Environmental life cycle assessment of production of the high intensity sweetener steviol glycosides from Stevia rebaudiana leaf grown in Europe: The SWEET project.

Purpose: There is an increasing interest in the use of non-nutritive sweeteners to replace added sugar in food and beverage products for reasons of improving consumer health. Much work has been done to understand safety of sweeteners, but very little on sustainability. To address that gap, this study presents the results of a life cycle assessment (LCA) of production of rebaudioside A 60%, 95% pure (RA60) steviol glycoside mix from Stevia rebaudiana leaf grown in Europe. Methods: An attributional cradle-to-factory-gate life cycle assessment was conducted on growing of stevia leaves and extraction of steviol glycosides in Europe. Primary data were used from a case study supply chain. Results are reported in impact categories from the ReCiPe 2016 (H) method, with focus given to global warming potential, freshwater eutrophication, water consumption, and land use. Impacts are expressed both in terms of production mass and sweetness equivalence, a common metric for understanding high intensity sweetener potency. Sweetness equivalence of RA60 is typically 200 to 300 times that of sugar. Comparison of environmental impact is made to sugar (sucrose) produced from both cane and beets. The research is part of the EU project SWEET (sweeteners and sweetness enhancers: impact on health, obesity, safety, and sustainability). Results and discussion: Global warming potential for production of RA60 was found to be 20.25 kgCO2-eq/kgRA60 on a mass basis and 0.081 kgCO2-eq/kgSE on a sweetness equivalence basis. Field production of stevia leaves was found to be the main source of impact for most impact categories, and for all four focus categories. Extraction of the RA60 was the main source of impact for the others. Leaf processing and seedling propagation were minor contributors to life cycle impact. Removal of international transport from the supply chain reduced global warming potential by 18.8%. Compared with sugar on a sweetness equivalence basis, RA60 has approximately 5.7% to 10.2% the impact for global warming potential, 5.6% to 7.2% the impact for land use, and is lower across most other impact categories. Conclusion: This is the first LCA of steviol glycoside mix RA60 produced from leaf in Europe. The results indicate that RA60 can be used to reduce environmental impact of providing a sweet taste by replacing sugar across all impact categories. However, it is important to note that specific formulations in which RA60 is used will have a bearing on the final environmental impact of any food or beverage products. For solid foods, this requires further research. Supplementary Information: The online version contains supplementary material available at 10.1007/s11367-022-02127-9.

Prevention of breast cancer in the context of a national breast screening programme.

Breast cancer is not only increasing in the west but also particularly rapidly in eastern countries where traditionally the incidence has been low. The rise in incidence is mainly related to changes in reproductive patterns and lifestyle. These trends could potentially be reversed by defining women at greatest risk and offering appropriate preventive measures. A model for this approach was the establishment of Family History Clinics (FHCs), which have resulted in improved survival in younger women at high risk. New predictive models of risk that include reproductive and lifestyle factors, mammographic density and measurement of risk-associated single nucleotide polymorphisms (SNPs) may give more precise information concerning risk and enable better targeting for mammographic screening programmes and of preventive measures. Endocrine prevention using anti-oestrogens and aromatase inhibitors is effective, and observational studies suggest lifestyle modification may also be effective. However, referral to FHCs is opportunistic and predominantly includes younger women. A better approach for identifying older women at risk may be to use national breast screening programmes. Here were described pilot studies to assess whether the routine assessment of breast cancer risk is feasible within a population-based screening programme, whether the feedback and advice on risk-reducing interventions would be welcomed and taken up, and to consider whether the screening interval should be modified according to breast cancer risk.

Evaluation of computer-aided detection (CAD) prompting techniques for mammography.

Computer-aided detection (CAD) systems, in which abnormalities are automatically detected and their locations presented to the radiologist as prompts, are increasingly being used to improve reader performance. The performance of CAD systems can be evaluated in two ways: by measuring the performance of the algorithms, or by monitoring the performance of readers using the system. All aspects of evaluation need careful consideration to avoid potential bias. This paper examines a variety of different approaches to evaluation and discusses their relative strengths and weaknesses.

Vitamin E supplementation and oxidative damage to DNA and plasma LDL in type 1 diabetes.

