to assess the role of the following urinary proteins in diagnosing renal impairment at an early stage: high-molecular-mass proteins (transferrin, IgG, albumin) as markers of glomerular damage, and low-molecular-weight proteins and parenchymal enzymes [α1-acid glycoprotein (AGP), α1-microglobulin (α1M), retinol-binding protein (RBP), β2-microglobulin (β2M), lysozyme (LZ), and N-acetyl-β-d-glucosaminidase (NAG)] as indicators of tubular disorder. Thirty MM patients (nine at disease onset and 21 previously treated) were included in the study. No correlation was found between the urinary proteins and the phase or the stage of the disease. By the Spearman test, Bence Jones proteinuria correlated significantly with the 24 h proteinuria (p=0.01) and β2M (p=0.02), and weakly with the α1M. Serum creatinine concentrations and urea correlated with most of the analytes evaluated: RBP correlated well with urea (p=0.004) and creatinine (p=0.004); IgG (p=0.006) albumin (p=0.009), AGP (p=0.04), and NAG (p=0.02) correlated with serum creatinine. Significant statistical correlation was found between all the analytes except LZ and the creatinine clearance. Twelve of the 30 MM patients (40%) showed abnormal values of urinary proteins. Four of these patients showed overt renal failure with significant modification of the serum parameters and of creatinine clearance, three showed an isolated decrease of creatinine clearance, and five did not present any alteration of serum or urinary parameters. This testifies to the utility of urinary proteins in highlighting renal damage even in cases where the customary serum indicators of renal disorder are normal. In conclusion, our results demonstrate that AGP, RBP, NAG, transferrin, and IgG are good indicators of renal damage. They do not correlate with the severity of the disease, but they seem to be helpful in identifying a subset of patients with initial renal dysfunction.

This study describes a simple and relatively rapid method of purifying Reed-Sternberg (R-S) cells and their morphologic variants from the lymph nodes of patients affected by Hodgkin's disease. Our initial studies defined the optimal procedure for a quantitative disaggregation of Hodgkin's lymph nodes and the densities of R-S cells in several donors. These preliminary steps were helpful in the development of strategies for selectively concentrating R-S cells by density gradient centrifugation. We layered a single-cell suspension over Percoll of appropriate density, centrifuged these samples for 15 minutes, and collected a fraction enriched in R-S cells. Most of the R-S cells were distributed between densities of 1.060 and 1.072, with a peak at approximately 1.066 g/mL. R-S cells are denser than many mononuclear cells present in the lymph nodes of Hodgkin's patients and lighter than reactive cells such as eosinophils, mast cells, and neutrophils. However, the ranges of densities of these cell types overlap, making purification of R-S cells by isopyknic centrifugation impossible. Nevertheless, when this enriched fraction is further processed by velocity sedimentation in order to take advantage of the larger size of R-S cells as compared with all other cells, a substantial purification is achieved. We used three different velocity-sedimentation chambers to find the optimal conditions for obtaining the highest purity with a high final yield. The cells isolated by this method are viable, appear to be morphologically normal, and have been further characterized biologically.

We have analyzed the distribution and prognostic significance of terminal deoxynucleotidyl transferase (TdT) and adenosine deaminase (ADA) in connection with conventional cytology, cytogenetics, response to therapy, and survival. The study population consisted of 78 patients with AML, 44 patients with Ph1 + CML in chronic phase, and 35 adult patients with Ph1 + CML in blastic phase, among which 5 cases presented as Ph1 + acute leukemia. Nine percent of the AML cases were positive for TdT and were characterized by a high percentage of blast cells in bone marrow, myeloid features by cytochemistry and absence of the Philadelphia chromosome. The median ADA values of the TdT+ AML cases were several times higher than those obtained for the TdT- cases. The survival calculated for the two groups of AML cases subdivided according to ADA levels was significantly longer (p less than 0.025) for the patients with low levels of ADA (less than 250 U/10(8) cells). In chronic phase of CML, TdT was absent and ADA values were increased over normal controls only in cases with early signs of transformation. In blastic phase, 31% of the 35 cases were positive for TdT, and ADA values were significantly higher (p less than 0.001) in TdT+ than TdT- cases. The survival calculated from the onset of transformation was significantly longer for the TdT+ acute phase (10.4 mo) compared to the TdT- patients (4.8 mo; p less than 0.025). Four cases presenting as Ph1 + acute leukemia were TdT+ and had elevated levels of ADA; 3 of them responded to ALL therapy, reverting to a stable phase of CML.

