Effects of ABO incompatibility in allogeneic hematopoietic stem cell transplantation.

INTRODUCTION: The impact of ABO mismatch on outcomes following allo-HSCT remains controversial. In this study, our aim is to define the effect of ABO mismatch on post-transplant outcomes, engraftment kinetics and complications in a large cohort. PATIENTS AND METHODS: We retrospectively identified 1000 patients who underwent allo-HSCT from either bone marrow or peripheral blood stem cells at our center between 1988 and 2016. P<0.05 was considered statistically significant. RESULTS: Five hundred and ninety (59%) patient-donor pairs were ABO matched, 164 (16.4%) were ABO major mismatched (MM), 191 (19.1%) were ABO minor MM, and 55 (5.5%) were ABO bi-directionally MM. ABO matched pairs were more common in transplants from related donors (P<0.001) and using bone marrow as a stem cell source (P<0.001). In minor ABO MM transplantations, mild delayed hemolytic reaction occurred more frequently compared to major and bidirectional ABO MM transplantations (47% vs 35% and 18%, P<0.001). Neutrophil engraftment was slightly delayed in ABO MM patient-donor pairs when compared ABO matched donor pairs according to median engraftment time in all group (167/410, 41% vs 204/590, 35%, P=0.046). Pure red cell aplasia was diagnosed in 6 patients (1%). Higher risk of death was shown in ABO MM transplants compared to ABO matched transplants in overall survival (OS) analysis (HR:1.201, 95% CI:1.004-1.437, P=0.045). The non-relapse mortality (P=0.546) and cumulative incidences of acute graft versus host disease (aGVHD) and chronic (c) GVHD were comparable between ABO MM and ABO matched patient-donor pairs (for aGVHD, P=0.235; for cGVHD, P=0.137). CONCLUSION: ABO MM transplants were associated with decreased OS and slightly delayed neutrophil engraftment. NRM and the risk of GVHD were not related to ABO incompatibility.

Extracorporeal photochemotherapy in mycosis fungoides.

OBJECTIVES: Extracorporeal photo-chemotherapy (ECP, photopheresis) is an approved treatment modality for mycosis fungoides (MF). Our aim is to present our ECP data for MF. METHODS: We retrospectively evaluated 50 MF patients who received ECP for clinical activity, toxicity, and response and outcome rates, and we compared these with combination therapies. RESULTS: The overall response rate (ORR) was 42% (21/50), while the median time to response was 11months (range, 3-48months). Ten of the responders (48%) had 3 or more treatment lines prior to ECP. Eight patients (16%) had adverse events related to ECP. The overall survival (OS) of 50 patients was 72months (range, 3-211). There was no statistically significant difference in the OS in early-stage vs late-stage patients (77 vs 69months, P=0.077). The stage 3 and 4 patients received an average of 31 cycles compared to 55 cycles in stage 1 and 2 patients (P=0.006). The increased extent of ECP was not correlated with the response. Combined treatment with ECP significantly improved the OS (84months vs 62months, P=0.005). DISCUSSION: A low frequency of side effects and improved OS observed in combination therapy makes ECP a favorable option for treating MF.

Older age and capacity of colony forming unit in autologous peripheral derived hematopoietic cells.

BACKGROUND: Aging decreases bone marrow cellularity and alters the frequencies of stem cells. Aged hematopoietic stem cells can differ from their younger counterparts in functional capacity. STUDY DESIGN AND METHODS: We aimed to evaluate the relation between the age and the ability of colony forming capacity of peripheral blood-derived hematopoietic cell products collected for autologous stem cell transplantation (AHSCT). RESULTS: Elderly patients could be mobilized with lower total collected CD34+ cells. Colony forming capacity did not differ between young and old patients. CONCLUSION: This results can be translated into clinic as higher numbers of AHSCT candidates over age 60.

Impact of age and diagnosis on viability during centrifugation and cryopreservation of peripheral blood stem cell products.

