Radioimmunotherapy versus autologous hematopoietic stem cell transplantation in relapsed/refractory follicular lymphoma: a Fondazione Italiana Linfomi multicenter, randomized, phase III trial.

BACKGROUND: Optimal consolidation for young patilents with relapsed/refractory (R/R) follicular lymphoma (FL) remains uncertain in the rituximab era, with an unclear benefit of autologous stem cell transplantation (ASCT). The multicenter, randomized, phase III FLAZ12 (NCT01827605) trial compared anti-CD20 radioimmunotherapy (RIT) with ASCT as consolidation after chemoimmunotherapy, both followed by rituximab maintenance. PATIENTS AND METHODS: Patients (age 18-65 years) with R/R FL and without significant comorbidities were enrolled and treated with three courses of conventional, investigator-chosen chemoimmunotherapies. Those experiencing at least a partial response were randomized 1 : 1 to ASCT or RIT before CD34+ collection, and all received postconsolidation rituximab maintenance. Progression-free survival (PFS) was the primary endpoint. The target sample size was 210 (105/group). RESULTS: Between August 2012 and September 2019, of 164 screened patients, 159 were enrolled [median age 57 (interquartile range 49-62) years, 55% male, 57% stage IV, 20% bulky disease]. The study was closed prematurely because of low accrual. Data were analyzed on 8 June 2023, on an intention-to-treat basis, with a 77-month median follow-up from enrollment. Of the 141 patients (89%), 70 were randomized to ASCT and 71 to RIT. The estimated 3-year PFS in both groups was 62% (hazard ratio 1.11, 95% confidence interval 0.69-1.80, P = 0.6662). The 3-year overall survival also was similar between the two groups. Rates of grade ≥3 hematological toxicity were 94% with ASCT versus 46% with RIT (P < 0.001), and grade ≥3 neutropenia occurred in 94% versus 41%, respectively (P < 0.001). Second cancers occurred in nine patients after ASCT and three after radioimmunotherapy (P = 0.189). CONCLUSIONS: Even if prematurely discontinued, our study did not demonstrate the superiority of ASCT versus RIT. ASCT was more toxic and demanding for patients and health services. Both strategies yielded similar, favorable long-term outcomes, suggesting that consolidation programs milder than ASCT require further investigation in R/R FL.

Role of a conserved glycine triplet in the NSS amino acid transporter KAAT1.

K+-coupled amino acid transporter 1 (KAAT1) belongs to the NSS family of solute transporters and it is expressed in the midgut and in salivary glands of Manduca sexta larvae. As more than 80% of family members, KAATI shows a stretch of three glycines (G85-G87) that according to the structure of the prototype transporter LeuT, is located close to the access of the permeation pathway. In this work the role of the triplet has been investigated by alanine and cysteine scanning methods in protein heterologously expressed in Xenopus laevis oocytes. All the mutants were functional but the surface expression level was reduced for G85A and G87A mutants and unaffected for G86A mutant. All presented altered amino acid uptake and transport associated currents in the presence of each of the cations (Na+, K+, Li+) that can be exploited by the wt. G87A mutant induced increased uncoupled fluxes in the presence of all the cations. Cross-linking studies, performed by the treatment of cysteine mutants with the oxidative complex Cu(Il)(l,10-phenanthroline)3, showed that limiting the flexibility of the region by covalent blockage of position 87, causes a significant reduction of amino acid uptake. Na+ protected G87C mutant from oxidation, both directly and indirectly. The conserved glycine triplet in KAAT1 plays therefore a complex role that allows initial steps of cation interaction with the transporter.

Molecular physiology of the insect K-activated amino acid transporter 1 (KAAT1) and cation-anion activated amino acid transporter/channel 1 (CAATCH1) in the light of the structure of the homologous protein LeuT.

K-activated amino acid transporter 1 (KAAT1) and cation-anion-activated amino acid transporter/channel 1 (CAATCH1) are amino acid cotransporters, belonging to the Na/Cl-dependent neurotransmitter transporter family (also called SLC6/NSS), that have been cloned from Manduca sexta midgut. They have been thoroughly studied by expression in Xenopus laevis oocytes, and structure/function analyses have made it possible to identify the structural determinants of their cation and amino acid selectivity. About 40 mutants of these proteins have been studied by measuring amino acid uptake and current/voltage relationships. The results obtained since the cloning of KAAT1 and CAATCH1 are here discussed in the light of the 3D model of the first crystallized member of the family, the leucine transporter LeuT.

