Post Wash Research Articles

#336 : Improving Success of Intrauterine Insemination

Background and Aims: IUI is one of the most used methods to treat various forms of infertility. The aims were to compare the success rate of IUI with that of timed intercourse, review the methods used to time the IUI with ovulation, identify the factors to determine IUI outcome, compare the success rate of double IUI vs single IUI. Method: This is a cross-sectional prospective study conducted at a Private Fertility Centre, Chattogram, Bangladesh over a period of 40 months from 996 patients who underwent IUI. Variables such as age of women, cause of infertility, drugs used for COH, number of mature follicles, endometrial thickness post wash sperm count, type of IUI (single IUI and double IUI) were recorded and statistically analyzed. Results: The age for females and males were 18-41 and 23-52 respectively. The mean endometrial thickness was 8.9 mm ± 0.2 mm. Pregnancy rate for total motile sperm count less than 10 million was 7.2%. The successful conception rate for females were the age 28 was 9.6% compared to the whole group at 12%. Among 996 patients, 109 patients (10.94%) had successful conception. 209 patients (20.98%) underwent double IUI out of which 40 patients (19.14%) had a successful conception while 787 patients (79.01%) underwent single IUI out of which 69 patients (8.76%) had successful conception. Conclusion: IUI is currently a successful treatment modality for infertility especially as invasive treatment for couples with unexplained infertility, male factor infertility, anovulation, female less than 30 years of age. The numbers of motile sperms, appropriate endometrial thickness, have a remarkable impact on the outcome of IUI. Double IUI increases the success rate significantly. Acknowledgment: The authors are grateful to all who participated in the preparation of this manuscript.

Estimation of follicular growth—widely used, seldom studied

We assessed whether estimation of follicular growth, rather than actual measurement of follicular size on the day of hCG trigger, affected pregnancy rates in intrauterine insemination (IUI) cycles. Patient and cycle characteristics were extracted from an existing database. Comparisons were made between the pregnant (defined as a positive beta hCG) and non-pregnant groups for the following variables: patient’s age, number of previous IUI cycles, type of ovarian stimulation, endometrial thickness, number of follicles measuring 14 mm and above, pre and post wash sperm parameters, cycle day when IUI was done and number of days between last ultrasound scan and ovulation trigger. A total of 7302 cycles were included in the final analysis. In 4055 cycles (55.5%) the hCG trigger was on the day of the last ultrasound, in 2285 cycles (31.3%) the hCG trigger was 1 day after the last ultrasound, in 850 (11.6%) it was 2 days after the last ultrasound and in 112 (1.5%) it was 3 or more days after the last ultrasound. Sperm parameters, younger maternal age, and the number of follicles above 14 mm were all associated with pregnancy. No association was found between positive pregnancy test rates and the time from last ultrasound to hCG trigger. Planning IUI based on the estimation of follicular growth 1–4 days before trigger, does not affect pregnancy rates.

P-721 Prediction model of pregnancy after intrauterine insemination: accessibility, relevance for clinical use and patient counseling

