The effect of MgSO4 addition and the increasing doses of calcium and phosphorus during ending drying period on the occurrence of hypocalcaemia and hypophosphataemia in dairy cows

2013 ◽  
Vol 16 (4) ◽  
pp. 655-662 ◽  
Author(s):  
R. Bodarski ◽  
S. Kinal ◽  
J. Preś ◽  
M. Słupczyńska ◽  
J. Twardoń
Keyword(s):  
Blood Serum ◽  
Magnesium Sulphate ◽  
Group Iv ◽  
Dry Period ◽  
Urine Ph ◽  
Inorganic P ◽  
Group Iii ◽  
Group I ◽  
Salt Addition ◽  
Group Ii

Abstract The aim of the presented study was the estimation of optimal Ca and P levels applied before calving together with anionic salt addition, as an element of hypocalcaemia and hypophosphataemia prevention. The experiment was carried out during the dry period on 48 cows with similar milk yield in the previous lactation. Cows were divided into four groups. In group I (control) the amount of minerals was in accordance to NRC standards. In experimental groups (groups II-IV), two weeks before calving, cows received 140 g/day/head of hydrated magnesium sulphate to achieve dietary cation- anion difference at the level of about 50 mEq/kg DM. In groups II and III cows received calcium carbonate (100 g/day) 10 days a.p. (antepartum) (group II), or 5 days a.p. (group III), while cows in IV group received dicalcium phosphate (100 g/day) for 5 days a.p. Application of MgSO4 × 7H2O significantly affected the urine pH of cows from group III and IV 4-5 d. before calving - 6.45 and 6.81, respectively. The acidification of urine was observed after calving in group IV (7.13). In cows from group II (100 CaCO3 10 days a.p.) urine pH decline was not found (7.97-7.75). In that group the incidences of hypophosphatemia were noted (blood serum inorganic P level 1.41-1.46 mmol/l). Addition of magnesium sulphate prevented hypocalcaemia occurrence - 4-5 d. before calving the concentration of ionized Ca in blood serum was 1.11, 1.13 and 1.16 mmol/l (respectively for group II, III and IV). Reproductive functions were significantly improved after the application of CaCO3 and CaHPO4 for 5 days a.p. in comparison with control and group II - progesterone concentration in the blood serum on the 45th day of lactation was 1.396 - 1.409 vs 0.799 - 0.401. The correlation between progesterone and inorganic P level in serum was almost significant. Based on the obtained results a treatment optimal in prevention of hypocalcaemia and hypophosphataemia is the application of 50 g CaCO3 and 50 g of CaHPO4 for the last 5 days of the dry period together with MgSO4 × 7H2O given for 14 days a.p.

scholarly journals Effect and mechanisms of reproductive tract infection on oxidative stress parameters, sperm DNA fragmentation, and semen quality in infertile males

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Kang-Sheng Liu ◽  
Xiao-Dong Mao ◽  
Feng Pan ◽  
Rui Fang An
Keyword(s):  
Reproductive Medicine ◽  
Reproductive Tract ◽  
Group Iv ◽  
Semen Parameters ◽  
Group Iii ◽  
Reproductive Tract Infection ◽  
Group I ◽  
Sperm Dna ◽  
Group Ii ◽  
Elastase Level