OBJECTIVE: To determine the effect of 400 IU/day of the antioxidant vitamin E on the susceptibility of plasma LDL and lymphocyte DNA to oxidative damage in type 1 diabetes. RESEARCH DESIGN AND METHODS: We studied 42 patients with type 1 diabetes and 31 age- and sex-matched control subjects in a randomized prospective double-blind placebo-controlled trial by using 400 IU/day of oral vitamin E for 8 weeks. Measurements were made of single-strand breaks in lymphocyte DNA at baseline and after hydrogen peroxide-induced stress (comet assay) and of copper-induced LDL oxidization and plasma antioxidant profiles. RESULTS: Plasma LDL and lymphocyte DNA were more resistant to induced oxidative change in the type 1 diabetes group than in control subjects. Vitamin E supplementation reduced LDL oxidizability in the control subjects but not in the type 1 diabetes group and had no effect on oxidative DNA damage in either group. The type 1 diabetes group had a significantly poorer plasma antioxidant profile with lower mean serum concentrations of alpha-tocopherol and most carotenoids than control subjects. CONCLUSIONS: Plasma LDL and lymphocyte DNA appear to be more resistant to oxidative change in type 1 diabetic subjects than in control subjects, and there was no evidence of oxidatively induced DNA or LDL change in type 1 diabetes. This study does not support the hypothesis of oxidative damage in these patients, and a dose of vitamin E (400 IU/day) that reduced LDL oxidative susceptibility in control subjects did not do so in patients with type 1 diabetes.

Measurement of cellular repair activities for oxidative DNA damage.

Two related assays capable of determining cell extract repair activities for different oxidative lesions in DNA are described. Both assays measure the incorporation of radiolabeled nucleotides during repair of an oxidatively damaged template in a cell-free system. The assays differ in the type of oxidative damage present in the DNA. In one, singlet oxygen is used to generate predominantly 8-oxo-2'-deoxyguanosine lesions. In the other, hydroxyl radicals are used to generate a broad spectrum of damage including oxidized bases and strand breaks. Assay conditions were adjusted to ensure that radiolabel incorporation was directly proportional to cell extract repair activity. These assays represent sensitive tools for investigating the regulation of repair systems for oxidative DNA damage.

Number of axons in the corpus callosum of the Mature macaca nemestrina: increases caused by prenatal exposure to ethanol.

The effects of prenatal exposure to ethanol on the number of callosal axons were examined. Pregnant Macaca nemestrina were treated with ethanol (1.8 g/kg b.wt.) 1 day per week during the first 6 weeks (Et6) or full 24 weeks (Et24) of gestation. Control macaques were intubated with an isocaloric amount of sucrose water (Ct). The mid-sagittal area of the corpus callosum in 4- to 5-year-old offspring was examined in magnetic resonance (MR) images and in fixed brains. The number of callosal axons was determined by using electron microscopy. In both MR images and fixed brains of macaques treated with ethanol, the corpus callosum was 26% larger than in the controls. The rostral portion was particularly affected by ethanol; it was 55% larger in Et6- and Et24-treated macaques. Axonal size and myelin thickness were unaffected by ethanol, but ethanol-treated macaques had more callosal axons (13.7 x 10(7)) than did controls (9.4 x 10(7) axons). The increase in the rostral corpus callosum suggests that parietal and frontal cortices are particularly susceptible to ethanol. The altered callosal connectivity may be a component of the structural abnormalities that underlie executive processing problems associated with fetal alcohol syndrome.

Analysis of facial shape in children gestationally exposed to marijuana, alcohol, and/or cocaine.

The association between fetal marijuana and/or alcohol exposure and facial features resembling fetal alcohol syndrome was investigated in a sample of 80 children. Standardized lateral and frontal facial photographs were taken of 40 children, 5 to 7 years of age, whose mothers reported frequent use of marijuana during the first trimester of pregnancy and 40 children whose mothers reported no use of marijuana during pregnancy. The marijuana-exposed and unexposed children were group-matched on alcohol exposure prior to and during pregnancy, sex, race, and age at the time of assessment. The photographs were assessed clinically by a study staff dysmorphologist and morphometrically by computerized landmark analysis. Fetal alcohol syndrome-like facial features were not associated with prenatal marijuana exposure in this study sample. No consistent patterns of facial features were identified among the marijuana-exposed group. Maternal consumption of two or more ounces of alcohol per day, on average, in early gestation was found to be associated with fetal alcohol syndrome-like facial features identified both clinically and morphometrically. Cocaine use reported by 13 of the 80 women was independently associated with mild facial dysmorphic features of hypertelorism and midfacial flattening. The results demonstrate the usefulness of this diagnostic technique for quantifying anomalies apparently unique to fetal alcohol syndrome and for targeting clusters of anomalies in new conditions for future evaluation.

Demystified...recombinant antibodies.

