Pretreatment morning urine osmolality and oral desmopressin lyophilisate treatment outcome in patients with primary monosymptomatic enuresis.

PURPOSE: To determine the association between urine osmolality (Uosm) in patients with primary monosymptomatic enuresis (PMNE) and response to desmopressin (dDAVP) lyophilisate. METHODS: This was a prospective cohort study that included 419 children with enuresis seen in outpatient clinic between October 2017 and October 2019. Patient workup included symptom checklist, 48 h frequency/volume chart, kidney and bladder ultrasound, uroflow, urinalysis and culture, spot urine Ca/creatinine, and first-morning Uosm. Patients < 5 years, with secondary enuresis, or loss of follow-up were excluded. Oral dDAVP lyophilisate was recommended to all with PMNE and normal bladder capacity. After 1 month of therapy, initial success was assessed according to ICCS. Significant predictor variables for complete response were identified and analyzed using correlation coefficients and binary logistic regression. RESULTS: There were 48 patients with PMNE who received dDAVP and were followed for treatment success. Partial and complete responses were achieved for 14 (29.2%) and 20 cases (41.7%), respectively. Older age and lower Uosm were found to be significantly in favor of complete response to dDAVP lyophilisate, P = 0.007 and 0.033, respectively. ROC analysis determined the Uosm of ≤ 814 mOsm/kg as a cut-off value for complete success (sensitivity 65% and specificity 75%, AUC = 68.2%). The odds ratio for complete success for selected cut-off value was 5.57 (95% CI 1.588-19.551, P = 0.007). CONCLUSION: High pretreatment morning Uosm (> 814 mOsm/kg) might be suggestive of an alternative treatment to dDAVP lyophilisate in PMNE because of the higher risk of treatment failure.

Predicting posterior urethral obstruction in boys with lower urinary tract symptoms using deep artificial neural network.

PURPOSE: To assess the prediction model for late-presenting posterior urethral valve (PUV) in boys with lower urinary tract symptoms (LUTS) using artificial neural network (ANN). MATERIALS AND METHODS: 408 boys aged 3-17 years (median 7.2 years) with LUTS were examined and had bladder diary, ultrasound, uroflowmetry, urine, and urine culture. Cystoscopy was recommended when peak flow rate (Qmax) was persistently ≤ 5th percentile in patients who were unresponsive to urotherapy and pharmacological treatment (oxybutynin). With four-layered backpropagating deep ANN, the probability of finding PUV was estimated using noninvasive, quantitative parameters (age, Qmax, time to Qmax, voided volume, flow time, voiding time, average flow rate). RESULTS: There were 97 patients with low Qmax and 74 were unresponsive. In 41, cystoscopy was performed and PUV was diagnosed in 37 (9.1%). In multivariate analysis, significant variables in favor of PUV were urgency (OR = 3.96, 95% CI = 1.30-12.03, p = 0.015), increased voiding frequency (OR = 3.81, 95% CI = 1.03-14.11, p = 0.045), and weak stream/intermittency (OR = 8.30, 95% CI = 2.49-27.63, p = 0.001). The ANN dataset included 87 uroflows of children with PUV and 114 uroflows classified as normal. The best performance was with two hidden layers with four neurons each. The best test accuracy was 92.7% and AUROC was 98.0%. With cutoff value of 0.8, sensitivity was 100.0%, specificity 89.7%, positive predictive value 80.0%, and negative predictive value 100.0%. CONCLUSIONS: With ANN, we accurately predicted 92.7% of late-presenting PUV using uroflow. Considering the high frequency of PUV in boys with LUTS, especially in cases of urgency, increased voiding frequency, and weak stream or intermittency, accurate prediction could lead to timely treatment.

POMT1-associated walker-warburg syndrome: a disorder of dendritic development of neocortical neurons.

