Delayed diagnosis in Hirschsprung disease.

INTRODUCTION: Diagnostic delay in Hirschsprung disease is uncommon. Different definitions have been proposed but that of a diagnosis achieved after 12 months of age seems to be the most reliable and resorted to. Some authors reported a worse outcome in case of delay. Our study aims at providing the most relevant features of a series of patients who received a delayed diagnosis of Hirschsprung disease. MATERIALS AND METHODS: All consecutive patients admitted to our Center with a delayed diagnosis of Hirschsprung diseases between January 2017 and July 2023 have been retrospectively enrolled. Demographic data, phenotype, genotype, surgical complications, and outcome were assessed and compared to those of literature. A number of variables were also compared to those of a series of patients admitted during the same study period without a delayed diagnosis. RESULTS: A total of 45 patients were included (16.4% out of a series of 346 patients with data regarding age at diagnosis). Male to female ratio was 3.1:1. Median age at diagnosis was 41 months with a wide variation (range between 17 months and 58 years). All patients but 2 suffered from classic rectosigmoid aganglionosis. Normal meconium passage (58%) was reported in a significantly higher number of patients compared to what observed in a series without diagnostic delay (p = 0.0140). All other variables (associated anomalies, preoperative enterocolitis, complications, and functional outcome) proved not to have statistically significant differences compared to a series of patients without a diagnostic delay. CONCLUSIONS: The results of our study underline that a significant percentage of patients are basically missed in the neonatal period mostly due to mild symptoms. Overall outcome does not differ from that of patients without diagnostic delay. Nonetheless, we underline the importance of a throughout investigation of all patients with meconium delay/failure and that of adopting a low threshold for performing rectal suction biopsies in constipated children to avoid misdiagnosis to serve the best for our patients.

A unicentric cross-sectional observational study on chronic intestinal inflammation in total colonic aganglionosis: beware of an underestimated condition.

BACKGROUND: Inflammatory Bowel Diseases (IBD) are known to occur in association with Hirschsprung disease (HSCR). Most of cases are represented by Crohn Disease (CD) occurring in patients with Total Colonic Aganglionosis (TCSA) with an estimated prevalence of around 2%. Based on these considerations and on a number of provisional data belonging to our Center for Digestive Diseases, we developed a unicentric cross-sectional observational study aimed at describing phenotype, genotype, pathology and metagenomics of all patients with TCSA and Crohn-like lesions. RESULTS: Out of a series of 62 eligible TCSA patients, 48 fulfilled inclusion criteria and were enrolled in the study. Ten patients did not complete the study due to non-compliance or withdrawal of consent and were subsequently dropped out. A total of 38 patients completed the study. All patients were tested for chronic intestinal inflammation by a combination of fecal calprotectine (FC) or occult fecal blood (OFB) and underwent fecal metagenomics. Nineteen (50%) tested positive for FC, OFB, or both and subsequently underwent retrograde ileoscopy. Fourteen patients (36.8%) presented Crohn-like lesions, occurring after a median of 11.5 years after surgery (range 8 months - 21.5 years). No statistically significant differences regarding demographic, phenotype and genotype were observed comparing patients with and without lesions, except for need for blood transfusion that was more frequent in those with lesions. Faecal microbiome of patients with lesions (not that of caregivers) was less biodiverse and characterized by a reduction of Bacteroidetes, and an overabundance of Proteobacteria. FC tested negative in 3/14 patients with lesions (21%). CONCLUSIONS: Our study demonstrated an impressive 10-folds higher incidence of chronic inflammation in TCSA. Up to 50% of patients may develop IBD-like lesions postoperatively. Nonetheless, we failed in identifying specific risk factors to be used to implement prevention strategies. Based on the results of our study, we suggest screening all TCSA patients with retrograde ileoscopy regardless of FC/OFB values. The frequency of endoscopic assessments and the role of FC/OFB screening in prompting endoscopy is yet to be determined.

The effectiveness of subgingival irrigant ozone-based as adjuvant for non-surgical periodontal therapy in the treatment of chronic periodontitis: a review.