Abstract Study question The aim of this study is to develop a prognostic model for the outcome of intrauterine insemination that is useful for daily clinical practice and to develop an app Summary answer Our predictive model is able to make a good distinction between patients with a good or poor pregnancy chance What is known already IUI is a common, accessible and easily accepted method of treating infertile couples with a success rate varying largely between 5% and 70%. The factors that contribute to the successful IUI rate are numerous including the woman's age, type of infertility, the number of follicles obtained by ovarian stimulation, the estradiol concentration on the day of human chorionic gonadotrophin administration, the sperm parameters (concentration, motility), duration of infertility, endometrial thickness, the number of IUI cycles. Several studies have already tried to establish a success rate formula, but no prediction model is largely used. Study design, size, duration This retrospective cohort study has been conducted at an university level fertility clinic from Oradea, Romania. Patients eligible to participate in the study were 426 infertile couples who underwent IUI treatment between January 2015 and October 2020. Participants/materials, setting, methods The 426 couples were classified into three groups: 1. male infertility factor 2. female infertility factor (minimal/mild endometriosis, ovulation disorders, tubal disease, with a single patented Fallopian tube), 3. Unexplained infertility. All couples underwent IUI with/or without ovarian stimulation and we documented several factors: age, type of infertility, infertility duration, number of procedures, ovarian stimulation, number of follicles, endometrial thickness, ovulation induction, day of induction, associated pathology, tubal patency, post wash sperm count. Main results and the role of chance Our pregnancy rate was 10.3%, according to this the couples were divided intro two groups the negative and the positive result group. We found 4 factors to be significantly associated to a positive result: age of the partners (p-value = 0.012), infertility duration (p-value = 0.018), endometrium thickness (p-value = 0.035) and semen preprocess concentration (p-value = 0.046). We calculated a probability function for a woman to get pregnant after IUI procedure: Prob = 1/(1+e-z) Z = -3.36998 + 0.364*Endometrium (mm) – 0.04567*Female Age (years) – 0.049956*Age of partner (years)– 0.17299*Duration of infertility (years) + 0.018562*prewash count (mil) – 0.9933*A (motility) + 2.29085*B(motility)+ 0.03203*post wash count For example, a patient with a score of 1.507 will have a probability of 81.86% of becoming pregnant. Our predictive model in general has a performance that is close to 0.7 AUROC -0.687 (AUROC between 0.7-0.8 has a fair performance) with a value p-0.001 (high statistical significance, CI confidence interval - 99.9%), meaning it is able to make a good distinction between couples with a good pregnancy chance and those with a poor pregnancy chance after IUI. Limitations, reasons for caution In statistical models fertility treatment chances are never 0 or 100%, and the outcome of IUI is a subject to chance and from this point of view the trick would be in repeating the cycles. Another limitation is the small cohort sample and the fact that it needs external validation. Wider implications of the findings Developing an application program that could be used easily by the clinicians in their daily clinical practice and also by the patients in order to better understand their chances at a successful procedure of assisted human reproduction that could also mean the next step after IUI, the in vitro fertilization. Trial registration number not applicable

IMPACT OF SPERM DNA FRAGMENTATION ON PRE- AND POST-WASH SEMEN SAMPLES AND PREGNANCY OUTCOME WITH INTRACYTOPLASMIC SPERM INJECTION

The spermatozoa nuclear component especially it’s DNA integrity is essential for normal fertilization, pregnancy & embryo development. Currently sperm DNA fragmentation (SDF) testing is one advanced sperm function test used along other conventional tests to evaluate male infertility.We aim to assess the impact of SDF on pre- & post-wash semen parameters and to investigate effect of various SDF cut-off values on pregnancy outcome with Intracytoplasmic sperm injection (ICSI). Retrospective, Chart review. The charts of 2277 patients who underwent ICSI were reviewed. 481 infertile men who had SDF testing were included. Patients demographic & laboratory data along with ICSI cycle outcome were included. Semen samples & SDF were analyzed according to WHO 5thedition guidelines & sperm chromatin dispersion (Halosperm kit, Halotech, Madrid, Spain) respectively. Patients were divided according to SDF cut-offs; <20, 20-30 & >30%. Variables were reported as mean ± SE and compared using Kruskal Wallis Test. P value of <0.05 was considered statistically significant The mean patient age was 36.88 ± 0.362, while mean SDF was 25.41± 0.72. The pre & post wash semen parameters in patients with SDF cut-offs <20, 20-30 & >30% were compared (Table 1). Significantly worse results were obtained with increasing SDF cut-offs for pre- & post-wash sperm concentration, total motility & post-wash normal morphology. SDF was significantly inversely correlated with pre wash parameters (concentration: r -.140, p=o.oo3; total Motility: r -.282, p<0.001; normal morphology: r -.143, p=0.003) and post wash parameters (concentration: r -.156, p=o.oo1; total Motility: r -.218, p<0.00; normal morphology: r -.140, p=0.002) The clinical pregnancy rate decreased with increasing SDF cut-offs (44.4% with SDF <20%, 31.4% with SDF 20-30% and 24.3% with SDF >30), though the result was not statistically significant SDF negatively impacts pre-& post-processing semen parameters for patients undergoing ICSI. This may influence ICSI pregnancy rate, though the impact was not statistically significant in this cohort.Tabled 1ParametersTotalSDF <20SDF 20-30SDF >30P ValuePre wash Concentration (M/ml)38.94 ± 1.6945.24 ± 2.7935.82 ± 2.6130.57 ± 2.870.016Pre wash Total Motility %38.61 ± 0.944.18 ± 1.2837.82 ± 1.6229.26 ± 1.68<0.001Pre wash Normal Morphology %12.49 ± 0.6112.01 ± 0.7814.53 ± 1.3511.18 ± 1.270.121Post Wash Concentration (M/ml)13.93 ± 0.6315.96 ± 0.9513.854 ± 1.129.95 ± 1.16<0.001Post Wash Total Motility %4.21 ± 0.275.24 ± 0.453.99 ± 0.432.45 ± 0.47<0.001Post Wash normal Morphology %12.31 ± 0.6512.12 ± 0.8913.36 ± 1.3111.53 ± 1.370.010 Open table in a new tab

