Research of a holiday kind: A clinical trial gone awry: the Chocolate Happiness Undergoing More Pleasantness (CHUMP) study

The randomized controlled trial is the “gold standard” for evaluating the benefits and harms of interventions. The Chocolate Happiness Undergoing More Pleasantness (CHUMP) study was designed to compare the effects of dark chocolate, milk chocolate and normal chocolate consumption on happiness. Although the intention-to-treat analysis showed that participants who received either dark or milk chocolate were happier than those who received no additional chocolate, the actual-consumption analysis showed that there were no differences between any of the groups. The reason for this result is that many participants switched groups mid-study because of their personal chocolate preferences. Although the CHUMP study was pleasurable, it demonstrated the difficulties associated with performing a truly blinded clinical trial.The randomized controlled trial is the “gold standard” for testing the beneficial and harmful effects of interventions. There has been a growing concern in the literature about the potential for bias during randomization of participants in clinical trials.^1–5 I designed the Chocolate Happiness Undergoing More Pleasantness (CHUMP) study to compare the effects of dark chocolate, milk chocolate and “normal” chocolate consumption on happiness. The CHUMP study was a double-blinded clinical trial, and it demonstrated the difficulties associated with performing a truly blinded clinical trial.     

Transformation of Bacillus subtilis in chocolate milk: evidence for low frequency of establishment of cells transformed under non-selective conditions

Transformation of naturally competent Bacillus subtilis with plasmid was carried out in chocolate milk without antibiotics. Transformed cells were enumerated during the entire growth phase in chocolate milk. When DNA was added to aliquots of a batch culture after different times of incubation, transformation events were detected at all different growth stages. When DNA was added to a batch culture together with the inoculum, transformed cells were detected at the onset of exponential growth. However, apparently no or only limited growth of these transformed cells was observed. To clarify, whether the limitation of growth was due to suppression by non-transformed cells, different proportions of B. subtilis cells either carrying or not carrying the plasmid were mixed and inoculated into chocolate milk without antibiotic. Our results indicate that suppression appears to be of minor importance. Instead, plasmid-bearing cells appear to suffer from a prolonged lag-phase. However, the failure to exhibit significant growth of cells which had taken up the plasmid in chocolate milk appears to be due to failure of these cells to establish themselves as permanently transformed under non-selective conditions.     

Chocolate Milk Consequences: A Pilot Study Evaluating the Consequences of Banning Chocolate Milk in School Cafeterias

Objectives

Currently, 68.3% of the milk available in schools is flavored, with chocolate being the most popular (61.6% of all milk). If chocolate milk is removed from a school cafeteria, what will happen to overall milk selection and consumption?

Methods

In a before-after study in 11 Oregon elementary schools, flavored milk–which will be referred to as chocolate milk–was banned from the cafeteria. Milk sales, school enrollment, and data for daily participation in the National School Lunch Program (NSLP) were compared year to date.

Results

Total daily milk sales declined by 9.9% (p<0.01). Although white milk increased by 161.2 cartons per day (p<0.001), 29.4% of this milk was thrown away. Eliminating chocolate milk was also associated with 6.8% fewer students eating school lunches, and although other factors were also involved, this is consistent with the notion of psychological reactance.

Conclusions

Removing chocolate milk from school cafeterias may reduce calorie and sugar consumption, but it may also lead students to take less milk overall, drink less (waste more) of the white milk they do take, and no longer purchase school lunch. Food service managers need to carefully weigh the costs and benefits of eliminating chocolate milk and should consider alternative options that make white milk more convenient, attractive, and normal to choose.     

The effects of low fat chocolate milk on postexercise recovery in collegiate athletes

Spaccarotella, KJ and Andzel, WD. The effects of low fat chocolate milk on postexercise recovery in collegiate athletes. J Strength Cond Res 25(12): 3456-3460, 2011-Drinking chocolate milk between exercise sessions may improve recovery. The purpose of this study was to examine the effects of low fat chocolate milk vs. a carbohydrate-electrolyte beverage (CE) on recovery between preseason practice sessions among 5 male and 8 female Division III soccer players. The study used a randomized crossover design: between morning and afternoon practices, athletes received either an amount of chocolate milk that provided 1 g carbohydrate per kilogram body weight or an equal volume of CE (mean volume of 615 ± 101 ml). After their afternoon practice, they completed a shuttle run to fatigue. Data were analyzed using the Wilcoxon paired rank-sign test (for shuttle run time) and the paired samples t-test (for dietary intake). No significant differences in run time were reported for the group. For the men only, there was a trend of increased time to fatigue with chocolate milk compared with the CE (exact p = 0.03). Low fat chocolate milk may therefore be as good as a CE at promoting recovery between training sessions during preseason.     