Exposure to ionizing radiations (Rx) has been implicated as a causative factor of chronic myelogenous leukemia (CML). We performed a retrospective study of 443 consecutive CML patients, looking for a history of significant exposure to Rx, and evaluated the clinical and hematological characteristics in order to find any difference between radiation-related CML patients and those with de novo CML. We identified 406 patients without known exposure to mutagens (group I) and 37 patients with prior significant exposure to Rx (group II). In comparison to patients of group I, those of group II showed particular clinical and hematological features: significantly lower incidence of bulky splenomegaly (p<0.05) and hyperleukocytosis (WBC>100×109/l;p<0.05); significantly higher incidence of anemia (Hb<10 g/dl;p<0.01). Patients with radiation-related CML had a significantly better survival than those with de novo CML (median survival 61 months vs 42 months;p<0.05). In conclusion, this study of a large cohort of CML patients indicates that the subgroup of patients with a history of significant exposure to ionizing radiation has particular clinical and hematological features at onset (lower tumor burden, higher frequency of anemia) and a better survival.

Six lambs were anesthetized and connected in venouvenous mode to a Membrane Lung for Extracorporeal CO2removal. The animals underwent several hours periods of continuous positive pressure ventilation (CPPV), at 5 cmH2O positive end expiratory pressure (PEEP), alternated with several hours periods of low frequency positive pressure ventilation (5 cmH2O PEEP, 2 b.p.m.) with extracorporeal CO2removal (LFPPV-ECCO2R). During LFPPV-ECCO2R compared with CPPV, cardiac output increased by 26%, pulmonary vascular resistances and systemic vascular resistances decreased by 28% and 22% respectively. The renal function improved significantly during LFPPV-ECCO2R compared with CPPV, i.e. urinary flow, creatinine clearance and osmolar clearance increased by 50%, 37% and 52% respectively. In these experiments LFPPV-ECCO2R, a form of completely artificial ventilation, seems to prevent hemodynamic and renal complications of CPPV.

Summary Six cases are reported of variant Ph′ translocations found among 240 patients with Ph′-positive CML. Five cases had a three-chromosome rearrangement involving, in addition to chromosomes 9 and 22, chromosomes 7, 4, 2(two), and 3 respectively, and one case had a two-chromosome rearrangement 22/5. A review of the literature revealed that three- and two-chromosome variant Ph′ translocations are observed with equal frequency. It is postulated that all variant translocations are indeed three-chromosome rearrangements, that the specific event for the formation of the Ph′ chromosome is the reciprocal translocation 9/22, and that the transposition of regions 9q34 and 22 (q11→qter), plays a major role in the development of CML.

Fifty cases of Hodgkin's disease in intravenous drug users (IVDU) have been collected by the Italian Cooperative Group on AIDS-Related Tumors (G.I.C.A.T.). Ninety-two per cent of the patients were males; the median age was 26 years. Persistent generalized lymphadenopathy (PGL) at onset was present in 54% of patients, AIDS in 9%, ARC in 9% while 28% were simply HIV-positive. The initial median absolute number of CD4 lymphocytes was 264/mmc. Opportunistic infections were diagnosed in 20% of patients. In most patients the histological pattern was that of mixed cellularity and lymphocytic depletion (76%). In almost half the initial symptom was a persistent lymph node enlargement due to PGL. In the majority of patients (58%) only a clinical staging and bone marrow biopsy could be performed due to the presence of opportunistic infections, rapid disease progression or refusal of pathologic staging procedures. One patient presented with a Waldeyer's ring involvement, but no other unusual presentations were observed. After MOPP alternated or followed by ABVD or MOPP alone, 15/29 CR (52%) and 14/29 PR (48%) were observed. The median duration of CR was 14 months, while the median survival of CR has not been reached; the median survival of patients treated with chemotherapy with CD4 values at presentation {geq}400/mmc was significantly superior to that in those with CD4 < 400/mmc. The overall median survival was 16 months. Twenty-eight per cent of patients receiving chemotherapy + radiotherapy developed opportunistic as well as non-opportunistic infections (21%). Lethal hepatic toxicity was observed in 2 patients.In conclusion, Hodgkin's disease in IVDU was not found to be associated with unusual presentations, as previously reported for homosexuals. Complete remissions could be achieved in over 50% of patients, but in IVDU non-opportunistic infections in addition to opportunistic infections may also limit treatment administration. The presence of parenchymal functional impairment due to drug abuse, or drug abuse-related infections, such as pneumonia, endocarditis and hepatitis, should lead to the choice of antitumour agents with no or only minor potential liver, lung and cardiac toxicity.