The aim of this review was to establish the efficacy of Ozone therapy (OT) in Periodontal disease's (PD) tratment. An electronic search of publications was established from two electronic databases: PubMed and Web of Science. The search strategy used a combination of controlled vocabulary and freetext words. Inclusion and exclusion criteria were defined by the authors before the start of the study. The inclusion criteria were: all studies published in English language; studies in vivo on humans; studies analyzing the correlation between PD and OT. The search resulted in 102 titles. Only 9 articles were included in the quantitative analysis. By the analyzed studies a strictly connection is still not evincible. More clinical trials are recommended to investigate the role of ozone, in its various types of topical administration, as a valid subgingival irrigant to achieve the goal of periodontal tissue health.

Full-digital workflow to rehabilitate a high value aesthetic zone. A case report.

Replacing missing teeth in the aesthetic area is a clinical challenge which must be solved by a multidisciplinary approach in order to obtain an esthetic and functional implant-prosthetic rehabilitation. First therapeutic choice should be the less invasive option, in accordance with patients' expectations. The present clinical case presents the rehabilitation of the upper incisor group using a full-digital workflow to reduce operative time, costs and patient discomfort. The use of computer-guided-surgery and 3D technologies, as intraoral scanners or cone-beam-computed-tomography, allows the clinician to visualize all patient's information when planning the rehabilitation in order to obtain a more predictable and a less invasive surgery.

Clinical fitting of a cast metal post and core obtained by means of an intraoral optical scanning (IOS) and digital workflow.

Customization of post-and-cores using computer-aided-design and computer-aided-manufacturing (CAD-CAM) requires the scanning of a pattern and the subsequent digital design. This case report describes the production of a CAD-CAM customized post-and-core designed from an intraoral scan and milled from a metal block. The use of an intraoral scanner (IOS) for post-endodontic rehabilitation could lead to a faster and more efficient CAD-CAM customized post-and-core realization. The use of a high resistance material such as metal is paramount in cases with high loss of coronal structure. The patient has been treated with bisphosphonate (BP) for years. The risk of osteonecrosis of the jaw after extraction was high.

Genome Sequence of Rhizobium sullae HCNT1 Isolated from Hedysarum coronarium Nodules and Featuring Peculiar Denitrification Phenotypes.

The genome sequence of Rhizobium sullae strain HCNT1, isolated from root nodules of the legume Hedysarum coronarium growing in wild stands in Tuscany, Italy, is described here. Unlike other R. sullae strains, this isolate features a truncated denitrification pathway lacking NO/N2O reductase activity and displaying high sensitivity to nitrite under anaerobic conditions.

Carotid plaque instability is not related to quantity but to elemental composition of calcification.

BACKGROUND AND AIMS: Recent studies highlighted the role of calcification processes in the clinical progression of chronic cardiovascular diseases. In this study we investigated the relationship between the chemical composition of calcification and atherosclerotic plaque stability in carotid arteries. METHODS AND RESULTS: To this end, we characterized the calcification on 229 carotid plaques, by morphology, immunohistochemistry, transmission electron microscopy and energy dispersive X-ray microanalysis. Plaques were classified into two categories: unstable and stable. No significant differences were found in the incidence of the various risk factors between patients with and without carotid calcification, with the exception of diabetes. The energy dispersive X-ray microanalysis allowed us to identify two types of calcium salts in the atheromatous plaques, hydroxyapatite (HA) and calcium oxalate (CO). Our results showed that calcification is a common finding in carotid plaques, being present in 77.3% of cases, and the amount of calcium is not a factor of vulnerability. Noteworthy, we observed an association between HA calcification and unstable plaques. On the contrary, CO calcifications were mainly detected in stable plaques. CONCLUSIONS: The presence of different types of calcification in atheromatous plaques may open new perspectives in understanding the molecular mechanisms of atheroma formation and plaque instability.

Iron profile in Thoroughbreds during a standard training program.

