Early blowing in raw goats' milk cheese: gas production capacity of Enterobacteriaceae species present during manufacturing and ripening.

The aim of this study was to identify the main Enterobacteriaceae species responsible for early gas blowing during curdling and the first week of ripening in raw goats' milk cheese. Two batches of raw goats' milk cheese were selected. One of them showed early blowing within the first 24 h of cheese ripening while the other showed no alteration. Although initial levels of Enterobacteriaceae were similar in defective and non-defective cheese, their dynamics (growth and disappearance rates of the species detected) were different. Klebsiella oxytoca and Enterobacter cloacae were the main species in the defective curd, whereas Buttiauxela spp. was predominant in normal curd. Hafnia alvei was the prevailing isolated species for both normal and defective cheese throughout the ripening process. The highest gas production was rendered by K. oxytoca and H. alvei, mainly isolated from curd and cheese. However, other species relevant in milk or curd, like Pantoea ssp. or Buttiauxela spp. were considered as low gas producers. The analysis of digitalized images of cheese showed that most of the cheese eyes were formed before the first week of ripening, although this process continued during maturation.According to the species found in the defective and non-defective cheese, their proportions at different ripening stages, their ability to produce gas and eye formation, K. oxytoca might be considered the most likely responsible for early blowing in raw goats' milk cheeses; while H. alvei increased the eyes number in the later stages of the ripening period.

Improved detection of Escherichia coli and coliform bacteria by multiplex PCR.

BACKGROUND: The presence of coliform bacteria is routinely assessed to establish the microbiological safety of water supplies and raw or processed foods. Coliforms are a group of lactose-fermenting Enterobacteriaceae, which most likely acquired the lacZ gene by horizontal transfer and therefore constitute a polyphyletic group. Among this group of bacteria is Escherichia coli, the pathogen that is most frequently associated with foodborne disease outbreaks and is often identified by ß-glucuronidase enzymatic activity or by the redundant detection of uidA by PCR. Because a significant fraction of essential E. coli genes are preserved throughout the bacterial kingdom, alternative oligonucleotide primers for specific E. coli detection are not easily identified. RESULTS: In this manuscript, two strategies were used to design oligonucleotide primers with differing levels of specificity for the simultaneous detection of total coliforms and E. coli by multiplex PCR. A consensus sequence of lacZ and the orphan gene yaiO were chosen as targets for amplification, yielding 234 bp and 115 bp PCR products, respectively. CONCLUSIONS: The assay designed in this work demonstrated superior detection ability when tested with lab collection and dairy isolated lactose-fermenting strains. While lacZ amplicons were found in a wide range of coliforms, yaiO amplification was highly specific for E. coli. Additionally, yaiO detection is non-redundant with enzymatic methods.

Salt influence on surface microorganisms and ripening of soft ewe cheese.

The effect of different brining treatments on salt uptake and diffusion during the first 30 d of ripening was determined in soft ewe cheese. Additionally, salt influence on surface microorganisms and physicochemical parameters was evaluated. Cheeses were placed into different brine solutions (14, 18 and 24°Bé) at 5 and 10 °C for 1, 2 or 3 h. Samples from rind, outer core and inner core were analysed at 0, 7, 15 and 30 d. Complete salt diffusion from rind to the inner core took about 15 d. The resulting salt gradient favoured the development of a pH gradient from the surface to the inner core. Salt concentration also had a significant effect on the growth of surface microorganisms (mesophiles, pseudomonads and halotolerants). However, mould and yeasts were not affected throughout ripening by the salt levels achieved. Brine salting by immersion for 3 h at 10 °C in 24°B brine was found to be the most suitable treatment to control pseudomonads in cheese rind, as spoilage microorganism.

Responsiveness of the VISA-P scale for patellar tendinopathy in athletes.

