Evaluation of the Safety,Tolerability, and Pharmacokinetics of Gammaplex<Superscript>®</Superscript> 10% Versus Gammaplex<Superscript>®</Superscript> 5% in Subjects with Primary Immunodeficiency

Purpose

This phase 3, multicenter, open-label, randomized, two-period, crossover bioequivalence trial evaluated the safety, tolerability, and pharmacokinetics of intravenous immunoglobulins (IVIGs) Gammaplex 5% and Gammaplex 10% in 33 adults and 15 children with primary immunodeficiency diseases (PIDs).

Methods

Eligible adults received five Gammaplex 5% infusions followed by five Gammaplex 10% infusions, or vice versa, stratified by a 21- or 28-day dosing regimen. Pediatric subjects received five Gammaplex 10% infusions only.

Results

The primary objective, to demonstrate the bioequivalence of Gammaplex 10% and Gammaplex 5% at the 28-day dosing interval, was met based on the Gammaplex 10%/Gammaplex 5% ratio of area under the concentration versus time curve (AUC_0–28) values. Throughout the study, total immunoglobulin G trough levels were well maintained, with total values generally ≥600 mg/dL (minimum level for study inclusion). At the dosing schedules and infusion rates used in this study, safety and tolerability were comparable and acceptable in adult and pediatric PID subjects treated with Gammaplex 10% and 5%.

Conclusions

In this study, the first direct comparison of 5% IVIG and 10% IVIG products in PID subjects, the pharmacokinetic analysis demonstrated bioequivalence of Gammaplex 10% and Gammaplex 5% at the 28-day dosing interval. The Gammaplex 10% formulation was safe and well tolerated in pediatric and adult PID subjects. Based on the results from this bridging study in PID subjects, Gammaplex 10% could be expected to have a therapeutic effect similar to the licensed Gammaplex 5%, which has demonstrated efficacy and tolerability in patients with PID and idiopathic thrombocytopenic purpura.

     

Inhibitory effects of the neurotensin<Superscript>8–13</Superscript> analogs Asp<Superscript>13</Superscript>-NT<Superscript>8–13</Superscript> and Asp<Superscript>12</Superscript>-NT<Superscript>8–13</Superscript> on mast cell secretion

Pretreatment of isolated mast cells with analogs of neurotensin 8–13 (NT^8–13), in which the amino acids Leu¹³ or Ile¹² are replaced with an aspartic acid (Asp¹³-NT^8–13 or Asp¹²-NT^8–13), inhibits the secretion of histamine in response to NT. A 10 min pretreatment with either analog (10 μM) inhibited NT-induced histamine release by 90% (Asp¹³-NT^8–13) or by 98% (Asp¹²-NT^8–13). At concentrations that are inhibitory, Asp¹³-NT^8–13 and Asp¹²-NT^8–13 alone elicit very little release (<5% at 10 μM). In the continued presence of the analogs, the inhibitory effect lasts for more than 45 min; removal of the analogs resulted in restoration of sensitivity to NT within 10 min. Pretreatment with analog Asp¹³-NT^8–13 resulted in a 39% inhibition of stimulation by substance P and a 52% inhibition of stimulation by histaminereleasing peptide (HRP). In contrast, pretreatment with analog Asp¹²-NT^8–13 gave no inhibition of release by SP or HRP. Neither analog inhibited histamine release in response to bradykinin (BK), NT^1–12, compound 48/80 (48/80), the calcium ionophore A23187, or anti-IgE stimulation of passively sensitized mast cells. Although Asp¹²-NT^8–13 and Asp¹³-NT^8–13 differ slightly in regard to the peptides they inhibit, both probably act at a step early in the stimulus-secretion coupling sequence; most likely before the rise in the level of free intracellular calcium that has been shown to accompany secretion in mast cells. It is suggested that these analogs exert their inhibitory effect on NT by competing with NT for a binding site on the mast cell membrane. The limited number of peptides inhibited by these analogs suggest that not all basic peptides act at the same site to stimulate secretion.     