Rheological characteristics of heat-induced custard pudding gels with high antioxidative activity

Custard pudding gels were prepared from fresh whole egg, milk and sugar. The effects of D-psicose (Psi), a non-calorie rare hexose, on the antioxidative activity and rheological properties of the custard pudding gels were investigated at different temperatures for comparison with those of control sugars (sucrose, Suc; D-fructose, Fru). The rheological behavior of the heat-induced pudding gels was evaluated by using breaking and creep tests. During the heat-induced gel formation, custard pudding containing Psi (15%, wt/wt) demonstrated a stronger breaking strength and higher viscoelasticity than those containing Fru and Suc. The thermodynamic parameters obtained from DSC indicated that the egg white (EW) proteins were made less thermally stable when heated in the presence of Psi than in the presence of Fru and Suc. These findings are consistent with enhanced aggregation of the EW solution in the presence of Psi. Furthermore, the Psi pudding gels possessed higher antioxidative activity than the control sugar pudding gels by using an analysis of the scavenging activity on DPPH radicals and the ferric-reducing antioxidative power. These results suggest that Psi favored cross-linking of Psi-protein molecules through the Maillard reaction which increased the formation of intermediate products to improve functionality. Custard pudding containing Psi could therefore be an effective functional sweet desert with high antioxidative activity and the outstanding gelling characteristics.     

Evidence for natural transformation of Bacillus subtilis in foodstuffs

The effect of foodstuffs on the natural transformation of Bacillus subtilis was investigated. As examples of complex food matrices milk with various fat contents as well as chocolate milk were used. The frequencies of transformation varied with the fat content and ranged between 3.8×10⁻⁴ and 1.4×10⁻³. Highest frequencies of about 3×10⁻³ were observed in chocolate milk with 1.5% fat. Development of competence was observed in chocolate milk, resulting in maximal transformation frequencies upon incubation for 10–12 h at 37°C.     

Rheological Characteristics of Heat-Induced Custard Pudding Gels with High Antioxidative Activity

Custard pudding gels were prepared from fresh whole egg, milk and sugar. The effects of D-psicose (Psi), a non-calorie rare hexose, on the antioxidative activity and rheological properties of the custard pudding gels were investigated at different temperatures for comparison with those of control sugars (sucrose, Suc; D-fructose, Fru). The rheological behavior of the heat-induced pudding gels was evaluated by using breaking and creep tests. During the heat-induced gel formation, custard pudding containing Psi (15%, wt/wt) demonstrated a stronger breaking strength and higher viscoelasticity than those containing Fru and Suc. The thermodynamic parameters obtained from DSC indicated that the egg white (EW) proteins were made less thermally stable when heated in the presence of Psi than in the presence of Fru and Suc. These findings are consistent with enhanced aggregation of the EW solution in the presence of Psi. Furthermore, the Psi pudding gels possessed higher antioxidative activity than the control sugar pudding gels by using an analysis of the scavenging activity on DPPH radicals and the ferric-reducing antioxidative power. These results suggest that Psi favored cross-linking of Psi-protein molecules through the Maillard reaction which increased the formation of intermediate products to improve functionality. Custard pudding containing Psi could therefore be an effective functional sweet dessert with high antioxidative activity and the outstanding gelling characteristics.     

Formation of guaiacol in chocolate milk by the psychrotrophic bacterium Rahnella aquatilis

AIMS: The aim of this study was to identify the causative agent of a smoky/phenolic taint in refrigerated full cream chocolate milk. METHODS AND RESULTS: Microbiological examination of spoiled and unspoiled milk samples from the same processor showed high numbers of the psychrotrophic coliform Rahnella aquatilis in the spoiled samples only. Gas chromatography/mass spectrometry (GC/MS) was used to identify and quantify the taint compound as guaiacol (2-methoxyphenol) in the spoiled milk. Challenge studies in UHT chocolate and white milks inoculated with the isolate and incubated at 4-5 degrees C and 8-9 degrees C for 6 d showed the production of guaiacol in chocolate milk only, which was confirmed and quantified by GC/MS. CONCLUSIONS: The results indicate that if present in refrigerated chocolate milk, Rah. aquatilis can produce guaiacol within the expected shelf-life of the product, even without temperature abuse. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report that the coliform Rah. aquatilis can produce guaiacol in refrigerated chocolate milk products.     