Abstract:  The presenting features of 356 previously untreated multiple myeloma (MM) patients grouped according to age were analyzed in order (a) to elucidate the possible differences in initial clinical and laboratory features between patients younger than 50 years and the older ones and (b) to statistically assess the prognostic value of the parameters considered, with particular emphasis on the prognostic impact of age. Patients were divided into two groups: group I included 61 patients aged less than 50, group II comprised 295 patients aged 50 or more. No significant differences were found between the two groups in terms of either clinical or laboratory initial characteristics. The treatments adopted and the response to therapy did not differ in the two groups. The prognostic value of presenting features was evaluated for the whole cohort by univariate and multivariate analysis, considering both the observed survival rates and survival rates corrected for the effect of other independent causes of death, using a Poisson model. In both models, calcium level (RR 2.33), performance status (RR 1.83), and creatinine (RR 1.69) maintained their independent negative prognostic value. In contrast, the impact of age was different in the two models. In fact, patients younger than 50 seem to have a better prognosis when the observed survival rates are considered, but they show an increased risk of death when the model takes into account the expected mortality of the underlying population. In conclusion, this study shows that the younger cohort of MM patients has no distinctive initial characteristics with respect to older patients. In multivariate analysis, creatinine levels, calcemia, and performance status show a relevant negative independent prognostic value. Regarding the prognostic impact of age, survival seems to be better among patients younger than 50 than in older patients when the observed survival rates are considered but is significantly worse when the mortality of the corresponding general population. is taken into account.

Summary We determined the nucleotide sequence of the SH gene and its flanking regions over a range of 380 nucleotides for three distinct mumps virus (MUV) isolates. Two isolates from the 1992 mumps epidemic in Western Switzerland and one MUV isolated in 1995 in the same geographic area have been analyzed and compared to 16 recently published SH nucleotide sequences and their presumed amino acid sequences. The nucleotide sequences from the 1992 MUV isolates were identical and closely related to two MUV strains from Eastern Switzerland and strains from the U.K. The MUV isolated in 1995 is clearly different from all other strains.

The peculiar clinical, histomorphological and biological characteristics of PMBCL are reviewed. Special emphasis is given to the frequent aggressive clinical behaviour of this lymphoma in which conventional prognostic factors seem inadequate to identify high risk cases. The need for new clinical and/or biological prognostic markers is stressed.

The interaction of phenalenonc and 2-methylindole in presence of stoechiomctric amounts of paratoluenesulfonic acid in methanol under argon, yields twelve compounds which have been spectroscopically characterised. Seven compounds or sets of compounds could be scpareted by flash chromatography. Phenalene 4. and phenalanone 8 result from a disproportionalion reaction of a key intermediate 18. 2-Methylindolylphenalene 5 and bis(2-methyl)indolylphenalene 6 and 7 exist under the form of a complex mixture of lauiomers. 2-Methylindolylphenalanones 9 and 10. 2-methylindolylphenalenones 11. 12. 1 3. and bis(2-methylindolyl)phenalenones 14 and 15. could also be isolated from the reaction mixture. In presence of air and light compounds 5, 6, and 7. oxidize into 11, 12, 13, 14, and 15 via the corresponding phenalenyl radical. The mechanism of formation and interconversion of these products is discussed and experimentally supported. The peculiar reactivity of the phenalenic derivatives is analysed and it is suggested that such structures are present in asphaltenes. An hypothesis about their genesis is proposed.