BACKGROUND: The viability of the hematopoietic stem cells infused to the patient is important for transplant outcome. STUDY DESIGN AND METHODS: We evaluated 31 peripheral blood stem cell product collected from 15 patients. We aimed to check the viabilities of the cells from patients with different age and diagnosis, in different stages of the cryopreservation procedure. RESULTS: We showed a markedly decreased viability rate after centrifugation and addition of DMSO. Percentages of viabilities were similar between young and old patients in each step. Type of hematological malignancy did not make a significant influence on the viability. CONCLUSION: High speed centrifugation has a negative impact on the viability.

A controlled trial of erythromycin and UDCA in premature infants during parenteral nutrition in minimizing feeding intolerance and liver function abnormalities.

OBJECTIVE: To compare the effectiveness of oral erythromycin versus ursodeoxycholic acid (UDCA) treatment in preventing feeding intolerance and liver function abnormalities. STUDY DESIGN: A prospective, double blind, randomized, controlled trial in which three groups of preterm infants (birth weight <1500 g) were randomized to erythromycin (12.5 mg kg(-1) per day), UDCA (5 mg kg(-1) every 6 h) or placebo treatment. During the period 352 infants were admitted to our unit of which 75 infants whose parents accepted participation were enrolled in the study. Full enteral feeding or intestinal failure-associated liver disease was considered as the primary outcome measures. RESULTS: Time to achieve full feeding after beginning the treatment was significantly shorter in the erythromycin group (P=0.014). γ-Glutamyl transpeptidase levels in the placebo group were significantly higher than in the intervention groups (P=0.001). GTT level was slightly lower in UDCA groups than erythromycin. CONCLUSION: Oral erythromycin was most effective in facilitating enteral feeding and UDCA was most effective in preventing cholestasis in very low birth weight infants. Prophylactic usage of UDCA could be considered in infants with prolonged parenteral nutrition.

Management of serious isolated gingival synechia in a newborn: case report and review of the literature.

Congenital synechia of the gums is a rare malformation. The fusion may be partial or complete and may be associated with other facial dysmorphologic malformations. The synechia can be fibrous or bony and may develop between the upper and lower alveolar ridges. The authors report a newborn with partial, but broad and continuous, synechia of the gums and discuss its management during the first days of life.

In vitro evaluation of gentamicin- and vancomycin-containing minitablets as a replacement for fortified eye drops.

OBJECTIVE: Ocular bioadhesive minitablets containing gentamicin and vancomycin were developed using different powder mixtures of pregelatinized starch and Carbopol (physical or cospray-dried mixtures). METHODS: Drug content, antimicrobial activity, and radical formation of the powders used for tablet preparation were evaluated immediately and 30 days after gamma sterilization. Tablet properties and in vitro drug release from the sterilized minitablets were determined. Storage stability of vancomycin and gentamicin in sterilized bioadhesive mixtures was examined by LC-UV/MS and a microbiological assay, respectively. A bioadhesive powder mixture containing only vancomycin was irradiated by X electron-magnetic radiation to evaluate vancomycin stability following sterilization through irradiation. RESULTS: The antimicrobial activity of gentamicin against Staphylococcus epidermidis was not altered in comparison to nonsterilized formulations. Only after an overkill dose of 50 kGy, the concentration of vancomycin decreases to an extent that was pharmaceutically significant. No significant difference in radiation stability between drug substance and product (i.e., powder mixture) was observed. A shift in stability profile was not observed at 6 weeks after irradiation. All other degradation products were present only in small quantities not exceeding 1.0%. The in vitro drug release from the minitablets prepared with physical powder mixtures of pregelatinized starch and Carbopol® 974P NF (96 : 4) was faster compared to the cospray-dried mixtures of starch with Carbopol® 974P NF (ratio: 95:5 and 85:15). The electron paramagnetic resonance signals of the radicals formed during sterilization were still visible after storage for 30 days. The slug mucosal irritation test indicated mild irritation properties of the bioadhesive powder mixtures although no tissue damage was observed.

Characterization and in vivo evaluation of ocular minitablets prepared with different bioadhesive Carbopol-starch components.