POSTWASH TOTAL MOTILE SPERM COUNT EFFECT ON PREGNANCY OUTCOMES IN INTRAUTERINE INSEMINATION

Intrauterine insemination (IUI) is a frequently used method to treat couples with infertility. There is evidence of decreased pregnancy rates with total motile sperm count (TMSC) less than 5 and 10 million (1,2), yet there has been efficacy of reported IUI success with TMSC under 1 million (2). As such, there remains to be a consensus on semen parameters for which to recommend IUI in the infertile population. Further defining TMSC and associated pregnancy rates allows for more precise patient counseling and may also delineate a clearer threshold for escalation of care to in vitro fertilization. This may decrease the time to pregnancy and the financial burden of offering infertility treatment that is less likely to be effective. The aim of this study was to determine a minimum threshold of TMSC on semen analysis to offer IUI cycles. A retrospective cohort study of all IUI cycles at a private practice infertility center from June 2014 to May 2018. This is a retrospective cohort study of all IUI cycles at a private practice infertility center from June 2014 to May 2018. We obtained female characteristics including age, gravida, parity, BMI, AMH, AFC, tubal patency, stimulation, number of dominant follicles, and ET. Semen was processed with a gradient centrifugation process and the following semen parameters were collected: post wash TMSC, concentration, motility, and wet mount morphology. Our primary outcome of interest was the presence or absence of clinical pregnancy after each cycle. Clinical pregnancy was defined as fetal cardiac activity on ultrasound. A total of 999 women underwent 2169 IUI cycles during this four-year period. The average couple underwent 2.17 IUIs. The overall clinical pregnancy rate was 19.8% per cycle while pregnancy rate of only the first IUI per patient was 19.2%. Overall, clinical pregnancy rate per couple was 40.2%. During the first IUI each couple underwent, there was an increase in clinical pregnancy with increasing TMSC, OR 0.44 for TMSC ≤1M, 0.56 for TMSC 2-5M, and 0.99 for TMSC 6-10M, compared to TMSC >10M with p-values of 0.27, 0.057 and 0.975 respectively. Using receiver operating characteristic curves, we did not identify a TMSC threshold to offer IUI but rather a linear increase of sensitivity and specificity to predict pregnancy with increased TMSC. The lowest TMSC resulting in pregnancy was 660,000. With TMSC 6-10M, pregnancy outcomes improved with morphology >4% OR 0.84, compared to morphology <4% OR 0.25. There were 39 multifetal gestations, 9.1% of all clinical pregnancies. Although no distinct threshold of TMSC was identified to offer IUI versus proceeding with ART, there is a positive correlation between TMSC and IUI success. There appears to be improved outcomes with morphology >4% when TMSC <10. A follow up study, using a larger sample size, is necessary to further characterize the upper limits of efficacy of IUI specimens. Well-defined pregnancy rates based on semen parameters will allow for more direct counseling of infertility patients and also allow for more targeted therapy recommendations based on pregnancy success rates.