Less extensive surgical treatment has become possible for limited vulvar cancer. However, primary closure is advisable only when the defect is minimal and tissue replacement is always needed in radical excisions. Local flaps are always considered the first choice because of their characteristics of similarity to the resected area. The V–Y island flaps from the medial side of thigh constitute one of the numerous options. Clinical experience and anatomical studies demonstrate that this procedure is extremely safe thanks to the rich subcutaneous vascular network. After illustrating the theoretical bases and the indications of this surgical procedure, we report our experience. The V–Y technique is reliable, ensures good healing, and provides almost normal sensation. We conclude that, in most instances, it can be considered a good option for vulvar reconstruction for its versatility and rapidity of execution.

We use methods of nonlinear dynamics to describe the effect of periodic inhibition on the patterns of action potentials generated by regular spiking rat cortical neuron in vitro. Both direct measurements and our mathematical model reveal that chaotic patterns of discharge can be evoked at certain frequencies of inhibitory stimulation. We use detailed biophysical simulation of a cortical neuron to explain the firing patterns in terms of known membrane ionic conductances.

The E ⇆ Z photoisomerization of the title compound (UA) (a naturally occurring sunscreen) has been studied in aqueous solution. At a UA concentration of 6mMand using 313nm excitation, φE→z= 0.52, φZ→E= 0.47 and the photostationary state is 34% E. Under these conditions, loss of UA is minimal. Low energy triplet quenchers fail to impede the isomerization, but the reaction can be induced by several triplet sensitizers. The ETfor UA is estimated to be approximately 55 kcal/mol.

Monosomy 7 is frequent in acute myeloid leukaemia (AML) and in preleukaemic dysmyelopoietic syndromes but often it is not the only chromosome anomaly associated with these conditions. We report 14 patients with “pure” monosomy 7 and their clinical and haematological data are analysed in order to clarify the possible implications of this chromosome anomaly. The following points are considered:1)In spite of the apparent variability of clinical forms in which monosomy 7 is found, several characteristics are common to all monosomy 7 patients, i.e. the presence of a preleukaemic phase and blood and marrow features suggesting the early involvement in the disease of all marrow cell lines. The different diagnoses associated with monosomy 7 are correlated with different steps of a unique myeloproliferative disease whose typical course can be reconstructed.2)Monosomy 7 has a negative prognostic value. When it is found in a preleukaemic disorder it indicates a high risk of progression to AML, while in AML it implies recurrent infections poor response to therapy and short survival.3)The significance of the lack of Colton blood group antigens in monosomy 7 patients is discussed, with particular regard to the fact that the patients in whom this lack was found are the only ones who had not received transfusions in the months before the tests were done.4)The finding of defective neutrophil chemotaxis in monosomy 7 patients is confirmed and the clinical importance of this fact is emphasized.5)The data on the 14 patients support the opinion that AML, in general, is heterogeneous in origin. It is postulated that monosomy 7 is a marker of a specific pathogenetic pathway of AML, which implies the beginning of the malignancy in a pluripotent stem cell.

The pKa = 13.41 of the title compound (5), determined by a kinetic method, is about 1.3 units lower than the pKa of (2-oxacyclopentylidene)pentacarbonylchromium(0) (1) in 50% MeCN−50% water at 25 °C. The acidifying effect of the methyl group is attributed to its stabilizing effect on the C&dbd;C double-bond resonance structure of the anion (5^-). The rate constant for deprotonation of 5 by OH^- is about the same as for deprotonation of 1, despite the higher acidity of 5. This means that the intrinsic rate constant for proton transfer from 5 is significantly lower than that from 1. This reduction in the intrinsic rate constant is mainly the result of the imbalanced nature of the transition state which deprives the transition state from the stabilizing effect of the methyl group. Based on precedents, the most likely mechanism for the hydrolysis of 5 involves rate-limiting protonation of 5^- concerted with metal−carbon bond cleavage. Even though the relatively small kinetic solvent isotope effect cannot rigorously exclude an alternative nucleophilic substitution mechanism, a comparison of the reactivity of 5 and 1 allows one to rule out this latter mechanism.

