Determining the In Vivo Efficacy of Plant-Based and Probiotic-Based Antibiotic Alternatives against Mixed Infection with Salmonella enterica and Escherichia coli in Domestic Chickens.

Restrictions on the use of antimicrobial compounds have led to a surge of interest in alternative solutions, such as natural, plant-based compounds. In our study, we investigated the efficacy of three feed supplements containing different additives, namely, probiotics (Lactobacillus spp., "Test substance A"), turmeric (Curcuma longa L., "Test substance B"), and fenugreek (Trigonella foenum graecum, "Test substance C"). In the experiment, we tested 180 birds of the Bábolna Tetra-SL laying hybrid breed that were infected with Salmonella enteritidis strains. The birds were randomly divided into six groups: three groups treated with the different additives, a negative control group, a positive control group, and an antibiotic-treated group using enrofloxacin. We examined the maturation and the time course of shedding of Salmonella; at the end of rearing, pathological and histopathological examinations were performed. When Salmonella was isolated from the cloacal swab samples, the enrofloxacin-treated group had a high number of animals shedding Salmonella by day 9, which was like the group treated with test material C. The greatest reduction in Salmonella shedding was observed in the groups treated with test materials A and B. In terms of pathological parameters, villus length and crypt depth were significantly better in the group treated with test material C compared to the positive and negative controls, and when comparing the body weight of the tested animals, the group treated with test material B had a significantly larger absorption surface area compared to the positive control group. Overall, the supplement with test material C proved to be the most effective. In the future, it is worthwhile to investigate the combination of the tested active substances for their possible synergistic effects and to perform a dose-response study to select the optimal dosage.

Investigation of Circulating MicroRNA Levels in Antibody-Mediated Rejection After Kidney Transplantation.

BACKGROUND: One of the most important possible complications determining long-term graft survival after kidney transplant is antibody-mediated rejection (ABMR). The criterion standard approach to recognize ABMR is currently the kidney biopsy with histopathologic analysis. However, this test has limitations because of difficulties in timing of sampling, the evaluability of histology because of the questionable representativeness of specimens, and the limited number of this intervention. Hence, new reliable, noninvasive biomarkers are required to detect the development of ABMR in time. METHODS: In this study, we analyzed the clinical data of 45 kidney transplant patients (mean age of 44.51 years, 20 male and 25 female subjects). These participants were recruited into 5 subcohorts based on their clinical status, histologic findings, and level of donor-specific anti-HLA antibodies. Circulating microRNAs (miR-21, miR-181b, miR-146a, miR-223, miR-155, miR-150) in plasma samples were quantified by quantitative polymerase chain reaction and their levels were correlated with the clinical characteristics in different subgroups. RESULTS: The relative expression of plasma miR-155 (P = .0003), miR-223 (P = .0316), and miR-21 (P = .0147) were significantly higher in patients who had subsequent histology-approved ABMR with donor-specific anti-HLA antibody positivity (n = 10) than in the "triple negative" group (n = 21), and miR-155 showed the highest sensitivity (90%) and specificity (81%) to indicate ABMR development based on receiver operating characteristic analysis. CONCLUSIONS: According to our preliminary data, plasma miR-155, miR-21, and miR-223 can indicate the development of ABMR after kidney transplant in correlation with classic clinical parameters. However, future studies with larger number of participants are necessary to further evaluate the diagnostic properties of blood miRNAs in prediction of this life-threatening condition.

A method for characterizing Cas9 variants via a one-million target sequence library of self-targeting sgRNAs.

Detailed target-selectivity information and experiment-based efficacy prediction tools are primarily available for Streptococcus pyogenes Cas9 (SpCas9). One obstacle to develop such tools is the rarity of accurate data. Here, we report a method termed 'Self-targeting sgRNA Library Screen' (SLS) for assaying the activity of Cas9 nucleases in bacteria using random target/sgRNA libraries of self-targeting sgRNAs. Exploiting more than a million different sequences, we demonstrate the use of the method with the SpCas9-HF1 variant to analyse its activity and reveal motifs that influence its target-selectivity. We have also developed an algorithm for predicting the activity of SpCas9-HF1 with an accuracy matching those of existing tools. SLS is a facile alternative to the much more expensive and laborious approaches used currently and has the capability of delivering sufficient amount of data for most of the orthologs and variants of SpCas9.

"Silent" iodine prophylaxis only partly corrects iodine deficiency: the Hungarian story (1970­2020) / A "silent" jódprofilaxis csak részben korrigálja a jódhiányt: a magyar történet (1970-2020).