Artificial chromosomes for antibiotic-producing actinomycetes.

Bacteria belonging to the order Actinomycetales produce most microbial metabolites thus far described, several of which have found applications in medicine and agriculture. However, most strains were discovered by their ability to produce a given molecule and are, therefore, poorly characterized physiologically and genetically. Thus, methodologies for genetic manipulation of actinomycetes are not available and efficient tools have been developed for just a few strains. This constitutes a serious limitation to applying molecular genetics approaches to strain development and structural manipulation of microbial metabolites. To overcome this hurdle, we have developed bacterial artificial chromosomes (BAC) that can be shuttled among Escherichia coli, where they replicate autonomously, and a suitable Streptomyces host, where they integrate site-specifically into the chromosome. The existence of gene clusters and of genetically amenable host strains, such as Streptomyces coelicolor or Streptomyces lividans, makes this a sensible approach. We report here that 100 kb segments of actinomycete DNA can be cloned into these vectors and introduced into genetically accessible S. lividans, where they are stably maintained in integrated form in its chromosome.

Early assessment of brain maturation by MR imaging segmentation in neonates and premature infants.

PURPOSE: We evaluated the impact of premature extrauterine life on brain maturation. PATIENTS AND METHODS: Twelve neonates underwent MR imaging at 40 (39.64 +/- 0.98) weeks (full term). Fifteen premature infants underwent 2 MR imaging examinations, after birth (preterm at birth) and at 40 weeks (41.03 +/- 1.33) (preterm at term). A 3D MR imaging technique was used to measure brain volumes compared with intracranial volume: total brain volume, cortical gray matter, myelinated white matter, unmyelinated white matter, basal ganglia (BG), and CSF. RESULTS: The average absolute volume of intracranial volume (269.8 mL +/- 36.5), total brain volume (246.5 +/- 32.3), cortical gray matter (85.53 mL +/- 22.23), unmyelinated white matter (142.4 mL +/-14.98), and myelinated white matter (6.099 mL +/-1.82) for preterm at birth was significantly lower compared with that for the preterm at term: the average global volume of intracranial volume (431.7 +/- 69.98), total brain volume (391 +/- 66,1), cortical gray matter (179 mL +/- 41.54), unmyelinated white matter (185.3 mL +/- 30.8), and myelinated white matter (10.66 mL +/- 3.05). It was also lower compared with that of full-term infants: intracranial volume (427.4 mL +/- 53.84), total brain volume (394 +/- 49.22), cortical gray matter (181.4 +/- 29.27), unmyelinated white matter (183.4 +/- 27.37), and myelinated white matter (10.72 +/- 4.63). The relative volume of cortical gray matter (30.62 +/- 5.13) and of unmyelinated white matter (53.15 +/- 4.8) for preterm at birth was significantly different compared with the relative volume of cortical gray matter (41.05 +/- 5.44) and of unmyelinated white matter (43.22 +/- 5.11) for the preterm at term. Premature infants had similar brain tissue volumes at 40 weeks to full-term infants. CONCLUSION: MR segmentation techniques demonstrate that cortical neonatal maturation in moderately premature infants at term and term-born infants was similar.

Assembly of large genomic segments in artificial chromosomes by homologous recombination in Escherichia coli.

We developed a method for the reconstruction of a 100 kb DNA fragment into a bacterial artificial chromosome (BAC). The procedure makes use of iterative rounds of homologous recombination in Escherichia coli. Smaller, overlapping fragments of cloned DNA, such as cosmid clones, are required. They are transferred first into a temperature-sensitive replicon and then into the BAC of choice. We demonstrated the usefulness of this procedure by assembling a 90 kb genomic segment into an E.coli-STREPTOMYCES: artificial chromosome (ESAC). Using this procedure, ESACs are easy to handle and remarkably more stable than the starting cosmids.

Simultaneous measurements of ionic currents and leucine uptake at the amino acid cotransporter KAAT1 expressed in Xenopus laevis oocytes.

The transport properties of the intestinal amino acid cotransporter KAAT1, heterologously expressed in Xenopus oocytes, were studied using simultaneous voltage-clamp and tritiated leucine uptake measurements. While addition of 1 mM leucine to oocytes kept at -80 mV in presence of Na(+) or K(+) caused an increase in holding current, in presence of Li(+) the current was reduced. Uptake measurements in voltage-clamp conditions showed that a comparable accumulation of amino acid occurred in all three ionic conditions and irrespective of the direction and amount of the current change. The ratio of moles of transferred charge to moles of transported amino acid ranges from 1.45 for K(+) to 3.52 for Li(+). A hypothetical interpretation involving the coexistence of two populations of transporters, one operating in the uncoupled mode and the other in the substrate transport mode is discussed.

