Product Optimization, Storage Quality and Sensory Acceptance of Low Calorie Beverage Developed from Bitter Gourd and Kiwifruit

Abstract Bitter gourd (Momordica charantia L.) fruit is good source of many nutraceutical compounds and possess antioxidant, anti-diabetic and hypoglycaemic activities. However, its utilization in the preparation of beverages is limited due to its bitter after taste. Therefore, to realize the functional and therapeutic benefits of bitter gourd, an attempt was made to optimize nutritious and low caloriebitter gourd based beverage by blending with kiwifruit (Actinidia deliciosa), a store house of bioactive compounds and substituting sugar with stevioside (steviol glycoside). The standard (sugar sweetened) bitter gourd (BG)-kiwifruit (K) blended beverage was developed by utilizing 30% fruit part of BG:K blended juice (80: 20) with 40oB TSS and 1.3% acidity. Further, to develop the low calorie beverage, sucrose (table sugar) was replaced with 25, 50, 75 and 100% equi-sweetness level of stevioside (steviol glycoside). Results revealed that 75% substitution of sucrose with stevioside resulted in shelf stable beverage with identical taste, good antioxidant potential (68.80%) and strong antimicrobial activity (26 mm ZOI) with reduced calorie values (28.5 Kcal/100g) compared to the sugar sweetened control sample (150.60 Kcal/100g). Hence, the developed beverage can be commercialized as low calorie beverage with additional health benefits of natural compounds of bitter gourd and kiwifruit with highest bioactivity.

Sperm DNA fragmentation testing: Summary evidence and clinical practice recommendations.

We herein summarise the evidence concerning the impact of sperm DNA fragmentation in various clinical infertility scenarios and the advances on sperm DNA fragmentation tests. The collected evidence was used to formulate 41 recommendations. Of these, 13 recommendations concern technical aspects of sperm DNA fragmentation testing, including pre-analytical information, clinical thresholds and interpretation of results. The remaining 28 recommendations relate to indications for sperm DNA fragmentation testing and clinical management. Clinical scenarios like varicocele, unexplained infertility, idiopathic infertility, recurrent pregnancy loss, intrauterine insemination, in vitro fertilisation/intracytoplasmic sperm injection, fertility counselling for men with infertility risk factors and sperm cryopreservation have been contemplated. The bulk evidence supporting the recommendations has increased in recent years, but it is still of moderate to low quality. This guideline provides clinicians with advice on best practices in sperm DNA fragmentation testing. Also, recommendations are provided on possible management strategies to overcome infertility related to sperm DNA fragmentation, based on the best available evidence. Lastly, we identified gaps in knowledge and opportunities for research and elaborated a list of recommendations to stimulate further investigation.

Role of male factor in early recurrent embryo loss: do antioxidants have any effect?

OBJECTIVE: To evaluate whether increasing antioxidant intake in men with high levels of DNA damage or lipid peroxidation improves gestational results in couples with history of recurrent embryo loss. DESIGN: Descriptive study (case series). SETTING: Early recurrent embryo loss program at the University of Antioquia, Medellín, Colombia. PATIENT(S): Seventeen men whose spouses had a history of two or more embryo losses before 12 weeks of gestation. INTERVENTION(S): Male partners with increased DNA fragmentation index (%DFI) or high thiobarbituric acid reactive substances (TBARS) were instructed to consume a diet rich in antioxidants or commercial multivitamins containing beta-carotene, vitamin C, vitamin E, and zinc for at least 3 months. MAIN OUTCOME MEASURE(S): Pregnancy outcome was recorded in the spouses of men with increased %DFI or TBARS who received antioxidant supplementation. RESULTS: Of the 17 men, 9 (53%) presented with an increased %DFI or TBARS. They were started on an antioxidant supplementation regimen. Of these nine men, six of their spouses became pregnant. All couples whose male partners accepted antioxidant supplementation achieved a successful pregnancy. CONCLUSIONS: Our study demonstrates the benefits of an increased intake of antioxidant-rich food or antioxidant supplements by men who show high levels of sperm DNA fragmentation or lipid peroxidation, which could result in an improvement in gestational outcomes in couples with history of recurrent embryo losses.

Age-related increase of reactive oxygen species in neat semen in healthy fertile men.