Összefoglaló. A szerzo a bevezetoben emlékeztet a több mint 50 évvel korábbi publikációjára (Orv Hetil. 1968; 109: 360-363) és annak utóéletére: az 1970-es években megállt a jódprevenció fejlodése. Ezt követoen ismerteti az utóbbi 50 év jódellátottságra vonatkozó fontosabb hazai eredményeit. A számszeru adatok szerint az iskolás gyermekek, várandós anyák, idosek és újszülöttek jódellátottságára, valamint az anyatej jódtartalmára vonatkozó sorozatvizsgálatok az ezredforduló elott egybehangzóan enyhe, illetve mérsékelt jódhiányra utaltak. Az utóbbi két évtizedben egyre többször váltak adekváttá az ugyanezen kategóriák jódellátottságát jelzo eredmények. A szabályozatlan jódozottsó-forgalom (fakultatív jódprevenció) ellenére, valószínuleg a sikeres felvilágosítási kampányok miatt, a lakosság jódellátottsága határozottan javult. Ezt a szituációt nevezik "silent" profilaxisnak. Ezzel a módszerrel nem lehet a jódhiányt teljesen felszámolni. A legnagyobb kockázatot a várandós anyák (közel felének!) hiányos jódpótlása jelenti az utódok agyfejlodésének veszélyeztetése miatt. A befejezésben az EUthyroid Consortium "Krakkói kiáltvány"-ának (2018) rövid ismertetése tartalmazza a teendoket. A kötelezo jódprevencióhoz a nem jódozott sót ki kell váltani jódozott sóval szinte minden élelmiszerben. A graviditás alatt megnövekedett igényt az élelmiszerekben lévo jód gyakran nem fedezi, ilyenkor több jódpótlásra van szükség. Az egészségügyi szerveknek meg kell valósítaniuk a jóddúsító program összehangolt, rendszeres monitorozását és kiértékelését a lakosság optimális jódellátottságának biztosításához. Orv Hetil. 2020; 161(50): 2107-2116. Summary. In the introduction, the author reminds the readers of his publication presented more than 50 years ago in the same journal ("Data to the present state of the goiter problem in Hungary", 1968) and of its afterlife: the development of iodine prevention stopped in the 1970s. Then the major Hungarian results are reviewed related to the iodine supply gained in the latter 50 years. Numerical data are presented showing mild or moderate iodine deficiency according to the results of a range of screening studies among schoolchildren, pregnant women, elderly people and newborns as well as by iodine content of breast milk before the millennium. In the same categories, the data indicating the level of iodine supply became increasingly adequate in the recent two decades. The iodine supply of the people improved markedly, in spite of unregulated iodized salt trade (facultative prevention), presumably because of the successful public-information campaigns. This situation is called "silent" prophylaxis. The total eradication of iodine deficiency is impossible with this method. The highest risk is the deficient iodine supply of pregnant women (almost the half!) due to the endangerment of the offspring's brain development. In the end, a brief review of the Krakow Declaration on Iodine of the EUthyroid Consortium comprises the round of the duties. To the mandatory iodine prevention, iodized salt should replace non-iodized salt in nearly all food productions. During pregnancy, the increased need for iodine is frequently not covered by food sources, more iodine supplement is needed. Health authorities should perform harmonized monitoring and evaluation of fortification programs at regular intervals to ensure optimal iodine supply to the population. Orv Hetil. 2020; 161(50): 2107-2116.

Sex influenced association of directly measured insulin sensitivity and serum transaminase levels: Why alanine aminotransferase only predicts cardiovascular risk in men?

