Busulfan Group Research Articles

Impact of busulfan versus treosulfan dose intensity in myelofibrosis undergoing hematopoietic cell transplantation.

One key aspect of allogeneic hematopoietic cell transplantation (HCT) is pretransplant conditioning, balancing risk for relapse versus non-relapse mortality. Conditioning regimens with different alkylators at different doses can influence outcome, but data are missing for myelofibrosis, a challenging cohort of patients usually presenting at older age and with comorbidities. We evaluated in a multicenter retrospective study the comparative efficacy and safety of busulfan versus treosulfan in combination with fludarabine for myelofibrosis patients undergoing HCT. This study included 1115 patients (busulfan, n = 902; treosulfan, n = 213) receiving first HCT between 2005 and 2021. Patients were generally balanced for key patient characteristics. Overall survival at 4 years was 62% for the busulfan group versus 58% for the treosulfan group (p = .22). Impact on outcome was dose-dependent. Overall survival was 65% (95% CI, 61%-69%) for reduced intensity busulfan versus 69% (95% CI, 54%-84%) for reduced intensity treosulfan, 53% (95% CI, 44%-63%) for higher intensity busulfan, and 55% (95% CI, 46%-63%) for higher intensity treosulfan. Incidence of relapse was similar across intensity groups. In multivariable analysis, the hazard for death (with reduced intensity busulfan as reference) was 0.88 (95% CI, 0.39-2.01) for reduced intensity treosulfan (p = .77), 1.42 (95% CI, 0.96-2.10) for higher intensity busulfan (0.08), and 1.61 (95% CI, 1.14-2.26) for higher intensity treosulfan (p = .006). In terms of non-relapse mortality, comparison was not significantly different, while the hazard ratio for higher intensity treosulfan was 1.48 (95% CI, 0.98-2.23; p = .06). Here, we showed comparable outcomes and improved survival in myelofibrosis undergoing HCT with reduced intensity busulfan or treosulfan.

Treosulfan is a safe and effective alternative to busulfan for conditioning in adult allogeneic HSCT patients: Data from a single center.

Type of conditioning regimen impacts the outcome of patients who undergo allogeneic HSCT since graft versus host disease (GVHD), infections, regimen related toxicities (RRT) are important causes of post-transplant mortality. Despite the RRT profile of busulfan, it is frequently used worldwide. Treosulfan has advantages in terms of dose of administration, lower incidence of sinusoidal obstruction syndrome and lower neurotoxicity. We retrospectively investigated outcomes of patients who underwent allogeneic HSCT with treosulfan or busulfan based conditioning regimens in our institution. Treosulfan was administered to 94 patients while 85 patients received busulfan. Our outcomes were RRT, chronic and acute GVHD, relapse related mortality (RRM), non-relapse mortality, and fungal infection. The clinical follow up data, regarding the primary and secondary endpoints of our study, of the patients who received treosulfan or busulfan based conditioning regimens were statistically analyzed. The median follow-up was 14 months for the treosulfan group while it was 11 months for the busulfan group (p = 0.16). RRT was 11.7% and 7.1% for treosulfan and busulfan respectively. The incidence of extensive chronic GVHD was less frequent in the treosulfan group compared to the busulfan group (15.7% vs. 32.1%) (p < 0.001). The incidence of acute GVHD (Grade 3 or higher) was 32.2% in the treosulfan group while it was 31.6% in the busulfan group. The RRM was 17% in the treosulfan group while it was 34% in the busulfan group. The non-relapse mortality was 35.5% and 29.4% in the treosulfan group and in the busulfan group respectively (p = 0.962). Treosulfan, with a lower RRM, lower chronic GVHD incidence and with a similar RRT profile appears to be a safe alternative to busulfan.

Impact of biotechnological transgenesis procedures on duck productivity

The use of poultry as a unique model of biological research was characterised by a high level of efficiency, however, methods for creating transgenic ducks, complicated by the structure of waterfowl eggshells, are of low efficiency. The purpose of the study was to determine the influence of various biotechnological procedures for creating transgenic ducks on their productive qualities and reproductive ability to identify the optimal method for creating transgenic poultry for further use in scientific, research, or economic purposes. Weighting, morphometric and statistical analysis of productive traits were used during the study. 40 ducks (4 experimental groups of animals and about 3,000 of their eggs) were studied. The lowest value of the egg productivity index was obtained in the group created by busulfan injection (79.5±11.8%), the highest – in the group created by sperm-mediated gene transfer (91.8±2.3%), the group of direct injection of transgenic construct – 89.0±2.0%, which indicates that this biotechnological method of introducing transgenic construct did not have a clear effect on this indicator. The weight of ducks in different experimental groups ranged from 1,323.50±65.36 g (using the sperm-mediated gene transfer) to 1,608.08±94.76 g (in the group created using busulfan). Ducks that received direct injections had an average weight of 1,480.42±35.01 g. In the control group, the average weight at sexual maturity was 139.5±9.67 g, in the busulfan group – 148.2±13.13 g, in the direct injection group – 143.16±7.25 g, and in the spermmediated gene transfer group – 140.67±13.13 g. It was found that the method of injection into the embryo of a recipient sterilised with busulfan and the introduction of donor blastodermal cells negatively affect the reproductive qualities of ducks. The practical significance of the study lies in the fact that as a result of the analysis of the productivity of ducks obtained by various methods of transgenesis, it was determined that the most effective of the evaluated methods is the transfection of DNA of the transgenic construct with sperm (Sperm-mediated gene transfer, SMGT)