OBJECTIVES: The aim of this study was to evaluate the modifications of some haematological parameters and iron profile during an 80-day training program and their relationship with athletic performance of performance horses. METHODS: During the training period, 20 Thoroughbreds were subjected to a standardised exercise test, previously standardised for gallopers to evaluate athletic performance. The test was performed immediately before the beginning of the training program (T0) and after 30 days (T30), 60 days (T60) and 90 days (T90). For each horse, their velocities at blood lactate concentrations of 2 and 4 mmol/L (V2 and V4) and a heart rate of 200 beats/min (V200) were calculated to assess the effect of the training program on athletic performance. Blood samples were collected at 20-day intervals over a period of 80 days from the beginning of the training program. RESULTS: One-way repeated measures analysis of variance (ANOVA) showed a statistically significant effect of days of training (time) on iron, ferritin, transferrin, total iron-binding capacity, transferrin saturation and unsaturated iron-binding capacity (P < 0.001 for all). CONCLUSIONS: The results showed the influence of the intensity and duration of exercise on iron metabolism. These findings may be useful for evaluating the athletic status of horses during training and to assess the degree of training adaptability, providing an opportunity to modify the training schedule to achieve the desired performance.

Changes in blood coagulation induced by exercise training in young athletic horses.

Prothrombin Time (PT), Activated Partial Prothrombin Time (APTT), Fibrinogen concentration (Fbg) and Platelet number (Plt) were evaluated in 20 young athletic horses during a training program. A standardized exercise test (SET) was performed every month for three months. The V4 variations (the speed, in m/min, reached at the blood lactate concentration of 4 mmol/l) obtained for each test were calculated to assess the effect of training program on athletic performance. Blood samples were collected at 20-day intervals over a period of 80 days from the beginning of the training program. The V4 (P < 0.001), PT (P < 0.001), APTT (P < 0.01), Fbg (P < 0.01) and Plt (P < 0.001) varied throughout the training period showing that the modifications of clotting mechanism in response to training period may be considered as a normal physiological response of the hemostatic system to training exercise.

Sulfate influx on band 3 protein of equine erythrocyte membrane (Equus caballus) using different experimental temperatures and buffer solutions.

The aim of this study was to assess the anion transport in equine erythrocytes through the measurement of the sulfate uptake operating from band 3 using different experimental temperatures and buffer solutions. Blood samples of six clinically healthy horses were collected via jugular vein puncture, and an emochrome-citometric examination was performed. The blood was divided into four aliquots and by centrifugation and aspiration the plasma and buffy coat were carefully discarded. The red blood cells were washed with an isosmotic medium and centrifuged. The obtained cell suspensions were incubated with two different experimental buffer solutions (buffer A: 115 mM Na2SO4, 10 mM NaCl, 20 mM ethylenediaminetetraacetic acid, 30 mM glucose; and buffer B: 115 mM Na2SO4, 10 mM NaCl, 20 mM ethylenediaminetetraacetic acid, 30 mM MgCl2) in a water bath for 1 h at 25 °C and 37 °C. Normal erythrocytes, suspended at 3% hematocrit, were used to measure the SO4= influx by absorption spectrophotometry at 425 nm wavelength. Unpaired Student's t-test showed a statistically significant decrease (P < 0.01) of rate constants in equine erythrocytes at 25 °C versus 37 °C using both experimental buffer solutions. Comparing the buffer A with buffer B unpaired Student's t-test showed statistically lower values (P < 0.0001) for A solution versus B solution both at 25 °C and at 37 °C. The greater inhibition of SO4 (=) influx measured in equine erythrocytes indicates the increased formation of the sulfydryl bonds in band 3 and the modulation of the sulfydryl groups, culminating in the conformational changes in band 3.

Eating-related environmental factors in underweight eating disorders and obesity: are there common vulnerabilities during childhood and early adolescence?

