A Replication-Defective Human Cytomegalovirus Vaccine Elicits Humoral Immune Responses Analogous to Those with Natural Infection.

Human cytomegalovirus (HCMV) can cause congenital infections, which are a leading cause of childhood disabilities. Since the rate of maternal-fetal transmission is much lower in naturally infected (HCMV-seropositive) women, we hypothesize that a vaccine candidate capable of eliciting immune responses analogous to those of HCMV-seropositive subjects may confer protection against congenital HCMV. We have previously described a replication-defective virus vaccine based on strain AD169 (D. Wang, D. C. Freed, X. He, F. Li, et al., Sci Transl Med 8:362ra145, 2016, https://doi.org/10.1126/scitranslmed.aaf9387). The vaccine, named V160, has been shown to be safe and immunogenic in HCMV-seronegative human subjects, eliciting both humoral and cellular immune responses (S. P. Adler, S. E. Starr, S. A. Plotkin, S. H. Hempfling, et al., J Infect Dis 220:411-419, 2019, https://doi.org/10.1093/infdis/171.1.26). Here, we further showed that sera from V160-immunized HCMV-seronegative subjects have attributes similar in quality to those from seropositive subjects, including high-avidity antibodies to viral antigens, coverage against a panel of genetically distinct clinical isolates, and protection against viral infection in diverse types of human cells in culture. More importantly, vaccination appeared efficient in priming the human immune system, inducing memory B cells in six V160 recipients at frequencies comparable to those of three HCMV-seropositive subjects. Our results demonstrate the ability of V160 to induce robust and durable humoral memory responses to HCMV, justifying further clinical evaluation of the vaccine against congenital HCMV.IMPORTANCEIn utero HCMV infection can lead to miscarriage or childhood disabilities, and an effective vaccine is urgently needed. Since children born to women who are seropositive prior to pregnancy are less likely to be affected by congenital HCMV infection, it has been hypothesized that a vaccine capable of inducing an immune response resembling the responses in HCMV-seropositive women may be effective. We previously described a replication-defective virus vaccine that has been demonstrated safe and immunogenic in HCMV-seronegative subjects. Here, we conducted additional analyses to show that the vaccine can induce antibodies with functional attributes similar to those from HCMV-seropositive subjects. Importantly, vaccination can induce long-lived memory B cells at frequencies comparable to those seen in HCMV-seropositive subjects. We conclude that this vaccine is a promising candidate that warrants further clinical evaluation for prevention of congenital HCMV.

Survey of nurses' knowledge and practice regarding medication administration using enteral tubes.

AIM AND OBJECTIVES: To identify the practice variation of the individual practitioners in medications' formulation modification for patients using enteral feeding tubing and to support health practitioners involved in this process. BACKGROUND: Blockage of enteral tubes is a common problem that can sometimes be resolved but may require replacement of the tube. Medications are a common culprit. DESIGN: A survey of 73 registered nurses' practices around medication administration via enteral feeding tubes. METHODS: A questionnaire study was undertaken within a district general hospital across a broad variety of wards to explore nurses' experiences of medication administration via enteral tubes. The study is reported in accordance with the squire 2.0 guidelines from the EQUATOR network. RESULTS: Seventy-three nurses responded. Twenty-six per cent reported never checking about drug modification for administration via a tube, 12% check every time and 61% when unsure about a new drug. The volume of fluid flushes administered after medication ranged from 7.5-150 ml. Seventy-one per cent of participants reported stopping feed when medications are required, varying from 1-60 min. Sixty per cent had experienced a blocked tube and 52% the tube being removed for these reasons. The clinical nurse specialist was the commonest first point of call to help. Staff named 15 medications as the most problematic to administer, lactulose and omeprazole were the top two. CONCLUSIONS: Practice varies significantly amongst nurses around medication administration. Theoretically, this may contribute to blocked tubes and excessive fluid administration to some patients. Barriers to medication administration were thematically grouped into: time, difficulty modifying medication, medication interactions and knowledge. Areas identified to support staff include training, devices to crush medications, medication suitability, multidisciplinary approach to streamline care and quick reference guides. RELEVANCE TO CLINICAL PRACTICE: Health professionals may use these results to reduce and ultimately avoid problems with administering medications through feeding tubes. Organisations may use these results to develop their local practice pathways for prescribing, dispensing and training around administration of medications through enteral tubes. In a community setting, this paper may improve the awareness of patients, caregivers and prescribers of the possible implications of tubing blockages.