Recombinant antibodies are important tools for biomedical research and are increasingly being used as clinical diagnostic/therapeutic reagents. In this article, a background to humanized antibodies is given, together with details of the generation of antibody fragments--for example, single chain Fv fragments. Phage antibody fragments are fast becoming popular and can be generated by simple established methods of affinity enrichment from libraries derived from immune cells. Phage display methodology can also be used for the affinity enrichment of existing antibody fragments to provide a reagent with a higher affinity. Here, phage antibodies are demystified to provide a greater understanding of the potential of these reagents and to engage clinicians and biomedical scientists alike to think about potential applications in pathology and clinical settings.

Superordinate category formation in pigeons: association with a common delay or probability of food reinforcement makes perceptually dissimilar stimuli functionally equivalent.

Training associated pairs of perceptually dissimilar stimulus classes with a common delay or probability of food reinforcement in pigeons. Then, different choice responses were trained to 1 component class in each pair. In a choice test, the untrained class in each pair occasioned the same response as did the choice-trained class. In a 3rd experiment, 2 classes had reinforcement delays of 1 s and 15 s, respectively, and 2 other classes had reinforcement probabilities of 0.1 and 0.9. Then, 1 choice response was reinforced to a class previously associated with a better condition of reinforcement (e.g., 1-s delay or 1.0 probability), and a different response was reinforced to a class previously associated with a worse condition of reinforcement (0.1 probability or 0-s delay). Testing with all classes suggested that categorization was based on the relative reinforcement or hedonic value and not on the parametric details of reinforcement.

Superordinate categorization via learned stimulus equivalence: quantity of reinforcement, hedonic value, and the nature of the mediator.

Three experiments examined superordinate categorization via stimulus equivalence training in pigeons. Experiment 1 established superordinate categories by association with a common number of food pellet reinforcers, plus it established generalization to novel photographic stimuli. Experiment 2 documented generalization of choice responding from stimuli signaling different numbers of food pellets to stimuli signaling different delays to food reinforcement. Experiment 3 indicated that different numbers of food pellets did not substitute as discriminative stimuli for the photographic stimuli with which the food pellets had been paired. The collective results suggest that the effective mediator of superordinate categories that are established via learned stimulus equivalence is not likely to be an accurate representation of the reinforcer, neither is it likely to be a distinctive response that is made to the discriminative stimulus. Motivational or emotional mediation is a more likely account.

The pigeon's recognition of drawings of depth-rotated stimuli.

Four experiments used a four-choice discrimination learning paradigm to explore the pigeon's recognition of line drawings of four objects (an airplane, a chair, a desk lamp, and a flashlight) that were rotated in depth. The pigeons reliably generalized discriminative responding to pictorial stimuli over all untrained depth rotations, despite the bird's having been trained at only a single depth orientation. These generalization gradients closely resembled those found in prior research that used other stimulus dimensions. Increasing the number of different vantage points in the training set from one to three broadened the range of generalized testing performance, with wider spacing of the training orientations more effectively broadening generalized responding. Template and geon theories of visual recognition are applied to these empirical results.

New avenues in 3-D computerised (stereopathological) imaging of breast cancer (review).

Multimodal methods of three-dimensional (3-D) imaging of breast cancer are described. These involve scanning confocal microscopy, using 50 MHz acoustic or near-infrared images, four-view (tetrahedral) radiography and x-ray projection microscopy. Computerised volume data from these techniques can be used to produce three-dimensional images of tissue ranging from 500 microns to approximately 4 mm in thickness. Preliminary findings indicate that stereoscopic images or 3-D computerised reconstructions are capable of advancing the understanding of the structure of ductal carcinoma in situ, lesions simulating microinvasive breast carcinoma, surgical clearance of high-grade calcifying ductal carcinoma in situ, and the 3-D growth patterns of invasive forms of breast carcinoma. In the future computerised image fusion techniques seem likely to be able to take advantage of multimodal imaging of breast cancers, thus correcting primary imaging artefacts, improving robustness, and combining complementary information. In addition, the use of computerised tetrahedral radiography may change the intraoperative assessment of breast cancers, which mostly depend at present upon subsequent laboratory procedures that take days to perform.

Immunoreactivity of antibodies to epidermal growth factor, transforming growth factors alpha and beta, and epidermal growth factor receptor in the premenopausal ovary.