We have analyzed the morphology and dendritic development of neocortical neurons in a 2.5-month-old infant with Walker-Warburg syndrome homozygotic for a novel POMT1 gene mutation, by Golgi methods. We found that pyramidal neurons frequently displayed abnormal (oblique, horizontal, or inverted) orientation. A novel finding of this study is that members of the same population of pyramidal neurons display different stages of development of their dendritic arborizations: some neurons had poorly developed dendrites and thus resembled pyramidal neurons of the late fetal cortex; for some neurons, the level of differentiation corresponded to that in the newborn cortex; finally, some neurons had quite elaborate dendritic trees as expected for the cortex of 2.5-month-old infant. In addition, apical dendrites of many pyramidal neurons were conspiciously bent to one side, irrespective to the general orientation of the pyramidal neuron. These findings suggest that Walker-Warburg lissencephaly is characterized by two hitherto unnoticed pathogenetic changes in the cerebral cortex: (a) heterochronic decoupling of dendritic maturation within the same neuronal population (with some members significantly lagging behind the normal maturational schedule) and (b) anisotropically distorted shaping of dendritic trees, probably caused by patchy displacement of molecular guidance cues for dendrites in the malformed cortex.

S-Adenosylhomocysteine hydrolase (AdoHcyase) deficiency: enzymatic capabilities of human AdoHcyase are highly effected by changes to codon 89 and its surrounding residues.

Recently, S-adenosylhomocysteine hydrolase deficiency was confirmed for the first time in an adult. Two missense mutations in codons 89 (A>V) and 143 (Y>C) in the AdoHcyase gene were identified [N.R.M. Buist, B. Glenn, O. Vugrek, C. Wagner, S. Stabler, R.H. Allen, I. Pogribny, A. Schulze, S.H. Zeisel, I. Baric, S.H. Mudd, S-Adenosylhomocysteine hydrolase deficiency in a 26-year-old man, J. Inh. Metab. Dis. 29 (2006) 538-545]. Accordingly, we have proven the Y143C mutation to be highly inactivating [R. Beluzic, M. Cuk, T. Pavkov, K. Fumic, I. Baric, S.H. Mudd, I. Jurak, O. Vugrek, A single mutation at tyrosine 143 of human S-adenosylhomocysteine hydrolase renders the enzyme thermosensitive and effects the oxidation state of bound co-factor NAD, Biochem. J. 400 (2006) 245-253]. Now we report that the A89V exchange leads to a 70% loss of enzymatic activity, respectively. Circular dichroism analysis of recombinant p.A89V protein shows a significantly reduced unfolding temperature by 5.5 degrees C compared to wild-type. Gel filtration of mutant protein is almost identical to wild-type indicating assembly of subunits into the tetrameric complex. However, electrophoretic mobility of p.A89V is notably faster as shown by native polyacrylamide gel electrophoresis implicating changes to the overall charge of the mutant complex. 'Bioinformatics' analysis indicates that Val(89) collides with Thr(84) causing sterical incompatibility. Performing site-directed mutagenesis changing Thr(84) to 'smaller' Ser(84) but preserving similar physico-chemical properties restores most of the catalytic capabilities of the mutant p.A89V enzyme. On the other hand, substitution of Thr(84) with Lys(84) or Gln(84), thereby introducing residues with higher volume in proximity to Ala(89) results in inactivation of wild-type protein. In view of our mutational analysis, we consider changes in charge and the sterical incompatibility in mutant p.A89V protein as main reason for enzyme malfunction with AdoHcyase deficiency as consequence.

S-Adenosylhomocysteine hydrolase deficiency: a second patient, the younger brother of the index patient, and outcomes during therapy.

S-Adenosylhomocysteine (AdoHcy) hydrolase deficiency has been proven in a human only once, in a recently described Croatian boy. Here we report the clinical course and biochemical abnormalities of the younger brother of this proband. This younger brother has the same two mutations in the gene encoding AdoHcy hydrolase, and has been monitored since birth. We report, as well, outcomes during therapy for both patients. The information obtained suggests that the disease starts in utero and is characterized primarily by neuromuscular symptomatology (hypotonia, sluggishness, psychomotor delay, absent tendon reflexes, delayed myelination). The laboratory abnormalities are markedly increased creatine kinase and elevated aminotransferases, as well as specific amino acid aberrations that pinpoint the aetiology. The latter include, most importantly, markedly elevated plasma AdoHcy. Plasma S-adenosylmethionine (AdoMet) is also elevated, as is methionine (although the hypermethioninaemia may be absent or nonsignificant in the first weeks of life). The disease seems to be at least to some extent treatable, as shown by improved myelination and psychomotor development during dietary methionine restriction and supplementation with creatine and phosphatidylcholine.