BACKGROUND: Patient-reported outcome measures are increasingly used in sports medicine to assess results after treatment, but interpretability of change for many instruments remains unclear. OBJECTIVE: To define the minimum clinically important difference (MCID) for the Victorian Institute of Sport Assessment scale (VISA-P) in athletes with patellar tendinopathy (PT) who underwent conservative treatment. METHODS: Ninety-eight athletes with PT were enrolled in the study. Each participant completed the VISA-P at admission, after 1 week, and at the final visit. Athletes also assessed their clinical change at discharge on a 15-point Likert scale. We equated important change with a score of ≥3 (somewhat better). Receiver-operating characteristic (ROC) curve analysis and mean change score were used to determine MCID. Minimal detectable change was calculated. The effect of baseline scores on MCID and different criteria used to define important change were investigated. A Bayesian analysis was used to establish the posterior probability of reporting clinical changes related to MCID value. RESULTS: Athletes with PT who showed an absolute change greater than 13 points in the VISA-P score or 15.4-27% of relative change achieved a minimal important change in their clinical status. This value depended on baseline scores. The probability of a clinical change in a patient was 98% when this threshold was achieved and 45% when MCID was not achieved. CONCLUSIONS: Definition of the MCID will enhance the interpretability of changes in the VISA-P score in the athletes with PT, but caution is required when these values are used.

Stereoselective syntheses of the antihistaminic drug olopatadine and its E-isomer.

Practical stereoselective synthetic routes to the antihistaminic drug olopatadine and its E-isomer have been developed, the key steps being a trans stereoselective Wittig olefination using a nonstabilized phosphorus ylide and a stereoselective Heck cyclization. The stereoselectivity of the Wittig reaction depends on both the phosphonium salt anion and the cation present in the base used to generate the ylide.

Effects of a Long-Term Adapted Judo Program on the Health-Related Physical Fitness of Children with ASD.

Physical fitness is one of the most important physical and mental health aspects for children with Autism Spectrum Disorder (ASD). This study aimed to test the effects of a long-term adapted judo program on the health-related physical fitness of children with ASD. The participants were recruited from various associations of families and schools for children with special needs. Twenty-one children were assigned to an experimental group and nineteen to a control group. The experimental group participated in a six-month adapted judo program consisting of 90 min of practice each week. Health-related physical fitness was measured using the indicators obtained from the ALPHA-fitness battery, the estimated VO2max and the waist/height ratio0.5. Changes within and between groups were analyzed using linear mixed models for repeated measures designs and test-retest reliability of tests requiring a maximum score using the Intraclass Correlation Coefficient (ICC). A judo program tailored for children with ASD can improve the cardio-metabolic health and cardiorespiratory fitness of its participants. The problems involved with administering physical aptitude tests that involve maximum effort or performance in children with ASD cast serious doubts on the reproducibility of their results.

Dorsomedial pallium lesions impair taste aversion learning in goldfish.

The present work shows that the dorsomedial telencephalic pallium of teleost fish, proposed as homologous to the amygdala of mammals, is involved in taste aversion learning (TAL). To analyze the behavioral properties of TAL in goldfish, in Experiment 1, we used a delayed procedure similar to that employed with mammals, which consists of the presentation of two flavors on different days, one followed by lithium chloride and the other by saline, both after a 10-min delay. The results showed that goldfish developed a strong aversion to the gustatory stimulus followed by visceral discomfort and that, as in mammals, this learning was rapidly acquired, highly flexible and maintained for a long time. Experiment 2 showed that dorsomedial pallium lesions and the ablation of the telencephalic lobes impaired the acquisition of taste aversion in goldfish, whereas damage to the dorsolateral pallium (hippocampus homologue) or cerebellar corpus did not produce significant changes in this learning. Experiment 3 showed that these TAL deficits were not due to a lesion-related disruption of taste discrimination; goldfish with telencephalon ablation were able to learn to distinguish between the two tested flavors in a differential conditioning procedure. These functional data demonstrate that the dorsomedial pallium in teleosts is, like the amygdala, an essential component of the telencephalon-dependent taste aversion memory system and provide further support concerning the homology between both structures.

Behavioural Improvements in Children with Autism Spectrum Disorder after Participation in an Adapted Judo Programme Followed by Deleterious Effects during the COVID-19 Lockdown.