Clinical Success of Intrauterine Insemination in A Private Fertility Centre

Background : IUI is one of the most frequently and widely used therapeutic modalities to treat various forms of infertility. Achieving pregnancy after IUI is presumably due to increase in the number of highly motile spermatozoa with a high proportion of morphologically normal spermatozoa near the site of fertilization. The objectives were to compare the success rates of IUI with that of timed intercourse, compare the success rate of IUI according to the protocols, review methods used to time the IUI with ovulation, identify the factors to determine IUI outcome.&#x0D; Materials and methods : This is a cross sectional prospective study conducted in Surgiscope Fertility Centre, Chattogram, Bangladesh over a period of 21 months from 596 patients who underwent IUI. Multiple variables were selected such patient parameters like age of woman and cause of infertility, parameters related to ovulation induction such as drugs used for COH, number of mature follicles and endometrial thickness and laboratory parameters such as post wash sperm count and type of IUI (Single or double IUI) which were recorded and statistically analysed.&#x0D; Results : The minimum age for females and males were 18 and 19 respectively and the maximum age for females and males were 40 and 66 respectively. The mean age for females was 27.3 +/- 0.4 years and the mean age for males was 35.9 +/- 0.5 years. Another variable was endometrial thickness where the mean was 8.9 mm ± 0.2 mm and median was 8.5 mm. Furthermore, successful pregnancies also depend on total motile sperm count taken post-wash. The total motile sperm count was 29.8 million ± 1.5 million and median was 30 million. Pregnancy rate for total motile sperm count less than 10 million was 8.6%. For women over 28 and total motile sperm count &lt;10 million, the success rate was only 2.7%.Most of the patients who underwent IUI were between the ages of 28-31, which was 175 (29.3%). The successful conception rate for females over the age 28 was 9.6% compared to the whole group at 12%. Among 596 patients, 71 patients (11.91%) had successful conceptions while 525 patients (88.09%) did not. There is a statistically significant relationship between number of IUI cycles completed and the rate of successful pregnancy. There is also a relationship between the number of inseminations and the success rate.&#x0D; Conclusion : IUI is a successful contemporary treatment especially as a good first line invasive treatment for couples with unexplained infertility, male factor infertility, anovulation, female less than 30 years of age. Percentage of actual number of motile sperms appear to have an important impact on outcome. Success rate of conception was also found to be statistically significant in patients with appropriate endometrial thickness. Careful and criteria-based selection of patients along with ovarian stimulation seem to be the ideal model for achieving clinical success in IUI.&#x0D; JCMCTA 2019 ; 30 (2) : 26-31

88 Effects of two different hatcher basket wash systems on microbial counts

Abstract The recently hatched chicks’ umbilicus provides a pathway for the introduction of pathogenic microorganisms. Consequently, sanitation of hatcher basket is crucial to the future success of the chick. The design of this study was to examine the effects of two hatcher basket wash systems on microbial loads and subsequent 7 d chick mortality. Treatment 1 consisted of a single stage wash system and treatment 2 was a Multi-stage wash system. On sampling days, 10 hatcher baskets randomly selected off the chick processing line were individually swabbed pre-washing. A 5.08 cm2 templet placed in the lower left hand corner of each hatcher basket ensured accurate sampling. Following pre-wash sampling each basket was labeled, placed into the wash system, and recovered immediately post washing. A second templet placed in the same location as the first and a post wash swab collected. Samples were collected for 10 non-consecutive days. Following each collection day, each swab was plated on a 3M-Petrifilm Count Plate and allowed to incubate for 48 h at 39.44oC. Following incubation, plates were read to determine total plate count (TPC per cm2). There were no significant (P .05) at either the prewash (993 vs 595) or post wash (38 vs 58). There were no significant differences (P &gt; .05) in 7 day mortality between treatment 1 vs 2 (0.95% vs 1.12%). In conclusion, both wash systems were effective in reducing microbial loads.

51 Effects of two different hatcher basket wash systems on microbial counts

Abstract The recently hatched chicks’ umbilicus provides a pathway for the introduction of pathogenic microorganisms. Consequently, sanitation of hatcher basket is crucial to the future success of the chick. The design of this study was to examine the effects of two hatcher basket wash systems on microbial loads and subsequent 7 d chick mortality. Treatment 1 consisted of a single stage wash system and treatment 2 was a Multi-stage wash system. On sampling days, 10 hatcher baskets randomly selected off the chick processing line were individually swabbed pre-washing. A 5.08 cm2 templet placed in the lower left hand corner of each hatcher basket ensured accurate sampling. Following pre-wash sampling each basket was labeled, placed into the wash system, and recovered immediately post washing. A second templet placed in the same location as the first and a post wash swab collected. Samples were collected for 10 non-consecutive days. Following each collection day, each swab was plated on a 3M-Petrifilm Count Plate and allowed to incubate for 48 h at 39.44oC. Following incubation, plates were read to determine total plate count (TPC per cm2). There were no significant (P .05) at either the prewash (993 vs 595) or post wash (38 vs 58). There were no significant differences (P &gt; .05) in 7 day mortality between treatment 1 vs 2 (0.95% vs 1.12%). In conclusion, both wash systems were effective in reducing microbial loads.