OBJECTIVES: The effects of advancing paternal age on the male reproductive system are well known, but its effects on fecundity remain controversial. Although oxidative stress is associated with poor semen quality and function, a relationship with advancing male age has not been established. The objective of this study was to analyze the relationship between male age and seminal reactive oxygen species (ROS) levels in men presenting for voluntary sterilization. METHODS: We prospectively evaluated 98 fertile men who were candidates for vasectomy. These were divided into 2 age groups: less than 40 years (n = 78) and 40 or more years (n = 20). We used 46 infertile patients as positive controls. Standard semen analysis, seminal leukocyte count and ROS levels were measured in all samples. Fertile men with leukocytospermia were excluded. RESULTS: The mean age of the men was 35.1 +/- 5.6 years. Men 40 years and older had significantly higher ROS levels compared with younger men (P <0.001). We observed a positive correlation between seminal ROS levels and age (r = 0.20; P = 0.040). In addition, ROS was negatively correlated with sperm concentration (r = -0.48; P <0.001) and motility (r = -0.21; P = 0.030). CONCLUSIONS: Reactive oxygen species levels are significant higher in seminal ejaculates of healthy fertile men older than 40 years. ROS levels in whole ejaculate are significantly correlated to age among fertile men. Because ROS are clearly implicated in the pathogenesis of male infertility, these data suggest that delayed fatherhood may reduce the chances of pregnancy as men become progressively less fertile with age.

Semen quality and oxidative stress scores in fertile and infertile patients with varicocele.

OBJECTIVE: To compare semen quality and levels of seminal oxidative stress among three groups: infertile men with varicocele, fertile men with varicocele, and healthy semen donors (controls) without varicocele. DESIGN: Prospective study. SETTING: Academic medical centers. INTERVENTION(S): None. PATIENT(S): Semen specimens were obtained from 21 infertile patients with varicocele, 15 fertile men with varicocele, and 17 healthy fertile men with normal semen characteristics. MAIN OUTCOME MEASURE(S): Principal component analysis was applied to nine semen characteristics to provide a standardized semen quality score. Reactive oxygen species (ROS) production and total antioxidant capacity (TAC) were measured by chemiluminescence assays to create an ROS-TAC score. RESULT(S): The mean semen quality scores of the infertile patients with varicocele were lower than those of the control subjects but similar to those of the fertile men with varicocele. Compared with the healthy subjects, the infertile men with varicocele had higher ROS levels but lower TAC levels. They also had significantly lower ROS-TAC scores compared with control subjects, but the scores were not significantly different than those seen in fertile men with varicocele. CONCLUSION(S): These findings not only provide us with valuable information regarding semen quality but also can serve as a warning that the fertility potential in fertile varicocele patients can decline due to oxidative stress.

Evaluation of acrosomal status and sperm viability in fresh and cryopreserved specimens by the use of fluorescent peanut agglutinin lectin in conjunction with hypo-osmotic swelling test.

OBJECTIVE: In this study, we evaluated whether the hypo-osmotic swelling test (HOST) can be used as a vital marker in combination with peanut agglutinin (PNA) - labeling in fresh and cryopreserved spermatozoa. MATERIALS AND METHODS: Human sperm populations were exposed to a hypo-osmotic medium for 60 minutes, and then incubated in a 1 microg/mL solution of the fluorescent dye Hoescht 33258 (H33258) for 10 minutes. Excess stain was removed by washing in phosphate-buffered saline (PBS) solution, and the pellet was resuspended in 100 microL of culture medium. Twenty microliters of this solution were subsequently smeared on a microscope slide, and fixed in ice-cold methanol to permeabilize the sperm membranes. The fixed smears were finally incubated in a 40-microg/mL FITC-PNA solution for 20 minutes. Simultaneous assessment of acrosome and viability scores was done in a fluorescent microscope equipped with appropriate filters and phase contrast illumination. The same slide was examined for FITC-PNA labeling, tail swelling, and for Hoechst-33258 staining by interchanging the filters and phase contrast optics. RESULTS: In fresh specimens, HOST was found to provide viability assessments comparable to those obtained using the H33258 method (r = 0.95). However, the results of HOST and H33258 were not correlated in cryopreserved specimens (r = 0.22). There was no alteration of PNA-labeling due to the HOST or H33258. CONCLUSIONS: FITC-PNA labeling in conjunction with the visualization of the morphological change induced by exposure to hypo-osmotic solution provides a simple but effective method for establishing the state of acrosomal membrane and viability in fresh human spermatozoa, but this technique is not reliable for cryopreserved ones.

Evaluation of acrosomal status and sperm viability in fresh and cryopreserved specimens by the use of fluorescent peanut agglutinin lectin in conjunction with hypo-osmotic swelling test