Three kinds of currents in the canine betaine-GABA transporter BGT-1 expressed in Xenopus laevis oocytes.

The cloned canine betaine-GABA cotransporter BGT-1 has been heterologously expressed in Xenopus laevis oocytes in order to characterize its electrophysiological properties. Voltage-clamp experiments on transfected oocytes reveal the presence of three types of membrane current which are absent in non-injected oocytes: (i) an organic substrate-independent current (uncoupled current); (ii) a transport-associated current, seen upon addition of betaine or GABA; (iii) presteady-state currents induced by voltage changes. The three kinds of current are analogous to those reported in structurally similar cotransporters. The transport-associated current is strictly dependent on the presence of Na(+). The good correlation between the amount of charge underlying the presteady-state currents and the transport-associated current indicates that both processes are due to the activity of the transporter.

Regulation of cardiac L-type Ca2+ channel by coexpression of G(alpha s) in Xenopus oocytes.

Activation of G(alpha s) via beta-adrenergic receptors enhances the activity of cardiac voltage-dependent Ca2+ channels of the L-type, mainly via protein kinase A (PKA)-dependent phosphorylation. Contribution of a PKA-independent effect of G(alpha s) has been proposed but remains controversial. We demonstrate that, in Xenopus oocytes, antisense knockdown of endogenous G(alpha s) reduced, whereas coexpression of G(alpha s) enhanced, currents via expressed cardiac L-type channels, independently of the presence of the auxiliary subunits alpha2/delta or beta2A. Coexpression of G(alpha s) did not increase the amount of alpha1C protein in whole oocytes or in the plasma membrane (measured immunochemically). Activation of coexpressed beta2 adrenergic receptors did not cause a detectable enhancement of channel activity; rather, a small cAMP-dependent decrease was observed. We conclude that coexpression of G(alpha s), but not its acute activation via beta-adrenergic receptors, enhances the activity of the cardiac L-type Ca2+ channel via a PKA-independent effect on the alpha1C subunit.

Effect of propranolol on fetal tachycardia in diabetic pregnancy.

Propranolol was administered during the last 20 days of pregnancy to a diabetic woman because of fetal tachycardia (heart rate approximately 200 beats/min). With a daily dose of 160 mg of propranolol, a fetal heart rat of 120 to 160 beats/min could be achieved. Blood concentration of propranolol was measured in the mother and infant after birth. The level in the neonatal blood was 20 percent of the maternal sample, which is definitely higher than expected from animal experiments. No undersirable effect of propranolol treatment was detected. Postpartum, the infant demonstrated paroxysmal supraventricular tachycardia, and propranolol was again essential in maintaining a normal rate.

Retinopathy of prematurity.

This review deals with retinopathy of prematurity (ROP), a disease characterized by retinovitreal neovascularization, eventually retinal detachment and blindness. Due to the increasing number of extremely premature newborns, it is becoming more frequent. ROP of all stages occurs in 25-35% of surviving premature newborns of gestational age up to approximately 35 weeks. Stages 3 or more occur in 5-10%, blindness in 3-5% of very immature babies. The incidence is inversely related to gestational age. Classification is internationally unified (ICROP) and describes 5 stages. Its pathogenesis has not yet been clarified. More or less proven risk factors are retinovascular immaturity, hyperoxia and possibly circulatory and respiratory instability. Prophylaxis consists in avoiding hyperoxia, and probably also in keeping the extremely premature newborn stable. Ophthalmologic examinations must be performed by ophthalmologists experienced in this field or under their direct responsibility and must be standardized. Treatment of ROP can be carried out at a certain stage by coagulation therapy.

Hepatitis C antibody and cryoglobulins in patients with cirrhosis and hepatocellular carcinoma.