Effects of Perilla frutescens Var. Acuta in Busulfan-Induced Spermatogenesis Dysfunction Mouse Model.

The leaves of Perilla frutescens var. acuta (PFA) are generally reported to have antioxidant, anti-allergic, anti-inflammatory, and antitumor effects and commonly used as a traditional medicine in East Asia. This study aimed to investigate the protective effect and antioxidant activity of PFA on busulfan-induced testicular dysfunction, histological damage, oxidative stress (OS), sperm quality, and hormone levels using a mouse model. C57BL/6 male mice were divided into four groups: control, busulfan-only treated, and varying concentrations of PFA (100 and 200 mg/kg) with busulfan. In the busulfan group, 40 mg/kg of busulfan was intraperitoneally injected to induce azoospermia. Mice were orally administered PFA for 35 consecutive days after busulfan administration. Samples were collected and assessed for testis/body weight, testicular histopathology, sperm quality, serum hormone levels, and OS to evaluate the effects of PFA treatment on spermatogenesis dysfunction induced by busulfan. The busulfan-induced testicular dysfunction model showed reduced testis weight, adverse histological changes, significantly decreased sex hormones and sperm quality, and attenuated OS. These results indicate that PFA treatment significantly increased testis weight, testis/body weight, epididymal sperm count, motility, and testosterone level compared with busulfan alone. PFA treatment also attenuated the busulfan-induced histological changes. Furthermore, compared with mice treated with busulfan alone, PFA supplementation upregulated the testicular mRNA expression of the antioxidant enzymes superoxide dismutase 1 (Sod1) and glutathione peroxidase 1 (Gpx1), with a decrease in malondialdehyde (MDA) production and an increase in SOD and GPx activities. This study shows that PFA exerts a protective effect against testicular damage by attenuating OS induced by busulfan. Our results suggest that PFA is a potentially relevant drug used to decrease the side effects induced by busulfan on testicular function and sperm during cancer chemotherapy.

CD47;Rag2;IL-2rγ triple knock-out mice pre-conditioning with busulfan could be a novel platform for generating hematopoietic stem cells engrafted humanized mice.

Humanized mouse models to recapitulate human biological systems still have limitations, such as the onset of lethal graft-versus-host disease (GvHD), a variable success rate, and the low accessibility of total body irradiation (TBI). Recently, mice modified with the CD47-SIRPA axis have been studied to improve humanized mouse models. However, such trials have been rarely applied in NOD mice. In this study, we created a novel mouse strain, NOD-CD47nullRag2nullIL-2rγnull (RTKO) mice, and applied it to generate humanized mice. Four-week-old female NOD-Rag2nullIL-2rγnull (RID) and RTKO mice pre-conditioned with TBI or busulfan (BSF) injection were used for generating human CD34+ hematopoietic stem cell (HSC) engrafted humanized mice. Clinical signs were observed twice a week, and body weight was measured once a week. Flow cytometry for human leukocyte antigens was performed at intervals of four weeks or two weeks, and mice were sacrificed at 48 weeks after HSC injection. For a long period from 16 to 40 weeks post transplantation, the percentage of hCD45 was mostly maintained above 25% in all groups, and it was sustained the longest and highest in the RTKO BSF group. Reconstruction of human leukocytes, including hCD3, was also most prominent in the RTKO BSF group. Only two mice died before 40 weeks post transplantation in all groups, and there were no life-threatening GvHD lesions except in the dead mice. The occurrence of GvHD has been identified as mainly due to human T cells infiltrating tissues and their related cytokines. Humanized mouse models under all conditions applied in this study are considered suitable models for long-term experiments based on the improvement of human leukocytes reconstruction and the stable animal health. Especially, RTKO mice pretreated with BSF are expected to be a valuable platform not only for generating humanized mice but also for various immune research fields.

Exploring Intercellular Dynamics: Ultra-Weak Biophoton Emission as a Novel Indicator of Altered Cell Functions and Disease in Oligospermia Mice.

