Preclinical Characterization of an Antibody-Drug Conjugate Targeting CS-1 and the Identification of Uncharacterized Populations of CS-1-Positive Cells.

Multiple myeloma is a hematologic cancer that disrupts normal bone marrow function and has multiple lines of therapeutic options, but is incurable as patients ultimately relapse. We developed a novel antibody-drug conjugate (ADC) targeting CS-1, a protein that is highly expressed on multiple myeloma tumor cells. The anti-CS-1 mAb specifically bound to cells expressing CS-1 and, when conjugated to a cytotoxic pyrrolobenzodiazepine payload, reduced the viability of multiple myeloma cell lines in vitro In mouse models of multiple myeloma, a single administration of the CS-1 ADC caused durable regressions in disseminated models and complete regression in a subcutaneous model. In an exploratory study in cynomolgus monkeys, the CS-1 ADC demonstrated a half-life of 3 to 6 days; however, no highest nonseverely toxic dose was achieved, as bone marrow toxicity was dose limiting. Bone marrow from dosed monkeys showed reductions in progenitor cells as compared with normal marrow. In vitro cell killing assays demonstrated that the CS-1 ADC substantially reduced the number of progenitor cells in healthy bone marrow, leading us to identify previously unreported CS-1 expression on a small population of progenitor cells in the myeloid-erythroid lineage. This finding suggests that bone marrow toxicity is the result of both on-target and off-target killing by the ADC.

Synthesis and in vitro evaluation of SG3227, a pyrrolobenzodiazepine dimer antibody-drug conjugate payload based on sibiromycin.

A novel pyrrolobenzodiazepine dimer payload, SG3227, was rationally designed based on the naturally occurring antitumour compound sibiromycin. SG3227 was synthesized from a dimeric core in an efficient fashion. An unexpected room temperature Diels-Alder reaction occurred during the final step of the synthesis and was circumvented by use of an iodoacetamide conjugation moiety in place of a maleimide. The payload was successfully conjugated to trastuzumab and the resulting ADC exhibited potent activity against a HER2-expressing human cancer cell line in vitro.

[Delayed umbilical cord clamping in preterm infants born before 37 weeks of gestation: A prospective observational study]. / Clampage tardif du cordon ombilical chez les enfants prématurés nés avant 37 semaines d'aménorrhée : étude observationnelle prospective.

INTRODUCTION AND OBJECTIVES: Many international studies have demonstrated that delayed umbilical cord clamping reduces neonatal morbidity. However, in France, delayed umbilical cord clamping is still not performed in many neonatal units. The aims of this study were to evaluate the feasibility of developing a protocol of delayed umbilical cord clamping in the maternity ward of the Toulouse university hospital and to evaluate the impact of this new protocol on neonatal mortality. METHODS: We conducted a prospective observational study including 123 preterm infants born before 37 weeks of gestation between June 2012 and June 2013 and hospitalized at birth. Delayed cord clamping was performed for at least 30s after birth; otherwise, it was evaluated as early cord clamping. We excluded twin-to-twin transfusion syndrome, congenital abnormalities, alloimmunization, and perinatal asphyxia. We analyzed the reasons why delayed umbilical cord clamping was not performed and then neonatal morbidity in our population. RESULTS: Delayed umbilical cord clamping was performed on 79 infants and 44 infants had early umbilical cord clamping. The two groups had similar baseline characteristics. Preterm infants in the delayed cord-clamping group had a higher level of hemoglobin during the first 24h of life (17.9g/dL versus 16.6g/dL, P=0.005), fewer of them required transfusion (14% versus 35%, P=0.03), and fewer presented late-onset sepsis (8% versus 26%, P=0.02) or bronchopulmonary dysplasia (9% versus 26%, P=0.03). There was no statistically significant increase of hyperbilirubinemia requiring phototherapy. DISCUSSION AND CONCLUSION: Implanting a new protocol of delayed umbilical cord clamping in our maternity ward proved to be possible without difficulty. The advantages of delayed umbilical cord clamping were observed in this prospective study. Today, delayed cord clamping has become a common practice in our maternity unit.