FLAVOR MODIFICATION OF MILK CHOCOLATE BY CONCHING IN A TWIN-SCREW, CO-ROTATING, CONTINUOUS MIXER

Short-time continuous conching of chocolate offers significant economic advantages over traditional long-time batch methods. High-shear continous mixers can affect conching in a matter of minutes as compared to hours for traditional systems. Milk chocolate conched in two twin-screw, co-rotating, continous mixers operating in series was compared by a variety of sensory methods to chocolate conched by a batch method. A significant difference (P < 0.05) in flavor was found between chocolates conched at 60C by the continuous and batch methods, yet there was no preference for either chocolate. Chocolate conched by the batch method (23 h at 60C) had stronger caramel flavor (P < .10) than chocolate conched continuously, but there were no significant differences (P < .10) in sweet, chocolate or milk flavor. In the continuous system, caramel flavor generally increased with conching temperature and residence time; although, at the highest temperature (95C) and the longest residence time (7.5 min) caramel flavor decreased. Increasing the temperature of continuous conching from 70 to 90C produced chocolate significantly (P < .10) more like chocolate conched in a batch system for 21.5 h at 60C.     

CONFIDENCE ELLIPSES APPLIED TO THE COMPARISON OF SENSORY PROFILES

Our experiment involved seven panels and six chocolates – five dark chocolates and one milk chocolate. The aim of the study was to compare the sensory profiles of the chocolates. A natural question to ask is “Did the panelists detect any differences among the five dark chocolates or did they systematically contrast them with the milk chocolate?” The scatter plot of the chocolates obtained by principal component analysis was useless to answer that question, because of the proximity of the points. To overcome that, we used confidence ellipses calculated using bootstrap. The originality of the study lies in the fact that we applied those ellipses to hierarchical multiple factor analysis (HMFA): among the seven panels, six were composed of trained professionals and the last one was composed of untrained students, and through that method, we managed to compare the two types of panels and balance the role of each trained panel. HMFA provides in a single scatter plot a representation of the six chocolates for each panel, the trained panels and all the panels. Confidence ellipses around each chocolate show that the combined panels – the six trained panels and also the untrained panel – differentiate the five dark chocolates. They also show how much larger the untrained panel's variability is than that of the trained panels, and how comparable are the trained panels' variability to each other.     

Evaluation of three newly developed direct plating media to enumerate Listeria monocytogenes in foods

LiCl-phenylethanol-moxalactam Agar (LPMA), ARS Modified McBride Agar, and Modified Vogel Johnson Agar were compared with previously tested plating media in the enumeration of Listeria monocytogenes from pasteurized whole milk, chocolate ice cream mix, Brie cheese, and raw cabbage. LPMA was most suitable for analyzing Brie cheese and cabbage. Gum base-nalidixic acid-tryptone-soya medium (previously tested) was most suitable for analyzing milk and chocolate ice cream mix.     

Evaluation of three newly developed direct plating media to enumerate Listeria monocytogenes in foods.

LiCl-phenylethanol-moxalactam Agar (LPMA), ARS Modified McBride Agar, and Modified Vogel Johnson Agar were compared with previously tested plating media in the enumeration of Listeria monocytogenes from pasteurized whole milk, chocolate ice cream mix, Brie cheese, and raw cabbage. LPMA was most suitable for analyzing Brie cheese and cabbage. Gum base-nalidixic acid-tryptone-soya medium (previously tested) was most suitable for analyzing milk and chocolate ice cream mix.     

