A phase 2 trial of ponatinib in Philadelphia chromosome-positive leukemias.

BACKGROUND: Ponatinib is a potent oral tyrosine kinase inhibitor of unmutated and mutated BCR-ABL, including BCR-ABL with the tyrosine kinase inhibitor-refractory threonine-to-isoleucine mutation at position 315 (T315I). We conducted a phase 2 trial of ponatinib in patients with chronic myeloid leukemia (CML) or Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph-positive ALL). METHODS: We enrolled 449 heavily pretreated patients who had CML or Ph-positive ALL with resistance to or unacceptable side effects from dasatinib or nilotinib or who had the BCR-ABL T315I mutation. Ponatinib was administered at an initial dose of 45 mg once daily. The median follow-up was 15 months. RESULTS: Among 267 patients with chronic-phase CML, 56% had a major cytogenetic response (51% of patients with resistance to or unacceptable side effects from dasatinib or nilotinib and 70% of patients with the T315I mutation), 46% had a complete cytogenetic response (40% and 66% in the two subgroups, respectively), and 34% had a major molecular response (27% and 56% in the two subgroups, respectively). Responses were observed regardless of the baseline BCR-ABL kinase domain mutation status and were durable; the estimated rate of a sustained major cytogenetic response of at least 12 months was 91%. No single BCR-ABL mutation conferring resistance to ponatinib was detected. Among 83 patients with accelerated-phase CML, 55% had a major hematologic response and 39% had a major cytogenetic response. Among 62 patients with blast-phase CML, 31% had a major hematologic response and 23% had a major cytogenetic response. Among 32 patients with Ph-positive ALL, 41% had a major hematologic response and 47% had a major cytogenetic response. Common adverse events were thrombocytopenia (in 37% of patients), rash (in 34%), dry skin (in 32%), and abdominal pain (in 22%). Serious arterial thrombotic events were observed in 9% of patients; these events were considered to be treatment-related in 3%. A total of 12% of patients discontinued treatment because of an adverse event. CONCLUSIONS: Ponatinib had significant antileukemic activity across categories of disease stage and mutation status. (Funded by Ariad Pharmaceuticals and others; PACE ClinicalTrials.gov number, NCT01207440 .).

Modulation of Intestinal Epithelial Defense Responses by Probiotic Bacteria.

Probiotics are live microorganisms, which when administered in food confer numerous health benefits. In previous studies about beneficial effects of probiotic bacteria to health, particularly in the fields of intestinal mucosa defense responses, specific probiotics, in a strain-dependent manner, show certain degree of potential to reinforce the integrity of intestinal epithelium and/or regulate some immune components. The mechanism of probiotic action is an area of interest. Among all possible routes of modulation by probiotics of intestinal epithelial cell-mediated defense responses, modulations of intestinal barrier function, innate, and adaptive mucosal immune responses, as well as signaling pathways are considered to play important role in the intestinal defense responses against pathogenic bacteria. This review summarizes the beneficial effects of probiotic bacteria to intestinal health together with the mechanisms affected by probiotic bacteria: barrier function, innate, and adaptive defense responses such as secretion of mucins, defensins, trefoil factors, immunoglobulin A (IgA), Toll-like receptors (TLRs), cytokines, gut associated lymphoid tissues, and signaling pathways.

Phase II study of treatment of advanced ovarian cancer with folate-receptor-targeted therapeutic (vintafolide) and companion SPECT-based imaging agent (99mTc-etarfolatide).