Introduction: Biophoton emission, the spontaneous release of photons from living cells, has emerged as an attractive field of research in the study of biological systems. Scientists have recently discovered that changes in biophoton emission could serve as potential indicators of pathological conditions. This intriguing phenomenon suggests that cells might communicate and interact with each other through the exchange of these faint but significant light signals. Therefore, the present study introduces intercellular relationships with biophoton release to detect normal and abnormal cell functions to further achieve cellular interactions by focusing on cell and cell arrangement in disease conditions. Methods: Twenty male mice were assigned to control and busulfan groups. Five weeks after the injection of busulfan, the testis was removed, and then the stereological techniques and TUNEL assay were applied to estimate the histopathology of the testis tissue sections. Results: The findings revealed that the ultra-weak biophoton emission in the control group was significantly lower than in the busulfan group. The oligospermia mice model showed that it significantly changed the spatial arrangement of testicular cells and notably decreased the testis volume, length of seminiferous tubules, and the number of testicular cells. The results of the TUNEL assay showed that the percentage of apoptotic cells significantly increased in the busulfan group. Conclusion: The ultra-weak biophoton emission from testis tissue was reduced in oligospermia mice. As a result, the decline of ultra-weak biophoton can indicate a change in cell arrangement, a decrease in intercellular interaction, and eventually disease.

Outcomes of Treosulfan Vs. Busulfan Based Conditioning Regimens in Adult Patients Undergoing Allogeneic HSCT: Real Life Data from a Single Center

Introduction The type of conditioning regimen has a great impact on the outcome of patients who undergo allogeneic HSCT since graft versus host disease (GVHD), infections, regimen related organ and tissue toxicities are among the most important causes of post-transplant mortality. Despite the regimen related toxicity profile of busulfan, supported by information from the results of clinical trials and real-world data, it is still commonly used worldwide. Treosulfan has advantages in terms of dose of administration, lower incidence of sinusoidal obstruction syndrome and lower neurotoxicity. We aimed to retrospectively investigate outcomes and compare results of patients who underwent matched and haploidentical allogeneic HSCT with treosulfan based or busulfan based reduced intensity conditioning, non-myeloablative conditioning and myeloablative conditioning regimens in our institution. Methods From September 2016 to November 2022, treosulfan was administered to 94 patients while 85 patients received busulfan. We analyzed outcomes as regimen related-toxicity, chronic and acute GVHD, overall survival, non-relapse mortality, relapse related mortality and fungal infection. The myeloablative dose of busulfan was 12.8 mg/kg while it was 9.6 mg/kg for the non myeloablative regimen which was combined with fludarabine. The myeloablative dose of treosulfan was 42 mg/m2 while a dose range of 30-36 mg/m2 was administered for the reduced intensity conditioning. Results The median follow up was 14 months for the treosulfan group while it was 11 months for the busulfan group (p = 0.16). Regimen related toxicity, which was monitored during the hospital stay and 3 months after discharge were 11.7% and 7.1% for treosulfan and busulfan respectively and were similar between two groups. Extensive chronic GVHD was encountered in 15.7% of the patients in the treosulfan group compared to 32.1% in the busulfan group (p &amp;lt; 0.001). Similarly, chronic skin GVHD was observed in 3.6% of the patients who received treosulfan while it was seen in 22.6% of the busulfan group (p &amp;lt; 0.001). Acute GVHD (grade 3 or higher) was encountered in 32.2% of the patients within the treosulfan group while it was encountered in 31.6% of the patients in the busulfan group. Overall survival was 47.5% and 36.5 % in the treosulfan group and busulfan groups respectively. The relapse-related mortality was 17 % in the treosulfan group while it was 34% in the busulfan group. Fungal infection was observed in 23.4% and 36.5% of the patients in treosulfan and busulfan groups respectively. Conclusion Treosulfan, with a lower chronic GVHD incidence and similar regimen related toxicity profile appears to be a safe alternative to busulfan and prospective randomized controlled trials are needed to confirm the results in our study. Keywords: Busulfan; treosulfan; conditioning regimen; stem cell transplantation

Comparable Outcome after Busulfan- or Treosulfan-Based Conditioning Regimen in Children Above 4 Years of Age with ALL Undergoing Allogeneic HSCT. Results from the Prospective International Forum-Trial