BACKGROUND: Non alcoholic fatty liver disease (NAFLD) is an independent cardiovascular (CV) risk factor which is closely associated with insulin resistance measured by both direct or indirect methods. Gender specific findings in the relationship between alanine aminotransferase (ALT) and CV disease, the prevalence of NAFLD and type 2 diabetes (T2DM) have been published recently. The aim of the present study was to explore the gender aspects of the association between insulin sensitivity, liver markers and other metabolic biomarkers in order to elucidate the background behind the sex influenced difference in both NAFLD, T2DM and their association with CV risk. PATIENTS AND METHODS: 158 female (47 normal and 111 impaired glucose intolerant) and 148 male (74 normal and 74 impaired glucose tolerant) subjects were included (mean age: 46.5 ± 8.31 vs. 41.6 ± 11.3, average Hba1c < 6.1 %, i.e. prediabetic population, drug naive at the time of the study). Subjects underwent a hyperinsulinemic normoglycemic clamp to determine muscle glucose uptake (M3), besides liver function tests and other fasting metabolic and anthropometric parameters were determined. RESULTS: Significant bivariate correlations were found between clamp measured M3 and all three liver enzymes (ALT, aspartate aminotransferase and gamma-glutamyl transferase) in both sexes. When data were adjusted for possible metabolic confounding factors correlations ceased in the male population but stayed significant in the female group. Feature selection analysis showed that ALT is an important attribute for M3 in the female but not in male group (mean Z: 3.85 vs. 0.107). Multiple regression analysis confirmed that BMI (p < 0.0001) and ALT (p = 0.00991) significantly and independently predicted clamp measured muscle glucose uptake in women (R(2) = 0.5259), while in men serum fasting insulin (p = 0.0210) and leptin levels (p = 0.0294) but none of the liver enzymes were confirmed as significant independent predictors of M3 (R(2) = 0.4989). CONCLUSION: There is a gender specific association between insulin sensitivity, metabolic risk factors and liver transaminase levels. This might explain the sex difference in the predictive role of ALT elevation for CV disease. Moreover, ALT may be used as a simple diagnostic tool to identify insulin resistant subjects only in the female population according to our results.

[Combined l-thyroxine and l-triiodothyronine replacement therapy in congenital hypothyroidism]. / Kombinált l-tiroxin és l-trijód-tironin hormonpótlás congenitalis hypothyreosisban.

INTRODUCTION: L-thyroxine replacement therapy is the treatment of choice for hypothyroidism. Recently, several studies suggested to complete it with l-triiodothyronine in acquired hypothyroidism. AIM: To study the role of combined l-thyroxine and l-triiodothyronine therapy in special cases with congenital hypothyroidism. METHOD: Data of 16 patients (age: 11.9 ± 6.3 years; mean ± SD) are presented who had high serum free thyroxine values or even above the upper limit of reference range (21.16 ± 2.5 pmol/l) together with nonsuppressed TSH levels (15.7 ± 5.7 mIU/l), and therefore received l-triiodothyronine in completion (0.18 ± 0.09 µg/kg) once a day. RESULTS: The combined replacement therapy resulted in a rapid improvement of the hormone parameters (TSH: 4.2 ± 3.15 mIU/l; free thyroxine: 16.55 ± 2.4 and free triiodothyronine: 7.4 ± 1.8 pmol/l). The efficiency of this combined therapy proved to be more evident (TSH: 4.33 ± 3.2 mIU/l; free thyroxine: 16.85 ± 3.1 and free triiodothyronine: 6.4 ± 0.85 pmol/l) in 10 patients treated for a longer period of time (duration of treatment: 2.9 ± 2.0 years). The dose of thyroxine substitution decreased from 2.6 ± 0.9 to 2.18 ± 0.6 µg/kg/day), the ratio of these hormones was between 5:1 and 19:1 and the quotient of free fractions was normalized (3.8 ± 0.4→2.6 ± 0.3) during the replacement therapy. CONCLUSIONS: According to the observation of the authors a serious disturbance of feed-back mechanism may develop in some (>5%) children with congenital hypothyroidism (increased TSH release despite elevated free thyroxine level) after normal function of the feed-back system for years. Hormone parameters of these patients improve, then become normal on combined therapy supporting the rationale for this treatment method.

Expression of the H- and L-subunits of ferritin in bone marrow macrophages of patients with osteoarthritis.