BACKGROUND: This report examines (99m)Tc-etarfolatide imaging to identify the presence of folate receptor (FR) on tumors of women with recurrent/refractory ovarian or endometrial cancer and correlates expression with response to FR-targeted therapy (vintafolide). PATIENTS AND METHODS: In this phase II, single-arm, multicenter study, patients with advanced ovarian cancer were imaged with (99m)Tc-etarfolatide before vintafolide treatment. Up to 10 target lesions (TLs) were selected based on Response Evaluation Criteria In Solid Tumors criteria using computed tomography scans. Single-photon emission computed tomography images of TLs were assessed for (99m)Tc-etarfolatide uptake as either FR positive or negative. Patients were categorized by percentage of TLs positive and grouped as FR(100%), FR(10%-90%), and FR(0%). Lesion and patient response were correlated with etarfolatide uptake. RESULTS: Forty-nine patients were enrolled; 43 were available for analysis. One hundred thirty-nine lesions were (99m)Tc-etarfolatide evaluable: 110 FR positive and 29 FR negative. Lesion disease control rate (DCR = stable or response) was observed in 56.4% of FR-positive lesions versus 20.7% of FR-negative lesions (P < 0.001). Patient DCR was 57%, 36%, and 33% in FR(100%), FR(10%-90%), and FR(0%) patients, respectively. Median overall survival was 14.6, 9.6, and 3.0 months in FR(100%), FR(10%-90%), and FR(0%) patients, respectively. CONCLUSIONS: Overall response to FR-targeted therapy and DCR correlate with FR positivity demonstrated by (99m)Tc-etarfolatide imaging. CLINICAL TRIAL NUMBER: NCT00507741.

The effect of NaCl substitution with KCl on proteinase activities of cell-free extract and cell-free supernatant at different pH levels and salt concentrations: Lactobacillus acidophilus and Lactobacillus casei.

The aim of this study was to investigate the effect of substitution of NaCl with KCl at different pH levels and salt concentrations on proteinase activity of cell-free extract and cell-free supernatant of the probiotics Lactobacillus acidophilus and Lactobacillus casei. de Man, Rogosa, and Sharpe broth aliquots were mixed with 2 pure salts (NaCl and KCl) and 2 salt concentrations at 2 concentration levels (5 and 10%), inoculated with Lactobacillus acidophilus or Lactobacillus casei, and incubated aerobically at 37°C for 22 h. The cultures were then centrifuged at 4,000×g for 30 min, and the collected cell pellets were used to prepare cell-wall proteinases and the supernatants used as a source of supernatant (extracellular) proteinases. The proteolytic activity and protein content of both portions were determined. After incubation of both portions with 3 milk caseins (α-, ß-, κ-casein), the supernatants were individually subjected to analysis of angiotensin-converting enzyme (ACE)-inhibitory activity and proteolytic activity using the o-phthalaldehyde method. Significant differences were observed in ACE-inhibitory and proteolytic activities between salt substitution treatments of cell-free extract and cell-free supernatant from both probiotic strains at the same salt concentration and pH level.

The effect of NaCl substitution with KCl on Akawi cheese: chemical composition, proteolysis, angiotensin-converting enzyme-inhibitory activity, probiotic survival, texture profile, and sensory properties.

The effect of partial substitution of NaCl with KCl on Akawi cheese with probiotic bacteria was investigated during 30 d of storage at 4 °C. Chemical composition, the survival of probiotic and lactic acid bacteria, proteolytic activity, and texture profile analysis were analyzed and sensory analysis was carried out to determine the effects of substitution. No significant differences were observed in moisture, protein, fat, and ash contents among the experimental Akawi cheeses at the same storage period. Significant differences were observed in water-soluble nitrogen and phosphotungstic-soluble nitrogen between experimental cheeses at the same of storage period. No significant difference was observed in the growth of Lactobacillus delbrueckii ssp. bulgaricus between experimental cheeses at the same storage period. However, the growth of Streptococcus thermophilus, Lactobacillus casei, and Lactobacillus acidophilus was significantly affected among experimental cheeses. A significant difference was observed in soluble Ca among experimental cheeses at the same storage period. In general, no significant differences existed in hardness and adhesiveness among experimental cheeses at the same storage period. No significant differences existed in sensory attributes, including creaminess, bitterness, saltiness, sour-acid, and vinegar taste among experimental Akawi cheeses at the same storage period.

The effect of substituting NaCl with KCl on Nabulsi cheese: chemical composition, total viable count, and texture profile.