Thermal pasteurization of ready-to-eat foods and vegetables: Critical factors for process design and effects on quality.

Increasing consumer desire for high quality ready-to-eat foods makes thermal pasteurization important to both food producers and researchers. To be in compliance with the Food Safety Modernization Act (FSMA), food companies seek regulatory and scientific guidelines to ensure that their products are safe. Clearly understanding the regulations for chilled or frozen foods is of fundamental importance to the design of thermal pasteurization processes for vegetables that meet food safety requirements. This article provides an overview of the current regulations and guidelines for pasteurization in the U.S. and in Europe for control of bacterial pathogens. Poorly understood viral pathogens, in terms of their survival in thermal treatments, are an increasing concern for both food safety regulators and scientists. New data on heat resistance of viruses in different foods are summarized. Food quality attributes are sensitive to thermal degradation. A review of thermal kinetics of inactivation of quality-related enzymes in vegetables and the effects of thermal pasteurization on vegetable quality is presented. The review also discusses shelf-life of thermally pasteurized vegetables.

Spatial variance of physicochemical properties within mangos and the effect of initial ripeness stage on the quality of fresh-cut mangos.

BACKGROUND: This study aimed to assess the spatial variation in physicochemical properties within individual mangos, as well as to investigate the influence of initial ripeness level on physicochemical characteristics of fresh-cut mangos. Individual mangos were evaluated at 12 specific flesh positions in the inner and outer sides. Mango cubes of 1.5 cm prepared from three firmness stages were monitored for changes during 9 days of storage at 5 °C. RESULTS: Mango fruit varied significantly in firmness and color based on spatial position, with the ripening direction from the inner flesh outward and from the stem end to blossom end. Limitations to fresh-cut mango quality were 'desiccation' (dried cut surface) and 'edge or tissue damage' (cut edge damage or brown and bruise-like appearance). Firmer texture and paler yellow of inner flesh were found in less mature mango fruit (P < 0.001). The optimal ripeness stage for fresh-cut mango products was 45 N, based on ease of handling, fresh appearance at the time of purchase and intermediate physicochemical properties (firmness, color and SSC/TA ratio). CONCLUSION: Spatial variance and initial ripeness stage affect fresh-cut mango quality. Therefore, they must be considered by fresh-cut mango processors in order to attain optimal product quality. © 2015 Society of Chemical Industry.

The effects of bruising and temperature on enzyme activity and textural qualities of tomato juice.

BACKGROUND: During harvest and transportation, processing tomatoes are exposed to elevated temperatures, compression and vibration in the harvester and truck, making them prone to bruising. The objective of this study was to determine how bruising and exposure to high temperatures affect pectin methylesterase (PME) activation and the textural quality of tomato juice. RESULTS: Tomatoes were both hand and mechanically harvested using current harvest practices. Mechanically harvested fruits were significantly softer, had greater PME activity and greater juice consistency than hand harvested fruits. In a controlled bruising study, whole tomatoes were exposed to various compressive forces at 21 or 40 °C and held for 0 or 4 h. Greater bruising force and higher temperature resulted in a decrease in firmness and an increase in PME activity. Consistency of tomato juice improved when tomatoes were exposed to 40 °C. Tomatoes subjected to a temperature range from 21 to 65 °C had activated PME at 40 °C and increased activity as temperature increased. Consistency increased at 35 °C but decreased with increasing temperature. CONCLUSION: Tomatoes harvested using current mechanical techniques are likely to be less firm and have increased PME activity; however, increased consistency of processed juice is observed. Tomatoes harvested at higher temperatures are also likely to have better consistency when processed.

Transcriptional signature of durable effector T cells elicited by a replication defective HCMV vaccine.

