Ethics in pre-ART genetics: a missed X-linked Menkes disease case.

Assisted reproductive technology (ART) has experienced dramatic progress over the last 30 years, and gamete donation is routine in fertility clinics. Major advances in genetic diagnostics are part of this development due to the ability to analyze multiple genes or whole genomes fast and to an affordable prize. This requires knowledge and capability to evaluate genetic variants correctly in a clinical setting. Here we report a Menkes disease case, born after ART, where genetic screening and variant scoring failed to identify an egg donor as carrier of this fatal X-linked disorder. The gene variant is a deletion of a single base pair leading to a frameshift and premature termination of the protein, predicted to result in no or severely diminished function. The variant would be classified as likely pathogenic (class 4) and should be readily detectable by molecular genetic screening techniques. We wish to highlight this case to prevent future similar cases. IVI Igenomix has developed and embarked on an ambitious screening program to detect and prevent a large number of inherited severe childhood disorders in ART pregnancies. The company has recently achieved ISO 15189 certification with competence to evaluate and deliver timely, accurate, and reliable results. Failure to identify a pathogenic variant in the ATP7A gene leading to birth of two boys with Menkes disease invokes the required procedures to screen and detect disease-causing gene variants. This calls for ethical and legal considerations in ART diagnostics to prevent fatal errors like the present.

Legal enforcement and development directions of health law in Ukraine.

OBJECTIVE: Introduction: The development of medical law should take place systematically based on scientific basis and guided by international experience. The article describes the tendencies and prospects of the medical law development of Ukraine in the context of globalization and European integration processes. The aim of this work is to investigate the problems of legal enforcement and directions of the medical law development in Ukraine. PATIENTS AND METHODS: Materials and methods: We analyzed national and international acts that regulate community rights for medical care, protection of patient rights, which were studied using content analysis and analytical experience, as well as generalization of court practice and statistical data. RESULTS: Review and conclusions: Violation of patient rights is a socially harmful act, impunity of which also affects social security. The quality control mechanism of medical care should be permanent and aimed at improving this quality. It is necessary to create a National Quality Control Agency that would be independent, not subordinate to the Ministry of Healthcare of Ukraine, with professional experts with experience in this direction. The problem of legal protection of patient and doctor rights, as well as provision of legal support to medical institutions and the creation of a regulatory framework to reform the health care system is relevant. It is necessary to create a register of medical lawyers. Creating a system for reporting and monitoring medical errors should be one of the priorities of health care reform in Ukraine.

Problems of transplantation of human organs and other anatomical materials and directions of their solution in Ukraine.

OBJECTIVE: Introduction: The focus of the work is the importance of transplantation of human organs and other anatomical materials as the method of treating and a way of saving people's lives and it also attaches particular importance to solving issues of transplantations in Ukraine. The aim: To find out the problems of human organ transplantation and other anatomical materials and determine the directions of their solutions in Ukraine. PATIENTS AND METHODS: Materials and methods: The assessment and analysis of domestic and foreign experience has been developed. In addition, data collected from Ukrainian and international organizations and the results of scientific research by scientists are used in the work. RESULTS: Review and conclusions: Investigating the legislation of Ukraine in the part of transplantation of human organs and other anatomical materials, conclusions were made on the necessity of its improvement, in particular, Article 16 of the Law of Ukraine «On the application of transplantation of anatomical materials to a person¼ dated by May 17, 2018, is proposed to be worded as follows: «Any adult may give written consent or disagreement to become a donor of anatomical materials in case of his death. In the absence of such a statement of disagreement with the transplantation of his organs or other anatomical materials, consent of relatives or others is not required. In this case, the deceased person is deemed to have given consent to the transplantation of his organs or other anatomical materials¼. It is proposed to make changes and significant additions to Art. 17 of the Law of Ukraine «On the application of transplantation of anatomical materials to a person¼ of May 17, 2018 regarding the creation of the Unified State Register of persons in need of transplantation, as well as to consolidate the procedure for its administration. In the context of urgent transformations, the necessity to significantly increase state funding for the provision of medical services, raise the awareness and informality of citizens about transplantation, as well as to change their misconceptions about this issue are sometimes put forward.

A 37-year-old Menkes disease patient-Residual ATP7A activity and early copper administration as key factors in beneficial treatment.