Viable manufacture of cell therapies throughthe integration of multiple unit processes onto a counter-flow centrifugation device

Background & Aim With growing proof of efficacy in varied indications, regenerative medicine has reached a positive inflection point in the investment of time and money by established industry leaders and disruptive startups. With rapid growth comes the need to devote resources to the engineering challenges that currently prevent the quick and cost-effective manufacture of therapies that maintain a consistent, high-level of quality and, in turn, can support commercial manufacturing. This is especially true when looking at patient specific cell therapies that require rapid change over of equipment and benefit little from traditional sterile barriers (i.e. filters and heat inactivation). Counter-flow centrifugation (CFC) presents an intriguing technology that, when implemented using closed and automated system, provides a platform for upstream and downstream processing of cellular therapies by incorporating multiple unit processes and mitigating the risk imposed with manual equipment transfers. Methods, Results & Conclusion The CFC technology under development by a partnership Hitachi Chemical Advanced Therapeutic Solutions (HCATS) and Invetech compounds the benefits of a fully integrated, closed processing kit with easily customizable procedures to integrate multiple unit processes onto a singular device. Increased integration of unit processes will be integral to addressing the dynamic challenges of commercial cell therapy manufacturing in a scaled-out model. The CFC device has shown the capability to perform platelet wash on apheresis collections with 99% efficiency and retain 100% of the mononuclear cells. Elutriation was able to recover 95% of all monocytes from the MNC population at 78% purity. For lymphocyte isolation, CD3/CD28 beads were incubated with the MNC fraction on the device post platelet wash where 48.6% fewer beads were required to conjugate with the T-Cell population. By incorporating multiple unit processes onto a device that also meets the need for rapid change-over between lots this device offers a unique solution to the emergent challenges of cell therapy manufacturing. With growing proof of efficacy in varied indications, regenerative medicine has reached a positive inflection point in the investment of time and money by established industry leaders and disruptive startups. With rapid growth comes the need to devote resources to the engineering challenges that currently prevent the quick and cost-effective manufacture of therapies that maintain a consistent, high-level of quality and, in turn, can support commercial manufacturing. This is especially true when looking at patient specific cell therapies that require rapid change over of equipment and benefit little from traditional sterile barriers (i.e. filters and heat inactivation). Counter-flow centrifugation (CFC) presents an intriguing technology that, when implemented using closed and automated system, provides a platform for upstream and downstream processing of cellular therapies by incorporating multiple unit processes and mitigating the risk imposed with manual equipment transfers. The CFC technology under development by a partnership Hitachi Chemical Advanced Therapeutic Solutions (HCATS) and Invetech compounds the benefits of a fully integrated, closed processing kit with easily customizable procedures to integrate multiple unit processes onto a singular device. Increased integration of unit processes will be integral to addressing the dynamic challenges of commercial cell therapy manufacturing in a scaled-out model. The CFC device has shown the capability to perform platelet wash on apheresis collections with 99% efficiency and retain 100% of the mononuclear cells. Elutriation was able to recover 95% of all monocytes from the MNC population at 78% purity. For lymphocyte isolation, CD3/CD28 beads were incubated with the MNC fraction on the device post platelet wash where 48.6% fewer beads were required to conjugate with the T-Cell population. By incorporating multiple unit processes onto a device that also meets the need for rapid change-over between lots this device offers a unique solution to the emergent challenges of cell therapy manufacturing.

A comparative study between povidone iodine and metronidazole for peritoneal lavage in cases of peritonitis

Background: Peritonitis secondary to hollow viscous perforation is one of the commonest emergencies encountered in general surgical practice. This randomized study aimed to compare the efficiency of Povidone iodine and Metronidazole in these cases by quantitative analysis of bacterial colony count in patients who have received peritoneal lavage with either of them.Methods: A total of 100 consecutive patients who presented with peritonitis secondary to hollow viscous perforation were enrolled in this study (February 2010 to February 2011) and systematically divided into 2 groups: peritoneal lavage with povidone iodine group and lavage with metronidazole group. The post-operative progress was assessed by comparing the pre-wash and post wash colony counts in both the groups, development of surgical site infection and duration of hospital stay.Results: The median age of the patients was 33 years (range: 12-75 years), with a male: female ratio of 3.8:1. Perforation of duodenal ulcer (67%) was the commonest cause of peritonitis with E. coli (65%) being the commonest organism isolated from the peritoneal fluid. Reduction in bacterial colony count was seen in both the groups of patients, however, there was significantly greater reduction in the patients who received lavage with Metronidazole (p=0.06). Incidence of surgical site infection was less in the Povidone iodine group though it did not reach statistical significance.Conclusions: The use of povidone iodine in peritoneal lavage significantly reduce the bacterial colony and may also result in fewer surgical site infections.