The effect of substituting NaCl with KCl on Nabulsi cheese characteristics was investigated. Nabulsi cheese was made and stored in 4 different brine solutions at 18%, including NaCl only (A; control); 3NaCl:1KCl (wt/wt; B); 1NaCl:1KCl (wt/wt; C); and 1NaCl:3KCl (wt/wt; D). Chemical composition, proteolysis, total viable count, and texture profile analysis were assessed at monthly intervals for 5 mo. No significant effect was found among experimental cheeses in terms of chemical composition or texture profile. Proteolytic activities were higher in cheeses kept in brine solutions that contained higher KCl (B, C, and D) compared with the control. At the end of the storage period, water-soluble nitrogen in Nabulsi cheeses stored in B, C, and D was higher than that in the control cheese (A). In addition, total viable count increased significantly after 1 mo of storage for all salt treatments. Hardness and gumminess generally decreased significantly during storage within the same salt treatment.

The effect of substitution of NaCl with KCl on chemical composition and functional properties of low-moisture Mozzarella cheese.

The effect of NaCl substitution with KCl on chemical composition, organic acids profile, soluble calcium, and functionality of low-moisture Mozzarella cheese (LMMC) was investigated. Functionality (meltability and browning), organic acids profile, and chemical composition were determined. Chemical composition showed no significant difference between experimental cheeses at same storage period, and same salt treatment. Meltability of LMMC salted with 3NaCl:1KCl, 1NaCl:1KCl, and 1NaCl:3KCl was higher compared with only NaCl (control). The amount of soluble Ca and P increased significantly during storage, with no significant difference between salt treatments. Organic acids profile did not differ between salt treatments at the same storage time.

Proteolysis of low-moisture Mozzarella cheese as affected by substitution of NaCl with KCl.

The proteolytic and ACE inhibitory activities of low-moisture Mozzarella cheese (LMMC) as affected by partial substitution of NaCl with KCl were investigated. Experimental LMMC were made and salted with 4 salt mixtures: NaCl only (control), 3NaCl:1KCl, 1NaCl:1KCl, and 1NaCl:3KCl, and then proteolytic activity and angiotensin-converting enzyme inhibitory activity were determined. Salt treatment significantly affected angiotensin-converting enzyme inhibitory activity and phosphotungstic acid-soluble N of LMMC during storage. Water-soluble N, trichloroacetic acid-soluble N, lactic acid bacteria population, and total free amino acids were unaffected during storage. Nonetheless, water-soluble N and trichloroacetic acid-soluble N increased significantly during storage within a salt treatment. Peptide profiles and urea-PAGE gels did not differ between experimental cheeses at the same storage time.

The effect of sodium chloride substitution with potassium chloride on texture profile and microstructure of Halloumi cheese.

The effect of partial substitution of NaCl with KCl on texture profile and microstructure of Halloumi cheese was investigated. Four batches of Halloumi cheese were made and kept in 4 different brine solutions (18%, wt/wt), including A) NaCl only, B) 3NaCl:1KCl, C) 1NaCl:1KCl, and D) 1NaCl:3KCl and then stored at 4°C for 56 d. The texture profile was analyzed using an Instron universal machine, whereas an environmental scanning electron microscope was used to investigate the effect of NaCl substitution on the microstructure of cheeses. No significant difference was found in hardness, cohesiveness, adhesiveness, and gumminess among experimental cheeses at the same storage day. Hardness, cohesiveness, and gumminess decreased significantly during storage period with the same salt treatment, whereas adhesiveness significantly increased. Environmental scanning electron microscope micrographs showed a compact and closed texture for cheeses at the same storage period. The microstructure of all cheeses became more closed and compact with storage period. Calcium content negatively correlated with hardness and Na and K contents during storage with the same salt treatment.

High success and low recurrence with shorter treatment regimen for multidrug-resistant TB in Nepal.