AbstractRecent years have seen a rising incidence of male infertility, mostly caused by the decline of sperm quality. The ratio of infertile males to infertile females has escalated from 3:7 in 2013 to current 5:5, which turns male infertility into the research focus of reproductive medicine. This study aimed to clarify the effect of reproductive tract infection by ureaplasma urealyticum (UU) and chlamydia trachomatis (CT) on the DNA integrity and routine semen parameters of infertile males. A retrospective study was performed. A total of 259 infertile males who were treated at the Andrological Laboratory Examination and Reproductive Medicine Center in our hospital were analyzed. qRT-PCR was used to examine the infection status of CT and UU. According to the eligibility criteria, we evaluated the semen parameters and biochemical data of 253 men. Based on the results of PCR, the subjects were divided into four groups: Group I (CT positive, 63 cases), Group II (UU positive, 60 cases), Group III (CT positive and UU positive, 62 cases), and Group IV (no infection, 68 cases). DNA fragmentation index (DFI), sperm count, vitality and morphology, elastase level, seminal plasma malondialdehyde (MDA), and total antioxidant capacity (TAC) were assessed. Compared to Group IV, three groups (Group I, Group II and Group III) showed difference in semen volume, proportion of sperm with normal morphology, sperm motility, progressive motility, and vitality (P < 0.05). Compared to Group IV, Group II and Group III showed difference in DFI (P < 0.05). Compared to Group IV, Group II and Group III showed difference in elastase level (P < 0.05). VCL, VSL, VAP, WOB, ROS, TM, HDS showed differences between groups of abnormal/normal WBC (*P < 0.01).UU infection significantly increased the level of seminal leukocytes only in Group II, but not in the other three groups, indicating that UU is a factor to increase the level of seminal leukocytes. Compared with the normal leukocyte group, there were significant differences in total motility, forward motility and normal sperm ratio between the two groups. The proportion of sperm with abnormal morphology (mostly in the head) showed obvious difference between groups of high and normal seminal leukocytic levels. At the same time, in this study, SCGE and SCD verified that leukocytes could damage sperm DNA by increasing ROS, which ultimately affects male fertility.

Effect of amino acid infusion on renal hemodynamics in humans: a dose-response study

1994 ◽  
Vol 267 (5) ◽  
pp. F703-F708 ◽  
Author(s):  
M. Giordano ◽  
P. Castellino ◽  
E. L. McConnell ◽  
R. A. DeFronzo
Keyword(s):  
Amino Acid ◽  
Dose Response ◽  
Renal Hemodynamics ◽  
Group Iv ◽  
Group V ◽  
Group Iii ◽  
Amino Acid Infusion ◽  
Group I ◽  
Basal Plasma ◽  
Group Ii

We evaluated the dose-response relationship between the plasma amino acid (AA) concentration and renal hemodynamics in eight normal subjects. After an overnight fast, a balanced 10% AA solution was infused for 180 min at five separate infusion rates: 0.5 (group I), 1.0 (group II), 2.0 (group III), 4.0 (group IV), and 6.0 (group V) ml.kg-1.min-1 on separate days. Basal plasma AA concentration was 1.87 +/- 0.1 mmol/l and increased to 2.26 +/- 0.1 (group I), 2.66 +/- 0.2 (group II), 3.79 +/- 0.5 (group III), 5.81 +/- 0.4 (group IV), and 7.41 +/- 0.4 mmol/l (group V). Basal glomerular filtration rate (GFR) and renal plasma flow (RPF) averaged 95 +/- 4 and 476 +/- 29 ml.1.73 m-2.min-1, respectively, and rose to 98 +/- 5 and 506 +/- 40 (group I) [P = not significant (NS)], 102 +/- 3 and 533 +/- 30 (group II) (P < 0.05 vs. basal), 110 +/- 4 and 567 +/- 29 (group III), 115 +/- 7 and 610 +/- 55 (group IV), and 117 +/- 7 and 614 +/- 66 ml.1.73 m-2.min-1 (group V) (P = NS vs. group IV). Basal plasma glucagon concentration averaged 68 +/- 10 pg/ml and increased to 74 +/- 10 (group I), 83 +/- 11 (group II) (P < 0.05 vs. basal), 100 +/- 14 (group III), 121 +/- 14 (group IV), and 229 +/- 35 pg/ml (group V) (P < 0.01 vs. basal). Increases in plasma growth hormone (GH) and insulin levels were observed only during groups IV and V.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Pathomechanism of Insulin Resistance in Men with Central Obesity: Correlation of GGT, GPx, hs-CRP and Plasma Total Cysteine

2013 ◽  
Vol 5 (2) ◽  
pp. 101 ◽  
Author(s):  
Ritawaty Ritawaty ◽  
Indriyanti Rafi Sukmawati ◽  
Ilhamjaya Patellongi ◽  
Ferry Sandra
Keyword(s):  
Oxidative Stress ◽  
Insulin Resistance ◽  
Amino Acid ◽  
Fatty Liver ◽  
Central Obesity ◽  
Group Iv ◽  
Group Iii ◽  
Group I ◽  
Group Ii ◽  
Hs Crp