Two-Phase Slug Flow Heat Exchanger for Microbial Thermal Inactivation Research

A continuous two-phase (air-liquid), slug flow, tubular heat exchanger was developed for microbial thermal inactivation research to give exposure times and temperatures in the range of high-temperature, short-time milk pasteurization as well as heat-treated sample volumes of at least 2 ml. The use of air to compartmentalize the liquid in the capillary tubing prevented the development of laminar flow, which enabled precise identification of the residence time of the fastest flowing particles in the heating, holding, and cooling sections of the instrument. Residence time distributions were quantitated by measuring the degree of spreading of radioactive tracers for water, whole milk, chocolate milk, cream, and ice-cream mix with holding temperatures from 50 to 72 C, holding times from 2 to 60 sec, and heating and cooling times of 1.7 sec each. For a holding time of 60 sec and a fastest particle velocity of 10.2 cm/sec, the velocity ratios of the fastest moving particle to the median particle were 1.05, 1.05, 1.10, and 1.13 for whole milk, chocolate milk, cream, and ice-cream mix, respectively. With shorter holding times, these velocity ratios were even closer to unity. These velocity ratios indicated that the instrument would be an effective tool for thermal inactivation research on microorganisms suspended in homogeneous fluids with a viscosity of 15 centipoises or less at the exposure temperature.     

Serum 25-hydroxyvitamin-D responses to multiple UV exposures from solaria: inferences for exposure to sunlight

We investigate the relationship between blood serum 25-hydroxyvitamin D (25(OH)D) and UV exposure from two artificial sources. We then use the results to test the validity of the action spectrum for vitamin D production, and to infer the production from summer and winter sunlight. The results are based on a two-arm randomised clinical trial of biweekly UV exposure for 12 weeks using two different types of dermatological booths: one emitting primarily UV-A radiation, and the other emitting primarily UV-B radiation (booth A and booth B respectively). In terms of the vitamin D production per unit erythema, one of the booths mimics summer noon sunlight, while the other mimics winter noon sunlight. Blood samples were taken before and after the exposures. For all participants, the phototherapy booth treatments arrested the usual wintertime decline in 25(OH)D, and for most the treatments from either booth resulted in significant increases. The increases were highly non-linear and there was a high degree of variability in 25(OH)D and its response to UV from person to person. By the end of the 12 week period, the mean increase was >30 nmol l(-1) from a cumulative exposure of 17 SED from the UV-A booth, and twice that for the UV-B booth for which the cumulative exposure was 268 SED. Assuming a logarithmic relationship between UV and vitamin D, the results for the two booths show no obvious inconsistency in the action spectrum for pre-vitamin D production. However, further measurements with similar exposures from each booth are required to confirm its validity. A model was developed to describe the increases in serum 25(OH)D resulting from the UV exposures, which differed markedly between the two booths. The deduced initial rate of increase of 25(OH)D was approximately 5 nmol l(-1) per SED. From the large increases in 25(OH)D from each booth, along with knowledge of the spectral distribution of sunlight and assuming the currently-accepted action spectrum for photo-conversion to pre-vitamin D, we infer that the production of 25(OH)D from sunlight should be possible throughout the year, although in winter the exposures necessary to maintain optimal levels of 25(OH)D would be impractically long. This finding is at variance with the commonly-held view that no vitamin D is produced at mid-latitudes in the winter. Further work is needed to resolve that inconsistency.     

Use of pulsed-field gel electrophoresis to link sporadic cases of invasive listeriosis with recalled chocolate milk.

Pulsed-field gel electrophoresis established the linkage between recalled chocolate milk and a multistate invasive listeriosis outbreak during a four-product recall period. Listeria monocytogenes isolates from four hospitalized patients and an environmental dairy sample displayed AscI restriction endonuclease digestion profiles identical to that of the chocolate milk isolate.     

OPTIMIZATION OF A CHOCOLATE PEANUT SPREAD USING RESPONSE SURFACE METHODOLOGY (RSM)

Response surface methodology was used to optimize formulations of chocolate peanut spread. Thirty-six formulations with varying levels of peanut (25-90%), chocolate (5-70%) and sugar (5-55%) were processed using a three-component constrained simplex lattice design. The processing variable, roast (light, medium, dark) was also included in the design. Response variables, measured with consumers (n = 60) participating in the test, were spreadability, overall acceptability, appearance, color, flavor, sweetness and texture/mouthfeel, using a 9-point hedonic scale. Regression analysis was performed and models were built for each significant (p < 0.01) response variable. Contour plots for each attribute, at each level of roast, were generated and superimposed to determine areas of overlap. Optimum formulations (consumer acceptance rating of ≥ 6.0 for all attributes) for chocolate peanut spread were all combinations of 29-65% peanut, 9-41% chocolate, and 17-36% sugar, adding up to 100%, at a medium roast. Verification of two formulations indicated no difference between predicted and observed values.     