Purpose: Total body irradiation (TBI) has proved to be the “gold standard” as part of the conditioning regimen before allogeneic hematopoietic stem cell transplantation (HSCT) in pediatric patients with acute lymphoblastic leukemia (ALL). Its superiority over chemo-based conditioning was recently demonstrated in the prospective, international, randomized phase III study, which enrolled 417 patients beyond the age of 4 years transplanted for ALL in complete morphological remission (CR) from either matched sibling (MSD) or matched unrelated donor (MD) (Peters et al, JCO 2021, FORUM study; EudraCT: 2012-003032-22;ClinicalTrials.gov: NCT01949129). The use of either of the two protocol-prespecified chemo-conditioning regimens resulted in significantly worse EFS. Given the unavailability of TBI in some regions/centres and contraindications to TBI in individual patients, we here compare the outcomes of patients who received busulfan (BU) - based regimen vs of those who were given a treosulfan (TREO) - based conditioning in FORUM centres in both randomizing and non-randomizing countries in the years 2013-2018. Patients and methods: Patients ≤ 18 years at diagnosis (median age at HSCT 9.9 years, range 4-19.5), in CR pre-HSCT and with an MSD or MUD were assigned to myeloablative conditioning with fludarabine (FLU), thiotepa (THIO) and either BU or TREO according to country preference. Children transplanted from MSDs received cyclosporine A only as graft-versus-host disease (GvHD) prophylaxis, whereas recipients of MUD HSCT also received short-term methotrexate and anti-thymocyte globulin (ATG). Further details of the transplant procedure have been previously described (Peters C, et al. J Clin Oncol 2021). Results:In addition to the 193 patients from the randomizing countries, the FORUM trial included 115 additional children enrolled in countries where randomization could not occur (for legal or technical reasons). Overall, 180 vs. 128 patients received BU/THIO/FLU vs TREO/THIO/FLU, respectively. There were no differences about the patients' gender, age, and remission status and slight differences regarding donor, stem cell source and MRD status pre-transplant between the two cohorts of patients (see Table 1 for details). Patient's outcomes were updated as of February 20 th, 2023, and median follow-up was 4.2 years (range 0.3 - 9.1). There were neither differences between the 3-year overall survival (OS) (0.71±0.03 for BU/THIO/FLU vs 0.72±0.04 for TREO/THIO/FLU) nor event-free survival (EFS) (0.61±0.04 for BU/THIO/FLU vs 0.55±0.04 for TREO/THIO/FLU, p=NS). Three-year cumulative incidence of relapse (CIR) and non-relapse mortality (NRM) were 0.31±0.03 and 0.08±0.02 following BU/THIO/FLU and 0.36±0.04 and 0.09±0.03 following TREO/THIO/FLU, respectively (p=NS). Only one case of secondary malignancy was observed in the TREO cohort and one case of fatal liver veno-occlusive disease (VOD) in the BU group. No statistical differences were observed regarding the cumulative incidence of both aGvHD and cGvHD between the two groups; 3-year GFRS was almost identical (0.42± 0.04 in the BU/THIO/FLU group and 0.43±0.04 in the TREO/THIO/FLU cohort). There were no differences in OS, EFS, CIR, NRM and GFRS in either CR1 or CR2 patients between the two conditioning regimens. Furthermore, there were no differences in outcomes between countries. Patients given BU/THIO/FLU had a faster leucocyte, neutrophil and platelet recovery as compared to those prepared with TREO/THIO/FLU. Conclusion: Comparable long-term outcomes were observed after BU/THIO/FLU or TREO/THIO/FLU in children with ALL undergoing allogeneic HSCT included in the FORUM trial. Therefore, either of the 2 regimens may be effectively and safely used worldwide in patients &amp;gt; 4 years having a contraindication to or treated in centres/countries unable to deliver TBI.

Treosulfan Versus Busulfan-based Conditioning in Pediatric Patients Undergoing Hematopoietic Stem Cell Transplantation: A Systematic Review and Meta-analysis.

It is unclear whether there is a difference in outcomes with treosulfan or busulfan-based conditioning in pediatric patients undergoing hematopoietic stem cell transplantation (HSCT). We reviewed the evidence on this topic through a systematic review and meta-analysis, the comparison between treosulfan and busulfan-based conditioning in pediatric patients undergoing HSCT for instance. Six studies were included. Meta-analysis showed that there was no difference in the incidence of acute graft versus host disease (odds ratio [OR]: 0.96; 95% CI: 0.57, 1.61), grade II to IV acute graft versus host disease (OR: 1.19; 95% CI: 0.83, 1.72), chronic GVHD (OR: 1.18; 95% CI: 0.70, 2.00), and veno-occlusive disease (OR: 0.92; 95% CI: 0.22, 3.85) between treosulfan and busulfan groups. Pooled analysis indicated marginally better survival with treosulfan-based conditioning (OR: 1.57; 95% CI: 1.00, 2.44), however, these results were unstable on sensitivity analysis. A meta-analysis found no difference in transplant-related mortality (OR: 0.70; 95% CI: 0.34, 1.42) between the two groups. Retrospective data from a heterogenous population indicates that there is no difference in the rate of GVHD after treosulfan versus busulfan-based conditioning for pediatric HSCT. A marginal improvement in survival was noted with treosulfan but the results remained unstable. Future randomized controlled trials are needed to provide better evidence.