SETTING: Nine drug-resistant TB centres, some of them supported by Damien Foundation in Nepal where >80% of multidrug-resistant/rifampicin-resistant TB (MDR/RR-TB) patients are treated. OBJECTIVE: To assess the uptake, effectiveness and safety of the 9-12-month shorter treatment regimen (STR) in MDR/RR-TB patients registered from January 2018 to December 2019. DESIGN: This was a cohort study involving secondary programme data. RESULTS: Of 631 patients, 301 (48.0%) started and continued STR. Key reasons for ineligibility to start/continue STR were baseline resistance or exposure to second-line drugs (62.0%), contact with extensively drug-resistant TB (XDR-TB) or pre-XDR-TB (7.0%) patients and unavailability of STR drugs (6.0%). Treatment success was 79.6%; unsuccessful outcomes were death (12.0%), lost to follow-up (5.3%), failure (2.7%) and not evaluated (0.7%). Unsuccessful outcomes were significantly associated with HIV positivity and patient age ⩾55 years, with adjusted relative risk of respectively 2.39 (95% CI 1.52-3.77) and 3.86 (95% CI 2.30-6.46). Post-treatment recurrence at 6 and 12 months was respectively 0.5% and 2.4%. Serious adverse events (SAEs) were seen in 15.3% patients - hepatotoxicity and ototoxicity were most common. CONCLUSION: STR had a modest uptake, high treatment success and low post-treatment recurrence. For proper detection and management of SAEs, improving pharmacovigilance might be considered. Availability of rapid diagnostic test for second-line drugs is crucial for correct patient management.

CADRE: Neuf centres de traitement de la TB pharmacorésistante, dont certains sont financés par Action Damien au Népal où >80% des patients atteints de TB multirésistante/résistante à la rifampicine (MDR/RR-TB) sont traités. OBJECTIF: Évaluer l'utilisation, l'efficacité et l'innocuité d'un schéma thérapeutique plus court (STR) de 9-12 mois chez les patients atteints de MDR/RR-TB enregistrés de janvier 2018 à décembre 2019. MÉTHODE: Étude de cohorte comprenant des données programmatiques secondaires. RÉSULTATS: Sur 631 patients, 301 (48,0%) ont démarré et poursuivi un STR. Les raisons principales d'inéligibilité à l'instauration/la poursuite d'un STR étaient une résistance initiale ou une exposition aux médicaments de deuxième intention (62,0%), un contact avec des patients atteints de TB ultrarésistante (XDR-TB) ou de pré-XDR-TB (7,0%) et la non-disponibilité des médicaments pour le STR (6,0%). Le taux de réussite thérapeutique était de 79,6%. Les résultats liés à la non-réussite thérapeutique étaient décès (12,0%), perte de vue (5,3%), échec thérapeutique (2,7%) et absence d'évaluation (0,7%). Les résultats liés à la non-réussite thérapeutique étaient significativement associés à l'infection par le VIH et aux patients âgés ⩾55 ans avec un risque relatif ajusté de 2,39 (IC 95% 1,52­3,77) et de 3,86 (IC 95% 2,30­6,46), respectivement. Le taux de récidive post-traitement à 6 et 12 mois était de 0,5% et 2,4%, respectivement. Des évènements indésirables graves (SAE) ont été observés chez 15,3% des patients, le plus souvent hépatotoxicité et ototoxicité. CONCLUSION: Le STR a été associé à une utilisation modérée, à une réussite thérapeutique élevée et à un faible taux de récidive post-traitement. Pour une détection et une prise en charge adéquates des SAE, l'amélioration de la pharmacovigilance peut être envisagée. La disponibilité de tests diagnostiques rapides pour les médicaments de deuxième intention est essentielle à une prise en charge adéquate des patients.

Functional relation between HTLV-II x and adenovirus E1A proteins in transcriptional activation.

The mechanism of cellular transformation by the human T-cell leukemia viruses (HTLV) is thought to involve a novel gene known as the x gene. This gene is essential for HTLV replication and acts by enhancing transcription from the HTLV long terminal repeat. The HTLV x gene product may also cause aberrant transcription of normal cellular genes, resulting in transformation of the infected cells. Although there is no evidence as yet for such a mechanism, it was shown that the HTLV-II x gene product can activate transcription from adenovirus E1A-dependent early promoters and therefore has the potential to activate cellular genes. It was also shown that the adenovirus and herpes pseudorabies immediate early proteins activate expression from the HTLV-I and HTLV-II long terminal repeats, though at lower levels than with the x gene product. These findings indicate possible common mechanisms of action for transcription-regulatory genes of distinct viruses.

HTLV x gene mutants exhibit novel transcriptional regulatory phenotypes.