Osteoarthritis is a disease characterized by an increase in the production of reactive oxygen species (ROS) in afflicted joints. Excess iron, due to its role in the production of ROS and crystal deposition in the joints, is implicated in the disease progression of osteoarthritis. Ferritin is a major regulator of the bioavailability of iron, and its functions are determined largely by the combination of H- and L-subunits present in its outer protein shell. The purpose of the study was to investigate the expression of the H- and L-subunits of ferritin in bone marrow macrophages of osteoarthritis patients. The cytokine profiles were assessed as cytokines play an important role in the expression of the ferritin subunits. The H-subunit of ferritin in the bone marrow macrophages was significantly higher (P value = 0.035) in the osteoarthritis patients compared with the controls (107.84; 69.25-167.94 counts/µm(2); n = 7 versus 71.07; 58.56-86.26 counts/µm(2); n = 19). A marginally significant increase (P value = 0.059) was shown for the expression of the L-subunit in the osteoarthritis patients compared with the controls (133.03; 104.04-170.10 counts/µm(2); n = 7 versus 104.23; 91.53-118.70 counts/µm(2); n = 19). The osteoarthritis and control groups had comparable C-reactive protein, as well as proinflammatory and anti-inflammatory cytokine concentrations. The major exception was for transforming growth factor-ß (TGF-ß), which was higher (P value = 0.014) in the plasma of the osteoarthritis patients (16.69; 13.09-21.28 ng/mL; n = 7 versus 8.60; 6.34-11.67 ng/mL; n = 19). Up-regulation of the ferritin subunits decreases the levels of bioavailable iron and provides protection against the unwarranted production of ROS and crystal deposition. A role for TGF-ß in the up-regulation of the expression of the H-subunit, and possibly the L-subunit, of ferritin is postulated in osteoarthritis.

Three-year efficacy and safety of LB03002, a once-weekly sustained-release growth hormone (GH) preparation, in prepubertal children with GH deficiency (GHD).

BACKGROUND: GH treatment currently requires daily sc injections, resulting in suboptimal compliance. A GH regimen with fewer injections may offer patients and caregivers a less arduous option. LB03002 is a novel sustained-release GH formulation for once-weekly dosing. PATIENTS AND METHODS: GH-deficient, GH-naive prepubertal children were randomized to four groups who received 0.2 mg/kg/wk LB03002 for 12 months, followed by 0.5 mg/kg/wk for another 24 months (n=13); 0.5 mg/kg/wk LB03002 for 36 months (n=13); 0.7 mg/kg/wk LB03002 for 12 months, followed by 0.5 mg/kg/wk for another 24 months (n=13); or daily GH 0.03 mg/kg/d for 24 months, switched to 0.5 mg/kg/wk LB03002 for 12 months (n = 12). RESULTS: Height velocity increased in all groups; the increase was less for the 0.2 mg/kg/wk LB03002 group at 12 (P = 0.008) and 24 months (P = 0.030), with no statistically significant differences at any time for the 0.5 mg/kg/wk and 0.7 mg/kg/wk LB03002 groups, vs. daily GH. Height sd score gain at 12 months was significantly (P = 0.023) less for the 0.2 mg/kg/wk group (1.05 ± 0.38) than daily GH (1.47 ± 0.29), but with no statistically significant difference for the 0.5 mg/kg/wk (1.37 ± 0.39) and 0.7 mg/kg/wk (1.50 ± 0.44) LB03002 groups vs. daily GH. There were no significant differences in height sd score gain between any groups at 24 and 36 months. Bone maturation did not differ for any LB03002 dose compared with daily GH. Serum IGF-I concentrations increased as expected, with no long-term differences between groups. Mean fasting glucose and glycosylated hemoglobin concentrations did not exceed normal ranges for any treatment group at any time. CONCLUSION: LB03002 at doses of 0.5 mg/kg/wk and 0.7 mg/kg/wk was shown to be effective and safe with once-weekly dosing in GH-deficient children, and 0.5 mg/kg/wk LB03002 was chosen as the optimal dose for long-term assessment.

Congenital disorders of the thyroid: hypo/hyper.

This article summarizes the ontogenesis and genetics of the thyroid with regards to its possible congenital dysfunction and briefly refers to the roles of the mother-placenta-fetal unit, iodine effect, and organic and functional changes of the negative feedback mechanism, as well as maturity and illness, in some forms of congenital hypo- and hyperthyroidism. This article also describes the published literature and the authors' data on the clinical aspects of congenital hypothyroidism, on the alternating hypo- and hyperthyroidism in the neonatal period, and on neonatal hyperthyroidism.

Congenital disorders of the thyroid: hypo/hyper.

This article summarizes the ontogenesis and genetics of the thyroid with regards to its possible congenital dysfunction and briefly refers to the roles of the mother-placenta-fetal unit, iodine effect, and organic and functional changes of the negative feedback mechanism, as well as maturity and illness, in some forms of congenital hypo- and hyperthyroidism. This article also describes the published literature and the authors' data on the clinical aspects of congenital hypothyroidism, on the alternating hypo- and hyperthyroidism in the neonatal period, and on neonatal hyperthyroidism.

Pharmacokinetic and pharmacodynamic profile of a new sustained-release GH formulation, LB03002, in children with GH deficiency.