Effect of Carica papaya Leaf Juice on Bone Marrow Aspirate of Busulfan-induced Chronic Bone Marrow Aplasia in Mice

Introduction: Aplastic anemia is a disorder characterized by peripheral pancytopenia and hypoplastic bone marrow. Papaya, botanical name Carica Papaya, is a tropical fruit with a wide range of benefits. In past couple of years, researches have been carried out on beneficial outcomes of papaya leaf juice on blood and bone marrow, particularly platelet count. Aim: Taking into consideration the beneficial effects of papaya leaves on bone marrow, current research was done to probe the effects of C.papaya leaf juice on bone marrow differential cell count and cellularity in myelosuppressed mice where busulfan was used to induce myelosuppression. Study Design: Experimental. Place and Duration of Study: our study was performed in Post Graduate Medical Institute, Lahore in one year time. Materials &amp; Methods : It was an animal experimental study of 23 days in which 36 adult healthy swiss albino mice weighing 30-35 g of either sex were segregated into three groups with 12 mice in each group. Busulfan at a dose of 14mg/kg was given orally to the busulfan group to induce myelosuppression and to the C.papaya +busulfan group on 8 occassions (0, 3,8,11,15,18,21&amp;23) for a duration of 23 days. To observe the myeloprotective effect of Carica papaya leaf juice 10ml/kg of juice was given to the C.papaya + busulfan group daily for 23 days. After the final dose of busulfan and Carica papaya leaf juice on day 23, the samples for bone marrow aspirate were collected on same day. Results: The current study has revealed that C.papaya leaf juice not only prevented a decrease in all parameters of bone marrow aspirate but also maintained their normal values close to normal. The megakaryopoiesis was however found to be significantly higher than the normal. Keeping in view these beneficial effects carica papaya leaf juice can be used in chemotherapy induced thrombocytopenia and other related clinical conditions. Conclusion: it can be concluded from this study that C.papaya can be used in all such conditions that lead to fall in chemotherapy induced thrombocytopenia because of its property to increase megakaryopoiesis. Keywords: C.papaya, Busulfan, Megakaryopoiesis, Bone marrow failure.

The Gonadotoxic Effects of Conditioning Chemotherapy Regimens Prior to Hematopoietic Stem Cell Transplantation [ID: 1376168

INTRODUCTION: Primary ovarian insufficiency secondary to high-dose chemotherapy in females undergoing the conditioning regimen before hematopoietic stem cell transplantation is a challenging late complication. Limited studies have investigated the ovarian toxicity of various conditioning regimens. This study aimed to explore the gonadotoxic effects of cyclophosphamide (CY) and busulfan (BU) in CD-1 mice strain. METHODS: Sixteen 1-month-old female CD-1 mice (n=4/group) were subcutaneously injected with solvent, BU, CY, or BU+CY, respectively. The doses (144 mg/kg BU and 300 mg/kg CY) and injection plan were adjusted to human doses. The ovaries and female reproductive tracts were harvested 1 week after the last administration. Follicle counting was performed. The levels of serum antimüllerian hormone (AMH) and 17-beta-estradiol were measured. Animal use approval was obtained from the Institutional Animal Care and Use Committee at the University of Nebraska Medical Center. RESULTS: All mice survived the injections and grossly appeared normal. The average ovary weight was significantly lower in BU (P=.0007) and BU+CY (P&lt;.0001) groups than in the control and CY groups. The primordial (P&lt;.0001) and primary follicles (P=.01) in treatment groups were decreased compared to the control group. The serum AMH level in the BU group was also significantly lower than the control and CY group (P=.003). CONCLUSION: We show that conditioning chemotherapy with CY and BU can reduce the ovarian reserve in CD-1 mice strain and may have a synergic effect. Future studies to investigate the mechanism of chemotherapy-related ovarian follicle death are needed to improve fertility outcomes in patients receiving conditioning chemotherapy.

Protective effect of loboob (a Persian traditional remedy) on sexual hormones, antioxidant activities and stereological changes of testis tissue on busulfan induced oligospermia in rats

Loboob as a traditional drug in Iranis known for its beneficial effects on busulfan-induced oligospermia. In this experimental study, protective effects of loboob (a Persian traditional remedy) on sexual hormones, antioxidant levels and stereological changes of testis tissue were evaluated in an oligospermia rat model induced by busulfan. Fifty male rats were randomly divided into five different groups: control, received no treatments; and the other groups administrated with a single dose of busulfan (10 mg/kg body weight). After 30 days, these groups were treated with 0, 35, 70 or 140 mg/kg/day of loboob for 60 days. Blood samples were collected for hormone and antioxidant enzyme assays. Unbiased stereology was performed on testis tissues to evaluate the volume of different parts of the testis and the number of various testis cells. Data indicated that FSH, LH and MDA were increased, and testosterone, catalase, SOD were decreased in the busulfan group, while treatment with loboob at 70 and 140 mg/kg significantly improved these parameters (P <0.05). Treatment with 70 and 140 mg/kg of loboob ameliorated the germinal epithelium volume, types A and B spermatogonia, spermatocytes, elongated and round spermatids, and Sertoli cells in the seminiferous tubules (P <0.05). High concentration of loboob also improved testis weight and volume, and leydig cell number (P <0.05). Thus, loboob is more effective for the recovery of seminiferous tubules and their cells than for the interstitial tissue. Loboob with various antioxidants, minerals and vitamins could overcome the side effects of busulfan.