The purpose of this study was to evaluate different bioadhesive ocular formulations based on drum dried waxy maize starch (DDWM), Amioca starch and Carbopol 974P. The concentrations of Carbopol 974P in the mixtures varied between 5 and 25% (w/w). The rheological properties of the non-sterilized and gamma-irradiated physical blends of Carbopol 974P with either DDWM or Amioca were compared to those of the corresponding co-spray dried Amioca starch/Carbopol powders. Higher viscosity or consistency values were measured for sterilized co-spray dried powder mixtures containing an amount of Carbopol 974P equal or above 15% (w/w) compared to the physical blends. Sustained release minitablets (2 mm, 6 mg), consisting of sodium fluorescein as model drug and the bioadhesive powders, were manufactured at a compression force of 1.25 kN. Afterwards, the tablets were sterilized with gamma-irradiation. The amount of Carbopol in the co-spray dried powder mixtures on the one hand and gamma-irradiation on the other hand had no significant influence on the crushing strength and friability of the minitablets evaluated. However, these two factors affected the in vitro release properties of the minitablets. The slowest release was obtained with tablets containing 25% Carbopol 974P, which unfortunately possess mucosal irritating properties. By using co-spray dried Amioca with 15% (w/w) Carbopol 974P, a slower release can be achieved compared to the physical mixtures of DDWM or Amioca starch with Carbopol 974P. Moreover, this ocular formulation is very promising and is preferred, as it did not cause any mucosal irritation and released the model drug for at least 12 h, after application in the fornix.

Finite element method based stress analysis of zone I and zone II sacral fractures.

BACKGROUND: Sacral bone fractures after direct traumas such as gunshot wounds and blunt sacral traumas are rarely, whereas those occurring after indirect traumas with vertical shear mechanisms ( car accidents or falls) are more frequently seen. METHODS: A cadaver sacrum was modelled 3-dimensionally using finite element software. Left sacroiliac joint was modeled to simulate a 75- kg man falling on one leg from a height of 5 meters. An impact load of 10 kN was transferred to the sacrum via intervertebral disc surface and two facets. Von Mises equivalent stress distribution was estimated. RESULTS: Von Mises stress distribution was calculated for sacral ala (430 MPa), S1 pedicle (225 MPa), and S1 lamina (35 MPa). Stress values were 200 Mpa, 130 Mpa, 105 Mpa, and 55 MPa on ventrolateral, dorsomedial, ventromedial and dorsolateral sides of S1 foramen respectively. Vertical shear injury caused stress distribution to concentrate at sacral ala and S1 pedicle (zone I). Local stress distribution around S1 foramen is concentrated ventrolaterally CONCLUSION: High incidence of zone I sacral fractures and low incidence of neurological deficits could be explained by concentration of stress at sacral ala. Zone II fractures might be due to by transfer of triggered stresses from sacral ala and S1 pedicle to ventrolateral side of S1 foramen.

Rheological evaluation of dispersions, prepared with different non- and gamma-irradiated bioadhesive cospray dried powder mixtures.

Various polymer mixtures were used to prepare gels: native drum dried waxy maize starch, Amioca starch, Carbopol 974P NF and powder mixtures obtained by cospray drying Amioca starch and Carbopol 974P NF, and by blending Carbopol 974P NF with Amioca starch or drum dried waxy maize starch. Oscillatory rheology was employed to investigate the influence of gamma-irradiation, but also of the different compositions (i.e. Carbopol 974P NF varied from 5 to 25% w/w in the powder mixtures tested) and the preparation procedures of the powder mixtures on the viscoelastic properties of the polymer dispersions. The rheological data showed that gamma-irradiation has a significant influence on the rheological behaviour of the polymers dispersed. More elastic properties were obtained when the concentration of Carbopol 974P NF was increased. The dispersions containing at least an amount of 15% Carbopol 974P NF (w/w) are mainly elastic. Contrary to cospray drying the powders, blending the powders leads to higher elastic and gel properties of the dispersed polymers.

The effect of freeze-drying with different cryoprotectants and gamma-irradiation sterilization on the characteristics of ciprofloxacin HCl-loaded poly(D,L-lactide-glycolide) nanoparticles.