Chronotropic effect of D-Ala<Superscript>2</Superscript>,Leu<Superscript>5</Superscript>,Arg<Superscript>6</Superscript>-enkephalin (dalargin) is associated with activation of peripheral κ-opioid receptors

Intravenous infusion of D-Ala²,Leu⁵,Arg⁶-enkephalin (dalargin) caused bradycardia in narcotized rats. This effect was not observed during opioid receptor blockade with naloxone, naloxone methiodide, and norbinaltorphimine. Dalargin and (-)-U-50,488 added to Krebs—Henseleit perfusion solution for isolated rat heart decreased heart rate. Ganglionic blocker hexamethonium potentiated the negative chronotropic effect of dalargin. The negative chronotropic effect of dalargin is probably associated with activation of cardiac κ-opioid receptors. It should be noted that dalargin caused tachycardia in some animals. This reaction was not observed after treatment with hexamethonium. The positive chronotropic effect of dalargin is probably related to modulation of the parasympathetic autonomic nervous system. Agonists and antagonists of δ-opioid receptors caused persistent bradycardia. We hypothesized that selective δ-opioid antagonists exhibit properties of partial δ-receptor agonists. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 140, No. 12, pp. 633–638, December, 2005     

Effect of interleukin-1β on NMDA-induced <Superscript>45</Superscript>Ca<Superscript>2+</Superscript> uptake by synaptosomes of rat brain cortex

The effect of interleukin-1β on presynaptic NMDA receptors was evaluated by studying NMDA-induced ⁴⁵Ca²⁺ uptake by synaptosomes from rat brain cortex. Interleukin-1β inhibited ⁴⁵Ca²⁺ uptake by synaptosomes. Our results indicate that interleukin-1β modulates presynaptic NMDA receptors and is probably involved in the regulation of synaptic transmission in the central nervous system. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 140, No. 12, pp. 645–646, December, 2005     

In vitro binding evaluation of <Superscript>177</Superscript>Lu-AMBA,a novel <Superscript>177</Superscript>Lu-labeled GRP-R agonist for systemic radiotherapy in human tissues

Members of the gastrin-releasing peptide (GRP) family and its analogs bombesin (BBN) have been implicated in the biology of several human cancers including prostate, breast, colon and lung. To date, three mammalian GRP/BBN receptor subtypes have been cloned and characterized: the neuromedin B receptor (NMBR), the GRP receptor (GRPR) and the BBN-receptor subtype 3 (BB₃). The fourth BBN receptor subtype, BB₄, has only been identified in amphibian and at present no mammalian equivalent of this receptor has been described. GRPR analogs have been used as carriers to deliver drugs, radionuclides and cytotoxins to target various cancer types that are GRPR positive. We investigated the in vitro binding properties of ¹⁷⁷Lu-AMBA, a novel radiolabelled BBN analog currently undergoing clinical trial as systemic radiotherapy for hormone refractory prostate cancer (HRPC) patients. Pharmacological analyses of the ¹⁷⁷Lu-AMBA was determined using in vitro binding studies using membrane target system containing specific receptor subtypes. We investigated the distribution of binding sites for ¹⁷⁷Lu-AMBA by receptor autoradiography on human neoplastic and non-neoplastic tissues. Pharmacological characterizations of ¹⁷⁷Lu-AMBA shows, high affinity towards NMB and GRP receptors, while little or no affinity towards BB₃ receptor. Among the 40 different types of non-neoplastic tissues tested seven of them showed limited but specific binding of ¹⁷⁷Lu-AMBA. Fourteen of 17 primary prostate cancers, six of 13 primary breast cancers expressed binding sites for ¹⁷⁷Lu-AMBA. Furthermore, no apparent differences in ¹⁷⁷Lu-AMBA-binding sites expression were observed between matched pairs (primary vs. secondary) of prostate and breast cancer tissues. These data represent the molecular basis for clinical applications of ¹⁷⁷Lu-AMBA for diagnosis and treatment of GRP-R and NMB-R positive tumors.     