Effect of N-acetyl-L-cysteine on Testicular Tissue in Busulfan-Induced Dysfunction in the Male Reproductive System.

This study aimed to evaluate the protective effect of N-acetyl-L-cysteine (NAC) as an antioxidant on busulfan-induced testicular dysfunction in mice and elucidate its possible mechanism of action. Thirty-two C57BL/6 male mice were randomly divided into four groups (n=8/group) as follows: (1) control group (oral administration of saline [0.1 mL daily] for 35 days); (2) NAC group (oral administration of NAC [10 mg/kg daily] for 35 days); (3) busulfan group (double intraperitoneal injections of 20 mg/kg; total dose of 40 mg/kg); and (4) busulfan+NAC group (after double intraperitoneal injections of 20 mg/kg; total dose of 40 mg/kg, NAC administration [10 mg/kg daily] for 35 days). The testes were removed, weighed, and subjected to sperm parameter analysis and morphology assessment. Reproductive hormone, serum/testicular reactive oxygen species (ROS) level, oxidative stress and antioxidant markers were evaluated. The testicular expression of Nrf2 and HO-1 was examined using RT-qPCR. Busulfan treatment significantly decreased testicular weight, sperm count, and serum testosterone levels. Atrophy and degeneration of germinal epithelium were observed in the busulfan group. NAC administration after busulfan treatment partially attenuated the deterioration of testis weight, sperm quality, serum hormones, histomorphometric changes, and oxidative and antioxidative status. NAC treatment resulted in a considerable improvement in Nrf2 and HO-1 mRNA expression levels. This study provides compelling evidence that NAC as a potent antioxidant has significant protective effects against busulfan-induced male reproductive impairment possibly through modification of the Nrf2/HO-1 signaling pathway.

Peptides from the croceine croaker (Larimichthys crocea) swim bladder attenuate busulfan-induced oligoasthenospermia in mice

Context The swim bladder of the croceine croaker is believed to have a therapeutic effect on various diseases. However, there is no research about its effect on mammalian spermatogenesis. Objective We investigated the swim bladder peptides (SBPs) effect on busulfan-induced oligoasthenospermia in mice. Materials and methods We first extracted SBP from protein hydrolysate of the croceine croaker swim bladder, and then five groups of ICR male mice were randomly assigned: control, control + SBP 60 mg/kg, busulfan, busulfan + SBP 30 mg/kg and busulfan + SBP 60 mg/kg. Mice received bilateral intratesticular injections of busulfan to establish oligoasthenospermia model. After treatment with SBP for 4 weeks, testis and epididymis were collected from all mice for further analysis. Results After treatment with SBP 30–60 mg/kg for 4 weeks, epididymal sperm concentration and motility increased by 3.9–9.6- and 1.9–2.4-fold than those of oligoasthenospermia mice induced by busulfan. Meanwhile, histology showed that spermatogenic cells decreased, leading to increased lumen diameters and vacuolization in the busulfan group. These features were reversed by SBP treatment. RNA-sequencing analysis revealed that, compared with the busulfan group, Lin28b and Igf2bp1 expression related to germ cell proliferation, increased with a >1.5-fold change after SBP treatment. Additionally, PGK2 and Cfap69 mRNAs associated with sperm motility, also increased with a >1.5-fold change. Furthermore, these findings were validated by quantitative real-time PCR and Western blotting. Discussion and conclusions This is the first reported evidence for the therapeutic effect of SBP on oligoasthenospermia. SBP may be a promising drug for oligoasthenospermia in humans.

Combined Analysis of Gut Microbiota and Plasma Metabolites Reveals the Effect of Red-Fleshed Apple Anthocyanin Extract on Dysfunction of Mice Reproductive System Induced by Busulfan.