[Does the paper speed in fetal heart monitoring during labour influence the variability in the interpretation by professionals?] / La vitesse de déroulement du rythme cardiaque fœtal en cours de travail a-t-elle un impact sur la variabilité d'interprétation par les professionnels ?

OBJECTIVE: Assessing inter- and intra- observer agreement in the reading of fetal heart rate (FHR) between two different paper speeds (1 and 2cm/min) using FIGO classification. MATERIAL AND METHODS: Single-centre experimental study consisting in reading 60minutes FHR tracings by six readers (3 midwives and 3 obstetricians) during 1cm and 2cm/min sessions within a period of three weeks. The reading guideline was based on FIGO classification. Inter- and intra-observer agreement was assessed thanks to Kappa coefficient (K) and percentage of agreement (PA) using the classification of FHR tracings drawn up by readers. RESULTS: Intra-observer agreement reached 60% between the two paper speeds, and PA ranged from 48 to 67%. Inter-observer agreement was poor to moderate (K=0.42 for 1cm/min sessions and K=0.38 for 2cm/min sessions). Inter-observer agreement was significantly higher for normal tracings (PA ranged from 55.2% for 2cm/min sessions to 57.4% for 1cm/min sessions). The preterminal category had the lowest concordance rates (PA=19% for 1cm/min sessions and 20, 7% for 2cm/min sessions). CONCLUSION: This study did not highlight significant differences in intra- and inter-observer variability between the two FHR paper speeds. The 1cm/min paper speed, which is commonly used in France, is more economical and gives a better bedside overview of FHR. Therefore, it should be recommended.

[Combined photodynamic therapy with verteporfin and intravitreal triamcinolone acetonide for choroidal neovascularization with pigment epithelium detachment in age-related macular degeneration]. / Traitement simultané par photothérapie dynamique et triamcinolone intravitréenne des néovaisseaux associés à un décollement de l'épithélium pigmentaire.

OBJECTIVE: To assess the advantages of a treatment combining dynamic phototherapy with intravitreally injected Kenacort (triamcinolone acetonide) administered on the same day for choroidal neovascularization with pigment epithelium detachment in age-related macular degeneration. METHOD: This prospective study involved 24 patients (26 eyes) with pigment epithelium detachment. Each patient underwent dynamic phototherapy according to standard parameters, followed 2 h later by intravitreal injection of 4 mg Kenacort (triamcinolone acetonide) performed in the operating room. The effectiveness of the treatment was assessed with an ETDRS vision test and optical coherence tomography before the operation and again 1, 3, and 6 months later. RESULTS: The group of 24 patients (26 eyes) included 17 women and seven men whose mean age was 77 years (+/-6). In half of the cases, it was the second eye affected by age-related macular degeneration. Major inflammatory reaction following intravitreal injection was resolved within 24 h by local anti-inflammatory treatment. Baseline visual acuity was 40 characters on the ETDRS (+/-15) and final vision was significantly improved to 43 characters (+/-19) after 3 months. Three patients (12%) had their vision improved by more than three lines. For 19 patients (79%), the treatment eliminated pigment epithelium detachment after 1 month, with no recurrence. DISCUSSION: No single recognized treatment exists for pigment epithelium detachment. Photodynamic therapy has not proven effective, according to the TAP study. For the last few years, several teams have recommended intravitreal injections of Kenacort (triamcinolone acetonide) as a complement to photodynamic therapy to treat different types of choroidal neovascularization. This combination has also been tried in the treatment of pigment epithelium detachment associated with age-related macular degeneration, although the protocol has not yet been clearly established. In order to decrease the cost of treatment by reducing the transport back and forth between the patient's home and the hospital, we have administered this dual treatment on the same day and evaluated the benefits for the patient. We have obtained very satisfactory anatomical results, making it possible to proceed with low-vision rehabilitation, and eventually improve visual acuity. CONCLUSION: The results of this study indicate that a level of efficiency at least as high as that of treatment at two different times for occult choroidal neovascularization complicated by pigment epithelium detachment can be achieved by a dual treatment the same day, with an accompanying reduction in cost.