BACKGROUND: Gamma glutamyltransferase (GGT) was reported recently to be associated with inflammation, oxidative stress and increased amino acid. However, role of GGT in insulin resistance pathomechanism is not exactly known. Therefore correlation of GGT with inflammation, oxidative stress and elevated amino acid, in men with central obesity need to be confirmed.METHODS: A cross-sectional study was designed. Men with central obesity were recruited and selected. Anthropometric parameters, creatinine, hs-CRP, fasting glucose, fasting insulin, glutathione peroxidase (GPx) activity, GGT, plasma total cysteine (tCys) and fatty liver were measured. Subjects were then divided in 4 groups based on waist circumference (WC) and fatty liver: Group I: WC ≤100 cm, without fatty liver; Group II: WC ≤100 cm, with fatty liver; Group III: WC >100 cm, without fatty liver; Group IV: WC >100 cm, with fatty liver. All biochemical characteristics in each group were then statistically analyzed.RESULTS: Seventy-two men with central obesity were selected. Numbers of subjects in each group were: Group I: n=33; Group II: n=5; Group III: n=17; Group IV: n=17. We found significant difference of HOMA-IR between Group I and IV, significant correlation between GGT and HOMAIR, and significant negative correlation between tCys with HOMA-IR in Group IV.CONCLUSION: GGT was significantly correlated with HOMA-IR in men with WC >100 cm and fatty liver. Further investigation with more subjects is necessary to determine clear GGT cut-off to distinguish subjects with fatty liver and insulin resistance.KEYWORDS: GGT, hs-CRP, GPx, tCys, HOMA-IR, insulin resistance

scholarly journals Analyses of the results in the treatment of locally advanced hypopharyngeal carcinoma with different therapeutic approach

2009 ◽  
Vol 56 (1) ◽  
pp. 35-40 ◽  
Author(s):  
A. Mikic ◽  
I. Pendjer ◽  
Z. Dudvarski ◽  
A. Trivic ◽  
M. Kreacic
Keyword(s):  
Neoadjuvant Chemotherapy ◽  
Postoperative Radiotherapy ◽  
Locally Advanced ◽  
Group Iv ◽  
Hypopharyngeal Carcinoma ◽  
Group Iii ◽  
Group I ◽  
Primary Radiotherapy ◽  
Group Ii ◽  
Number Of Authors

Introduction: treatment of locoregional advanced hypopharyngeal cancers of grades III and IV would be surgery and/or radiotherapy. An increasing number of authors has emphasized favorable effects of neoadjuvant chemotherapy and radiotherapy with potentiation. The aim: the authors presented therapeutical results of three-year survived patients with squamocellular cancers of the hypopharynx (SCC Hy), grades III and IV, in relation to specific modalities of therapeutical approach. Methods: a series of 207 patients with grades III and IV of SCC Hy, treated in the period 1982 to 2002, was analyzed. Group I consisted of 131 patients who had surgery and postoperative radiotherapy (SUR-RT). Group II included 16 patients who underwent induction chemotherapy, surgery and postoperative radiotherapy (IC-SURRT). Group III consisted of 60 patients, who had primary radiotherapy (RT). Group IV involved 29 patients who were treated, in the period 2003-2006, by neoadjuvant chemotherapy followed by radiotherapy with potentiation (CHT-RT). Results: three-year survival in Group I was 67.94%, Group II - 50%, Group III - 30%, and in Group IV was 51.72%. Five-year survival was analyzed only in group IIII. The longest survival was observed in Group I - 33.59%. Conclusion: the best therapeutical results were achieved in patient group that was primarily operated on and then had postoperative radiotherapy, while the worst survival was recorded in patients who had primary radiotherapy.

scholarly journals Microbiome signature and diversity regulates the level of energy production under anaerobic condition

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
M. Shaminur Rahman ◽  
M. Nazmul Hoque ◽  
Joynob Akter Puspo ◽  
M. Rafiul Islam ◽  
Niloy Das ◽  
...  
Keyword(s):  
Indicator Species ◽  
Energy Production ◽  
Environmental Temperature ◽  
Group Iv ◽  
Methanosarcina Barkeri ◽  
Spearman Correlation ◽  
Positive Correlation ◽  
Group Iii ◽  
Group I ◽  
Group Ii