A study of the acidity of various Fischer-type (alkoxyalkylcarbene)pentacarbonylmetal complexes and of their rates of deprotonation by OH^- and amines in 50% acetonitrile−50% water at 25 °C is reported. The pKa values of (CO)5M&dbd;C(OMe)CH3 for M = W (12.36) and M = Mo (12.81) are quite comparable to that for M = Cr (12.50) determined previously, and the pKa of (CO)5W&dbd;C(OMe)CH2Ph (10.18) is similar to that of the chromium analog (10.40) reported earlier. These results indicate that the stabilization of the negative charge of the anion by the (CO)5M moiety depends little on the metal. The pKa of (CO)5Cr&dbd;C(OEt)CH3 (12.98) is 0.48 units higher than that of the methoxy analog; this increase probably reflects an enhanced stabilization of the zwitterionic resonance structure of the carbene complex, (CO)5Cr^-C(&dbd;O⁺Et)CH3, by the stronger electron-donating effect of the ethyl group. Intrinsic rate constants for the deprotonation of the carbene complexes by OH^- (ko^OH) were estimated based on the Marcus equation, and intrinsic rate constants for the deprotonation of some of the carbene complexes by amines (ko^B) were determined from Brønsted plots. The dependence of ko^OH and ko^B on the metal is rather small and hardly significant, except possibly for the reaction of (CO)5W&dbd;C(OMe)CH3 with secondary amines, for which ko^B appears to be lower for the tungsten complex than for the chromium and molybdenum derivatives; ko^OH for (CO)5Cr&dbd;C(OEt)CH3 is the same as for the methoxy analog.

The insertion sequence IS1245 is a novel mycobacterial repetitive element identified in Mycobacterium avium. It encodes a transposase which exhibits a 64% amino acid similarity with IS1081, an insertion element present in the M. tuberculosis complex. The host range of IS1245 appears limited to M. avium as this element was not identified in M. intracellulare or in any other of 18 mycobacteria species tested. When IS1245 was used for restriction fragment length polymorphism (RFLP) analysis, human isolates characteristically presented a high number of copies (median, 16; range, 3 to 27) and a diversity of RFLP patterns comparable to that found by pulsed-field gel electrophoresis. Isolates from nonhuman sources differed both in number of copies and in RFLP pattern diversity: while swine isolates shared the characteristics of human strains, those from several avian sources exhibited a very low copy number of IS1245 and appeared clonal on the basis of RFLP.

A prospective study was conducted to assess (a) the long-term results and toxicity of the alternating MOPP/ABVD regimen in advanced Hodgkin's disease; (b) the prognostic value of pretreatment variables and of drug dose intensity.

Controversy still exists over the optimal management of early-stage Hodgkin's disease (HD); presentation features may have a different prognostic impact according to initial therapy, and long-term toxicity must be fully evaluated.