Menkes disease (MD) is a lethal disorder characterized by severe neurological symptoms and connective tissue abnormalities; and results from malfunctioning of cuproenzymes, which cannot receive copper due to a defective intracellular copper transporting protein, ATP7A. Early parenteral copper-histidine supplementation may modify disease progression substantially but beneficial effects of long-term treatment have been recorded in only a few patients. Here we report on the eldest surviving MD patient (37 years) receiving early-onset and long-term copper treatment. He has few neurological symptoms without connective tissue disturbances; and a missense ATP7A variant, p.(Pro852Leu), which results in impaired protein trafficking while the copper transport function is spared. These findings suggest that some cuproenzymes maintain their function when sufficient copper is provided to the cells; and underline the importance of early initiated copper treatment, efficiency of which is likely to be dependent on the mutant ATP7A function.

Efficiency of hydrated sodium calcium aluminosilicate to ameliorate the adverse effects of graded levels of aflatoxin B1 in broiler chicks.

The objective of this study was to evaluate the efficiency of a hydrated sodium calcium aluminosilicate (HSCAS) adsorbent to ameliorate the adverse effects of 0.5 to 2 mg of aflatoxin B1 (AFB1)/kg in broiler chicks. The study consisted of 8 dietary treatments, including 4 concentrations of AFB1 (0, 0.5, 1, and 2 mg/kg) with or without HSCAS (0.5%) fed to 8 replicate cages per diet (6 males chicks per cage) from 0 to 21 d of age. Cumulative feed intake, BW gain (P < 0.0001), and G:F (P = 0.004) of birds fed the 2 mg of AFB1/kg of diet were significantly lower in comparison with birds fed 0 to 1 mg of AFB1/kg. Relative liver weight was increased in the 2 mg of AFB1/kg group (P < 0.0001). Dietary HSCAS improved cumulative BW gain (main effect P = 0.06), particularly from 14 to 21 d of age (P = 0.037). Dietary HSCAS also reversed the increase in relative liver weight for birds fed AFB1 (P = 0.019). Dietary AFB1 negatively affected major serum parameters (albumin, total protein, globulin, phosphorus, glucose, alkaline phosphatase, and creatine phosphokinase), whereas supplementation with HSCAS partially alleviated the affected serum biochemistry. In addition, serum complement activity and liver gene expression were negatively affected by 2 mg of AFB1/kg. The HSCAS supplement increased the liver expression of catalase and superoxide dismutase (P < 0.05). Results from this study indicate that dietary supplementation with HSCAS can effectively improve BW gain and partially ameliorate aflatoxicosis for broiler chicks fed AFB1-contaminated feeds.

Growth, serum biochemistry, complement activity, and liver gene expression responses of Pekin ducklings to graded levels of cultured aflatoxin B1.

A 14-d study was conducted to evaluate the effects of cultured aflatoxin B1 (AFB1) on performance, serum biochemistry, serum natural antibody and complement activity, and hepatic gene expression parameters in Pekin ducklings. A total of 144 male Pekin ducklings were weighed, tagged, and randomly allotted to 4 dietary treatments containing 4 concentrations of AFB1 (0, 0.11, 0.14, and 0.21 mg/kg) from 0 to 14 d of age (6 cages per diet; 6 ducklings per cage). Compared with the control group, there was a 10.9, 31.7, and 47.4% (P < 0.05) decrease in cumulative BW gain with 0.11, 0.14, and 0.21 mg of AFB1/kg of diet, respectively, but feed efficiency was not affected. Increasing concentrations of AFB1 reduced cumulative BW gain and feed intake both linearly and quadratically, and regression equations were developed with r(2) ≥0.73. Feeding 0.11 to 0.21 mg of AFB1/kg reduced serum glucose, creatinine, albumin, total protein, globulin, Ca, P, and creatine phosphokinase linearly, whereas serum urea N, Cl, alkaline phosphatase, and aspartate amino transferase concentrations increased linearly with increasing AFB1 (P < 0.05). Additionally, 0.11 to 0.21 mg of AFB1/kg diets impaired classical and alternative complement pathways in the duckling serum when tested by lysis of rabbit, human type O, and horse erythrocytes, and decreased rabbit and horse agglutinins (P < 0.05). Liver peroxisome proliferator activated receptor α (PPARα) expression was linearly downregulated by AFB1 (P < 0.01). Results from this study indicate that for every 0.10 mg/kg increase in dietary AFB1, cumulative feed intake and BW gain decrease approximately 230 and 169 g per duckling from hatch to 14 d; and that AFB1 at very low concentrations can significantly impair liver function and gene expression, and innate immune dynamics in Pekin ducklings.