Is ICSI superior to conventional IVF in severe male factor: a 3 year prospective observational study

To study whether IVF (in-vitro fertilisation) yields better outcome in severe male factor infertility over Intracytoplasmic sperm injection (ICSI) in terms of live-birth ,obstetric outcome and congenital anomalies. This prospective observational study included 253 women whose male partner had TMC (Total motile count) <1million and underwent conventional IVF or ICSI from January 2014 to December 2016. All subjects were between 25-37 years. Records of prenatal care of mother and neonates were reviewed. Developmental wellbeing of children born after IVF/ICSI were followed for three years. The 253 male partners who had semen analysis on day of oocyte retrieval with <1million motile sperm and morphology>4% after wash, were included and divided into Group A (GrA) (n=121) in which conventional insemination was done and Group B (GrB)(n=132) where ejaculated ICSI was done. Antagonist protocol was performed. Total fertilization failure were 6/121(4.9%) in GrA and 3/132(2.2%) in GrB. Rest 244 patients with 295 cycles were analysed. Other female factors-PCOS, endometriosis, tubal factors and uterine defects were excluded. Both groups were comparable in terms of age of both partners, BMI, dose of gonadotropins, estradiol on day of oocyte retrieval, endometrial thickness and number of oocytes retrieved. Conventional IVF group (496/840, 59.01%) had significantly lower fertilization rate than ejaculated ICSI (591/910, 64.94% p=0.0119). although number of day 3 embryos were comparable in both groups (GrA 407/496, 82.05% vs GrB 467/591,79.01%). Pregnancy rate per embryo transfer in GrA was 42.2% and GrB was 47.61% (p=ns). Pregnancy per oocyte retrieval was also comparable-38.58% in conventional IVF and 42.85% in ICSI. Live birth rate per oocyte retrieval was also similar in both groups (GrA 30.6% vs GrB 26.4%). Missed abortion, though higher in ICSI group was not significantly different than conventional IVF (GrA 26.53% vs GrB 29.1%). Antenatally two patients in ICSI group had pre-eclamptic toxaemia (PET) while none had the condition in IVF group. One baby in ICSI group was terminated due to chromosomal defect and two babies born after ICSI had birth defects (one hypospadius and the other imperforate anus). In IVF group one baby had posterior urethral valve. Incidence of PET and congenital anomalies could be chance findings in this study. In our study conventional IVF is as good as ejaculated ICSI in terms of reproductive outcome, where male partner had <1million motile sperm in post wash sample. IVF could be the first choice over ICSI in severe male factor infertility especially in developing countries which will decrease the cost of ART.

Evaluation of the Behaviour of Violet Acid Pigment 43 in Formulations of Shampoos and Conditioners in Bleached Hair

Objective: The objective of this work was to evaluate the behavior of Acid Violet 43 pigment shampoos and conditioners formulations and when applied to hair through 10 formulations. Method: 10 formulations were developed, these being 5 shampoos and 5 conditioners. Among 5 shampoos, 4 were pigmented in different concentrations and 1 served as control. In the same way it was done for the conditioners, being 4 conditioners pigmented in different concentrations and 1 conditioner for control. The samples were subjected to organoleptic and physicochemical characterization for 15 days, being evaluated appearance, color and odor, behavior after centrifugation, determination of pH, viscosity and foam content (for shampoo). Result: The samples showed satisfactory results in the tests, because their organoleptic and physicochemical characteristics remained stable throughout the analysis period. As for the behavior of the pigment on hair shaft, hair samples were bleached and washed with the formulations developed. Prewash and post wash solutions (wash water) were prepared at the same concentrations and were also read in a spectrophotometer at 570 nm. For white, the depigmented products were used. It was observed that both the shampoos and the conditioners deposit pigment in the wires, however we verified by visual analysis that the conditioners are more effective. Conclusion: The wicks were also subjected to moisture content tests to verify if pigment interfered with the deposition of water in hair. Key words: Acid violet 43, Bleaching, Semi-permanent hair dye, Hair dye.

Enhanced post wash retention of combed DNA molecules by varying multiple combing parameters

Recent advances in genomics have created a need for efficient techniques for deciphering information hidden in various genomes. Single molecule analysis is one such technique to understand molecular processes at single molecule level. Fiber- FISH performed with the help of DNA combing can help us in understanding genetic rearrangements and changes in genome at single DNA molecule level. For performing Fiber-FISH we need high retention of combed DNA molecules post wash as Fiber-FISH requires profuse washing. We optimized combing process involving combing solution, method of DNA mounting on glass slides and coating of glass slides to enhance post-wash retention of DNA molecules. It was found that average number of DNA molecules observed post-wash per field of view was maximum with our optimized combing solution. APTES coated glass slides showed lesser retention than PEI surface but fluorescent intensity was higher in case of APTES coated surface. Capillary method used to mount DNA on glass slides also showed lesser retention but straight DNA molecules were observed as compared to force flow method.