Human cytomegalovirus (HCMV) is a leading infectious cause of birth defects and the most common opportunistic infection that causes life-threatening diseases post-transplantation; however, an effective vaccine remains elusive. V160 is a live-attenuated replication defective HCMV vaccine that showed a 42.4% efficacy against primary HCMV infection among seronegative women in a phase 2b clinical trial. Here, we integrated the multicolor flow cytometry, longitudinal T cell receptor (TCR) sequencing, and single-cell RNA/TCR sequencing approaches to characterize the magnitude, phenotype, and functional quality of human T cell responses to V160. We demonstrated that V160 de novo induces IE-1 and pp65 specific durable polyfunctional effector CD8 T cells that are comparable to those induced by natural HCMV infection. We identified a variety of V160-responsive T cell clones which exhibit distinctive "transient" and "durable" expansion kinetics, and revealed a transcriptional signature that marks durable CD8 T cells post-vaccination. Our study enhances the understanding of human T-cell immune responses to V160 vaccination.

Implementation of a weight loss program for Latino outpatients with severe mental illness.

To determine feasibility of implementation of a weight loss program for overweight Latinos with severe mental illness. In this quasi-experimental study, a 14-week behavioral weight loss course (extended) was implemented at one clinic. A one-time nutrition class (brief) was given at a sister clinic. Implementation feasibility was assessed by consent and participation rates. Weight was followed for 6 months. Consent rates were high [77 % (49/64) extended; 68 % (39/57) brief], and 88 % (43/49) of extended subjects participated and 88 % (38/43) completed follow-up. Weight loss did not differ between groups. A behavioral weight loss course is feasible to implement for this population.

Advanced preservation methods and nutrient retention in fruits and vegetables.

Despite the recommendations of international health organizations and scientific research carried out around the world, consumers do not take in sufficient quantities of healthy fruit and vegetable products. The use of new, 'advanced' preservation methods creates a unique opportunity for food manufacturers to retain nutrient content similar to that found in fresh fruits and vegetables. This review presents a summary of the published literature regarding the potential of high-pressure and microwave preservation, the most studied of the 'advanced' processes, to retain the natural vitamin A, B, C, phenolic, mineral and fiber content in fruits and vegetables at the time of harvest. Comparisons are made with more traditional preservation methods that utilize thermal processing. Case studies on specific commodities which have received the most attention are highlighted; these include apples, carrots, oranges, tomatoes and spinach. In addition to summarizing the literature, the review includes a discussion of postharvest losses in general and factors affecting nutrient losses in fruits and vegetables. Recommendations are made for future research required to evaluate these advanced process methods.

Texture improvement of fresh and frozen mangoes with pectin methylesterase and calcium infusion.

BACKGROUND: A major problem of mango products is texture loss. The effect of commercial pectin methylesterase (PME) and calcium infusion on improvement of the texture of both fresh and frozen-thawed mango cubes was investigated in the present study. RESULTS: The weight gain and moisture content of mango samples were greater at relatively high vacuum level (10 kPa). The PME activity of samples infused with PME and calcium at 10 kPa increased fourfold in comparison with that of control and water-infused samples. The combined effect of PME and calcium was found to improve the texture and microstructure of both fresh and frozen-thawed mangoes. Fresh mangoes infused with PME and calcium at 10 kPa showed significantly higher firmness than control fresh samples. Frozen-thawed mangoes infused with PME and calcium at 50 kPa and atmospheric pressure had superior texture and microstructure in comparison with control frozen-thawed samples. CONCLUSION: The results of the present analysis allow for a better appreciation of the role of PME, calcium and appropriate infusion conditions in improving the texture of both fresh and frozen-thawed mangoes.

Changes in pH, acids, sugars and other quality parameters during extended vine holding of ripe processing tomatoes.

BACKGROUND: Two important quality attributes of processing tomatoes are pH and titratable acidity. These and other quality attributes can be affected by tomato fruit maturity and over-maturity. We have determined the magnitude of these maturity effects in four processing tomato cultivars commonly grown in California. RESULTS: Allowing tomatoes to remain on the vine for up to 4 weeks after ripening resulted in an increase in fruit pH of between 0.01 and 0.02 unit per day for the four cultivars examined. The increase in pH was paralleled by a decrease in titratable acidity, due to a loss of citric acid. Glucose and fructose concentrations also declined with increasing maturity after ripening. Other quality parameters (color, lycopene, total pectin, pectin solubility, and Bostwick consistency) all showed little change. CONCLUSION: Vine holding of ripe fruit adversely affects quality, especially pH and titratable acidity. Recent problems with high tomato juice pH encountered by tomato processors in California could be the result of increased average fruit maturity at harvest.