Effects of Hydroxyapatite and Biostite<Superscript>®</Superscript> on Osteogenic Induction of hMSC

When isolated from the iliac crest human mesenchymal stem cells (hMSC) differentiate into osteoblast-like cells with appropriate stimulation in culture. This in vitro study tested the hypothesis that Biostite^® and hydroxyapatite (HA) affect proliferation and differentiation of hMSC into osteoblastic cells. Cell proliferation was determined by measuring ³H-thymidine incorporation into DNA and typical markers of osteoblastic phenotype were determined by RT-PCR assay. No differences emerged in cell proliferation cultures with Biostite^® or hydroxyapatite (HA), but gene expression analysis revealed higher expression of collagen, alkaline phosphatase (ALP), osteopontin and bone sialoprotein (BSP) in the presence of Biostite^®. TGFβ₂ production, as assessed by an Elisa kit, and Runx2 expression by RT-PCR, were greater in Biostite cultures, suggesting Biostite^® provides a better environment for hMSC differentiation into osteoblasts and is, potentially, a more promising bone-filling material than HA.     

Adenosine Triphosphoric Acid as a Factor of Nervous Regulation of Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>/2Cl<Superscript>−</Superscript> Cotransport in Rat Skeletal Muscle Fibers

Exogenous adenosine triphosphoric acid produces a biphasic effect on the resting membrane potential of muscle fibers in rat diaphragm. Depolarization of the sarcolemma observed 10 min after application of adenosine triphosphoric acid results from activation of Na⁺/K⁺/2Cl⁻ cotransport. The increase in chloride cotransport is related to activation of postsynaptic P2Y receptors and protein kinase C. Repolarization of the membrane develops 40 min after treatment with adenosine triphosphoric acid and after 50 min the resting membrane potential almost returns the control level. This increase in the resting membrane potential of the sarcolemma is probably associated with activation of the Na⁺/K⁺ pump and increase in membrane permeability for chlorine ions in response to longterm activity of Cl⁻ cotransport. Thus, adenosine triphosphoric acid co-secreted with acetylcholine in the neuromuscular synapse probably plays a role in the regulation resting membrane potential and cell volume of muscle fibers.     

Maternal–neonatal erythrocyte membrane Na<Superscript>+</Superscript>, K<Superscript>+</Superscript>-ATPase and Mg<Superscript>2+</Superscript>-ATPase activities in relation to the mode of delivery

Free radical production and high catecholamine levels are implicated in the modulation of Na(+), K(+)-ATPase, and Mg(2+)-ATPase activities. The aim of this study was to investigate the effect of the mode of delivery on the above-mentioned enzyme activities in maternal-neonatal erythrocyte membrane. Women with normal pregnancy (N = 30) were divided into two groups: Group A (N = 16) with normal labor and vaginal delivery, and Group B (N = 14) with scheduled cesarean section; 20 non-pregnant women were the controls. Blood was obtained from controls and mothers, pre- versus post-delivery, and from the umbilical cord (CB). Total antioxidant status (TAS), membrane enzyme activities, and catecholamine blood levels were measured with a commercial kit, spectrophotometrically, and by HPLC methods, respectively. The results showed that: TAS levels, catecholamine, and the membrane enzyme activities were similar in the two groups of mothers pre-delivery, whereas both enzyme activities were lower than those of controls. TAS levels were reduced whereas Na(+), K(+)-ATPase activities (0.35 +/- 0.03 vs. 0.65 +/- 0.06 micromol Pi/h x mg protein, P < 0.001), and catecholamine levels were increased post-delivery in mothers of Group A and unaltered in Group B (0.38 +/- 0.02 vs. 0.40 +/- 0.03 micromol Pi/h x mg protein, P > 0.05), at the same times of study. Mg(2+)-ATPase activities remained unaltered in both groups of mothers and newborns. Na(+), K(+)-ATPase activity was similarly lower in the CB of neonates than those of their mothers, pre-delivery. Our results suggest that: (a) during a normal vaginal delivery process, the low TAS and the increased levels of catecholamines may increase Na(+), K(+)-ATPase activity, post-delivery; (b) the low enzyme activities evaluated in mothers pre-delivery may be due to the high estrogen levels and those in newborns due to perinatal immaturity.     