Isolation of a candidate gene for Menkes disease that encodes a potential heavy metal binding protein.

Menkes disease is a lethal-X linked recessive disorder associated with copper metabolism disturbance. We have recently mapped two chromosome breakpoints related to this disease in a 1 megabase yeast artificial chromosome contig at Xq13.3. We now report the construction of a phage contig and the isolation of candidate partial cDNAs for the Menkes disease gene. The candidate gene expresses an 8 kb message in all investigated tissues, and deletions were detected in 16% of 100 unrelated Menkes patients. The deduced partial protein sequence shared the GMTCXXC motif with bacterial metal resistance operons, suggesting a potential heavy metal binding protein. These findings should lead to more accurate prenatal diagnosis of this severe disease and a better understanding of the cellular homeostasis of essential heavy metals.

Can tibio-femoral kinematic and kinetic parameters reveal poor functionality and underlying deficits after total knee replacement? A systematic review.

BACKGROUND: Extensive efforts have been made to understand joint kinematics and kinetics in total knee arthroplasty (TKA) in subjects with satisfactory outcomes during daily functional activities and clinical tests, but it remains unclear whether such movement characteristics hold the potential to indicate the underlying aetiology of unsatisfactory or bad TKA outcomes. PURPOSE: To investigate which kinematic and kinetic parameters assessed during passive clinical tests and functional activities of daily living are associated with poor functionality and underlying deficits after total knee replacement. METHODS: We focused on studies characterizing the kinematic or kinetic parameters of the knee joint that are associated with poor clinical outcome after TKA. Seventeen articles were included for the review, and kinematic and kinetic data from 719 patients with minimal follow up of 6 months were extracted and analyzed. RESULTS: Passive posterior translation at 90°flexionexhibited good potential for differentiating stable and unstable TKAs. Anterior-posterior (A-P) translation of the medial condyle at 0-30° and 30-60° flexion, A-P translation of the lateral condyle at 60-90°during closed chain exercises, as well asknee extension moment during stair ascent and descent, knee abduction moment during stair descent, knee internal rotation moment and plantar flexion moment during walking, 2ndpeak ground reaction force during stair ascent and walkingshowed the greatest promise as functional biomarkers for a dissatisfied/poor outcome knee after TKA. CONCLUSION: In this study, we systematically reviewed the state-of-the-art knowledge of kinematics and kinetics associated with functional deficits, and found 11 biomechanical parameters that showed promise for supportingdecision making in TKA.

Early TICI 2b or Late TICI 3-Is Perfect the Enemy of Good?

BACKGROUND AND PURPOSE: A Thrombolysis in Cerebral Infarction (TICI) score of 3 has been established as therapeutic goal in endovascular therapy (EVT) for acute ischemic stroke; however, in the case of early TICI2b reperfusion, the question remains whether to stop the procedure or to continue in the pursuit of perfection (i.e., TICI 2c/3). METHODS: A total of 6635 patients were screened from the German Stroke Registry. Patients who underwent EVT for occlusion of the middle cerebral artery (M1 segment), with final TICI score of 2b/3 were included. Multivariable logistic regression was performed with functional independence (modified Rankin Scale, mRS at day 90 of 0-2) as the dependent variable. RESULTS: Of 1497 patients, 586 (39.1%) met inclusion criteria with a final TICI score of 2b and 911 (60.9%) with a TICI score of 3. Patients who achieved first-pass TICI3 showed highest odds of functional independence (Odds ratio [OR] 1.71, 95% confidence interval [95% CI] 1.18-2.47). Patients who achieved TICI2b with the second pass (OR 0.53, 95% CI 0.31-0.89) or with three or more passes (OR 0.44, 95% CI 0.27-0.70) had significantly worse clinical outcomes compared to first-pass TICI2b. TICI3 at the second pass was by trend better than first-pass TICI2b (OR 1.55, 95% CI 0.98-2.45), but TICI3 after 3 or more passes (OR 0.93, 95% CI 0.57-1.50) was not significantly different from first-pass TICI2b. CONCLUSION: First-pass TICI2b was superior to TICI2b after ≥ 2 retrievals and comparable to TICI3 at ≥ 3 retrievals. The potential benefit in outcome after achieving TICI3 following further retrieval attempts after first-pass TICI2b need to be weighed against the risks.