The Hidden Burden of Community Enteral Feeding on the Emergency Department.

BACKGROUND: Enteral feeding tubes are associated with their most serious complications in the days and weeks after insertion, but there are limited published data in the literature on late complications and the implications for the healthcare service. METHODS: This is a retrospective observational study of attendances to a UK hospital emergency department (ED), with enteral tube complications as the primary reason for attendance. RESULTS: Over 24 months, 139 attendances were recorded. Dislodged tubes and blocked tubes accounted for the majority of complications and subsequent admissions, with a mixture of enteral tube types being associated with both. Thirty-five percent of patients were admitted, and the average healthcare cost per attendance was $1071. CONCLUSION: Enteral tube complications can place a hidden burden on the patient, the ED, and healthcare costs. More work on education and supporting caregivers to resolve problems themselves could reduce the burden on busy EDs.

Longitudinal complications associated with PEG: Rate and severity of 30-day and 1-year complications experienced by patients after primary PEG insertion.

BACKGROUND AND AIMS: Percutaneous Endoscopic Gastrostomy (PEG) feeding is utilised in patients with exceptionally poor oral intake but is associated with both short and long-term complications. This study reviews longitudinal PEG complications and compares key subgroups. METHODS: Single-centre retrospective observational study of all patients receiving PEG insertion between January 2016 and December 2018. RESULTS: 306 patients met the inclusion criteria. The mean age at insertion was 67 years. The majority were cared for in their own home (80.4%) by themselves or family (74.9%). 127 PEG tubes were inserted for dysphagia and 165 prophylactically prior to treatment for head and neck cancer. In the first 30 days 16.7% experienced a complication. The most frequently reported was peristomal pain (9.2%). In the first year, 35.6% experienced at least one complication, 12.4% two complications and 6.6% three complications and 6.5% required inpatient treatment for their complication. The most common was pain (14.4%) followed by site weeping, site infection and external overgranulation. Patients with dysphagia took longer to develop complications, had fewer complications and took longer to require management by members of the secondary care team than those with head and neck cancer. Discounting peristomal pain, there was no difference in total complications between patients caring for themselves when compared to those receiving professional input. CONCLUSION: One third of patients will experience a complication related to their PEG tube over 1 year, but the majority are managed in an outpatient setting. This study has implications for planning support services and consenting and counselling patients pre-PEG-insertion.

A conditionally replication-defective cytomegalovirus vaccine elicits potent and diverse functional monoclonal antibodies in a phase I clinical trial.

A conditionally replication-defective human cytomegalovirus (HCMV) vaccine, V160, was shown to be safe and immunogenic in a two-part, double-blind, randomized, placebo-controlled phase I clinical trial (NCT01986010). However, the specificities and functional properties of V160-elicited antibodies remain undefined. Here, we characterized 272 monoclonal antibodies (mAbs) isolated from single memory B cells of six V160-vaccinated subjects. The mAbs bind to diverse HCMV antigens, including multiple components of the pentamer, gB, and tegument proteins. The most-potent neutralizing antibodies target the pentamer-UL subunits. The binding sites of the antibodies overlap with those of antibodies responding to natural HCMV infection. The majority of the neutralizing antibodies target the gHgL subunit. The non-neutralizing antibodies predominantly target the gB and pp65 proteins. Sequence analysis indicated that V160 induced a class of gHgL antibodies expressing the HV1-18/KV1-5 germline genes in multiple subjects. This study provides valuable insights into primary targets for anti-HCMV antibodies induced by V160 vaccination.

Color, flavor, texture, and nutritional quality of fresh-cut fruits and vegetables: desirable levels, instrumental and sensory measurement, and the effects of processing.