Retention and sustained release of fragrance by Cyclodextrin functionalized cotton fabric modified using maleic anhydride

Abstractβ‐cyclodextrin(CD) was incorporated in cotton fabric following pad‐dry‐cure technique employing maleic anhydride as modifying agent under the dual influence of free radical polymerization catalyst ammonium peroxodisulfate [(NH4)2S2O8] and esterification catalyst tetrasodium pyrophosphate (Na4P2O7). β‐cyclodextrin(CD) can be esterified if treated with maleic anhydride (an unsaturated dicarboxylic acid) due to the presence of hydroxyl groups in β‐cyclodextrins and also such compound can form inclusion complex with essential oils. The retention of essential oil and its sustained release after repeated post wash treatments was determined quantitatively by UV–vis spectrophotometry.

Utilization of Calophyllum inophyllum shell and kernel oil cake for reducing sugar production

This study is aimed at fully utilizing fruit biomass of Calophyllum inophyllum for reducing sugar production. The effects of pretreatment conditions and post reaction wash on the lignin removal and enzymatic hydrolysis of shell were investigated. The oil cake was also subjected to solvent extraction followed by enzymatic hydrolysis. The results showed that the sequential acid/alkaline pretreatment of C. inophyllum shell resulted in better delignification than alkaline or acid only pretreatment. The reducing sugar yields obtained from sequential acid/alkaline pretreated shell and solvent extracted oil cake were 0.24g/g and 0.66g/g, respectively. The results suggested that the shell and oil cake of C. inophyllum could also be feedstocks for reducing sugar production.

Compatibility analysis of reactive dyes by exhaustion-fixation and adsorption isotherm on knitted cotton fabric

AbstractCompatibility analysis was done by exhaustion, fixation and adsorption isotherm upon three reactive dyes; Remazol Red RR, Remazol Yellow RR and Remazol Blue RR. The known solutions of each dye were tested in UV-visible spectrophotometer and the extinction coefficients were calculated by drawing absorbance versus concentration curve. Dyeing processes were carried out in 4 different dye concentrations (0.5%, 1%, 2% and 3%) for each of the dye. The post dye and post wash liquors were tested in UV-visible spectrophotometer and the dye concentrations of those solutions were calculated through the Beer-Lambert law. The exhaustion%, fixation% of all three dyes was compared between them and their adsorption isotherm models were drawn. In between Langmuir and Freundlich isotherm model, the last one had the supremacy. Among the dyes, Blue RR had better exhaustion performance although it was not pleasingly attached with the fiber during fixation. Red RR was excellent in both exhaustion and fixation stages. It was found that in deeper concentrations the compatibility among these three dyes would be low. But experimental data shows that introducing more electrolytes and reducing the temperature as well as the alkali amount can improve these dyes’ compatibility.

A novel polymer-coated nanoparticle (NP) for urothelium penetration and drug delivery.

e15543 Background: Up to 40% of patients with non-invasive bladder cancer (BC) will develop invasive disease progression despite locally-directed therapy. Overcoming the urothelial barrier is a challenge for intravesical drug delivery. We designed a biodegradable poly(lactide-co-glycolide) (PLGA) NP coated with a novel cell penetrating polymer, poly (guanidinium oxanorbornene) (PGON) for testing against BC in vitro and in vivo. We chose to deliver the HDAC inhibitor belinostat (bel) for its BC cell cytotoxicity and inhibition of invasion &amp; migration pathways; key mechanisms that enable progression of BC. Methods: Fluorophore (C6) or bel was encapsulated in PLGA using an oil/water nanoemulsion method, and surface coated with PGON, and then characterized for morphology, size, and loading. BC cell lines UM-UC-3 and T24 were treated with belinostat or NP-bel-PGON vs controls for assessment of cytotoxicity and acetyl-H4 histone expression over time. In vivo murine bladder and ex vivo human ureter were treated with NP-C6-PGON, and compared to NPs coated with chitosan or PEG for urothelial penetration using FACS analysis, tissue extraction, and fluorescence microscopy. UM-UC-3 murine flank xenografts were treated locally biweekly with NP-bel-PGON or controls and assessed for tumor size and acetyl-H4 expression. Results: C6 extraction of intravesically treated mouse bladder and ex-vivo human ureter showed uptake improved ten-fold in NP-C6-PGON compared to other NPs and corroborated by fluorescent microscopy. In vitro, NP-bel-PGON and bel had similar IC50 of ~2.0 μM in UM-UC-3 &amp; T24 lines, and no effect from PGON. Significantly, the NP-bel-PGON treated groups retained 30% of max H4 hyperacetylation whereas bel groups declined to basal at 12hr post wash, which supports a mechanism of intracellular NP release and activity. In vivo, xenografts treated with NP-bel-PGON showed tumor volume reduced 70% and had 2.5 fold higher intratumoral acetyl-H4 expression compared to vehicle three days post final treatment. Conclusions: NP-bel-PGON penetrates the urothelium, is taken up by BC cells, sustains HDAC inhibition, and causes tumor regression. These data demonstrate the potential for NP-bel-PGON as an intravesical nanotherapy of BC.