OBJECTIVE: This study aimed to examine whether there is an association between individual, social and family influences and dysfunctional eating patterns early in life and the likelihood of developing a subsequent underweight eating disorder (ED) or obesity. METHOD: The total sample comprised 152 individuals (underweight ED, n = 45; obese patients, n = 65; healthy controls; n = 42) from Barcelona, Spain. The Cross-Cultural Questionnaire (CCQ) was used to assess early eating influences as well as individual and family eating patterns and attitudes towards food. RESULTS: Even though a few shared eating influences emerged for both groups, unique factors were also observed. Whereas relationship with friends, teasing about eating habits by family members and the mass media were of specific relevance to the underweight ED group, the patient's own physical appearance, body dissatisfaction, teasing about eating habits by friends, teasing about body shape by family members and dysfunctional eating patterns were unique to obesity. CONCLUSIONS: Overlapping environmental risk factors provide evidence for integral prevention and intervention approaches that simultaneously tackle a range of weight-related problems. The unique factors might be important for targeting high-risk individuals.

Effect of storage time on haematological parameters in mullet, Mugil cephalus.

The aim of the present study was to assess the effect of storage time at +4 °C on red blood cell count (RBC), haematocrit (Hct), haemoglobin (Hb), white blood cell count (WBC), thrombocyte count (TC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) in mullet (Mugil cephalus) using an automatic method. After blood collection (T0 ), all samples were analyzed using both the manual and automatic method. To test the validation of the automatic method, a paired t-test was applied, and no statistical difference was observed. The samples were successively divided into four different aliquots and stored at +4 °C to assess the haematological parameters using the automatic method. The first aliquot was refrigerated for 6 h, the second one for 24 h, the third one for 48 h and the last one at for 72 h. One-way repeated-measures ANOVA showed a significant effect of storage time (P < 0.05) on Hb, WBC, TC, MCH and MCHC. These results suggest that haematological parameters can be assessed within 6 h from blood collection when samples are stored at +4 °C because long-term storage modifies the results of the analyses. Further studies on these parameters could be still needed in various fish species to validate an appropriate method for haematological analysis useful not only for the evaluation of the health status of animal living in captivity and in aquaculture but also to have reliability environmental haematological biomarkers.

ADP-induced platelet aggregation after addition of tramadol in vitro in fed and fasted horses plasma.

Adenosine diphosphate (ADP)-induced platelet aggregation in fed and fasted horses after addition of tramadol hydrochloride was evaluated in vitro. On 10 horses citrated blood samples were collected 2h after feeding (fed animals) and 21 h after feeding (fasted animals). Final concentrations of ADP 1 and 0.5 µM, and tramadol hydrochloride (1, 15, 30, 45 and 60 min after the addition of tramadol) were used to determine the maximum degree and initial velocity of platelet aggregation. Repeated measures multifactor analysis of variance (MANOVA) was used to evaluate the effect of feeding/fasting condition, ADP concentration and addition of tramadol. Findings showed statistical differences (P≤0.05) on studied parameters after addition of tramadol to different ADP concentrations in fed and fasted horses. The clinical relevance of these results is that tramadol provides many advantages as a therapeutic option; in fact, it is an inexpensive and a relatively new analgesic in equine veterinary medicine. Further investigations would be appropriate to compare the effects of different opioids but also using different concentrations of tramadol associated with other drugs in order to have substances which can regulate the functional activity of the platelets and to extend the knowledges on equine platelet aggregation.

Influence of transportation on serum concentrations of acute phase proteins in horse.

The modifications of Haptoglobin (Hp), Serum Amyloid A (SAA), Fibrinogen (Fbg) and White Blood Cells (WBCs) were evaluated in 15 Saddle Italian horses. Ten horses were transported covering a distance of about 320 km within 4 h with an average speed of 80 km/h (experimental group) and five horses were not subject to transportation (control group). Blood was collected via jugular venipuncture before the transportation (T0), immediately after the transportation (T1), 12 (T12), 24 (T24) and 48 (T48)hours after the transportation in experimental group and at the same time point in control group. For each parameter statistical analysis of different groups and sampling time was performed using a two-way analysis of covariance, with the data before the transportation (T0) as the covariate, by the GLM procedure of SAS. For all parameters the interaction (Group × Time) was tested and it was resulted no significant. The application of statistical analysis showed significant differences between the control group and horses subjected to transportation (P<0.01), and the influence of sampling time (P<0.05) on Hp, SAA and WBCs. These modifications appeared to be innovative showing that equine Hp, generally considered as moderate acute phase protein, increases more rapidly than the SAA after transportation-induced stress.