More Retrieval Attempts are Associated with Poorer Functional Outcome After Unsuccessful Thrombectomy.

PURPOSE: In mechanical thrombectomy, it has been hypothesized that multiple retrieval attempts might the improve reperfusion rate but not the clinical outcome. In order to assess a potential harmful effect of a mechanical thrombectomy on patient outcome, the number of retrieval attempts was analyzed. Only patients with a thrombolysis in cerebral infarction (TICI) score of 0 were reviewed to exclude the impact of eventual successful reperfusion on the mechanical hazardousness of repeated retrievals. METHODS: In this study 6635 patients who underwent endovascular thrombectomy (EVT) for acute large vessel occlusion (LVO) from the prospectively administered multicenter German Stroke Registry were screened. Insufficient reperfusion was defined as no reperfusion (TICI score of 0), whereas a primary outcome was defined as functional independence (modified Rankin scale [mRS] 0-2 at day 90). Propensity score matching and multivariable logistic regressions were then performed to adjust for confounders. RESULTS: A total of 377 patients (7.8%) had a final TICI score of 0 and were included in the study. After propensity score matching functional independence was found to be significantly more frequent in patients who underwent ≤ 2 retrieval attempts (14%), compared to patients with > 2 retrieval attempts (3.9%, OR 0.29, 95% CI 0.07-0.73, p = 0.009). After adjusting for age, sex, admission NIHSS score, and location of occlusion, more than two retrieval attempts remained significantly associated with lower odds of functional independence at 90 days (OR 0.2, 95% CI 0.07-0.52, p = 0.002). CONCLUSION: In patients with failure of reperfusion, more than two retrieval attempts were associated with a worse clinical outcome, therefore indicating a possible harmful effect of multiple retrieval attempts.

[Long and short QT syndromes : Emergency treatment and secondary prophylaxis]. / Long- und Short-QT-Syndrome : Notfalltherapie und Sekundärprophylaxe.

Long QT syndrome (LQTS) is a rare inherited or acquired channelopathy associated with a relevant mortality if left untreated. Therapy can reduce the sudden cardiac death (SCD) rate significantly. Of 17 subtypes, LQTS1-3 are the most common. Clinical presentation ranges from asymptomatic patients to torsade de pointes (TdP) and SCD. Emergency therapy includes defibrillation, administration of magnesium, betablockers and temporary pacing and sedation. Secondary prevention is based on betablocker therapy and implantation of an implantable cardioverter-defibrillator (ICD), if appropriate. Short QT syndrome (SQTS) is a rare channelopathy that manifests as SCD in 34%. So far 250 cases with mutations in 8 genes have been reported. ICDs are the only reliable protection against SCD. Drug therapy is based on hydroquinidine. Further therapeutic options exist for certain subtypes of both diseases. Patients should be referred to specialized centers.

Intestinal mucin dynamics: response of broiler chicks and White Pekin ducklings to dietary threonine.