Sorghum stem extract modulates Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>-ATPase,ecto-5′-nucleotidase,and acetylcholinesterase activities

Sorghum stem (Sorghum bicolor) has been in use in traditional medicine systems for the management of neurodegenerative conditions. However, there is dearth of information on the scientific basis for its use in the treatment of such conditions. This study sought to assess the antioxidant activity and effects of phenolic extract from sorghum stem (Sorghum bicolor) on some cholinergic (acetylcholinesterase (AChE)) and purinergic (Na⁺/K⁺-ATPase and ecto-5′-nucleotidase) enzymes associated with neurological conditions. Phenolic-rich extract was prepared using methanol: 1 N HCl (1:1, v/v) mixture and characterized using high-performance liquid chromatography-diode array detector (HPLC-DAD). In vitro tests were used to investigate the effects of the phenolic extract on AChE, Na⁺/K⁺-ATPase, and ecto-5′-nucleotidase activities. Furthermore, the hydroxyl (OH) radical scavenging and Fe²⁺-chelating abilities of the extract were investigated. HPLC-DAD analysis revealed the presence of some phenolic acids such as caffeic acid (120.58 mg/g), ferulic acid (76.45 mg/g), gallic acid (17.48 mg/g), chlorogenic acid (16.25 mg/g), and flavonoids such as kaempferol (15.98 mg/g), rutin (51.07 mg/g), quercetin (263.16 mg/g), and quercitrin (89.21 mg/g) in the phenolic extract. The extract significantly inhibited AChE and ecto-5′-nucleotidase activities in a dose dependent manner with IC₅₀?=?24.88 μg/ml and IC₅₀?=?37.49 μg/ml, respectively, and increased Na⁺/K⁺-ATPase activity in a dose dependent manner. Furthermore, the phenolic extract also scavenged OH radicals and was able to chelate Fe²⁺ in a dose dependent manner with IC₅₀?=?54.27 μg/ml and 18.47 μg/ml, respectively. This study revealed the antioxidant and modulatory effects of phenolic extracts from sorghum stem on some cholinergic and purinergic enzymes.     

Distinct modulation of microglial amyloid β phagocytosis and migration by neuropeptides<Superscript>i</Superscript>

Microglial activation plays an integral role in the development and course of neurodegeneration. Although neuropeptides such as bradykinin (BK), somatostatin (SST), and endothelin (ET) are known to be important mediators of inflammation in the periphery, evidence of a similar function in brain is scarce. Using immunocytochemistry, we demonstrate the expression of receptors for BK (B1, B2 subtypes), ET (ETA, ETB subtypes) and SST (SST 2, 3, 4 subtypes) in primary microglia and microglial cell lines. Exposure of BV2 and N9, as well as primary microglial cells to BK or SST increased Aβ uptake in a concentration-dependent manner, whereas endothelin decreased Aβ uptake. This was caused by increased phagocytosis of Aβ since the rate of intracellular Aβ degradation remained unaffected. All neuropeptides increased chemotactic activity of microglia. In addition, BK reduced Aβ-induced expression of proinflammatory genes including iNOS and COX-2. ET decreased the Aβ-induced expression of monocyte chemoattractant protein 1 and interleukin-6. These results suggest that neuropeptides play an important role in chemotaxis and Aβ clearance and modulate the brain's response to neuroinflammatory processes.     

A randomized,double-blind comparison of OROS<Superscript>®</Superscript>hydromorphone and controlled-release morphine for the control of chronic cancer pain

Background

Long-acting opioid formulations are advocated for maintaining pain control in chronic cancer pain. OROS^® hydromorphone is a sustained-release formulation of hydromorphone that requires dosing once daily to maintain therapeutic concentrations. The objective of this study was to demonstrate the clinical equivalence of immediate-release and sustained-release formulations of hydromorphone and morphine for chronic cancer pain.

Methods

200 patients with cancer pain (requiring ≤ 540 mg/d of oral morphine) participated in this double-blind, parallel-group trial. Patients were randomized to receive hydromorphone or morphine (immediate-release for 2–9 days, sustained-release for 10–15 days). Efficacy was assessed with the Brief Pain Inventory (BPI), investigator and patient global evaluations, Eastern Cooperative Oncology Group performance status, and the Mini-Mental State Examination. The primary endpoint was the 'worst pain in the past 24 hours' item of the BPI, in both the immediate-release and sustained-release study phases, with treatments deemed equivalent if the 95% confidence intervals (CI) of the between-group differences at endpoint were between -1.5 and 1.5. No equivalence limits were defined for secondary endpoints.