The public health lockdown prompted by the novel coronavirus (COVID-19) pandemic, which included school closures that may have potentially serious consequences for people with disabilities or special educational needs, disrupted an ongoing adapted judo training intervention in children with Autism Spectrum Disorder (ASD). The purpose of this study was to compare repetitive behaviours, social interaction, social communication, emotional responses, cognitive style and maladaptive speech scores across four time-points: baseline, after an eight-week control period, after an eight-week judo intervention and after an eight-week lockdown period due to COVID-19. The sample consisted of 11 children diagnosed with ASD according to the criteria of the Diagnostic and Statistical Manual of Mental Disorders-Fifth Edition (DSM-V), with an intelligence quotient (IQ) range between 60 and 70. Significant improvements were shown following the judo intervention period compared to the baseline and control periods. However, the same values significantly declined during the COVID-19 lockdown period resulting in values lower than those recorded at baseline, and following the control period and the judo intervention. The decline in psychosocial and behavioural scores are likely due to the stress caused by the sudden halt in activity and the increase in sedentary practices associated with the lockdown.

Corrigendum: Diversity and Local Coadaptation of Escherichia coli and Coliphages From Small Ruminants.

[This corrects the article DOI: 10.3389/fmicb.2020.564522.].

Diversity and Local Coadaptation of Escherichia coli and Coliphages From Small Ruminants.

Bacteriophages are highly specific predators that drive bacterial diversity through coevolution while striking tradeoffs among preserving host populations for long-term exploitation and increasing their virulence, structural stability, or host range. Escherichia coli and other coliform bacteria present in the microbiota of milk and during early ripening of raw milk cheeses have been linked to the production of gas, manifested by the appearance of eyes, and the development of off-flavors; thus, they might cause early blowing and cheese spoilage. Here, we report the characterization of coliphages isolated from manure from small ruminant farms and E. coli strains isolated from goat and sheep raw milk cheese. Additionally, the virulence and host range of locally isolated and laboratory collection phages were determined by comparing the susceptibility of E. coli strains from different sources. In agreement with the high genetic diversity found within the species E. coli, clustering analysis of whole-cell protein revealed a total of 13 distinct profiles but none of the raw milk cheese isolates showed inhibition of growth by reference or water-isolated coliphages. Conversely, 10 newly isolated phages had a broad host range (i.e., able to lyse ≥50% of bacterial hosts tested), thus exhibiting utility for biocontrol and only one cheese-isolated E. coli strain was resistant to all the phages. Whereas there was a high positive correlation between bacterial susceptibility range and lysis intensity, the phages virulence decreased as range increased until reaching a plateau. These results suggest local gene-for-gene coevolution between hosts and phages with selective tradeoffs for both resistance and competitive ability of the bacteria and host-range extension and virulence of the phage populations. Hence, different phage cocktail formulations might be required when devising long-term and short-term biocontrol strategies.

Quaternary structural transitions in the DeoR-type repressor UlaR control transcriptional readout from the L-ascorbate utilization regulon in Escherichia coli.

UlaR is a DNA binding protein of the DeoR family of eubacterial transcriptional repressors which maintains the utilization of the L-ascorbate ula regulon in a repressed state. The availability of L-ascorbate in the growth medium releases UlaR-mediated repression on the ula regulon, thereby activating transcription. The molecular details of this induction by L-ascorbate have remained elusive to date. Here we have identified L-ascorbate 6-phosphate as a direct effector of UlaR; using a combination of site-directed mutagenesis, gel retardation, isothermal titration calorimetry, and analytical ultracentrifugation studies, we have identified the key amino acid residues that mediate L-ascorbate 6-phosphate binding and constructed the first model of regulation of a DeoR family member, establishing the basis of the ula regulon transcription control by UlaR. In this model, specific quaternary rearrangements of the DeoR-type repressor are the molecular underpinning of the activating and repressing forms. A DNA-bound UlaR tetramer establishes repression, whereas an L-ascorbate-6-phosphate-induced breakdown of the tetrameric configuration in favor of an UlaR dimeric state results in dissociation of UlaR from DNA and allows transcription of ulaG and ula ABCDEF structural genes. Despite the fact that similar changes have been described for other unrelated repressor factors, this is the first report to demonstrate that specific oligomerization changes are responsible for the activating and repressing forms of a DeoR-type eubacterial transcriptional repressor.

Telencephalon ablation impairs goldfish allocentric spatial learning in a "hole-board" task.