This study provides a valuable insight into the localization of growth factors in paraffin sections of human ovarian tissue. Antibodies to epidermal growth factor (EGF), transforming growth factors alpha and beta (TGF alpha and beta) and epidermal growth factor receptor (EGFR) were applied to paraffin sections of 16 cases of formalin-fixed normal or benignly abnormal ovarian tissue. All growth factor antibodies reacted with theca, but not granulosa cells, whilst the antibody to EGFR reacted with both types of follicular cells and was weakly reactive in ovarian stroma. There were no discernible qualitative changes in reactivity during the follicular cycle. These immunohistochemical findings generally support previously published molecular and biochemical data from tissue culture. One exception is in the observation of immunoreactivity to EGF in theca and granulosa cells. This may be due to differences in sensitivity of the methods in use. The possibility of a cross-reaction of the anti-EGF antibody with TGF alpha is also discussed. This study provides evidence for both paracrine and autocrine roles for growth factors in folliculogenesis.

Outcomes of suction curettage and mifepristone abortion in the United States. A prospective comparison study.

A prospective, nonconcurrent cohort analysis of 178 mifepristone/misoprostol and 199 suction curettage abortion subjects, ages > or = 18 years, with intrauterine pregnancies < or = 63 days estimated gestational age, was conducted to compare the outcomes of suction curettage abortion to those of medical abortion. The medical abortion subjects received 600 mg of mifeprisone orally, followed by 400 micrograms of oral misoprostol 2 days later. Surgical abortion subjects underwent electronic vacuum aspiration. All subjects were followed for 2 weeks or until the absence of clinical bleeding. Outcome measures included a successful abortion (complete abortion without a surgical intervention), duration of bleeding, and morbidity. Overall, 18.3% medical and 4.7% surgical patients failed their primary procedure and received an unanticipated suction curettage (RR 3.93, 95% CI 1.87, 8.29). Four mifepristone subjects required curettage for acute bleeding, nine to manage ongoing pregnancy, and five for incomplete abortion. Fourteen mifepristone and eight surgical subjects required curettage for persistent bleeding. The median time delay for therapeutic curettage was significantly longer in the medical abortion group (35 versus 8 days; Mann-Whitney U = 17.0, p = 0.008). Medical subjects experienced significantly longer bleeding (mean difference = 9.6 days, 95% CI 6.8, 12.4). No significant change in hemoglobin occurred in either group. Mifepristone patients reported significantly greater pain (77.1% vs 10.5%; RR 7.4, 95% CI 4.7, 11.5), and nausea or vomiting (68.6% vs 0.6%; RR 117.9, 95% CI 16.7, 834.7). Women receiving mifepristone/misoprostol are more likely to require an unplanned surgical intervention than women who undergo suction curettage. They experience more discomfort with their procedure and in the follow-up interval, bleed for a longer period, and remain at risk for surgical completion curettage for several weeks.

PIP: A prospective, nonconcurrent cohort analysis was conducted to compare the outcomes of suction curettage abortion to those of medical abortion in the US. The study included 178 mifepristone/misoprostol and 199 curettage abortion subjects, ages 18 years or older, with intrauterine pregnancies of 63 or fewer days. Medical abortion subjects received 600 mg of mifepristone orally, followed by 400 mcg of oral misoprostol 2 days later. Surgical abortion subjects underwent electronic vacuum aspiration. Results showed that 18.3% of medical and 4.7% of surgical patients failed their primary procedure and received an unanticipated suction curettage. Medical subjects experienced significantly longer bleeding; however, no significant change in hemoglobin occurred in either group. While, mifepristone patients reported significantly greater pain, nausea or vomiting. Thus, women receiving mifepristone/misoprostol are more likely to require an unplanned surgical intervention than women who undergo curretage. Medical abortion patients have more discomfort, they bleed longer, and remain at risk for surgical completion curettage for several weeks.

Measuring quality of life in children with obstructive sleep disorders.