Results

Least-squares mean differences (95% CI) between groups were 0.2 (-0.4, 0.9) in the immediate-release phase and -0.8 (-1.6, -0.01) in the sustained-release phase (intent-to-treat population), indicating that the immediate-release formulations met the pre-specified equivalence criteria, but that the lower limit of the 95% CI (-1.6) was outside the boundary (-1.5) for the sustained-release formulations. BPI 'pain now PM' was significantly lower with OROS^® hydromorphone compared with controlled-release morphine (least-squares mean difference [95% CI], -0.77 [-1.49, -0.05]; p = 0.0372). Scores for other secondary efficacy variables were similar between the two sustained-release treatments. At endpoint, > 70% of investigators and patients rated both treatments as good to excellent. The safety profiles of hydromorphone and morphine were similar and typical of opioid analgesics.

Conclusion

Equivalence was demonstrated for immediate-release formulations of hydromorphone and morphine, but not for the sustained-release formulations of OROS^® hydromorphone and controlled-release morphine. The direction of the mean difference between the treatments (-0.8) and the out-of-range lower limit of the 95% CI (-1.6) were in favor of OROS^® hydromorphone.

Trial registration

ClinicalTrials.gov: NCT0041054

     

T<Superscript>+</Superscript> NK<Superscript>+</Superscript> IL-2 Receptor γ Chain Mutation: a Challenging Diagnosis of Atypical Severe Combined Immunodeficiency

Purpose

All reported patients with hypomorphic X-linked severe combined immunodeficiency (X-SCID) due to c.664C>T (p.R222C) mutations in the gene (IL2RG) encoding the common γ chain (γc) have presented with opportunistic infections within the first year of life, despite the presence of nearly normal NK and T cell numbers. Reporting five children of one extended family with hemizygous mutations in IL2RG, we explore potential diagnostic clues and extend our comprehension of the functional impact of this mutation.

Methods

Whole exome sequencing (WES); detailed immune phenotyping; cytokine-induced STAT phosphorylation; B, T, and NK cell activation; and quantification of sjTRECs in five Arab children with c.664C>T (p.R222C) IL2RG mutation.

Results

The mean age at clinical presentation with respiratory tract infection or diarrhea was 6.8 (range: 2–12) months. None of the children presented with opportunistic infections. Diagnostic clues were early onset in the first year of life, and a suggestive family history associated with reduced naïve CD4 T cells and absent switched memory B cells. Number and phenotype of NK cells and innate-like lymphocytes were normal. The diagnosis was made by WES and corroborated by absent STAT phosphorylation and reduced functional response after IL-2 and IL-21 stimulation. Four patients underwent successful hematopoietic stem cell transplantation.

Conclusions

As early diagnosis and treatment are important, a high index of suspicion in the diagnosis of c.664C>T (p.R222C) X-SCID is needed. This requires prompt genetic testing by next generation sequencing in order to avoid unnecessary delays in the definite diagnosis since immunological work up may not be discriminating. Assays directly testing cytokine signaling or cytokine-dependent functions are helpful in confirming the functional impact of the identified hypomorphic variants.

     

Dimethyl amiloride,a Na<Superscript>+</Superscript>–H<Superscript>+</Superscript> exchange inhibitor,and its cardioprotective effects in hemorrhagic shock in in vivo resuscitated rats