Busulfan is currently an indispensable anti-cancer drug, but the side effects on male reproductive system are so serious. Meanwhile, red-fleshed apples are natural products with high anthocyanin content. In this research, we analyzed the effect of red-fleshed apple anthocyanin extract (RAAE) on busulfan-treated mice. Compared with the busulfan group, main plasma biochemical indicators were significantly improved after RAAE treatment. Compared with BA0 (busulfan without RAAE) group, total antioxidant capacity(T-AOC) and the activity of superoxide dismutase (SOD) and glutathione catalase (GSH-Px) in RAAE treatment groups were obviously increased, while the activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were significantly decreased. Malondialdehyde (MDA) was significantly decreased in the RAAE groups. In addition, we found RAAE alleviated busulfan-disrupted spermatogenesis through improving genes expression which are important for spermatogenesis, such as DDX4, PGK2, and TP1. Furthermore, we found that RAAE increased beneficial bacteria Akkermansia and Lactobacillaceae, and significantly depleted harmful bacteria Erysipelotrichia. The correlation studies indicated that RAAE ameliorated busulfan-induced rise in LysoPC levels through regulating gut microbial community and their associated metabolites. In conclusion, this study extends our understanding of the alleviated effect of RAAE on busulfan-induced male reproductive dysfunction through regulating the relationships between gut microbiota and metabolites.

Effects of melatonin on both testicular regeneration and recovery of spermatogenesis in busulfan-treated rats

Testicular damage is one of the most hazardous effects of chemotherapy as it is frequently associated with oligozoospermia and azoospermia. This study aimed at evaluating the protective effect of melatonin in a rat model of busulfan-induced testicular injury. Rats were divided into four groups: control, melatonin, busulfan, busulfan plus melatonin. After 15 days, the semen was collected from the epididymis and testes were assessed. Sperm removed from cauda epididymis and analyzed for sperm count and viability. Testis tissues were also removed, fixed in formalin and were embedded in paraffin. Sections of testis tissue were stained with hematoxylin-eosin for histological examination and prepared for TUNEL (Terminal deoxynucleotide transferase dUTP Nick End Labeling) assay to detect apoptosis and PCNA (proliferating cell nuclear antigenassay) to detect proliferation cells. Serum and testes supernatants were separated to detect testosteron level and oxidative stress parameters. In histological examination, degenerative changes in seminiferous tubules were observed in the experimental groups. In biochemical examination, the total oxidant status (TOS) levels in Busulfan group were significantly higher than in the control group while the total antioxidant status (TAS) levels of all the groups were similar. In conclusion, the beneficial properties of melatonin treatment by its potent anti-oxidants may reduce adverse effects of chemotherapy in the reproductive system in a rodent system.

Effect of Carica papaya Leaf Juice on Blood Cell Count of Busulfan-Induced Chronic Bone Marrow Aplasia in Mice

Introduction: Aplastic anemia is a syndrome of bone marrow failure comprising of peripheral pancytopenia and marrow hypoplasia. Recently, research has shown papaya leaf juice to increase blood cells counts.&#x0D; Aims &amp; Objectives: This research was planned to investigate the effect of papaya leaf juice on blood cell counts in busulfan induced myelosuppressed mice.&#x0D; Place and duration of study: Post Graduate Medical Institute, Lahore from September 2015 to December 2015. Material &amp;Methods: This animal experimental study comprised of 36 adult healthy Swiss albino mice weighing 30-35 g, divided randomly into three equal groups. Busulfan was given orally (14mg/kg) to induce myelosuppression to busulfan group and Carica papaya+ busulfan group on 8 occasions over a period of 23 days. Carica papaya leaf juice (10ml/kg) was given to the Carica papaya+ busulfan group daily for 23 days to observe its role in preventing myelosuppression. Normal group was given distilled water in an equivalent amount. One day after the final dose of busulfan and Carica papaya, the animals were sampled for blood cell counts. Data was analyzed using SPSS version 20. A p-value &lt; 0.05 and below was considered as significant.&#x0D; Results: The Carica papaya leaf juice not only prevented significant fall in hemoglobin (10.86 ± 1.455 g/dl), RBC (9.54 ± 0.954× 106/mm3) and platelet count (958.92 ± 176.22 x103/mm3) vs (9.70 ± 0.904,11.24 ± 0.952 g/dl), RBC count (6.5025 ± 1.43,10.43 ± 0.234× 106/mm3) and platelet count (627.83 ± 151.63, 3881.08 ± 83.13 x103/mm3) in busulfan group but also kept them close to normal range. Total leukocyte count in C.papaya treated group improved significantly to 8116.67± 1540.85/mm3 vs (4516.67±789.51, 6666.67±798.10/mm3) busulfan treated and normal groups respectively.&#x0D; Conclusion: The Carica papaya leaf juice possesses significant myeloprotective activity. It may be a good candidate for prevention of chemotherapy induced pancytopenia

Protective effects of honey compound syrup on busulfan-induced azoospermia in male rats

Objective: To evaluate the protective effects of honey compound syrup on sperm count and testis tissue in rats. Methods: Thirty rats were randomly assigned to five groups. The control group received 1 mL normal saline with dimethyl sulfoxide intraperitoneally; the busulfan group received busulfan 10 mg/kg body weight at the first and twenty-first days of the experiment via intraperitoneal injection; the last three groups received busulfan 10 mg/kg body weight to induce azoospermia, and then received 1.0, 1.5, or 2.0 mg/kg honey compound syrup, respectively, after induction of azoospermia. After administration, the testis and epididymis of all rats were removed. Then, reproductive organ weight and sperm parameters (sperm concentration, epididymal sperm reserve and daily sperm production) were measured. After hematoxylin-eosin staining, seminiferous tubule cells and diameters were assessed. Results: Busulfan damaged the testis tissue and impaired spermatogenesis. Administration of honey compound syrup in three doses improved testis tissue and spermatogenesis. The protective effects of honey compound syrup may relate to the antioxidant properties of honey and other compounds in this syrup. Conclusions: Administration of honey compound syrup could be an ameliorative agent for the side effects of chemotherapy drugs such as busulfan on the male reproductive system.