The human T-cell leukemia viruses, HTLV-I and HTLV-II, contain a gene, termed x, with transcriptional regulatory function. The properties of the x proteins were analyzed by constructing mutant genes containing site-directed deletions and point mutations. The results demonstrate that the amino terminal 17 amino acids of the x protein constitute part of a functional domain that is critical for the transcriptional activating properties of the protein. Within this region, substitution of a leucine residue for a proline residue results in major changes in the trans-activation phenotype of the protein. The mutant HTLV-II x protein, though incapable of activating the HTLV-II long terminal repeat, will block trans-activation of the HTLV-II long terminal repeat by the wild-type protein. The altered phenotype of this mutant suggests a potential negative regulatory function of the x protein.

U3 sequences from HTLV-I and -II LTRs confer pX protein response to a murine leukemia virus LTR.

Human T-cell leukemia virus (HTLV) types I and II are unusual among replication-competent retroviruses in that they contain a fourth gene (chi) necessary for replication. The chi gene product, p chi, transcriptionally transactivates the viral long repeat (LTR), and is thus a positive regulator. To investigate p chi transactivation, sequences from the U3 regions of the LTRs of HTLV-I and -II were inserted into the Moloney murine leukemia virus (M-MuLV) LTR by recombinant DNA techniques. Transient expression assays of the chimeric LTRs indicated that the HTLV sequences conferred to the M-MuLV LTR responsiveness to HTLV p chi protein. M-MuLV enhancers were not required for function of the chimeric LTRs. Infectious recombinant M-MuLVs containing chimeric LTRs were also generated. These viruses showed higher infectivity when assayed in mouse cells expressing HTLV-II p chi protein compared to normal mouse cells. Thus the HTLV sequences were able to confer p chi responsiveness to infectious M-MuLV. The generation of a virus dependent on a transactivating protein for its replication has implications for the evolution of the human T-cell leukemia viruses.

The x gene is essential for HTLV replication.

The human T-cell leukemia viruses (HTLV) are associated with T-cell malignancies in man and will transform normal human T cells in vitro. The mechanism of malignant transformation by HTLV is unknown but appears to be distinct from that of other classes of retroviruses, which induce malignant transformation through viral or cellular oncogenes. Recently a new gene, termed x, was identified in HTLV. This gene has been hypothesized to be the transforming gene of HTLV because of its conservation within the HTLV class of retroviruses. By in vitro mutagenesis of the HTLV-II x gene, it is now demonstrated that the presence of a functional x gene product is necessary for efficient HTLV transcription. Therefore, these studies provide direct evidence for an important function of the x gene in HTLV replication. The functional analogies between the x gene and transcriptional regulatory genes of some DNA viruses suggest that these viruses share similar mechanisms for cellular transformation.

HTLV-II transactivation is regulated by the overlapping tax/rex nonstructural genes.

The human T-cell leukemia virus (HTLV) types I and II have two nonstructural genes that are encoded in overlapping reading frames. One of these genes, known as tax, has been shown to encode a protein responsible for enhanced transcription (transactivation) from the viral long terminal repeats (LTRs). Genetic evidence indicates that the second nonstructural gene of HTLV-II, here designated rex, acts in trans to modulate tax gene-mediated transactivation in a concentration-dependent fashion. The rex gene may regulate the process of transactivation during the viral life cycle.

Effect of exopolysaccharides on the proteolytic and angiotensin-I converting enzyme-inhibitory activities and textural and rheological properties of low-fat yogurt during refrigerated storage.

The aim of this study was to examine the influence of using exopolysaccharide (EPS) producing strain of Streptococcus thermophilus on the viability of yogurt starters, their proteolytic and angiotensin-I converting enzyme-inhibitory activities, and on the textural and rheological properties of the low-fat yogurt during storage at 4 degrees C for 28 d. The use of an EPS-producing strain of S. thermophilus did not have influence on pH, lactic acid content, or the angiotensin-I converting enzyme-inhibition activity of low-fat yogurt. However, EPS showed a protective effect on the survival of Lactobacillus delbrueckii ssp. bulgaricus. Presence of EPS reduced the firmness, spontaneous whey separation, yield stress, and hysteresis loop area but not the consistency and flow behavior index of low-fat yogurt.