Stable human calcitonin analogues with high potency on bone together with reduced anorectic and renal actions.

Various derivatives of human calcitonin have been synthesized and their biological characteristics compared with those of existing calcitonins. The acute effects of these analogues in reducing serum calcium levels and suppressing appetite were assessed in rats. A calcitonin analogue, PO-1 (CGNLSTCMLGKLSQELHKLQTYPQTAIGVGAP-NH2), having both the N- and C-terminal ten amino acid sequences those of human calcitonin, and the 12 amino acid central region that of salmon calcitonin, was found to have equal effectiveness with salmon calcitonin and elcatonin for reducing serum calcium levels. Strong hypocalcemic activity was also exhibited by PO-23 ([cyclo-Asp1, Lys7]-[des-Gly2]-[Leu8]-PO-1) and PO-29 ([Asp15, Asn17 , Phe19, His20]-PO-23). PO-23 was prepared by replacing the N-terminal Cys-Cys S-S bond of PO-1 with a ring structure composed of an Asp-Lys peptide bond to enhance physicochemical stability. PO-29 was prepared by modifying the central area of the PO-23 molecule to more closely mimic human calcitonin. When tested in vitro, human calcitonin analogues with a [cyclo-Asp1, Lys7] structure showed biological activities on osteoclast-like cells comparable to those of existing calcitonins (salmon calcitonin and elcatonin) in keeping with their relative potencies for in vivo hypocalcemic action. Acute anorectic activity in rats was strong with salmon calcitonin and elcatonin but relatively reduced with human calcitonin analogues having a [cyclo-Asp1, Lys7] structure. The activities of these analogues on kidney cells were also weaker than that of salmon calcitonin or elcatonin. These results suggest that stable human calcitonin analogues with a [cyclo-Asp1, Lys7] structure suppress bone resorption to a degree similar to that of salmon calcitonin or elcatonin with weaker activities on non-osseous tissues which might be related to adverse reaction.

Early experience and reinforcer quality in delayed flavour-food learning in the rat.

The ability of hungry rats to associate flavours with the consequences of ingesting glucose solutions was studied in three experiments. Experiment 1 used a procedure in which on some days one flavour, e.g. cinnamon, was presented and followed after 20 min by 20% glucose, while on other days a second flavour, e.g. wintergreen, was presented, but not followed by any event. During this training, subjects who received quinine-tainted glucose increased their consumption of the predictive flavour relative to groups given no quinine, but quinine tainting did not affect conditioned preference for the predictive flavour in choice tests. With the aim of discovering whether prior experience of a variety of foods improves ability to learn new flavour-calorie associations, Experiment 2 and used a similar procedure to compare subjects raised on a varied diet ("supermarket" rats) with controls previously given only chow. Contrary to expectation, the supermarket rats showed some impairment both on this delay task and, in Experiment 3, on one using a "mixtures" procedure. This involved presenting a mixture of one cue flavour with glucose-quinine on some days and a mixture of a second cue with an equally palatable saccharin solution on other days. Acquisition was particularly rapid in control subjects, reaching asymptote after only two flavour-glucose pairings. It was concluded that neither a decrease in palatability, as in Experiment 1, nor prior experience with a range of foods, as in Experiments 2 and 3, improve a rat's ability to associate a new flavour with calories.

Effect of age on circulating immunoreactive and bioactive parathyroid hormone levels in women.