Treosulfan-induced myalgia in pediatric hematopoietic stem cell transplantation identified by an electronic health record text mining tool

Treosulfan is increasingly used as myeloablative agent in conditioning regimen prior to allogeneic hematopoietic stem cell transplantation (HSCT). In our pediatric HSCT program, myalgia was regularly observed after treosulfan-based conditioning, which is a relatively unknown side effect. Using a natural language processing and text-mining tool (CDC), we investigated whether treosulfan compared with busulfan was associated with an increased risk of myalgia. Furthermore, among treosulfan users, we studied the characteristics of given treatment of myalgia, and studied prognostic factors for developing myalgia during treosulfan use. Electronic Health Records (EHRs) until 28 days after HSCT were screened using the CDC for myalgia and 22 synonyms. Time to myalgia, location of pain, duration, severity and drug treatment were collected. Pain severity was classified according to the WHO pain relief ladder. Logistic regression was performed to assess prognostic factors. 114 patients received treosulfan and 92 busulfan. Myalgia was reported in 37 patients; 34 patients in the treosulfan group and 3 patients in the busulfan group (p = 0.01). In the treosulfan group, median time to myalgia was 7 days (0–12) and median duration of pain was 19 days (4–73). 44% of patients needed strong acting opiates and adjuvant medicines (e.g. ketamine). Hemoglobinopathy was a significant risk factor, as compared to other underlying diseases (OR 7.16 95% CI 2.09–30.03, p = 0.003). Myalgia appears to be a common adverse effect of treosulfan in pediatric HSCT, especially in hemoglobinopathy. Using the CDC, EHRs were easily screened to detect this previously unknown side effect, proving the effectiveness of the tool. Recognition of treosulfan-induced myalgia is important for adequate pain management strategies and thereby for improving the quality of hospital stay.

Busulfan-containing conditioning regimens in allogeneic hematopoietic stem cell transplantation for acute lymphoblastic leukemia: A Taiwan observational study.

BackgroundAllogeneic stem cell transplantation (allo‐HSCT) is the ultimate cure for acute lymphoblastic leukemia (ALL).AimThis study was performed to compare the outcomes of ALL patients receiving busulfan (Bu) with cyclophosphamide (Cy)‐based or total body irradiation (TBI)‐based regimen in a Chinese population.MethodsWe enrolled 224 adult patients with ALL who received allo‐HSCT at National Taiwan University Hospital between 1997 and 2016.ResultsThe median age at transplantation was 33 years. Before allo‐HSCT, 75.9% of patients attained first or late complete remission. A total of 141 patients (62.9%) received Bu/Cy‐based conditioning, either myeloablative (MA) or reduced‐intensity stem cell transplantation (RIST), and 83 patients received a TBI‐based regimen (MA‐TBI). Patients receiving the MA‐Bu regimen had longer relapse‐free survival (RFS) than those receiving the MA‐TBI regimen (median, 24.1 vs. 6.7 months, p = .044). There was no difference in overall survival (OS, MA‐Bu vs. MA‐TBI vs. RIST‐Bu: 39.4 vs. 28.2 vs. 13.1 months, p = .276), treatment‐related mortality (TRM), or incidences of grade 3–4 acute graft‐versus‐host disease (GvHD). Among patients receiving identical GvHD prophylactic regimens, there was no difference between MA‐Bu and MA‐TBI groups regarding the incidence of grade 3–4 acute GvHD, grade 2–4, and all‐grade chronic GvHD. In subgroup analysis, patients receiving oral busulfan had comparable RFS and OS to the intravenous busulfan group (p = .436 and p = .236, respectively), but a higher TRM (25% vs. 9.8%, p = .016). In the multivariable analysis, disease status before allo‐HSCT was the only risk factor impacting RFS and OS.ConclusionIn summary, patients receiving Bu/Cy‐based or TBI‐based regimens as conditioning had similar results in terms of OS, TRM, and acute GvHD, whereas the use of myeloablative Bu/Cy resulted in a better RFS. A Bu‐based regimen could be an alternative conditioning choice for patients who are ineligible to receive TBI. Prospective and randomized controlled trials are warranted to validate the long‐term outcomes.