Light speed invariance and nonconservation of simultaneity of separate events are often considered as peculiar properties of Einstein's special relativity (SR). On the contrary it is possible to formulate SR so that simultaneity be conserved and light speed be no longer invariant.Vice versait is possible to formulate prerelativistic, or Galilean physics (GP), so that simultaneity be no longer conserved and light speed be invariant. Simultaneity conservation (or not) and light speed invariance in both GP and SR depend on the arbitrary way in which the clocks of reference systems are synchronized. This arbitrariness leads to infinite possible coordinate transformations (T) in GP, the simplest and most convenient of which are the GalileanT. Similarly there are infinite possibleTin SR and the LorentzTis one of the two most convenient ones. The other suitableTis that of Ives-Tangherlini which implies absolute simultaneity and a noninvariant speed of light. This allows a revival of the ether, although no longer considered as a classical fluid, but as the electromagnetic field radiated by all particles of the universe. This ether should produce the slowing down of clock rate and longitudinal shortening of the moving rods which are the peculiar properties of SR. L'invarianza della velocità della luce e la non conservazione della simultaneità di eventi separati sono spesso considerati come proprietà peculiari della relatività speciale (SR) di Einstein. Al contrario è possibile formulare la SR in modo che la simultaneità sia conservata e la velocità della luce non risulti invariante; viceversa è possibile formulare nella fisica Galileana (GP) una teoria prerelativistica che non conservi la simultaneità e nella quale la velocità della luce risulti invariante; dimostreremo infatti che, tanto nella GP quanto nella SR, la conservazione o meno della simultaneità e dell'invarianza della velocità della luce dipendono dal modo, arbitrario, con cui si sincronizzano gli orologi dei sistemi di riferimento. Da tale arbitrarietà derivano, in GP, infinite possibili trasformazioni (T) delle coordinate di cui le più semplici e convenienti sono le trasformazioni galileane; similmente sono infinite le possibiliTdi coordinate in SR di cui le lorenziane sono fra le più convenienti, oltre le più conosciute. Un'altra notevoleTè quella di Ives-Tangherlini caratterizzata dalla simultaneità assoluta e dalla varianza della velocità della luce. Инвариантность скорости света и несохранеие одновременности разделенных событий часто рассматриваются как характерные свойства специальной теории относительности Эинштеина. Напротив, можно сформулировать специальную теорию относительности так, что одновременность сохраняется, а скорость света не является инвариантом. И наоборот, можно сформулировать дорелятивистскую ияи галилееву физику так, что одновременность не сохраняется, а скорость света является инвариантом. Сохранение одновременности (или нет) и инвариантность скорости света в галилеевой физике и в специальной теории относительности зависят от произвольного способа, с помощью поторого синхронизируются часы в системах отсчета. Эта произвольность приводит к бесконечному числы возможных преобразований координат в галилеевой физике, наиболее простыми и удобными преобразованиями являются преобразования Галилея. Аналогично, существует бесконечное число возможных преобразований в специальной теории относительности, преобразования Поренца являются однимииз двух наиболее удобных. Другое удобное преобразование представляет преобразование Ивса-Тангерлини, которое подразумевает абсолютную одновременность и неинвариантность скорости света. Это преобразование приводит к возрождению эфира, который уже не рассматривается как классическая жидкость, а как электромагнитное поле, излучаемое всеми частицами вселенной. Этот эфир вызывает замедление хода часов и продольное укорочение движущихся стержней, которые представляют характерные свойства спецальной теории относительности.

Tenascin-C binds to cell surface and matrix proteoglycans and to heparin. Two heparin binding regions have recently been localized per tenascin-C monomer, one in the C-terminal fibrinogen-like domain and the other in fibronectin type III repeats 3-5. Here we show that a single region in each subunit is necessary and sufficient for heparin binding by whole tenascin-C at physiological ionic strength. First, native tenascin-C was bound to heparin-agarose and digested with Pronase. A 29-kDa fragment retained on the heparin column was recognized by a monoclonal antibody against the fibrinogen-like domain. In contrast, small fragments labeled by an antibody against fibronectin type III repeats 2-5 were released. Second, mild tryptic digestion of tenascin-C yielded two related fragments of 180 and 170 kDa. The latter missed part of the fibrinogen domain and had lost affinity for heparin, in contrast to the former. Finally, chick tenascin-C constructs were recombinantly expressed in human cells. Whereas the complete protein and a mutant lacking fibronectin type III repeats 1-5 bound to heparin-agarose, recombinant tenascin-C missing the C-terminal fibrinogen-like globe did not. Thus, whole chick tenascin-C contains one essential heparin binding region per subunit, located in the fibrinogen-like domain within 10 kDa from the C terminus.

The reservoir and transmission route of Mycobacterium kansasii are largely unknown. In addition, culturing of M. kansasii from human sources is not proof of disease because it may represent colonization rather than infection. Unfortunately, investigation of the epidemiology and pathogenicity of M. kansasii is complicated by evidence of heterogeneity within the species. A comprehensive study by detailed genotypic analysis of a large collection of M. kansasii isolates (n = 276) from various geographical sources within Europe was conducted. Five defined subtypes of M. kansasii were identified; of these subtypes, type I represents the most common isolate from humans. Although phylogenetic analysis confirmed its relationship to the other M. kansasii types, significant sequence divergence was found at the 16S-23S intergenic spacer. Analysis of the chromosomal polymorphism of type I demonstrated a marked clonal structure for this particular organism. Because M. kansasii is becoming a significant pathogen among immunodeficient hosts, future epidemiological and pathogenicity studies should take into consideration both the heterogeneity within the species and the apparent clonality of the most prevalent M. kansasii isolates infecting humans.