Stimulation of the Na⁺–H⁺ exchanger plays an important role in the pathway of myocardial dysfunction and injury following hemorrhagic shock. Inhibition of the Na⁺–H⁺ exchanger appears to be a new pharmacological tool for myocardial protection. Despite the extensive research that has been done on the role of the Na⁺–H⁺ exchanger in ischemia reperfusion, little is known about the role of the exchanger following hemorrhagic shock. The purpose of this study was to examine the protective effects of blocking the cardiac Na⁺–H⁺ exchanger, using 20 μM dimethyl amiloride (DMA), a specific Na⁺–H⁺ exchanger blocker, on myocardial contractile function after ex vivo perfusion of isolated rat heart following 1 h of hemorrhagic shock. Sprague–Dawley rats were assigned to hemorrhage + DMA, hemorrhage, sham hemorrhage + DMA and sham hemorrhage groups (n = 6 per group). Hearts were perfused with a balanced salt solution for 60 min. In the DMA treated group, 20 μM DMA was added for the first 5 min of the 60-min ex vivo heart resuscitation. The results showed that inhibition of the Na⁺–H⁺ exchanger for 5 min on ex vivo perfusion of the isolated hearts following hemorrhagic shock using 20 μM DMA improved myocardial contractile function. Blocking the Na⁺–H⁺ exchanger on ex vivo perfusion of isolated hearts using 20 μM DMA has protective effects on myocardial contractile function.     

The effect of Neuragen PN<Superscript>®</Superscript> on Neuropathic pain: A randomized,double blind,placebo controlled clinical trial

Background

A double blind, randomized, placebo controlled study to evaluate the safety and efficacy of the naturally derived topical oil, "Neuragen PN^®" for the treatment of neuropathic pain.

Methods

Sixty participants with plantar cutaneous (foot sole) pain due to all cause peripheral neuropathy were recruited from the community. Each subject was randomly assigned to receive one of two treatments (Neuragen PN^® or placebo) per week in a crossover design. The primary outcome measure was acute spontaneous pain level as reported on a visual analog scale.

Results

There was an overall pain reduction for both treatments from pre to post application. As compared to the placebo, Neuragen PN^® led to significantly (p < .05) greater pain reduction. Fifty six of sixty subjects (93.3%) receiving Neuragen PN^® reported pain reduction within 30 minutes. This reduction within 30 minutes occurred in only twenty one of sixty (35.0%) subjects receiving the placebo. In a break out analysis of the diabetic only subgroup, 94% of subjects in the Neuragen PN^® group achieved pain reduction within 30 minutes vs 11.0% of the placebo group. No adverse events were observed.

Conclusions

This randomized, placebo controlled, clinical trial with crossover design revealed that the naturally derived oil, Neuragen PN^®, provided significant relief from neuropathic pain in an all cause neuropathy group. Participants with diabetes within this group experienced similar pain relief.

Trial registration

ISRCTN registered: ISRCTN13226601

     

CD3<Superscript>+</Superscript>ICOS<Superscript>+</Superscript> T cells show differences in the synthesis of nitric oxide,IFN-γ, and IL-10 in patients with pulmonary tuberculosis or in healthy household contacts

Evidence indicates that more than 90 % of infected individuals never develop active tuberculosis. This fact highlights the relevance of the immune response in tuberculosis control. The inducible co-stimulator (ICOS) is a regulator of the function, differentiation, proliferation, and activation of T cells. Moreover, T cells synthesise nitric oxide (NO), interferon gamma (IFN-γ), and interleukin (IL)-10, which help regulate the immune response to tuberculosis. Therefore, we assessed the synthesis of NO, IFN-γ, and IL-10 in CD3⁺ICOS⁺ T cells from healthy individuals, household contacts (HHC), and patients with active pulmonary tuberculosis (PTB), previously stimulated with the antigen H37Rv. Our results indicated a significant increase in both the percentage of ICOS⁺ cells and CD3⁺ICOS⁺ T cells producing NO, IFN-γ, and IL-10 in cells obtained from patients with PTB (p < 0.01). In addition, a high mitochondrial membrane potential (ΔΨ _m) in CD3⁺ICOS⁺ T cells was observed in the cells from HHC and from PTB patients, and is associated with the activation of T cells. In conclusion, results show that the CD3⁺ICOS⁺ T cells obtained from PTB patients are the main producers of NO, IFN-γ, and IL-10. In addition, our results imply that NO is a modulator of ICOS expression of T cells from PTB patients.     