The color, flavor, texture, and the nutritional value of fresh-cut fruit and vegetable products are factors critical to consumer acceptance and the success of these products. In this chapter, desirable and undesirable quality attributes of fresh-cut fruit and vegetable products are reviewed. Both instrumental and sensory measurements for determining these critical quality attributes are discussed. The advantages and disadvantages of sensory and instrumental quality measurements are described. A review of typical unit operations involved in the production of fresh-cut products is presented. The effects of fresh-cut processing techniques and treatments on sensory quality, including the appearance, texture, flavor (taste and aroma) of vegetables, and fruits are detailed.

Combined enzymatic and colorimetric method for determining the uronic acid and methylester content of pectin: Application to tomato products.

A simple procedure for determining the galacturonic acid and methanol contents of soluble and insoluble pectins, relying on enzymatic pectin hydrolysis and colorimetric quantification, is described. Pectin samples are incubated with a commercial pectinase preparation, Viscozyme, then the galacturonic acid content of the hydrolyzed pectin is quantified colorimetrically using a modification of the Cu reduction procedure originally described by Avigad and Milner. This modification, substituting the commonly used Folin-Ciocalteau reagent for the arsenic containing Nelson reagent, gives a response that is linear, sensitive, and selective for uronic acids over neutral sugars. This method also avoids the use of concentrated acids needed for the commonly used m-phenylphenol method. Methanol, released by the action of the pectin methylesterase found in the Viscozyme, is quantified using alcohol oxidase and Purpald. This combined enzymatic and colorimetric procedure correctly determined the galacturonic acid and methanol content of purified, soluble citrus pectin. Application of the procedure to water insoluble pectins was evaluated with water insoluble material from apples and oranges. In both cases good agreement was obtained between this method and commonly used methods based on chemical pectin hydrolysis. Good agreement between these procedures was also found in the analysis of both soluble and insoluble pectins from several tomato products.

Nonenzymatic degradation of citrus pectin and pectate during prolonged heating: effects of pH, temperature, and degree of methyl esterification.

The underlying mechanisms governing nonenzymatic pectin and pectate degradation during thermal treatment have not yet been fully elucidated. This study determined the extent of nonenzymatic degradation due to beta-elimination, acid hydrolysis, and demethylation during prolonged heating of citrus pectins and its influence on physicochemical properties. Solutions of citrus pectins, buffered from pH 4.0 to 8.5, were heated at 75, 85, 95, and 110 degrees C for 0-300 min. Evolution of methanol and formation of reducing groups and unsaturated uronides were monitored during heating. Molecular weight and viscosity changes were determined through size exclusion chromatography and capillary viscometry, respectively. Results showed that at pH 4.5, the activation energies of acid hydrolysis, beta-elimination, and demethylation are 95, 136, and 98 kJ/mol, respectively. This means that at this pH, acid hydrolysis occurs more rapidly than beta-elimination. Furthermore, the rate of acid hydrolysis is diminished by higher levels of methyl esterification. Also, citrus pectin (93% esterified) degrades primarily via beta-elimination even under acidic conditions. Acid hydrolysis and beta-elimination caused significant reduction in relative viscosity and molecular weight.

Reactivation of broccoli peroxidases: structural changes of partially denatured isoenzymes.

UNLABELLED: Structural changes involved in the reactivation of peroxidases (PODs) from broccoli and horseradish (HRP) following heat denaturation were investigated by using circular dichroism and absorption spectroscopy. Cooling heat-treated enzymes resulted in rapid refolding of the secondary structure into an inactive structural species, similar in conformation to the native enzyme. Reassociation of heme to the refolded peroxidase, as well as molecular rearrangement of the structure around the heme, occurs during incubation at approximately 25 degrees C and results in the return of biological activity. The secondary structure of neutral broccoli POD (N) is relatively heat labile, resulting in a rapid loss of activity, but the level of reactivation is high because the structure at the heme pocket is relatively stable. Acidic broccoli POD and HRP are more heat stable than N, but have a low degree of reactivation. Loss of activity is due primarily to alteration of the structure at the heme pocket. Effects of bovine serum albumin and pH on reactivation of PODs are also discussed. KEYWORDS: Peroxidase; reactivation; horseradish; broccoli; circular dichroism; absorption spectroscopy.