Progress in understanding the basis of resistance to isoniazid (INH) and rifampin (RMP) has allowed molecular tests for the detection of drug-resistant tuberculosis to be developed. Consecutive isolates (n = 95) of Mycobacterium tuberculosis, from a Spanish reference laboratory investigating outbreaks of multidrug-resistant tuberculosis, were coded and sent to two external laboratories for genotypic analysis of INH and RMP resistance by PCR-single-strand conformation polymorphism (SSCP) analysis of specific regions of four genes: part of the coding sequence of katG and the promoter regions of inhA and ahpC for INH and the RMP resistance region of rpoB. After correction for the presence of outbreak strains and multiple isolates from single patients, RMP resistance was detected successfully by PCR-SSCP in > 96% of the RMP-resistant strains. PCR-SSCP had a sensitivity of 87% for INH resistance detection, and mutations in katG, inhA, katG-inhA, ahpC, and katG-ahpC were identified in 36.8, 31.6, 2.6, 13.2, and 2.6%, respectively, of the unique strains. Specificity was 100%. Molecular detection of resistance to the two main antituberculous drugs, INH and RMP, can be accomplished accurately by using a strategy which limits analysis to four genetic regions. This may allow the expedient analysis of drug resistance by reference laboratories.

In order to define normal reference limits of a power law fitting method for evaluating whole blood viscosity, 110 healthy subjects were studied. We found that whole blood viscosity and blood viscosity standardized for hematocrit and plasma viscosity influence were significantly different in adult males, adult females and subjects over 60 years old (p<0.0001). In our opinion, it was also possible to distinguish blood viscosity differences into red blood cells aggregability and deformability components.

Abstract: The application of molecular techniques to investigate strain relatedness may help define the local epidemiology ofMycobacterium avium infection, and, by identifying false isolates (i.e. neither pathogens nor colonizers) resulting from contamination, may serve as a tool for quality control in the laboratory. For this purpose, isolates from all patients (n=129) withMycobacterium avium infections identified over a two-year period were investigated by pulsed-field gel electrophoresis (PFGE). Of 38 PFGE patterns identified, 34 corresponded to unique strains or to isolates present in no more than two or three individuals. One prevalent strain was identified among HIV-infected patients and three patterns were related to culture contamination events. PFGE (i) established the diversity ofMycobacterium avium strains in a community; (ii) identified the existence of a unique strain that may account for one-fifth ofMycobacterium avium isolated from HIV-infected patients locally; (iii) documented the extent and resolution of a suspected pseudo-outbreak; and (iv) uncovered an additional unsuspected contamination event.

There is growing concern that tuberculosis is spread in Europe in the way that it is in the USA. We have used DNA "fingerprinting" in a systematic evaluation of tuberculosis cases notified in our community to uncover foci of transmission. An IS6110 probe was used to test all isolates from culture-confirmed tuberculosis cases (163 patients) notified in 1991-92 in the Canton of Berne. In total, 45 patients (27 6%), potentially linked on the basis of restriction fragment length polymorphism, were investigated epidemiologically. The largest group (n=22) included members of a defined social group (drug addicts, homeless persons, alcoholics), from whom tuberculosis spread to the general population. A key patient developed multidrug-resistant tuberculosis during the surveillance period. This population study showed that (i) extensive transmission of Mycobacterium tuberculosis is now taking place in Europe in the same social setting as in the USA; (ii) there is definite "spillover" to the general population; (iii) the dimensions of the problem cannot be recognised easily by routine public health service activities because of the complexity of the transmission network; and (iv) multidrug-resistant tuberculosis develops in this setting.