OBJECTIVE: To assess: (1) the role of hepatitis C virus (HCV), hepatitis B virus (HBV) and alcoholism as risk factors for hepatocarcinoma (HCC) in patients with liver cirrhosis, and (2) the presence of cryoglobulins in HCV + patients with and without HCC. PATIENTS: 82 cirrhotic patients, 41 with and 41 without HCC, who were admitted consecutively to our General Medicine Division from January 1992 to June 1994, were studied. RESULTS: The prevalence of HBV markers, anti-HCV and alcoholism in patients with liver cirrhosis and HCC was 39%, 63.4% and 24%, and in cirrhotics without HCC it was 31.7%, 51.2% and 63.4%, respectively. Cryoglobulins were present in 87.5% of the patients with HCC (78.5% anti-HCV +) and in 57.8% of the patients without HCC (81.8% anti-HCV +). The cryoglobulins, as characterized in 11 cases, were type III in nine cases and type II in two. CONCLUSIONS: Patients with cirrhosis, especially when associated with HCV and HBV infection, are at high risk for HCC and therefore require careful follow-up. Moreover, a strong association between HCV and the cryoglobulins in cirrhotics with and without HCC was evident, thus supporting the possible role of this virus in stimulating lymphocytes to produce cryoglobulins.

Comparison of sequence of cDNA clone with other genomic and cDNA sequences for human C-reactive protein.

A clone for C-reactive protein (CRP) has been isolated from a human liver cDNA library; this clone harbors a plasmid, pC81, which has an insert of 1631 bp. When compared to genomic and cDNA sequences published to date now, pC81 has revealed homologies and differences that might help to clarify the structure of this gene and the presence of allelic variants in man.

Comparison between transcutaneous PO2 and pulse oximetry for monitoring O2-treatment in newborns.

213 paired tcpO2/paO2-data and 186 paired tcSO2/SaO2-data measured in 25 newborns (10 term, 15 prematures) were compared. The correlation coefficient for tcpO2/paO2 was 0.796, for tcSO2/SaO2 0.944. Sensitivity for discriminating between normo- and hypoxemia (paO2 less than 50 torr) was 82% for the tcpO2- and 88% for the tcSO2-method. Positive predictive values for discriminating between normo- and hypoxemia were 88% for both methods. Sensitivity for discrimination between normo- and hyperoxemia (paO2 greater than 100 torr) was 85% for the tcpO2- and 100% for the tcSO2-method. Positive predictive values for the discrimination between normo- and hyperoxemia were 58% and 25% for tcpO2- and tcSO2 respectively. Pulse oximetry proved to be less cumbersome than the tcpO2-method. However, as tcpO2, it could not be used in some very immature newborns and in those with circulatory instability. In conclusion, these preliminary results show a similar discrimination between normoxemia and hypo-hyperoxemia for both methods. A better sensitivity of pulse oximetry for hyperoxemia is counteracted by a lesser positive predictive value.

[Critical organs and external irradiation in gynaecological cancers: can water be used as contrast agent to make easier the delineation of the small intestine?]. / Organes critiques et irradiation externe des cancers gynécologiques: l'eau peut-elle être utilisée comme produit de contraste pour faciliter la délinéation de l'intestin grêle?

PURPOSE: To validate the use of water as contrast agent for the delineation of the small intestine on the planning CT of external beam in patients treated with conformal radiotherapy for gynaecological tumours. PATIENTS AND METHODS: From March to September 2003, 20 patients received an external irradiation for a gynaecological carcinoma (13 with cervix carcinoma, seven with endometrial carcinoma) in the radiotherapy department of the Centre G.F. Leclerc of Dijon. The protocol of opacification of the small intestine consisted in administration of a "negative" contrast agent: water. The protocol commonly used for the bladder filling, i.e. absorption of 500 cm(3) of water from 60 to 30 min before the CT-scan, was applied for the evaluation of the visualisation of the small intestine in the 12 first patients (group I). For the last eight patients (group II), the absorption of the same amount of water was fractionated, every 10 min within half an hour before the start of the examination. RESULTS: The small bowel identification was possible in 100% of cases without any need of administration of a "positive" contrast agent. In overall, the identification of the small intestine was considered as easy in 14 patients (70%) and as difficult in two patients (10%). In group I, the delineation was considered as easy in 50% of cases, moderately easy in 33% of cases and none easy in 17% of cases. Conversely, no difficulty was encountered for the definition of the small bowel in all patients of group II. CONCLUSIONS: Water is an efficient "negative" contrast agent for the differentiation of the small bowel from the colon on the planning abdomino-pelvic CT. Nevertheless, the delineation was really made easier only when the fractionated protocol of water absorption within half an hour before CT was used.

[Acute and chronic cerebrovascular diseases. A therapeutic approach]. / Vasculopatie encefaliche acute e croniche. Approccio terapeutico.