Ten-year comparison of the influence of organic and conventional crop management practices on the content of flavonoids in tomatoes.

Understanding how environment, crop management, and other factors, particularly soil fertility, influence the composition and quality of food crops is necessary for the production of high-quality nutritious foods. The flavonoid aglycones quercetin and kaempferol were measured in dried tomato samples (Lycopersicon esculentum L. cv. Halley 3155) that had been archived over the period from 1994 to 2004 from the Long-Term Research on Agricultural Systems project (LTRAS) at the University of California-Davis, which began in 1993. Conventional and organic processing tomato production systems are part of the set of systems compared at LTRAS. Comparisons of analyses of archived samples from conventional and organic production systems demonstrated statistically higher levels (P < 0.05) of quercetin and kaempferol aglycones in organic tomatoes. Ten-year mean levels of quercetin and kaempferol in organic tomatoes [115.5 and 63.3 mg g(-1) of dry matter (DM)] were 79 and 97% higher than those in conventional tomatoes (64.6 and 32.06 mg g(-1) of DM), respectively. The levels of flavonoids increased over time in samples from organic treatments, whereas the levels of flavonoids did not vary significantly in conventional treatments. This increase corresponds not only with increasing amounts of soil organic matter accumulating in organic plots but also with reduced manure application rates once soils in the organic systems had reached equilibrium levels of organic matter. Well-quantified changes in tomato nutrients over years in organic farming systems have not been reported previously.

Characterization of the temperature activation of pectin methylesterase in green beans and tomatoes.

Low-temperature blanching of vegetables activates the enzyme pectin methylesterase (PME), which demethylates cell wall pectins and improves tissue firmness. This temperature activation of PME has been investigated by measuring the formation of methanol in intact tissue of green beans and tomatoes. Rates of methanol formation at temperatures of 35-65 degrees C were obtained by measuring the release of methanol from thin slices of tomato pericarp or green bean pod material. Activation energies of 112 and 97 kJ mol(-1) were calculated for PME activity in green beans and tomatoes, respectively. These activation energies indicate that the rate of pectin demethylation at 65 degrees C will be nearly 100 times that at 25 degrees C. PME activity was also determined titrimetrically using a solubilized form of the enzyme and purified pectin at temperatures from 30 to 60 degrees C. Under these conditions, much lower activation energies of 37 and 35 kJ mol(-1) were obtained for green beans and tomatoes, respectively. Methanol accumulation during heating of whole intact green beans was also determined and yielded an activation energy similar to that obtained with sliced beans. Whole green beans held at room temperature did not accumulate any methanol, but sliced or homogenized beans did. If whole beans were first heated to 45 degrees C and then cooled, methanol accumulation was observed at room temperature. These results indicate that two factors contribute to the observed high rate of pectin de-esterification during low-temperature blanching: (1) An irreversible change, causing PME to become active, occurs by heating to > or = 45 degrees C. (2) The high activation energy for pectin de-esterification means that the rate of de-esterification increases substantially with increasing temperature.

Effects of Peach Cultivar on Enzymatic Browning Following Cell Damage from High-Pressure Processing.

Peach cultivars contribute to unique product characteristics and may affect the degree of browning after high-pressure processing (HPP). Nine peach cultivars were subjected to HPP at 0, 100, and 400 MPa for 10 min. Proton nuclear magnetic resonance (1H NMR) relaxometry, light microscopy, color, polyphenol oxidase (PPO) activity, and total phenols were evaluated. The development of enzymatic browning during refrigerated storage occurred because of damage during HPP that triggered loss of cell integrity, allowing substrates to interact with enzymes. Increasing pressure levels resulted in greater damage, as determined by shifts in transverse relaxation time (T2) and by light micrographs. Discoloration was triggered by membrane decompartmentalization but limited by PPO activity, which was found to correlate to cultivar harvest time (early, mid, and late season). Outcomes from the microstructure, 1H NMR ,and PPO activity evaluation were an effective means of determining membrane decompartmentalization and allowed for prediction of browning scenarios.