Most of the literature dealing with the effect of the pituitary function on the serum proteins and polysaccarides is concerned with the changes caused by total hypophysectomy (cf. Bernasconi, 1956) or by treatment with some of the purified pituitary hormones, primarily somatotrophin and corticotrophin. The observations reported on the effect of injections of pituitary anterior lobe extracts or of implantation of the total gland in animals are, however, relatively few.Goldberg(1938) observed an increase in the serum protein level in normal and hypophysectomized dogs which were treated with a pituitary hormone preparation. This increase was mainly due to an increase both of the albumin and globulin concentrations. Podhradszky(1942) observed that the hypoalbuminemia, which occurred in dogs after hypophysectomy, was further aggravated by the implantation of pituitary glands into the same animals, while the influence of this procedure on total protein content of serum was found to be variable.

Variations in the concentration of total serum protein and of serum protein fractions after hypophysectomy have been observed by different investigators. For fractionation some workers have used salt precipitation (Goldberg, 1938; Levin & Leathem, 1941, 1942; Podhradszky, 1942; Levin, 1943; Moore et al., 1944: Baker & Miller, 1951), while others have used electrophoretic separation (Li, 1944; Moore et al., 1944; Pedersen & Carstensen, 1947). With the first method a globulin increase was always observed, associated with hypoalbuminemia. With the second method this finding was confirmed and the hyperglobulinemia was found to depend upon an increase either of the α-fraction (Moore et al., 1944) or of the γ-fraction (Pedersen & Carstensen, 1947). The total serum protein was generally reduced.Observations on the behaviour of the serum glycoproteins in hypophysectomized animals have been reported by Shetlar et al.(1955), who found a decreased concentration of protein bound hexose, and by Boas &

In serum of patients with cancers the variations of the electrophoretic pattern of proteins (hypoalbuminemia, hyper-alpha-2- and hyper-gamma-globulinemia) are accompanied by a deviated behaviour of the protein-bound carbohydrates: hyperglycoproteinemia (with an almost parallel increase of hexose, hexosamine and sialic acid levels) caused by an elevation of the carbohydrate percentage in the alpha-1 and alpha-2-globulins.As to evaluate the clinical interest of such alterations, which do not show any difference in the various neoplastic localisations, the Authors examinate their importance in the diagnosis of cancers, study their behaviour regarding to diffusions and metastatisation of the disease, and follow their changes in respect to therapy.

Abstract:  It is well established that in acute lymphoblastic leukemia (ALL) patients l-asparaginase (l-Ase) provokes thrombotic events reducing coagulation inhibitors in both adults and children. A tight correlation between thrombotic and hemorrhagic complications and ALL has also been hypothesized because of the high incidence of disseminated intravascular coagulation (DIC) found during the early period of chemotherapeutic treatment apart from l-Ase. All the authors reporting on this subject, however, consider the remission induction phase of treatment the most risky, if not exclusive, for the development of a thrombotic event, in particular if it includes the administration of l-Ase. We report here two cases of ALL patients who experienced a cerebral sinus thrombosis in a later phase of treatment, demonstrating that the thrombotic risk is surely exacerbated by chemotherapy but is not exclusive to the remission induction period.

in an Italian cooperative study on AIDS and cancer a diagnosis of Hodgkin's disease was established in 36 HIV-positive patients. the series was characterized by a high proportion of drug abusers, a high proportion of mixed cellularity and lymphocytic depletion subtypes and short survival. It is still unclear if HIV infection promotes the development of Hodghn's disease or only modifies the course of the disease. According to the authors, however, patients who are HIV seropositive and have biopsyproven Hodgkin's disease should be considered as fulfilling the criteria for AIDS.

An immunohistochemistry method using formalin-fixed tissues, a direct immunofluorescence method using cryostat sections, an enzyme-linked immunosorbent assay (ELISA), and a PCR method were compared for diagnosis in a litter of weaned pigs that had been experimentally inoculated with wild-type porcine epidemic diarrhea virus (PEDV) and killed between 6 and 60 h after onset of diarrhea. The immunohistochemistry method proved to be as reliable as direct immunofluorescence for diagnosis of PEDV in tissues collected postmortem. The good reliability of ELISA for investigating clinical samples was confirmed, whereas the PCR method used was ineffective.