Oxidative stress and band 3 protein function in Liza aurata and Salmo irideus erythrocytes: effect of different aquatic conditions.

The aim of this study was to assess the effect of the different aquatic conditions on anion transport of fish erythrocytes through the measurement of the sulphate uptake operating from band 3, the determination of reduced glutathione (GSH) and oxidized glutathione (GSSG). To this purpose, blood samples of 30 Liza aurata and 30 Salmo irideus fishes were collected via caudal vein and washed with physiological buffer. Successively, erythrocytes suspended at 3% haematocrit were used to measure the SO(4) (=) influx by atomic absorption spectrophotometry at 425 nm wavelength and the GSH concentration using an immunodiagnostic assay intended for the quantitative determination of glutathione in ethylenediaminetetraacetic acid in blood. All results were analyzed using unpaired Student's t-test and a P < 0.05 was considered statistically significant. The results of this study showed statistically significant differences about the sulphate uptake and the glutathione levels in S. irideus with respect to L. aurata (P < 0.0001). In conclusion, the different aquatic conditions play an important role on anion transport in fish erythrocytes, showing that environmental differences induce perturbations in erythrocyte membrane and should be evaluated to prevent physiological damages in fishes.

Influence of temperature and humidity on rumen pH and fatty acids in dairy cows.

The aim of this study was to investigate the variations of rumen pH and fatty acids (acetic acid, propionic acid, iso-butyric acid, n-butyric acid, iso-valerianic acid, n-valerianic, caproic acid and total fatty acids) in 245 early lactating dairy cows under different temperature and humidity conditions. The animals were divided into six groups and rumen fluid was collected by rumenocentesis on 22 dairy cows in April (Group A), 33 in May (Group B), 43 in June (Group C), 48 in July (Group D), 36 in September (Group E) and 60 in October (Group F). One-way analysis of variance (ANOVA), followed by the Bonferroni's test, showed a significant effect of environmental variations on all studied parameters (P < 0.0001). Changes in studied parameters can be explained in relation to the microbial population and shift in the optima for rumen conditions associated with variations of environmental conditions. We can affirm that the microbial assemblages that underlie energy and protein supply to wild ruminant are evident especially in relation to temperature and humidity conditions.

Electrophoretic serum protein fraction profile during the different physiological phases in Comisana ewes.

The aim of this study was to evaluate the influence of different physiological phases on serum total proteins and their fractions of ten Comisana ewes housed in Mediterranean area. From each animal, blood samples were collected at different physiological phases: late pregnancy, post-partum, early, mid-, end lactation and dry period. On all samples serum total proteins were determined by the biuret method, and albumin, α-globulins, ß(1) -globulins, ß(2) -globulins and γ-globulins concentrations were assessed using an automated system. One-way repeated measures analysis of variance was applied to determine the significant effect of different physiological phases on the parameters studied. During the late pregnancy and post-partum, total proteins, ß1- and ß2-globulins and γ-globulins showed the highest values. Starting from post-partum, α-globulins increased to reach their peaks in mid-lactation. Early lactation was characterized by low γ-globulins values. The increase in serum albumin concentration and the drop in some globulin fractions determined the significant increase in albumin/globulin ratio. The obtained results contributed to improve the knowledge on electrophoretic profile during the different physiological phases in ewes, confirming that pregnancy and lactation periods affect the protein metabolism. Particularly, serum protein fractions pattern could give information about dehydration, plasma volume expansion and hepatic function, which occur during the different physiological phases. Dynamics of the protein profile - from pregnancy to dry period - which are provided by our results, could be considered as guidelines for the management strategies to guarantee the nutritional needs of these animals during the different physiological phases and to avoid a decline of productive performance and consequently an economic loss.

Effect of different workload and hydrocortisone in vitro on platelet aggregation in athletic horse.