Association of tuberculous endometritis with infertility and other gynecological complaints of women in India.

Endometrial biopsy samples derived from 393 patients with assorted gynecological complaints were investigated for mycobacterial infection. By employment of four different techniques, mycobacterial pathogens were detected irrespective of the nature/type of clinical complaint. Mycobacterium tuberculosis was the predominant pathogen detected among the samples investigated.

Bifurcation of the inferior dental nerve canal: an anatomical study.

The aims of this study were to find the incidence of bifurcation of the inferior dental nerve (IDN) canal, to describe the characteristics of this variant, and to examine the sensitivity and specificity of dental panoramic tomography to identify it. We classified bifurcations by size and position relative to the main canal and the lower third molar using cone-beam computed tomography (CT) and dental panoramic tomography. In our study of 281 patients, 106 (38%) had bifurcations, and in one quarter, these were classified as large accessory canals. Bifurcations were most commonly found posterior to the lower third molar (n=64, 57%) or within 2mm of the roots of the third molar (n=40, 38%). The sensitivity and specificity of dental panoramic tomography to identify all bifurcations was 11% (95% CI: 5.67 to 17.97) and 91% (95% CI: 85.58 to 94.68), respectively; this was 33% (95% CI: 15.63 to 55.32) and 94% (95% CI: 90.34 to 96.50), respectively, for large bifurcations. Our use of cone-beam CT suggested an incidence of bifid canals of 38%, with a variation in size and distribution in relation to the lower third molar. It also showed that the sensitivity of panoramic radiography to identify them was poor.

Characterization of the BCR promoter in Philadelphia chromosome-positive and -negative cell lines.

The t(9;22) Philadelphia chromosome translocation fuses 5' regulatory and coding sequences of the BCR gene to the c-ABL proto-oncogene. This results in the formation of hybrid BCR-ABL mRNAs and proteins. The shift in ABL transcriptional control to the BCR promoter may play a role in cellular transformation mediated by this rearrangement. We have functionally localized the BCR promoter to a region 1 kb 5' of BCR exon 1 coding sequences by using a chloramphenicol acetyltransferase reporter gene assay. Nucleotide sequence analysis of this region revealed many consensus binding sequences for transcription factor SP1 as well as two potential CCAAT box binding factor sites and one putative helix-loop-helix transcription factor binding site. No TATA-like or "initiator" element sequences were found. Because of low steady-state levels of BCR mRNA and the high GC content (78%) of the promoter region, definitive mapping of transcription start sites required artificial amplification of BCR promoter-directed transcripts. Overexpression from the BCR promoter in a COS cell system was effective in demonstrating multiple transcription initiation sites. In order to assess the effects of chromosomal translocation on the transcriptional control of the BCR gene, we determined S1 nuclease protection patterns of poly(A)+ RNA from tumor cell lines. No differences were observed in the locations and levels of BCR transcription initiation sites between those lines that harbored the t(9;22) translocation and those that did not. This demonstrates that BCR promoter function remains intact in spite of genomic rearrangement. The BCR promoter is structurally similar to the ABL promoters. Together, this suggests that the structural fusion of BCR-ABL and not its transcriptional deregulation is primarily responsible for the transforming effect of the t(9;22) translocation.

Comparison of the trans-activation capabilities of the human T-cell leukemia virus type I and II chi proteins.

The mechanism of cellular transformation by the human T-cell leukemia viruses (HTLVs) is thought to involve a novel retrovirus gene known as chi. The chi gene is essential for HTLV replication and acts by enhancing transcription from the viral long terminal repeat. By using the HTLV type I and II chi gene-coding regions inserted into a highly efficient expression vector, we directly compared the efficiencies of the two chi proteins to trans activate the HTLV type I and II long terminal repeats. We demonstrate that the two chi proteins have different patterns of trans activation. The patterns were highly reproducible in all mammalian cells tested. A different pattern of activation was observed in avian cells. These results suggest that the mechanism of trans activation involves specific cellular factors that are highly conserved throughout mammalian species but different in avian cells. Understanding the mechanism of trans activation by the chi gene product may provide insights into mechanisms of cellular transformation by HTLV.