OBJECTIVE: To validate a disease-specific health-related quality of life (HRQOL) instrument for children with obstructive sleep disorders (OSDs). DESIGN: Prospective cohort study using a 6-item health-related instrument (OSD-6). SUBJECTS: One hundred caregivers of patients with OSDs secondary to adenotonsillar hypertrophy (age range, 2-12 years) from 2 tertiary care, pediatric otolaryngology practices. INTERVENTION: The OSD-6 was administered on initial presentation and 4 to 5 weeks after adenotonsillectomy. A subset of patients repeated the OSD-6 within 3 weeks after presentation to assess test-retest reliability. MAIN OUTCOME MEASURES: Test-retest reliability, internal consistency, construct validity, and responsiveness to clinical change of the OSD-6 score. RESULTS: Test-retest reliability was good (intraclass correlation coefficient = 0.74). Median OSD-6 score was 4.5 (0- to 6-point scale) with higher scores indicating poorer quality of life (QOL). Construct validity was demonstrated by the moderate correlation between OSD-6 score and global adenoid and tonsil-related QOL (R = -0.62), strong correlation between the OSD-6 change score and change in global adenoid and tonsil-related QOL (R = -0.63), and the moderate correlation between the change score and parent estimate of clinical change (R = 0.40). The mean change in OSD-6 score after adenotonsillectomy was 3.0 (95% confidence interval, 2.7-3.4). The mean standardized response was 2.3 (95% confidence interval, 1.9-2.7) indicating the instrument's large responsiveness to clinical change. The change score was very reliable (R = 0.85). CONCLUSIONS: The OSD-6 is a reliable, responsive, easily administered instrument. It is valid for detecting change after adenotonsillectomy in children with OSDs. Arch Otolaryngol Head Neck Surg. 2000;126:1423-1429

Detection and measurement of DNA adducts in the cervix of smokers and non-smokers.

Smoking has been associated, on epidemiologic grounds, with an increased risk of cervical neoplasia. We have investigated this association, using laboratory-based methods. A 32P post-labeling assay was performed on 97 cervical biopsies to detect and measure DNA adducts (additional products formed by the covalent binding of potential chemical carcinogens to nuclear DNA). The specimens were taken from both normal cervices as well as the histologically normal regions of cervices with invasive and intraepithelial neoplasia. A detailed smoking history was obtained from each patient and correlated with an assay of cotinine level in urine. Characteristic smoking-related DNA adducts were found, and a significant difference in their levels was detected between current and non-current smokers (P = 0.017, Mann-Whitney test). There was also a highly significant trend in median adduct levels between the three tissue types (P < 0.002). We conclude that the finding of smoking-related cervical DNA damage is suggestive of a causal association between smoking and cervical neoplasia.

Expression of cytochrome P450 enzymes in the cervix. An immunohistochemical study.

The expression of cytochrome P450 isoenzymes (subfamilies) CYP1A, CYP2B, CYP2C, CYP3A and CYP4A in the histologically normal cervix was explored using a panel of polyclonal antibodies. There was variation in the intensity of immunohistochemical reaction between the isoenzymes and between the various components of the cervix. Half the subjects tested were smokers and had increased urinary cotinine levels. Statistical analysis revealed no significant differences between smokers and nonsmokers in the expression of these isoenzymes. The implications of these observations in relation to cervical carcinogenesis are discussed.

Maternal marijuana use during lactation and infant development at one year.

Prenatal marijuana exposure is associated with adverse perinatal effects. Very little is known about the effect of postnatal marijuana exposure on infant development. Postnatal exposure can result from maternal use of marijuana during lactation. Delta-9-tetrahydrocannabinol (THC) transfers and concentrates in the mother's milk and is absorbed and metabolized by the nursing infant. The present study investigated the relationship between infant exposure to marijuana via the mother's milk and infant motor and mental development at one year of age. One hundred and thirty-six breast-fed infants were assessed at one year of age for motor and mental development. Sixty-eight infants were exposed to marijuana via the mother's milk. An additional 68 infants were matched to the marijuana-exposed infants on pre- and postpartum maternal alcohol and tobacco use. Marijuana exposure via the mother's milk during the first month postpartum appeared to be associated with a decrease in infant motor development at one year of age.

Magnetic resonance imaging and spectroscopy in fetal ethanol exposed Macaca nemestrina.

Magnetic resonance imaging (MRI) and proton magnetic resonance spectroscopy (1H-MRS) offer noninvasive ways to observe structural and biochemical changes which might serve as valuable diagnostic markers for detecting brain damage from prenatal ethanol teratogenesis. Cranial MR imaging and spectroscopy were performed on 20 nonhuman primates (Macaca nemestrina) with known prenatal ethanol exposures and well-documented cognitive and behavioral levels of performance. The choline: creatine ratio detected by 1H-MRS in the brain increased significantly with increasing duration of in utero ethanol exposure. These signal alterations occurred in the absence of gross structural brain anomalies (detectable by MRI) and were significantly correlated with alcohol-related cognitive and behavioral dysfunction. These observations are consistent with reports of elevated choline: creatine ratios associated with various neurologic insults and disease states. The association observed between brain choline:creatine ratios and in utero ethanol exposure suggest a role for 1H-MRS in elucidating mechanisms of ethanol teratogenicity.