In the present study, the influence of freeze-drying with several cryoprotective agents and gamma (gamma)-irradiation sterilization on the physicochemical characteristics of ciprofloxacin HCl-loaded poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles was evaluated. Nanoparticles were prepared by W/O/W emulsification solvent evaporation followed by high-pressure homogenization. They were freeze-dried in the presence of 5.0% (w/v) mannitol, trehalose or glucose, with 5.0% (w/v) or 15.0% (w/v) dextran as cryoprotectants. The nanoparticles were irradiated at a dose of 25 kGy using a 60Co source. The following physicochemical properties of the formulations were investigated: the ratio of particle size before (initial) and after freeze-drying, the ease of reconstitution of the nanoparticle suspensions and the drug-release profiles of irradiated and non-irradiated nanoparticles. The antibacterial activity against Pseudomonas aeruginosa was measured. The freeze-drying process induced a significant increase in particle size when no cryoprotectant was employed. Similar results were observed when cryoprotectants were added to the formulation. Only when mannitol was used was no significant size increase measured. Moreover, for formulations with dextran, reconstitution after freeze-drying was difficult by manual agitation and particle size could not be determined because of aggregation. After gamma-sterilization no significant difference in mean particle size was observed, but reconstitution was more difficult and drug release was influenced negatively. Ciprofloxacin HCl incorporated in the nanoparticles was still effective against the micro-organism selected after freeze-drying and gamma-sterilization.

In vitro cytotoxicity of mitoxantrone-incorporated albumin microspheres on acute promyelocytic leukaemia cells.

In the present study, the preparation and characterization of bovine serum albumin (BSA) microspheres and the evaluation of the in vitro cytotoxicity of these microspheres on acute promyelocytic leukaemia (HL-60) cells were described. Mitoxantrone (MTZ)-incorporated microspheres were evaluated for particle size, drug loading, release characteristics and surface morphology. The biological effect of MTZ released from BSA microspheres was determined on an in vitro cultured HL-60 cell line, showing that, after encapsulation, MTZ still retains cytotoxic activity. For this purpose, methyl-thiazol-tetrazolium (MTT) assay was used to evaluate the in vitro cytotoxicity of MTZ-loaded microspheres. Particle size of BSA microspheres was determined between 17.61-20.38 microm and they were smooth and spherical in shape. Encapsulation efficiency of the drug-loaded microspheres was between 22.26-60.50%. For MTZ-containing microspheres, the cell death ratios were greater than 80% for all formulations. This study demonstrate that BSA microspheres were well suited for the controlled release of MTZ and were promising for anti-cancer chemotherapy.

In vitro evaluation and intra-articular administration of biodegradable microspheres containing naproxen sodium.

The dispersion of non-steroidal antiinflammatory drugs (NSAIDs) into biodegradable polymeric matrices have been accepted as a good approach for obtaining a therapeutic effect in a predetermined period of time meanwhile minimizing the side effects of NSAIDs. In the present study, it was aimed to prepare Naproxen Sodium (NS), (a NSAID) loaded microsphere formulation using natural Bovine Serum Albumin (BSA) and synthetic biodegradable polymers such as poly(lactide-co-glycolic acid) (PLGA) (50:50 MW 34,000 and 88,000 Da) for intra-articular administration, and to study the retention of the drug at the site of injection in the knee joint. NS incorporated microspheres were evaluated in vitro for particle size (the mean particle size; for BSA microspheres, 10.0 +/- 0.3 microm, for PLGA microspheres, 9.0 +/- 0.2 and 5.0 +/- 0.1 microm for MW 34,000 and 88,000 Da, respectively), yield value, drug loading, surface morphology and drug release. For in vivo studies, monoarticular arthritis was induced in the left knee joints of rabbits by using ovalbumin and Freund's Complete Adjuvant as antigen and adjuvant. A certain time (4 days) is allowed for the formation of arthritis in the knee joints, then the NS loaded microspheres were injected directly into the articular cavity. At specific time points, gamma scintigrams were obtained to determine the residence time of the microspheres in knee joints, in order to determine the most suitable formulation. This study indicated that PLGA, a synthetic polymer, is more promising than the natural type BSA microspheres for an effective cure of mono-articular arthritis in rabbits.