Although levels of serum immunoreactive parathyroid hormone (iPTH) increase with age in women, this could be caused by retention of non-biologically active PTH fragments by the aging kidney. In 102 normal women, aged 30 to 89 yr, serum iPTH increased with age by 58% (r = 0.33, p less than 0.001) with antiserum GP-1M (which has midmolecule specificity) and 43% (r = 0.32, p less than 0.001) with antiserum CH-12M (which may have whole molecule specificity); urinary cAMP/GFR excretion increased by 29% (r = 0.22, p less than 0.05). The results of these assays were validated by comparison with serum levels of biologically active PTH (BioPTH) in immunoextracts of serum followed by renal adenylate cyclase assay in a selected subgroup of 25 of the women. Serum BioPTH correlated with serum iPTH assessed by antiserum GP-1M (r = 0.48, p less than 0.05) and antiserum CH-12M (r = 0.48, p less than 0.05) but not with urinary cAMP. The data are consistent with an increase of parathyroid function with aging: clearly, we do not find decreased parathyroid function as would be expected if age-related bone loss was not mediated, in part, by PTH.

[Anti-inflammatory activity of 5 copper complexes]. / Activité anti-inflammatoire de cinq complexes du cuivre.

The anti-inflammatory activity of five copper complexes is shown in the cotton wad granuloma test in Rats. The activity due to copper seems to be only modulated by the ligands in the complexes studied.

Value of high resolution real-time ultrasonography in secondary hyperparathyroidism.

Thirty-two patients were treated surgically for symptomatic secondary or tertiary hyperparathyroidism, and 27 of these patients had high resolution (10 mHz) real-time ultrasonography before parathyroidectomy. This preoperative localization study identified one or more enlarged hyperplastic parathyroid glands in all but one patient who had not had a previous parathyroid operation, and in five of six patients who did have previous parathyroid operations. In both of the patients in whom no parathyroid glands were identified by ultrasonography the only abnormal enlarged parathyroid glands were those situated within the superior mediastinum. When large glands are not observed by ultrasonography in patients with severe secondary hyperparathyroidism, the glands are usually situated in the superior mediastinum, behind the trachea or esophagus, or deeply within the neck. The size of the parathyroid glands correlated positively with the serum parathyroid hormone level and with the severity of the secondary hyperparathyroidism. Thus, the preoperative identification of parathyroid glands by ultrasonography not only localizes the site of most hyperplastic parathyroid glands (70 percent of patients), but also detects those patients who have enlarged parathyroid glands, elevated serum parathyroid hormone levels, and severe secondary hyperparathyroidism. These are the patients who are thus unlikely to respond to further medical therapy.

The pathogenesis of infantile malignant osteopetrosis: bone mineral metabolism and complications in five infants.

Bone mineral metabolism was studied in five infants aged 8 to 22 months with severe osteopetrosis. There were findings consistent with biochemical osteomalacia. These included hypocalcemia, hypophosphatemia, high serum acid phosphatase and alkaline phosphatase activity, high levels of serum parathyroid hormone, and high urinary cyclic AMP. Serum 1,25(OH)2 vitamin D3 level was high in the one patient tested. Radiographs in all infants revealed rachitic changes in the metaphyses. However, dense bones on radiographs, calcium balance studies, and radio-calcium absorption studies demonstrated markedly positive calcium balance. Iliac crest bone biopsies showed increased quantity of woven bone with abundant numbers of osteoclasts, excessive amounts of osteoid, myelofibrosis, and a decreased number of Howship's lacunae. The wide bands of unmineralized osteoid did not take up tetracycline. In vitro bone resorbing activity due to osteoclast activating factor from cultured stimulated leukocytes was normal. Bone turnover however, was now as evidenced by low urinary hydroxyproline levels. We interpret these findings as indicating there is decreased bone remodeling and resorption in spite of increased humoral stimuli and osteoclasts. Since calcitonin levels were normal for age, the most likely cause of the impaired bone remodeling sequence was defective osteoclast function. We postulate that there may be a common genetic defect in phagocyte cells, including monocytes, neutrophils and osteoclasts, which accounts for the abnormalities of mineral metabolism and previously reported hematologic, neurologic, and infectious complications.