OBJECTIVE: In this study, we evaluated whether the hypo-osmotic swelling test (HOST) can be used as a vital marker in combination with peanut agglutinin (PNA) - labeling in fresh and cryopreserved spermatozoa. MATERIALS AND METHODS: Human sperm populations were exposed to a hypo-osmotic medium for 60 minutes, and then incubated in a 1 µg/mL solution of the fluorescent dye Hoescht 33258 (H33258) for 10 minutes. Excess stain was removed by washing in phosphate-buffered saline (PBS) solution, and the pellet was resuspended in 100 µL of culture medium. Twenty microliters of this solution were subsequently smeared on a microscope slide, and fixed in ice-cold methanol to permeabilize the sperm membranes. The fixed smears were finally incubated in a 40-µg/mL FITC-PNA solution for 20 minutes. Simultaneous assessment of acrosome and viability scores was done in a fluorescent microscope equipped with appropriate filters and phase contrast illumination. The same slide was examined for FITC-PNA labeling, tail swelling, and for Hoechst-33258 staining by interchanging the filters and phase contrast optics. RESULTS: In fresh specimens, HOST was found to provide viability assessments comparable to those obtained using the H33258 method (r = 0.95). However, the results of HOST and H33258 were not correlated in cryopreserved specimens (r = 0.22). There was no alteration of PNA-labeling due to the HOST or H33258. CONCLUSIONS: FITC-PNA labeling in conjunction with the visualization of the morphological change induced by exposure to hypo-osmotic solution provides a simple but effective method for establishing the state of acrosomal membrane and viability in fresh human spermatozoa, but this technique is not reliable for cryopreserved ones.

Identification of male factor infertility using a novel semen quality score and reactive oxygen species levels.

PURPOSE: To determine whether patients with male factor infertility can be accurately identified by calculating a novel semen quality score and measuring levels of reactive oxygen species during routine infertility screening. METHODS: Semen samples from 133 patients and 91 healthy donors were evaluated with manual and computer-assisted semen analysis. A principal component analysis model was employed to calculate a semen quality score. In brief, this score was calculated by base 10 logarithms multiplied by varying weights given to 9 sperm parameters. Reactive oxygen species levels were measured using chemiluminescence assay. RESULTS: The semen quality score had a sensitivity of 80.45% and accuracy of 77% at a cutoff of 93.1 in identifying patients with male factor infertility. The area under the receiver operating characteristic curves for the semen quality score was 84.28% (95% CI: 65.22%-100%). Reactive oxygen species levels [log10 (reactive oxygen species +1)] were significantly higher in male factor infertility patients. Reactive oxygen species had a sensitivity of 83.47% and specificity of 60.52% with an accuracy of 75% at a cutoff of 1.25 in identifying male factor infertility patients. The area under the receiver operating characteristic curve for reactive oxygen species levels was 78.92% (95% CI: 72.60%-85.23%). Semen quality scores were significantly and negatively correlated with reactive oxygen species levels in the donors and the male factor infertility patients. CONCLUSIONS: The semen quality score and reactive oxygen species levels in semen samples appear to be strongly associated with male factor infertility. Because both of these parameters are more sensitive than individual sperm parameters in identifying male factor infertility, they should be included in routine infertility screening.

Identification of male factor infertility using a novel semen quality score and reactive oxygen species levels

OBJETIVO: Determinar se pacientes portadores do fator de infertilidade masculina podem ser precisamente identificados através do cálculo de um novo escore de qualidade de sêmen e pela medida de espécies reativas de oxigênio durante uma avaliação rotineira de infertilidade. MÉTODOS: Amostras de sêmen de 133 pacientes e de 91 doadores saudáveis foram avaliadas através de análise manual e computadorizada de sêmen. Um modelo de análise do componente principal foi empregado para calcular o escore de qualidade de sêmen, utilizando logaritmos base 10, multiplicados por ponderações variáveis de 9 parâmetros espermáticos. Os níveis de espécies reativas de oxigênio foram medidos através de testes de quimiluminescência. RESULTADOS: O escore de qualidade de sêmen apresentou sensibilidade de 80.45% e precisão de 77% para um "cutoff" de 93.1 na identificação do fator de infertilidade masculina. A área sob a curva "receiver operating characteristic" para o escore de qualidade de sêmen foi de 84.28% (95% intervalo de confiança: 65.22%-100%). Os níveis de espécies reativas de oxigênio [log10 (espécies reativas de oxigênio +1)] foram siginificativamente mais elevados nos pacientes portadores de fator de infertilidade masculina. A medica de espécies reativas de oxigênio apresentou sensibilidade de 83.47% e especificidade de 60.52% com uma precisão (definida como pacientes portadores do fator de infertilidade masculina com diagnóstico positivo e doadores corretamente excluídos) de 75% para um "cutoff" de 1.25 na identificação de pacientes portadores do fator de infertilidade masculina. A área sob a curva "receiver operating characteristic" para níveis de espécies reativas de oxigênio foi de 78.92% (95% intervalo de confiança: 72.60%-85.23%). Os escores de qualidade de sêmen correlacionaram negativamente com os níveis de espécies reativas de oxigênio tanto nos doadores e nos pacientes portadores do fator de infertilidade masculina. CONCLUSÕES: O escore de qualidade de sêmen e os níveis espécies reativas de oxigênio nas amostras de sêmen parecem associar-se fortemente com o fator de infertilidade masculina. Na medida em que os dois parâmetros mostraram-se mais sensíveis que parâmetros espermáticos individuais na identificação do fator de infertilidade masculina, deveriam ser incluídos na avaliação rotineira de infertilidade.