The effectiveness of a specific treatment for ictus in its acute phase is highly controversial. A series of variables is identified in 1503 cases that would appear to have a particular effect on the reduction of mortality. On these, specific treatment with Buflomedil coincides with a reduction in lethal outcome of 4% over the two-year period out of 380 cases. In their turn chronic postictal and cerebral ischaemia states (a total of 105 cases) respond positively to prolonged treatment with Buflomedil in a percentage varying from 57% in postictal chronic states to 70-73% in chronic cerebral ischemias.

A mydriatic eye-drop combination without systemic effects for premature infants: a prospective double-blind study.

Eye drops used for diagnostic mydriasis may produce systemic side effects in preterm infants. Studies on the pupil dilating and systemic effect of various mydriatic agents yielded conflicting results. We conducted a prospective randomized double-blind study on the systemic effect of two mydriatic eye-drop combinations. Thirty-nine preterm infants were randomly assigned to two groups. An eye-drop combination of 2.5% phenylephrine and 0.5% tropicamide (group D) was compared with the combination of 0.5% cyclopentolate and 0.5% tropicamide (group F). Either eye-drop combination was followed by 0.5% tropicamide given 20 minutes later. Heart rate (HR) and the systolic, mean, and diastolic blood pressure (BP) were recorded before and after eye-drop instillation and after ophthalmoscopy. A control session with NaCl eye drops was added for each infant. A significant increase of BP and HR peak values was observed within 7 to 10 minutes after the cyclopentolate/tropicamide combination only. On the other hand, the mydriatic effect of the phenylephrine/tropicamide combination was significantly superior to that of the cyclopentolate/tropicamide combination. We recommend the combination of 2.5% phenylephrine and 0.5% tropicamide to achieve a sufficient diagnostic mydriasis without systemic side effects in preterm infants.

[Evaluation of published recommendations for screening studies of retinopathy of prematurity]. / Uberprüfung publizierter Empfehlungen für Screening-Untersuchungen bei Retinopathia praematurorum.

PURPOSE: Evaluation of different and partially contradictory guidelines for screening for retinopathy of prematurity published in Germany. PATIENTS: The data on 1219 preterm infants examined in Bern (n = 900) and Berlin (n = 319) were analyzed. A total of 680 preterm infants (56%) had a birth weight below 1500 g. The remaining infants were examined because oxygen had to be supplied or surgery done before or around the estimated delivery time. RESULTS: Stage 3 retinopathy was found in 88/1219 (7.2%) preterm infants. Only 5/88 infants weighed more than 1500 g at birth. Three of these infants had a birth weight below 2000 g and needed supplemental oxygen for a prolonged period of time. The other 2 infants had birth weights of more than 2000 g and were severely ill. Of 1219 preterm infants, 37 (3%) developed threshold retinopathy (according to the criteria of the multicenter trial); 35/37 infants weighed less than 1230 g at birth. The remaining 2 children (1650 g and 2185 g birth weight) were severely ill. CONCLUSIONS: Preterm infants should be screened for retinopathy of prematurity: (1) if the birth weight is below 1500 g; (2) if the birth weight is below 2000 g and oxygen supply was necessary for more than 30 days; (3) if infants are very sick or must undergo multiple surgery before term. The first examination should be scheduled for the 6th week postnatal and not before 31 weeks postmenstrual age.

[Severe pelvic injuries: indications and techniques of skeletal fixation]. / I traumi complessi del bacino: indicazioni e tecniche della stabilizzazione scheletrica.

The aim of this work is to assess the role of immediate Hoffmann's external fixation frame and delayed posterior internal fixation in the management of severe pelvic injuries. We have also analysed the role of selective arteriography and embolization for controlling pelvic fractures hemorrhage and the relationship between embolization and external fixation in the management of severe bleeding. We reviewed 112 patients with severe pelvic injuries admitted to our hospital from 1986 to 1998 (71 males, 41 females). The average age was 39.8 years, mean ISS 31.2. Unstable pelvic ring fracture was present in 59 (52.6%) patients at admission in the emergency room: hemorrhagic shock in 43 patients (38.3%). Fourteen (12.5%) patients died. Eight deaths (57%) were directly caused by hemorrhagic shock, two secondary to major caval lesion, three brain lesions and two respiratory failure. The last patient died secondary to MOF. Twenty-three patients (20.5%) with unstable fracture underwent pelvic fracture ring fixation surgery: eleven (9.8%) Hoffmann's anterior external fixation frame (hemorrhage control in all patients) and eight (7.1%) delayed internal posterior fixation. Six (5.3%) patients underwent arteriography and embolization: hemorrhage was stopped in all cases and no patient died in this group.