OBJECTIVE: LB03002 is a novel, sustained-release recombinant human GH, developed for once-a-week s.c. injection. To evaluate the suitability for long-term GH replacement therapy in children with GH deficiency (GHD), the present study assessed the pharmacokinetic (PK) and pharmacodynamic (PD) profiles of LB03002 at three doses. STUDY DESIGN AND PATIENTS: The randomised, comparator-controlled, assessor-blinded, phase II study assessed 37 (24 boys, 13 girls) pre-pubertal, GH-naïve children with GHD, in 11 European centres, for PK and PD analyses. GH, IGF1 and IGFBP3 concentrations were measured following the last daily GH dose and the first and 13th once-a-week administration of LB03002 at doses of 0.2, 0.5 or 0.7 mg/kg. RESULTS: GH C(max) values after the three doses of LB03002 were increased up to fourfold, with a clear dose proportionality. For each LB03002 dose, GH area under the concentration versus time curve did not increase from the first to 13th (month 3) administration, indicating no accumulation of circulating GH. IGF1 C(max) showed a progressive increase during LB03002 administration. Conversely, IGFBP3 showed a rapid increase in C(max). IGF1 SDS were fully normalised after 3 months of treatment, whereas IGFBP3 SDS were already in the normal range for all the three LB03002 dosages after 1 week. CONCLUSIONS: At the doses used, LB03002 has a suitable profile for long-term treatment to promote growth in children with GHD. The quantitative changes in IGF1 and IGFBP3 indicate adequate stimulation of the IGF system by LB03002 and the pattern of increase is comparable with that seen in GHD children in a standard IGF1 generation test using daily GH.

Thyroid dysfunction in the offspring of mothers with autoimmune thyroid diseases.

UNLABELLED: An integrated three-compartment thyroid model exists during gestation: pregnancy influences thyroid function in several ways, the placenta plays an active role in TRH-thyroid and iodide transport and metabolism, and the fetus develops its own hypothalamic-pituitary-thyroid axis. This commentary reviews some basic and new data, especially in the maternal-offspring relationship in the case of maternal autoimmune thyroid diseases. CONCLUSION: The careful control of maternal autoimmune thyroid disease is essential in order to avoid pathological changes in the offspring.

Sex steroid replacement during and after the induction of puberty.

The main aim of sex steroid replacement during induction of puberty is to obtain a balance between healthy psychological and somatic development and optimal final height and peak bone mass (PBM). At present, the age at which it is considered appropriate to begin inducing puberty is younger than it was in the past. This trend aims for optimisation of prepubertal and pubertal growth through earlier onset of growth hormone (GH) therapy and administration of higher doses during puberty as necessary. Adequate initiation of puberty should ensure not only optimal final height and psychological balance, but also normal body proportions, appropriate bone formation and preservation of fertility for children with GH and gonadotropin deficiencies.

[Incidence of thyroid disorders in children in Hungary]. / A gyermekkori pajzsmirigybetegségek elófordulása hazánkban.

Author gives information to the home tendencies of incidence of some important thyroid disorders in childhood mainly on the basis of his studies done with his coworkers. Results are compared with the standpoints raised since Chernobyl accident. It is verified on the basis of cca one million neonatal TSH screening that the frequency of congenital hypothyroidism in Hungary corresponds to those of European countries in the last 17 years. Changes connected with Chernobyl has not occurred. It deals with incidence of Graves' disease as the main form of hyperthyroidism relying upon TSH receptor antibody positivity. The yearly distribution of the 176 proved cases shows a slow increasing trend in the same period of time (17 yrs). In this radiation does not play a role. The paper treats of the topic of iodine deficiency with of high priority. Author draws the attention again to the fact that the iodine nutrition in significant part of the population is behind of optimum in harmony with the results of some other national studies also according to their own three thousand urinary iodine tests and more than four thousand thyroid sonographies. This iodine deficiency should be regarded as a potential risk from the viewpoint of radiation hazard! While enlargement of thyroid was often found by sonography, the formation of nodules only rarely in contrast to those of the Chernobyl area. The frequency of thyroiditis, especially in puberty increases too, but the degree of it falls far behind from those of published around of Chernobyl. The incidence of differentiated thyroid carcinoma--according to the available data--moderately increases as well, without any connection between the residence and the geographical distribution of radiation pollution. The epidemiological, clinical, and histological characteristics of the Hungarian children are similar to the so-called sporadic (non-radiation induced) form and different from the cases of the Chernobyl area.