Effect of olive oil- and corn oil-enriched diets on the tissue mineral content in mice.

The mineral content (zinc, iron, magnesium, and calcium) in the liver, spleen, and thymus of male Balb/C mice was analyzed. Animals were fed, over 21 d, diets enriched with corn oil (FCO diet) or olive oil (FOO diet) (5% addition to standard pellet, w/w). Olive oil with predominant oleic acid (C18:1, n-9) had a quite different composition than corn oil, in which linoleic acid (C18:2, n-6) prevails. The zinc and magnesium tissue concentrations were not changed in either group. The calcium concentration in liver as well as the calcium concentration in spleen increased in mice fed both the FCO and FOO diets. Furthermore, mice fed both the FOO and FCO diets had increased spleen iron concentration. Mice fed the FCO diet had increased thymus calcium concentration compared to controls. The results show the effect of diets with unsaturated, particularly polyunsaturated fatty acids, on the calcium and iron concentration in some organs.

On the role of T lymphocytes in stimulation of humoral immunity induced by peptidoglycan-monomer linked with zinc.

Effects of peptidoglycan linked with zinc (PGM-Zn) were investigated on plaque-forming cell (PFC) generation to sheep red blood cell (SRBC) and SRBC-unrelated antibody production in primary and secondary immune response in mice depleted in vivo of CD4+ and/or CD8+ T lymphocytes. PGM-Zn in nondepleted mice stimulated the PFC generation and IgM or IgG and IgG1 production in primary and secondary reaction. Single depletion of CD4 or CD8+ T cells did not change this ability. The effects of PGM-Zn after CD8+ depletion were even greater than those in nondepleted mice. Depletion of both T cell subsets, however, completely abrogated immunostimulatory effects of PGM on PFC generation (primary and secondary response), as well as on primary SRBC-unrelated antibody production, leaving only the increase of IgG in secondary response unchanged. Immunostimulatory effects and isotype switching to IgG1 and IgG2a correlated with the changes in splenic CD4+, CD8+, CD5+ cells, pointing to the regulatory role of these cells and/or their cytokines in PGM-Zn-induced immunostimulation. Altogether the data suggest that PGM-Zn may potentiate the costimulatory signals coming from activated T cells and act on B cells without the T cell help.

Tissue zinc dynamics during the immune reaction in mice.

Owing to the importance of zinc for the functioning of the immune system, the role of endogenous Zn, located both in lymphoid and nonlymphoid organs, was investigated during the standard humoral and cellular types of immune response. For this purpose, the dynamics of hepatic, thymic, splenic, and renal Zn content was determined in mice sensitized with (a) sheep red blood cells and (b) semiallogeneic lymphocytes during the local host vs graft reaction (HVGR). The data obtained by ion-coupled plasma spectrometry revealed that the humoral type of immunity is characterized by a significant increase of Zn concentration in the liver and in the thymus. Simultaneously, linear regression analysis showed that the generation of plaque-forming cells in the individual mouse was highly positively correlated with Zn concentration in the liver (r=0.897), and spleen (r=0.833), and negatively with Zn concentration in the thymus (r=-0.624). Similar relationships between the intensity of local immune reaction and tissue Zn levels were found in local HVGR at the fifth day in the liver and spleen (r=0.861 and r=0.695, respectively), at the seventh day in the thymus (r=-0.797), and at the tenth day in the liver (r=-0.859). The data emphasize the necessity of Zn for the development of normal immune response and point to the existence of a Zn-dependent hepato-thymic axis during the humoral and cellular types of immune reactivity.

Immunosuppressive and antiproliferative effects of somatostatin analog SMS 201-995.