The objective of the present study was to evaluate the platelet response to different workload in athletic horse and the influence of hydrocortisone on this response. For this purpose, six Sella Italiana gelding athletic horses, ranging in age from 8 to 10 years and clinically healthy, were subjected to four different exercises: horse-walker, treadmill, obstacle course of 1.00 m and obstacle course of 1.25 m. From all the horses, citrated plasma samples were collected at rest, immediately after exercise and 30 min after the end of the exercise to assess the maximum degree of platelet aggregation and the initial velocity of aggregation (slope) after platelet activation with 10 microM ADP and 20 microM ADP. Successively, the effect of 20 minutes pre-incubation with hydrocortisone in ADP-induced aggregation was determined. One-way repeated measures analysis of variance (ANOVA) was applied to determine significant differences due to experimental conditions (P < 0.05) and, for each sampling time, one-way analysis of variance was applied to determine significant differences due to workload (P < 0.05). Bonferroni's test was applied for post-hoc comparison. Our results revealed a statistically significant effect of experimental conditions only after obstacle course of 1.00 m and 1.25 m. These findings showed an inhibition of platelet aggregation during different workload in athletic horses that probably represents a protective endothelial mechanism through the production of nitrix oxide, and a kinetic measurement of the reaction that changes in relation to percentage of aggregation. The mechanism of hydrocortisone had no effect on platelet receptor expression in vitro in athletic horses, so it might be useful to evaluate the effect of different anti-inflammatory steroids in order to confirm, or note any differences, with drugs that have the same mechanism of action.

Modifications of some acute phase proteins and the white blood cell count in thoroughbreds during training.

The concentrations of plasma fibrinogen and serum haptoglobin and the white blood cell count (WBC) were assessed in 17 thoroughbreds. All horses were trained for 80 days, six days a week, with one day of rest per week. Blood samples were collected from each animal by jugular venepuncture, at 06.00, every 20 days for a period of 80 days, and the fibrinogen and haptoglobin concentrations and WBC were determined. One-way repeated measures analysis of variance, followed by the Bonferroni test, was used to determine significant differences. Training was associated with a significant increase in the concentrations of plasma fibrinogen (P<0.01) and serum haptoglobin (P<0.0005), but had no significant effect on the WBC count.

Analysis of serum proteins in clinically healthy goats (Capra hircus) using agarose gel electrophoresis.

BACKGROUND: Electrophoretic patterns of serum proteins provide useful information on pathological conditions in ruminants. Their reference values, however, are dissimilar to those of other species. Reference values for goats using agarose gel as the supporting matrix have not been reported. OBJECTIVE: The aim of this study was to evaluate serum concentrations of total protein and protein fractions (albumin and globulins) by means of agarose gel electrophoresis (AGE) in goats in order to establish electrophoretic reference intervals and to evaluate potential changes associated with aging. METHODS: Blood was collected from 105 clinically healthy Girgentana goats by means of jugular venipuncture. Serum protein concentrations were assessed by AGE. Three age groups were compared: 1-1.5 years, 2-4 years, and 5-12 years. RESULTS: Values (mean ± SD) were determined for concentrations of total protein (72.26 ± 6.40 g/L), albumin (31.80 ± 4.00 g/L), α-globulins (6.40 ± 1.23 g/L), ß(1) -globulins (10.50 ± 2.58 g/L), ß(2) -globulins (5.18 ± 1.60 g/L), and γ-globulins (18.65 ± 5.90 g/L) and for albumin/globulin (A/G) ratio (0.82 ± 0.20). One-way ANOVA showed statistically significant age-related differences for total protein and α-globulin concentrations and A/G ratios. Age influenced protein concentrations with the 5-12-year-old group having higher total protein and α-globulin concentrations and lower albumin concentration and A/G ratios than the 2-4-year-old group. CONCLUSIONS: This study provides reference values for total protein concentrations and protein fractions obtained by AGE in goats. Some values vary with age. Age-specific reference intervals are reported in order to provide clinicians with an additional diagnostic aid.