Application of rapid read-out cleaning indicators for improved process control in hospital sterile services departments

Heightened awareness of the importance of cleaning has led to an emphasis on automated systems for the decontamination of re-usable medical devices. The authors have previously described an enzymatic indicator system, based on thermostable adenylate kinases (tAK), for quantitative monitoring of automated cleaning processes within hospital sterile services departments (SSDs). To evaluate tAK indicators for routine process monitoring across a range of SSDs with different cleaning chemistries and different automated washer disinfectors (AWDs). tAK indicator devices and alternative industry test indicators were included in five independent cleaning cycles in each of eight different AWDs. Residual tAK post wash was determined by a coupled luciferase assay using a modified hygiene monitoring system. In all cases, with the exception of a single test, the alternative indicators showed that cleaning had been adequate. They were not able to discriminate between the performance of different processes. In contrast, the tAK indicators were able to resolve differences in the performance of processes across the different SSDs. Where the tAK indicators identified cleaning to the limits of detection of the assay, this demonstrated a log10 enzyme removal factor of >5.69. The results suggest that tAK indicators are suitable for providing improved process control for automated cleaning processes, being able to distinguish between wash performance in different hospital settings and between individual process runs. This technology is believed to be a useful addition to routine AWD performance qualification when used as a daily or weekly test.

Does centrifugation and semen processing with swim up at 37°C yield sperm with better DNA integrity compared to centrifugation and processing at room temperature?

AIM:To evaluate whether semen processing at 37°C yield sperm with better DNA integrity compared to centrifugation and processing at room temperature (RT) by swim-up method.SETTINGS:This study was done at tertiary care center attached to Reproductive Medicine Unit and Medical College.DESIGN:Prospective pilot study.PATIENTS:Normozoospermic men (n = 50) undergoing diagnostic semen analysis.MATERIALS AND METHODS:Normozoospermic samples (World Health Organization, 2010 criteria) after analysis was divided into two aliquots (0.5 mL each); one was processed at 37°C and the other at RT by swim-up method. DNA fragmentation of both samples post wash was calculated by acridine orange method.STATISTICAL ANALYSIS USED:The values of sperm DNA fragmentation were represented as mean and standard error (mean ± SEM) of the mean. Paired t-test was used for calculating the sperm DNA integrity difference between post wash at RT and 37°C.RESULTS:Statistically significant difference was not observed in post wash sperm DNA fragmentation values at 37°C compared to RT.CONCLUSION:Our data represents that there was no significant difference in sperm DNA fragmentation values of samples processed at 37°C and at RT. Hence, sperm processing at 37°C does not yield sperm with better DNA integrity compared to centrifugation and processing at RT.

Two step meso-acidophilic bioleaching of chalcopyrite containing ball mill spillage and removal of the surface passivation layer

Meso-acidophilic bacterial leaching of ball mill spillage (containing chalcopyrite >80%) was carried out in an innovative two-step bioleaching method. The major drawback of meso-acidophilic bioleaching limiting industrial application is the passivation phenomenon over the ore surfaces in iron–sulfur rich environments. In the present study, we present a novel wash solution that efficiently removed the passivation layer. FTIR characterization of the bioleached sample indicated that the residues could be further leached to recover extra copper after wash solution application. XRD study indicated accumulation of sulfates (SO4-) of Na, K, Fe and oxy hydroxides of iron [FeO(OH)] in the form of jarosite outlining the passivation layer. SEM, FESEM-EDS studies indicated severe corrosion effects of the wash solution on the passivation layer. Two step bioleaching of the ore sample yielded 32.6% copper in 68days in the first interlude and post wash solution application yielded 10.8% additional copper.