The present work analyzes the involvement of telencephalon of goldfish in spatial strategies, using a procedure analogue to the hole-board task. With this aim, goldfish with sham operation or telencephalon ablation were trained to find a baited feeder within a twenty-five feeder matrix, which maintained stable spatial relationships relative to five peripheral landmarks. After training, different types of probe tests were conducted. Although both groups learned the task, probe trials showed that whereas the sham animals used the whole configuration of cues to implement map-like strategies the telencephalic animals used a guidance strategy based on cues located in the vicinity of the baited feeder. These results confirm the role of teleost fish telencephalon in the use of allocentric strategies obtained with other spatial procedures, and indicate that the hole-board task described here for goldfish is a useful tool to assess the neural bases of spatial cognition in teleost fish.

Predictors of therapeutic outcome and process.

This article presents an intensive study of the outcome and process of the psychotherapeutic treatments conducted at the Blanquerna Psychotherapy Service in Barcelona. It analyzes in detail the relation between outcome (improvement) and process (therapeutic alliance). Specifically, it discusses (a) the therapeutic process in terms of the relation between therapeutic alliance and psychotherapy outcome and (b) the effects of the temporary weakening of the therapeutic alliance and temporary symptomatic relapse on therapeutic outcome and process. Two hundred thirty-nine clients were included in the study, and the assessment instruments were the CORE Outcome Measure and the short version of the Working Alliance Inventory. In addition to good outcome of psychotherapy and maintenance of therapeutic change after treatment, the results also demonstrated the significance of different aspects of the therapeutic alliance as predictors of the therapeutic outcome and process.

Enantioselective spirocyclizations from tryptophanol-derived oxazolopiperidone lactams.

A straightforward synthetic route to enantiopure spiro[indole-3,3'-indolizidines] is reported. The key step is a Lewis acid promoted cyclization of a Na-tosyltryptophanol-derived oxazolopiperidone lactam in the presence of Et3SiH.

Épulis congénito gigante: importancia del diagnóstico prenatal / Congenital giant cell epulis: Importance of the prenatal diagnosis

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Development of different spatial frames of reference for orientation in small-scale environments.

BACKGROUND: Previous studies show that there is a developmental transition in the frames of reference children use to orientate from a body-centered to an allocentric strategy. However, there is no agreement concerning the age at which they begin to integrate and flexibly use both strategies in small-scale environments. METHOD: 6-10-year-old children and adults were trained to locate a hidden object in an arm-maze placed within a small-scale model, which maintained stable relationships with the frames of reference provided by the experimental room and by the subject (Experiment 1), and in a situation of inconsistency between the frame provided by the small-scale model and the other two (Experiment 2). RESULTS: When the frames of reference provided by the room and by the subject conflict with that of the manipulative space, the performance deteriorates compared to the situation when multiple frames of reference can be used cooperatively to locate the goal. The flexible use of the information provided by the model (i.e., the cues surrounding the maze and the geometrical features) emerged at 10 years. CONCLUSIONS: Through development, children acquire new spatial abilities and increasing flexibility in the conjoint use of egocentric and allocentric frames of reference in small-scale environments.

Cross-cultural adaptation of VISA-P score for patellar tendinopathy in Spanish population.

STUDY DESIGN: Clinical measurement. OBJECTIVES: To adapt the VISA-P questionnaire into Spanish and to assess its psychometric properties. BACKGROUND: Health status questionnaires and scales to report outcomes are increasingly used in medical research and clinical practice. Validated versions of these tools are necessary to avoid bias during use in different languages and cultures. METHODS: We followed international recommendations to perform cross-cultural adaptation. The Spanish VISA-P (VISA-P-Sp) questionnaire and the Short Form (36) Health Survey (SF-36) were administered to 150 individuals: 40 healthy students, 40 professional players in sports requiring jumping, 40 athletes with patellar tendinopathy, and 30 athletes with knee injuries other than patellar tendinopathy. Participants were assessed at baseline and after 1 week. Athletes with tendinopathy also completed questionnaires and other knee measures (the Kujala Scoring Questionnaire and the Cincinnati Knee Rating Scale) after physiotherapy treatment, which consisted of rest, ice, eccentric exercise, electrotherapy, and manual therapy. RESULTS: The VISA-P-Sp showed high reliability for both temporal stability (intraclass correlation coefficient [ICC] = 0.994; 95% CI: 0.992, 0.996) and internal consistency (Cronbach α = 0.885). Based on a factor analysis, a 2-factor solution explained 76.1% of the variance. The VISA-P-Sp score in the tendinopathy group was significantly correlated with scores on other knee scales (Kujala score [Spearman rho = 0.897; P<.001] and Cincinnati scale [Spearman rho = 0.782, P<.001]) and with SF-36 physical components score (Spearman rho>0.6, P<.001). The standardized size effect was 1.14, and the standardized response mean was 1.17. CONCLUSION: The VISA-P-Sp questionnaire proved to be a valid and reliable instrument, sensitive to clinical changes and comparable to the original English-language version.