Mucin dynamics may be particularly sensitive to a Thr deficiency due to the high concentration and structural importance of Thr in the mucin protein backbone. Intestinal mucin secretion, expression of mucin gene (MUC2), and histological characteristics were investigated in male broilers and White Pekin ducklings offered diets containing 3.3, 5.8, or 8.2 g of Thr/kg in 4 studies. Seventy-two birds of each species were fed a standard broiler starter diet from 1 to 14 d of age followed by assignment to 3 dietary treatments in a randomized complete block design for a 7-d feeding trial in experiment 1 (broilers) and experiment 2 (ducklings). The dietary treatments consisted of an isonitrogenous corn-soybean meal-based diet with the addition of crystalline amino acids and graded levels of Thr. Dietary treatments contained 3.3, 5.8, or 8.2 g of Thr/kg. Dietary formulation and experimental design for experiments 3 (broilers) and 4 (ducklings) were similar to experiments 1 and 2 except that birds were fed 3.3 or 8.2 g of Thr/kg for durations of 7 or 14 d. For chicks, increased dietary Thr resulted in higher levels of intestinal crude mucin excretion in experiment 1 (P=0.04) but not in experiment 3, whereas intestinal sialic acid excretion increased in experiment 3 (P=0.02) but not in experiment 1. Furthermore, there was no effect of Thr on intestinal goblet cell density or MUC2 mRNA abundance for broilers. For ducklings, there was an increase in intestinal crude mucin excretion in both experiments (P<0.05) as dietary Thr increased, although there was no effect of Thr on intestinal sialic acid excretion. There was a tendency for an increase in intestinal goblet cell density (cells/microm of villus length; P=0.09) as dietary Thr increased in experiment 2. For experiment 4, intestinal MUC2 mRNA abundance increased (P=0.03) as dietary Thr increased for the 14-d feeding trial but not for the 7-d feeding trial. The data establish a link between dietary Thr and intestinal crude mucin dynamics in chicks for experiment 1 and ducklings for both experiments.

Influence of baking enzymes on antimicrobial activity of five bacteriocin-like inhibitory substances produced by lactic acid bacteria isolated from Lithuanian sourdoughs.

AIM: To evaluate the effect of four different baking enzymes on the inhibitory activity of five bacteriocin-like inhibitory substances (BLIS) produced by lactic acid bacteria (LAB) isolated from Lithuanian sourdoughs. METHODS AND RESULTS: The overlay assay and the Bioscreen methods revealed that the five BLIS exhibited an inhibitory effect against spore germination and vegetative outgrowth of Bacillus subtilis, the predominant species causing ropiness in bread. The possibility that the observed antibacterial activity of BLIS might be lost after treatment with enzymes used for baking purposes was also examined. CONCLUSIONS: The enzymes tested; hemicellulase, lipase, amyloglucosidase and amylase had little or no effect on the majority of the antimicrobial activities associated with the five BLIS studied. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests a potential application in the sourdough baking industry for these antimicrobial producing LAB strains in the control of B. subtilis spore germination and vegetative outgrowth.

Immunity gene pedB enhances production of pediocin PA-1 in naturally resistant Lactococcus lactis strains.

Heterologous production of the antilisterial bacteriocin pediocin PA-1 in lactococci is an attractive objective to increase the safety of dairy products. In a previous paper, we developed a system for the heterologous production of the bacteriocin pediocin PA-1 in pediocin-resistant lactococcal hosts through a leader exchange strategy. The system was based on 3 genes, 1 encoding the fusion between the lactococcin A leader and propediocin PA-1, and the other 2 encoding the lactococcin A secretion machinery. In this study, we investigated whether the addition of the pediocin PA-1 immunity gene (pedB) to this system has any effect on pediocin production. Introduction of the plasmid(s) carrying the genes described above into nisinproducing and non-nisinproducing lactococcal hosts led to a significant increase in the production of pediocin compared with the equivalent pedB-devoid systems. In addition, we obtained a nisin-producing strain with the ability to secrete pediocin PA-1 at a level equivalent to that of the parental strain Pediococcus acidilactici 347, which represents a notable improvement over our previous systems.

Garlic diallyl disulfide and diallyl trisulfide mitigates effects of pro-oxidant induced cellular stress and has immune modulatory function in LPS-stimulated porcine epithelial cells.