AbstractThe microbiome of the anaerobic digester (AD) regulates the level of energy production. To assess the microbiome diversity and composition in different stages of anaerobic digestion, we collected 16 samples from the AD of cow dung (CD) origin. The samples were categorized into four groups (Group-I, Group-II, Group-III and Group-IV) based on the level of energy production (CH4%), and sequenced through whole metagenome sequencing (WMS). Group-I (n = 2) belonged to initial time of energy production whereas Group-II (n = 5), Group-III (n = 5), and Group-IV (n = 4) had 21–34%, 47–58% and 71–74% of CH4, respectively. The physicochemical analysis revealed that level of energy production (CH4%) had significant positive correlation with digester pH (r = 0.92, p < 0.001), O2 level (%) (r = 0.54, p < 0.05), and environmental temperature (°C) (r = 0.57, p < 0.05). The WMS data mapped to 2800 distinct bacterial, archaeal and viral genomes through PathoScope (PS) and MG-RAST (MR) analyses. We detected 768, 1421, 1819 and 1774 bacterial strains in Group-I, Group-II, Group-III and Group-IV, respectively through PS analysis which were represented by Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria, Spirochaetes and Fibrobacteres phyla (> 93.0% of the total abundances). Simultaneously, 343 archaeal strains were detected, of which 95.90% strains shared across four metagenomes. We identified 43 dominant species including 31 bacterial and 12 archaeal species in AD microbiomes, of which only archaea showed positive correlation with digester pH, CH4 concentration, pressure and temperature (Spearman correlation; r > 0.6, p < 0.01). The indicator species analysis showed that the species Methanosarcina vacuolate, Dehalococcoides mccartyi, Methanosarcina sp. Kolksee and Methanosarcina barkeri were highly specific for energy production. The correlation network analysis showed that different strains of Euryarcheota and Firmicutes phyla exhibited significant correlation (p = 0.021, Kruskal–Wallis test; with a cutoff of 1.0) with the highest level (74.1%) of energy production (Group-IV). In addition, top CH4 producing microbiomes showed increased genomic functional activities related to one carbon and biotin metabolism, oxidative stress, proteolytic pathways, membrane-type-1-matrix-metalloproteinase (MT1-MMP) pericellular network, acetyl-CoA production, motility and chemotaxis. Importantly, the physicochemical properties of the AD including pH, CH4 concentration (%), pressure, temperature and environmental temperature were found to be positively correlated with these genomic functional potentials and distribution of ARGs and metal resistance pathways (Spearman correlation; r > 0.5, p < 0.01). This study reveals distinct changes in composition and diversity of the AD microbiomes including different indicator species, and their genomic features that are highly specific for energy production.

scholarly journals Assessment of Outcome of Intrathecal Analgesia in Multiparous Women Undergoing Vaginal Delivery

2020 ◽  
Vol 5 (1) ◽  
pp. 153-155
Author(s):  
Sanjay Melville Masih ◽  
Rakesh Kumar Gupta
Keyword(s):  
Vaginal Delivery ◽  
Group Iv ◽  
Normal Vaginal Delivery ◽  
Secondary Outcome ◽  
Group Iii ◽  
Group I ◽  
Significant Difference ◽  
Multiparous Women ◽  
Group Ii ◽  
Intrathecal Analgesia

Background: The present study was conducted to assess the outcome of intrathecal analgesia in multiparous women undergoing vaginal delivery. Subjects and Methods: The present study was conducted among 80 multiparous women ages ranged 18- 40 years. All patients received 0.5 ml of intrathecal injection of 2.5 mg bupivacaine 0.5% and 1 ml dexamethasone 4 mg plus a 0.5 ml adjuvant. The adjuvants in group I patients were 100 μg morphine, 25 μg fentanyl in group II, 5 μg dexmedetomidine in group III and normal saline in group IV (control group). The primary and secondary outcome was the duration of pain relief, the analgesia onset time, the maximum level of sensory block, the visual analogue scale (VAS) was recorded. Results: The mean duration of analgesia in group I was 182.4 minutes, in group II was 170.5 minutes, in group III was 200.4 minutes and in group, IV was 140.2 minutes. The onset of analgesia was 3.9 minutes, 2.8 minutes, 2.7 minutes and 4.5 minutes in group I, II, III and IV respectively. S1 sensory regression time (minutes) was 181.4, 157.4, 185.3 and 130.6 in group I, II, III and IV respectively. Modified Bromage scale 5 minutes, 15 minutes and 30 minutes after IT in all groups was 0.0. VAS was 1.3, 1.6, 1.6 and 3.1 with significant differences in all groups (P< 0.05). There was non- significant difference in mean age, weight, height and gestational age between all groups (P> 0.05). There was a non- significant difference in APGAR 1, 5, umbilical pH immediately after delivery, neonatal HR after 5minute, 15 minutes and 30 minutes in all groups (P> 0.05). Conclusion: The authors found that dexmedetomidine is a safe and effective adjuvant to intrathecal bupivacaine-dexamethasone in multiparous women undergoing normal vaginal delivery.