Osteomalacia and celiac disease: response to 25-hydroxyvitamin D.

In this 54 year old woman with celiac disease, osteomalacia developed while she was on a gluten-free diet which had caused regression of her steatorrhea. She was not responsive to large doses of parenterally administered dihydrotachysterol and calcium, but she was responsive to the oral administration of 25-hydroxyvitamin D3 (25-OHD3). The data suggest that 25-OHD3 is the treatment of choice for patients with vitamin D deficiency due to intestinal malabsorption.

1,25 dihydroxycholecalciferol effects in chronic dialysis. A double-blind controlled study.

1,25 dihydroxycholecalciferol [1,25(OH)2D3] was studied in a double-blind controlled fashion in patients on chronic dialysis. Serum calcium was unchanged in 16 patients on vitamin D3 (D3) (400 to 1200 IU/day). In 15 patients on 1,25(OH)2D3 (0.5 to 1.5 microgram/day), serum calcium increased from 9.05 +/- .15 to 10.25 +/- .20 mg/dl (p less than 0.001), returning to 9.37 +/- .16 mg/dl (p less than 0.001) in the post control period. Patients on D3 showed no reversible decrease in immunoreactive parathyroid hormone levels, but patients on 1,25(OH)2D3 did, from a control of 1077 +/- 258 to 595 +/- 213 microliter equivalents/ml (p less than 0.01), and returned to 1165 +/- 271 microliter equivalents/ml (p less than 0.005). Nine of 12 patients on D3 who underwent serial iliac-crest biopsies showed histologic deterioration, and six of seven who received 1,25(OH)2D3 were improved or unchanged (p less than 0.025). Bone mineral and calcium decreased in patients on D3 (p less than 0.05) but not in those on 1,25(OH)2D3. Hypercalcemia occurred in five of 15 patients. We conclude that 1,25(OH)2D3 has a calcemic effect in chronic dialysis patients, decreases levels of immunoreactive parathyroid hormone, and is associated with histologic improvement in bone disease. Thus, 1,25(OH)2D3 is a valuable adjunct to the management of renal osteodystrophy but requires monitoring of serum calcium to avoid hypercalcemia.

The effect of 25-hydroxy vitamin D3 in the osteomalacia of chronic renal failure.

1. Five patients with the osteomalacia of chronic renal failure were given 50--100 nmol of 25-hydroxy vitamin D3 intravenously per day for 24--28 days. 2. In all five patients, during administration of 25-hydroxy vitamin D3 there was a substantial rise in the plasma concentration of 25-hydroxy vitamin D from initially abnormally low values. 3. Significant improvement in bone mineralization, intestinal calcium absorption and muscle strength occurred in the three patients with the greatest rise in plasma 25-hydroxy vitamin D.

Effects of phosphorus supplementation on serum parathyroid hormone and bone morphology in osteoporosis.

The effect of phosphorus (inorganic phosphate) supplementation was studied in seven postmenopausal women with osteoporosis. Prior to supplementation, all chemical parameters studied in serum and urine were normal. Bone density was below the fifth percentile for age in all but one patient, and the percentage of bone surface involved in resorption was higher than normal. During administration of the phosphorus supplement, fasting serum concentrations of calcium and immunoreactive parathyroid hormone showed no significant changes, while serum phosphorus, urinary calcium, and tubular reabsorption of phosphorus decreased. In four patients studied by balance techniques, calcium balance became positive or less negative. Bone-forming surface decreased and bone-resorbing surface increased in all patients. Bone-resorbing surface was highly correlated with total phosphorus intake. Density of the distal radius changed variably, while density of the midradius increased slightly in all patients.