The objective of the current study was to determine if garlic-derived diallyl disulfide (DADS) and diallyl trisulfide (DATS) could mitigate oxidative and endotoxin stress, using an intestinal porcine epithelial cell (IPEC-J2) model. The experiment was arranged as a 2 × 2 × 2 factorial of DADS + DATS (0 or 18 µM), pro-oxidant stressor (hydrogen peroxide at 0 or 100 µM), and endotoxin stressor (lipopolysaccharide [LPS] at 0 or 10 µg/mL) with 8 replicates per treatment. Cells were incubated with DADS + DATS for 18 h, LPS for 6 h, then with hydrogen peroxide for 3 h. Gene expression was measured by RT-PCR for cytokines, interleukin 8 (IL-8) and tumor necrosis factor α (TNF-α), and tight junction proteins, claudin 1 (CL-1), occludin (OC), and zonula occludens 1 (ZO-1). Trans-epithelial electrical resistance (TEER), the antioxidant enzyme catalase, and apical secretion of IL-8 protein into the incubation medium was also measured. There was an increase ( < 0.01) in TNF-α and IL-8 gene expression due to LPS, although there was no effect of hydrogen peroxide or DADS + DATS. Furthermore, there was a tendency for an increase ( = 0.08) in ZO-1 gene expression due to DADS + DATS. Treatment with DADS + DATS and hydrogen peroxide did not affect TEER, although there was a decrease ( = 0.02) in TEER with LPS incubation. Treatment of cells with hydrogen peroxide reduced catalase activity ( < 0.01), which was restored with pre-incubation of DADS + DATS ( < 0.10). There was an increase ( < 0.01) in IL-8 secretion due to LPS, which was further augmented ( < 0.01) by pre-incubation with DADS + DATS. Based on the results from the current study, DADS + DATS can ameliorate oxidative effects of hydrogen peroxide, as well as alter IL-8 secretion in LPS-treated IPEC-J2 cells.

Ability of garlic-derived diallyl disulfide and diallyl trisulfide supplemented by oral gavage to mitigate effects of an acute postweaning feed and water deprivation event in nursery pigs.

Compounds in garlic have been shown to contain anti-inflammatory, antioxidant, and immune modulatory properties that may be able to mitigate the effects of nursery pig stressors. The objective of the current experiment was to determine if oral gavage of garlic-derived diallyl disulfide (DADS) and diallyl trisulfide (DATS) could mitigate the effects of a 24-h postweaning feed + water deprivation event in nursery pigs. Pigs (6.0 ± 0.05 kg and 21 d old) were allotted to 4 treatments in a randomized complete block design at weaning with 8 replicate pens per treatment that consisted of with or without a 24-h postweaning feed + water deprivation event and with or without an oral gavage containing 3.6 mg DADS + DATS/kg BW. Growth performance and morbidity were recorded throughout the experiment, and on 1, 6, and 21 d after weaning, 1 pig per pen was selected, blood was collected, the pig was euthanized, and a segment of the distal ileum was subsequently excised for morphological and gene and protein expression measurements. Mucosal gene expression was conducted by reverse transcription PCR for immune, antioxidant, and cellular integrity markers. Furthermore, activity of mucosal superoxide dismutase was measured by colorimetric assay. Immediately following the feed + water deprivation event, there was a decrease ( < 0.01) in growth performance and an increase ( = 0.01) in serum cortisol. The feed + water deprivation event tended ( = 0.10) to decrease ileal villus height and supplementation of DADS + DATS by oral gavage increased ( = 0.03) villus height 1 d after weaning. Supplementation of DADS + DATS by oral gavage decreased ( = 0.03) and tended to decrease ( = 0.08) gene expression of on 6 and 21 d after weaning, respectively. Furthermore, at 1 d after weaning, ileal mucosa SOD activity was decreased ( = 0.01) by the feed + water deprivation and increased ( = 0.04) by oral supplementation of DADS + DATS. Expression of the tight junction genes and were reduced ( ≤ 0.05) due to the feed + water deprivation event 1 d after weaning. Results from the current study show that an acute feed + water deprivation event can impact growth performance, intestinal characteristics, and antioxidant status in nursery pigs, which can be partially mitigated by oral supplementation of garlic compounds DADS + DATS.

The efficacy of dietary xylanase and phytase in broiler chickens fed expeller-extracted camelina meal.