Adaptive dose response pilot study of intravaginal Prostaglandin E2 application in free farrowing sows during parturition

2020 ◽  
Author(s):  
Ramona Bill ◽  
Heiko Nathues ◽  
Alexander Grahofer
Keyword(s):  
Prostaglandin E2 ◽  
Total Duration ◽  
Group Iv ◽  
Control Group ◽  
Birth Process ◽  
Group Iii ◽  
Beneficial Effects ◽  
Group I ◽  
Group Ii ◽  
Meconium Staining

Abstract Background The duration of birth is an important factor influencing the survival of piglets and the health of sows. A prolonged parturition is usually treated with oxytocin, even though several undesirable side effects are described. Therefore, the aim of this study was to evaluate the safety and efficacy of Prostaglandin E2 (PGE2) of different concentrations as an intravaginal applied gel in sows. MethodsTwelve sows were randomly allocated to one of four treatment groups: Group I (control group) application of placebo gel; Group II application of 2.0 mg PGE2; Group III application of 1.0 mg PGE2; Group IV application of 0.5 mg PGE2. The gel was administered intravaginal after the birth of the fourth piglet. Total duration of birth (time between first piglet and last placenta), piglet interval and placenta expulsion duration (time between first and last placenta) were recorded, and each piglet was scored for meconium staining and vitality. Furthermore, stillborn piglets were categorized into ante-partum and intra-partum deaths. Results Although no significant differences between the groups were detected, a beneficial tendency in several parameters was observed in group III. The duration of birth was 284 min (average of 14.3 piglets per litter), whereas in all other groups it was more than 400 min (average of 18.0 piglets per litter). The piglet interval was 10.1 min in group II compared to 10.2 min in group III, 16 min in group IV and 21.4 min in group I. In group III the placenta expulsion duration was 119 min in contrast to the other groups with more than 266 min. Severe meconium staining in more than 10% of piglets was observed in group II and IV. Moreover, piglets of group II showed oedematous and haemorrhagic umbilical cords, lethargy and anoxia, and intra-partum deaths were recorded. Conclusion Although the sample size is limited in this study, it is assumed that an application of 1mg PGE2 intra-partum instead of lower or higher dosages has the most beneficial effects on the birth process in sows and on the vitality of piglets. Further investigation is necessary to confirm the positive effect of PGE2 in daily practise.

Is there a difference in prevalence of anxiety and depression symptoms among patients with different stages of hypertension?

2017 ◽  
Vol 41 (S1) ◽  
pp. S414-S414 ◽  
Author(s):  
A. Mermerelis ◽  
S.M. Kyvelou ◽  
V. Akke ◽  
C. Papageorgiou ◽  
C. Stefanadis ◽  
...  
Keyword(s):  
Depression Scale ◽  
Group Iv ◽  
Anxiety And Depression ◽  
Depression Symptoms ◽  
Study Cohort ◽  
Hypertensive Patients ◽  
Group Iii ◽  
Group I ◽  
Significant Difference ◽  
Group Ii