Neuropsychology of learning and memory in teleost fish.

Traditionally, brain and behavior evolution was viewed as an anagenetic process that occurred in successive stages of increasing complexity and advancement. Fishes, considered the most primitive vertebrates, were supposed to have a scarcely differentiated telencephalon, and limited learning capabilities. However, recent developmental, neuroanatomical, and functional data indicate that the evolution of brain and behavior may have been more conservative than previously thought. Experimental data suggest that the properties and neural basis of learning and memory are notably similar among teleost fish and land vertebrates. For example, lesion studies show that the teleost cerebellum is essential in classical conditioning of discrete motor responses. The lateral telencephalic pallium of the teleost fish, proposed as homologous to the hippocampus, is selectively involved in spatial learning and memory, and in trace classical conditioning. In contrast, the medial pallium, considered homologous to the amygdala, is involved in emotional conditioning in teleost fish. The data reviewed here show a remarkable parallelism between mammals and teleost fish concerning the role of different brain centers in learning and memory and cognitive processes. These evidences suggest that these separate memory systems could have appeared early during the evolution of vertebrates, having been conserved through phylogenesis.

Desarrollo de diferentes marcos de referencia espacial para la orientación en entornos a pequeña escala / Development of different spatial frames of reference for orientation in small-scale environments

Background: Previous studies show that there is a developmental transition in the frames of reference children use to orientate from a body-centered to an allocentric strategy. However, there is no agreement concerning the age at which they begin to integrate and flexibly use both strategies in small scale environments. Method: 6-10-year-old children and adults were trained to locate a hidden object in an arm-maze placed within a small-scale model, which maintained stable relationships with the frames of reference provided by the experimental room and by the subject (Experiment 1), and in a situation of inconsistency between the frame provided by the small scale model and the other two (Experiment 2). Results: When the frames of reference provided by the room and by the subject conflict with that of the manipulative space, the performance deteriorates compared to the situation when multiple frames of reference can be used cooperatively to locate the goal. The flexible use of the information provided by the model (i.e., the cues surrounding the maze and the geometrical features) emerged at 10 years. Conclusions: Through development, children acquire new spatial abilities and increasing flexibility in the conjoint use of egocentric and allocentric frames of reference in small-scale environments (AU)

Antecedentes: estudios previos muestran una transición durante el desarrollo en las estrategias que los niños utilizan para orientarse, aunque no hay consenso en la edad de inicio para emplear conjuntamente estrategias alocéntricas y egocéntricas en entornos a pequeña escala. Método: niños de 6-10 años y adultos fueron entrenados para encontrar un objeto escondido en un laberinto radial ubicado en una maqueta que mantiene una relación constante con los marcos de referencia proporcionados por la habitación experimental y por el participante (experimento 1), y en una situación de inconsistencia entre el marco de referencia del entorno a pequeña escala y los de la habitación y el sujeto (experimento 2). Resultados: cuando los marcos de referencia de la habitación y el sujeto entran en conflicto con el del espacio manipulativo, la ejecución empeora respecto a una situación en que múltiples marcos de referencia cooperan para localizar la meta. Desde los 10 años los niños usan la información del espacio manipulativo, tanto las claves que rodean al laberinto como la geometría de dicho entorno. Conclusiones: durante el desarrollo los niños adquieren nuevas habilidades espaciales y mayor flexibilidad en el uso conjunto de marcos de referencia egocéntricos y alocéntricos en entornos a pequeña escala(AU)