A study was conducted to investigate the effects of dietary phytase (PHY) and xylanase (XYL) on growth performance, nutrient utilization, and intestinal characteristics in broilers fed corn-soybean meal-based diets with added expeller-extracted camelina meal (CM). The corn-soybean meal-based diets without or with CM was formulated to contain 2.9 or 2.7 g/kg non phytate phosphorus, respectively. A total of 384 male Ross 708 broilers were allocated to 8 dietary treatments in a randomized complete block design with 6 replicates per treatment, from 7 to 21 d post hatching. The experiment consisted of a 2 × 2 × 2 factorial arrangement of treatments with 2 dietary CM levels (zero or 100 g/kg), 2 dietary levels of XYL (zero or 800 unit/kg), and 2 dietary levels of PHY (zero or 4,000 unit/kg). Chromic oxide was included in the diets as an indigestible marker. Growth was measured throughout the experiment and excreta were collected on d 18 to 21 post hatching for measurement of nutrient and energy retention. On d 21 post hatching, broilers were euthanized by CO2 asphyxiation, ileal digesta was collected for nutrient and energy digestibility measurements, and the left tibia was removed for bone ash measurement. Furthermore, duodenal digesta was collected and a segment of the mid jejunum was excised for viscosity and morphology measurement, respectively. Broilers fed 100 g/kg CM diets showed lower (P < 0.001) BW gain and G:F than those fed zero g/kg CM diets for 7 to 14 d and 7 to 21 d post hatching periods, respectively. Phytase supplementation improved (P < 0.05) all the growth parameters measured regardless of CM addition. Phytase supplementation resulted in an increase (P < 0.05) in jejunum villus height but there was no impact of CM or XYL. Additionally, duodenal digesta viscosity increased (P < 0.001) with added CM. Tibia ash and weight increased (P < 0.001) with PHY supplementation. There was a CM × PHY interaction (P < 0.05) on percentage tibia ash, with greater PHY effect in zero g/kg CM diets compared with 100 g/kg CM diets. Phytase supplementation increased (P < 0.05) ileal DM, N, energy, P, and neutral detergent fiber (NDF) digestibility. In diets with 100 g/kg CM, addition of PHY had a greater effect (CM × PHY; P < 0.05) on ileal P digestibility compared with diet without CM. There was a CM × PHY interaction (P < 0.05) for ileal digestible energy (IDE) in which PHY increased the IDE in birds fed CM supplemented diets. Both ileal digestibility and retentions of DM, N, energy, and IDE, AME, and AMEn were decreased (P < 0.05) by CM supplementation. Retention of ether extract and NDF decreased (P < 0.05) with CM supplementation. Nitrogen retention, AME, and AMEn decreased (CM × XYL; P < 0.05) with XYL supplementation in zero g/kg CM diets. There were CM × XYL interactions (P < 0.01) for P and Ca retentions in which XYL increased retentions in 100 g/kg CM diets. In conclusion, PHY was efficacious at improving P digestibility and retention of birds fed low P corn-soy based diets without or with CM. However, regardless of CM addition, XYL did not improve nutrient utilization and growth performance of broiler chickens. The present data also show that the mechanism underlying the detrimental effects of CM inclusion on nutrient utilization is not mediated through gut morphology and goblet cell density. The data also show that the villus height increase occurring with PHY addition might be a contributor to the improvements in ileal DM, N, and energy digestibility obtained with PHY supplementation.

Comparison of goblet cell staining methods in jejunal mucosa of turkey poults.

This study compared the intestinal goblet cell density of turkey poults at 2 different ages using Alcian blue-periodic acid-Shiff (AB-PAS) and mucicarmine stains. Neutral mucins are stained with periodic acid-Shiff whereas acidic mucins are stained with Alcian blue. Mucicarmine and AB-PAS are specific to the mucins of epithelial origin. Mucicarmine has only been used for the assessment of goblet cells in human specimens, and it may have advantages for use in animals as a result of the methodological simplicity of staining as compared to AB-PAS. A mid-section of jejunum was taken from 80 turkey poults at 21 and 28 d, and fixed in 10% buffered formalin for 48 h. Each fixed tissue was dehydrated with ethanol, cleared with Sub-X, placed in paraffin wax, prepared on 2 slides, cleared and hydrated. The 2 slides were randomly assigned to 2 treatments which consisted of AB-PAS and mucicarmine stains in a completely randomized design. Goblet cell counts were taken from four villi per slide and the villus height was measured and averaged. There was no difference in the goblet cell density between the staining methods AB-PAS and mucicarmine at 21 or 28 d posthatch. These results show that both staining methods are viable for assessment of goblet cell density in turkey poults.

Evaluation of aplastic anemia with indium chloride In 111 scanning.

Twenty-one patients (12 male, nine female) with diverse causes of aplastic anemia underwent indium chloride in 111 bone marrow scintigraphy. Good correlation of the scan interpretation with other clinical values, including the peripheral blood counts and bone marrow biopsy, was found in 25 of the 26 studies. These encouraging results with a noninvasive procedure that permits evaluatin of the entire bone marrow organ suggest a broader use for indium bone marrow scintigraphy in aplastic anemia.