IntroductionWhether anxiety and depression are associated with hypertension and to what extent is not clear.AimsThe aim of the present study was to assess any differences in the prevalence of anxiety and depression among different groups of hypertensive patients.MethodsThe study cohort comprised of 127 patients (75 male, mean age 54 ± 14) who underwent assessment of their blood pressure levels and were divided in four groups: group I (normotensives, n = 34), group II (stage 1 HTN, n = 33), group III (stage 2 HTN, n = 30) and group IV (stage 3 HTN, n = 30). The evaluation of anxiety disorder was made by means of Hospital Anxiety Depression Scale (HADS), while the evaluation of depression was made with the Beck Depression Inventory (BDI). Statistical analysis was done with SPSS for windows. P-value was set at 0.05 for differences to be considered significant.ResultsComparing the four groups of patients there was a significant difference both in BDI (8.6 ± 7.0 vs. 11.6 ± 10.4 vs. 27.1 ± 5.8 vs. 32.4 ± 3.9, P < 0.0001) and HADS (10.2 ± 7.2 vs. 9.7 ± 7.0 vs. 16 ± 4.7 vs. 27 ± 5.1, P < 0.0001). We proceeded to comparison among the 4 groups and there was a significant rise in the BDI and HADS in three of the four groups group II > group III > group IV, P < 0.0001.ConclusionThese data suggest that there is a clear burden of anxiety and depression as the levels of BP increase. This finding is of important clinical significance as it could contribute to a different approach of hypertensive patients. A larger cohort study could enlightened the mechanisms involved.Disclosure of interestThe authors have not supplied their declaration of competing interest.

scholarly journals Remifentanil and Propofol Sedation for Retrobulbar Nerve Block

2002 ◽  
Vol 30 (4) ◽  
pp. 433-437 ◽  
Author(s):  
V. Rewari ◽  
R. Madan ◽  
H. L. Kaul ◽  
L. Kumar
Keyword(s):  
Adverse Effects ◽  
Respiratory Depression ◽  
Study Groups ◽  
Group Iv ◽  
Control Group ◽  
Group Iii ◽  
Group I ◽  
Retrobulbar Block ◽  
Significant Movement ◽  
Group Ii

We studied remifentanil and propofol for analgesia and sedation during the placement of an ophthalmic block. Eighty ASA I or II patients undergoing elective cataract surgery under a retrobulbar block in a rural camp setting were included in the study. Patients were randomly divided into four groups and received different drug combinations as follows: Group I—remifentanil 1 μg/kg, Group II—remifentanil 0.5 μg/kg and propofol 0.5 mg/kg, Group III— remifentanil 1 μg/kg with propofol 0.5 mg/kg and Group IV—saline 0.1 ml/kg. Patients were observed for degree of movement, sedation, pain, recall and respiratory depression. No patient in the study groups reported pain or displayed movement whereas most of the patients in the control group had significant pain during the placement of the block. Also, seven (35%) patients in the control group showed significant movement which may have led to failure of block in two patients and retrobulbar haemorrhage in one patient. Incidence of significant respiratory depression was maximum in Group III patients (60%), followed by Group I (20%) and least in Group II (5%). All patients in the study groups remained cooperative and obeyed commands except four patients in group III (OAA/S-4). Postoperatively, other than the control group, recall was maximum in Group I (55%) and least in Group II (5%). Hence, a combination of remifentanil 0.5 μg/kg with propofol 0.5 mg/kg as a bolus was considered to provide excellent relief of pain and anxiety with least adverse effects for the placement of ophthalmic blocks.

Droplet Digital PCR Reliably Detects a Single Copy of BCR-ABL1

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 2784-2784
Author(s):  
Jaspal Kaeda ◽  
Simone Bonecker ◽  
Frauke Ringel ◽  
Michaela Schwarz ◽  
Bernd Dörken ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Chronic Phase ◽  
Digital Pcr ◽  
Group Iv ◽  
Droplet Digital Pcr ◽  
Pcr Assay ◽  
Group Iii ◽  
Group I ◽  
Increased Sensitivity ◽  
Group Ii