Analysis of cocoa products for ochratoxin A and aflatoxins

Eighty-five samples of cocoa products sampled in Canada were analysed for ochratoxin A (OTA) and aflatoxins in 2011–2012. Inclusion of the aflatoxins in this survey required additional method development. Chocolate was extracted with methanol–water plus NaCl, while for cocoa two successive extractions with methanol and methanol–water were made. Extracts were cleaned on an AflaOchra immunoaffinity column (IAC). Determination was by reversed phase high performance liquid chromatography (HPLC). Detection of the aflatoxins was with a post-column photochemical reactor and of OTA by fluorescence detection. Mean limits of quantification (LOQ) of chocolate and cocoa powders were 0.16 ng/g (OTA) and 0.07 ng/g (aflatoxin B₁), respectively. Survey results showed that the incidences of OTA above the LOQ in natural cocoa were 15/15 (mean 1.17 ng/g), 20/21 for alkalized cocoa (mean 1.06 ng/g), 9/9 for baking chocolate (mean 0.49 ng/g), 20/20 for dark chocolate (mean 0.39 ng/g), 7/10 for milk chocolate (mean 0.19 ng/g), 5/5 for cocoa liquor (mean 0.43 ng/g), and 0/5 for cocoa butter. These results confirm our previous work with OTA. In the same samples, incidences of aflatoxin B₁ above the LOQ were 14/15 for natural cocoa (mean 0.86 ng/g), 20/21 for alkalized cocoa (mean 0.37 ng/g), 7/9 for baking chocolate (mean 0.22 ng/g), 16/20 for dark chocolate (mean 0.19 ng/g), 7/10 for milk chocolate (mean 0.09 ng/g), 4/5 for cocoa liquor (mean 0.43 ng/g), and 0/5 for cocoa butter. Both aflatoxins and OTA were confirmed by HPLC-MS/MS when OTA or aflatoxin levels found were above 2 ng/g in cocoa.     

The effect of benzene on the activity of adenosine deaminase in tissues of rats

Swiss Albino (Rat rattus norvegicus) rats were intraperitoneally injected with a 100 mg kg(-1) dosage of benzene, a toxic and carcinogenic agent widely used for industrial purposes. Changes in the adenosine deaminase (ADA) activity in the liver, kidney and serum of rats were investigated at 0, 2, 4, 8, 16, 32 and 64 h following injection. Serum physiological was administered to each control group. Enzyme activities were measured spectrophotometrically. Our purpose was to further investigations of some diseases caused by benzene, and present evidence of variations in the activity of ADA enzyme effected by benzene. While benzene caused significant inhibitions in ADA activity in the liver at 16 and 32 h and at 0.05 probability level, no significant inhibition or activation occurred at other test periods (hours). ADA activity did not present any significant variation in the kidneys. It was observed that ADA activity displayed similar patterns in the control groups. Comparisons of ADA activities in the two groups showed a statistically significant decrease between 4(th) and 64(th) hours (p< 0.05), demonstrating a direct correlation between benzene and its effects on ADA enzymes.     

Studies on the Thermal Destruction of Escherichia coli in Milk and Milk Products

Escherichia coli (ATCC no. 9637) was shown to have mean D values (slope of destruction rate curve) of 28.2, 5.1, 1.3, 0.00195, 0.00100, 0.00055, 0.00029, and 0.00016 min in milk at temperatures of 125, 130, 135, 168, 170, 172, 174, and 176 F, respectively. The mean D values for this organism in chocolate milk were 32.2, 10.4, 2.6, 0.00265, 0.00133, 0.00069, 0.00035, and 0.00028 min at respective temperatures of 125, 130, 135, 168, 170, 172, 174, and 176 F. Mean D values of 34.4, 10.0, 3.5, 0.00093, 0.00080, 0.00068, 0.00043, and 0.00036 min were found in cream for respective temperatures of 125, 130, 135, 168, 170, 172, 174, and 176 F. In ice cream mix the organism was found to have mean D values of 39.3, 15.2, 5.1, 0.00147, 0.00120, 0.00078, 0.00070, and 0.00053 min for temperatures of 125, 130, 135, 170, 172, 174, 176, and 178 F, respectively. The slopes of the thermal death time curves were found to be 10.2, 10.2, 10.0, and 10.3 F for this organism in milk, chocolate milk, 40% fat cream, and ice cream mix, respectively.