The effects of long acting somatostatin analog SMS 201-995 were examined in vivo on: 1) lymphoid morphostasis and functional reactivity of cells obtained from SMS treated donors, 2) on humoral, and 3) cellular type of immunity; and in vitro on: 1) blastic transformation of lymphocytes stimulated by activators of different transmembrane pathways (CD2 by PHA and CD3/TCR by anti-CD3 monoclonal antibody and by allogeneic cells) and 2) on growth and secretory activity of several hybridoma cell lines. The data have shown that SMS in vivo decreases the proportion of CD4+, CD5+ and Ig+ cells in spleen. The reactivity of these cells to Con A was suppressed, but their spontaneous blastic transformation was increased. SMS suppressed also the plaque forming cells generation and proliferation of cells in popliteal lymph nodes during the local host versus graft reaction. The former immunosuppression was abrogated with the use of growth hormone, while in the latter, the time dependent changes in spleen composition were also noticed. The data obtained in vitro revealed that SMS may inhibit only the CD2-induced blastogenesis (in early and late interval after the use of PHA). SMS inhibited also the spontaneous growth and/or secretion of antibodies in some hybridoma cell lines.

Immunoregulating effects of peptidoglycan monomer linked with zinc in adult mice.

Since it is well known that both zinc ions and bacterial immunostimulants influence the function of the immune system, in the present study we investigated the immunomodulating activity of a new analog of peptidoglycan monomer (PGM), in which the basic molecule was linked to zinc (PGM-Zn). Its effects in BALB/c mice, aged 10-12 months, were compared with the effects of equimolar doses of PGM and ZnCl2. The treatment lasted 26 days (one i.p. injection every fifth day). The results showed that PGM-Zn may markedly enhance antibody production to sheep red blood cells, as well as spontaneous and concanavalin A (ConA)-induced blastogenesis. The generation of plaque-forming cells in individual mouse was positively correlated with the expression of class II antigens in the liver and negatively correlated with the total quantity of hepatic proteins. PGM-Zn also induced the appearance of peritoneal macrophages, which in cocultures with syngeneic splenocytes were less able to enhance the spontaneous, and particularly the ConA-induced blastic transformation. The enhancing activities of PGM-Zn were in some respects more closely correlated with the action of PGM, whereas the induction of suppressive macrophages resembled more the activity of ZnCl2. The data emphasize that PGM-Zn may both stimulate and inhibit immunoregulative pathways by mechanisms which are not identical to those of PGM or ZnCl2.

Fearful distortions.

Clinical observations suggesting that perceptual distortions can take place during episodes of fear are described, and two hypotheses are set out: (1) perceptual distortions occur during episodes of fear, and (2) such distortions decline after reduction of the pertinent fear. An experiment in which fearful subjects were asked to report their perceptions of a feared object during episodes of fear, and then again after fear-reduction, was carried out. Snake-phobic and spider-phobic subjects showed evidence of some distortions in the activity of the pertinent fear object, but no distortions of size. After the reduction of the relevant fear, the subjects reported significant declines in the activity of the pertinent animal. The two hypotheses received partial support.

Programmed versus spontaneous and stimulated labour.

The present study indicates advantages of programmed labour compared with labour which started spontaneously on or before term day. Programmed labour was shorter and the condition of the newborn, assessed according to Apgar and acid-base values of cord-blood, was better than in spontaneous or stimulated labour groups. There was no appreciable difference in the frequency of other complications such as prematurity, hyperbilirubinemia, and neuropathological symptoms, among groups studied.

The effects of lavour time on neonatal depression.

The effects of labour time on neonatal depression were investigated in 1072 cases. It was found that the everage Apgar score assessed at one minute post-parturition decreased significantly as a function of duration of labour. Additionally, the incidence of neonatal depression (Apgar indes of 7 or less) was found to increase significantly with increase significantly with increases in labour duration. Apgar indices taken five minutes after birth did not reveal any significant effects related to duration of labour. Further analysis of the labour conditions indicated that the effects of labour time on the condition of the infant was greater in spontaneius labours than induced or stimulated labours. In spotanious labours, the five-minute Apgar score continued to be influenced by duration of labour. After five minutes individual differences were clearly observable in all labours.