Abstract Data show 40% of chronic myeloid leukemia (CML) patients maintain complete molecular remission (CMR), i.e. failure to detect BCR-ABL1, consenting to termination of Imatinib mesylate (IM) therapy, following undetectable disease for ≥2 years by quantitative PCR (Q-PCR). These findings suggest majority of the patients experience molecular relapse. Furthermore, majority relapse in the first 6 months, implying Q-PCR assay sensitivity is suboptimum, to confidently identify patients for discontinuation of IM. Droplet digital PCR (ddPCR) is suggested to have sensitivity that is one log greater than the Taqman (Q-PCR) assay. If verified, ddPCR would enhance safe withdrawal of IM therapy from CML patients. Here we present data comparing ddPCR with Q-PCR. In total we assayed 161 samples, of these 6 were serial dilutions of the International reference (IR) BCR-ABL1 plasmid, the remainder were cDNA samples. The 161 samples comprised of 4 sample groups; I: CML chronic phase samples (n=118); II: CML samples post stem cell transplant (SCT) (n=22); III: normal control (NC) samples (n=16); IV: Serially diluted BCR-ABL1 1.04x10e2 copies/µl, IR ERM-AD623e (n=6). Group I comprised of 121 samples from 21 CML patients in chronic phase treated with IM. Group II comprised of 21 samples from 19 CML patients (2 samples each for 2 of these 19 patients) who had undergone SCT. Group II comprised of samples from normal adult blood donor volunteers. Finally, Group IV included 6 serially diluted samples, ranging 100 to 0.001 copies of the IR (1.04x10e2 copies/µl) plasmid (Sigma, Munich, Germany). All the Group I, III and IV samples were subjected to Q-PCR Taqman assay and ddPCR (Biorad, California, USA). The Group III samples were in addition subjected to nested PCR. Only those samples with cycle threshold (Ct) <37 in 2 or more replicates by Q-PCR were recorded positive. For ddPCR those samples with sum of ≥3 positive droplets with a minimum 5500 droplets per well were reported positive. All the PCR reactions were performed in triplicate in final volume of 20µl, which included 5µl of cDNA or reference plasmid. Among Group I, Q-PCR detected BCR-ABL1 in 57 of the 118 patient samples assayed, with a median of 16.72 transcripts (range 1.38-47450). In only 2 (2.25 and 4.96 transcripts) of these 57 samples, ddPCR failed to detect BCR-ABL1 transcripts. Q-PCR and ddPCR failed to detect BCR-ABL1 in 45 (38.1%) samples. For 14 (11.8%) of the samples ddPCR was positive, median 3.4 copies (range 3-15) but negative by Q-PCR. Among Group II, one of the 21 samples was excluded from the analysis because <5500 droplets were generated. Of the 21 samples 7 were negative by ddPCR. Nested PCR was negative for all 7 samples. Three samples were positive by all 3 technologies, nested PCR, Q-PCR and ddPCR. Remarkably, 5 of the 21 samples were negative by nested, but positive by ddPCR; median 18.0 copies (6.2-22.0). These 5 samples were not subjected to Q-PCR. In Group III all 16 NC samples were negative by Q-PCR and ddPCR. In Group IV, ddPCR did detect BCR-ABL1 in the serially diluted IR sample calculated to have 1 copy (26 positive droplets of the 106095 total droplets), but Q-PCR failed. However, the lower dilutions, calculated to contain 0.1, 0.01 and 0.001 copies were negative by ddPCR and Q-PCR. We assayed 161 samples by ddPCR and Q-PCR, of these 139 were from CML patients. In addition 21 of the samples were also subjected to nested PCR. Our data support the notion ddPCR is at least one log more sensitive than Q-PCR. Of the 140 patient samples assayed, 19 (13.5%) were positive by ddPCR but negative by Q-PCR. Only 2 of the 118 samples in Group I were negative by ddPCR but positive by Q-PCR. There was in insufficient sample to repeat these 2 assays. The increased sensitivity of ddPCR as implied by the clinical samples was supported by assays performed using the IR. The serial dilution equivalent to 1.0 BCR-ABL1 copy was reliably detected by ddPCR, but was negative by Q-PCR. In summary, these data suggest ddPCR is more sensitive. However, the clinical significance of this must be assessed in context of long-term clinical outcome of patients with detectable BCR-ABL1 by ddPCR and negative by Q-PCR. But, clearly increased sensitivity is likely to enhance safe withdrawal of IM therapy for CML patients in CMR. Furthermore, regular monitoring of these patients by ddPCR would enable early detection of molecular relapse and thereby minimize the risk of disease progression. Disclosures Le Coutre: Novartis: Honoraria.

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