Effects of two dietary exogenous multi-enzyme supplementation,Natuzyme<Superscript>®</Superscript> and beta-mannanase (Hemicell<Superscript>®</Superscript>), on growth and blood parameters of Caspian salmon (<Emphasis Type="Italic">Salmo trutta caspius</Emphasis>)

Recent increases in feed ingredient costs have motivated the fisheries industry to identify technologies that will improve feed utilisation and reduce the cost per pound of gain. The effects of two supplemental exogenous enzymes (Natuzyme® and Hemicell®) on the growth performance in Caspian salmon (Salmo trutta caspius) were examined over an 8-week feeding trial. After the experimental period, the survival rate ranged from 91.33?±?1.15 % in controls to 96.67?±?1.15 % in the group that received 0.5 g Natuzyme® kg^?1?+?0.5 g Hemicell® kg^?1 (NH) in their diet and there was a statistical difference between experimental and control groups (p?<?0.05). Growth rate was significantly higher in the NH group (1.01?±?0.01) than the other groups (Sig.?=?0.00). The best feed conversion rate (0.64?±?0.01) was in the NH group and it was significantly lower than the control group, the 0.5 g Natuzyme® kg^?1 group, and the 0.25 g Hemicell® kg^?1 group (Sig.?=?0.03). The best final body weight (80.68?±?5.27) was observed in the NH group. Also, WBC count (7,716.67?±?348.80 N/mm³) was significantly higher in the NH group compared to the control (6,916.67?±?194.10 N/mm³; p?<?0.05). No difference was observed in haematocrit%, haemoglobin, red blood cell, mean corpuscular volume, mean corpuscular haemoglobin, and mean corpuscular haemoglobin concentration (p?>?0.05). The results suggested that enzyme supplementation caused significant improvement on growth performance and feed utilisation in Caspian salmon.     

Comparative phase I randomized open-label pilot clinical trial of Gynophilus<Superscript>®</Superscript> (Lcr regenerans<Superscript>®</Superscript>) immediate release capsules versus slow release muco-adhesive tablets

Gynophilus^® (Lcr regenerans^®) is a live biotherapeutic product (LBP) that contains the live biotherapeutic microorganism Lactobacillus rhamnosus Lcr35^®, which is indicated to restore vaginal health. The aim of the study was to compare the safety, ease of use, and compliance of two formulations (immediate release: IR capsule and slow release: SR muco-adhesive tablets) as well as the colonization of Lcr35^® in healthy women. This phase I study (Comprigel) is a parallel, randomized, 4-arm, and open-label clinical trial evaluating an IR daily capsule formulation vs. a SR tablet administered every 3, 4, or 5 days for 21 days. Self-collected vaginal swabs were used to quantify Lcr35^® and characterize the composition and structure of the vaginal microbiota. Both LBPs were well-tolerated, and no severe adverse effects were reported. All groups had Lcr35^® vaginal concentrations over 10⁷ colony forming unit per milliliter of vaginal secretion on each day in the study. The new Gynophilus^® slow release tablets administered either every 3, 4, or 5 days provided vaginal concentrations that were not significantly different from those of classic Gynophilus^® (capsule) once-a-day regimen. The LBPs and the different regimens did not adversely influence the abundance of native Lactobacillus spp. and indeed tended to favor their growth and reduce colonization by non-Lactobacillus spp. This study illustrates that the SR muco-adhesive LBP tablet (Gynophilus^® SR) administered every 3 or 4 days as a safe, well-tolerated, and efficacious alternative to a more demanding IR daily capsule and could protect women’s healthy vaginal microbiome by promoting endogenous Lactobacillus spp.     

Improvement in the detection of enteric protozoa from clinical stool samples using the automated urine sediment analyzer sediMAX<Superscript>®</Superscript> 2 compared to sediMAX<Superscript>®</Superscript> 1

Detection of intestinal parasites from fecal samples is routinely performed by direct wet mount examination. This method requires skilled personnel, and it is time consuming. The aim of this work is to demonstrate the usefulness of the newer automated urinary sediment analyser sediMAX 2 for a fast detection of intestinal protozoa in stool samples. A total of 700 consecutively preserved samples consisting of 70 positives and 630 negatives were analyzed. SediMAX 2 takes digital images of each sediment sample, and analysis was conducted using a dilution of stool specimens, allowing determination of typical morphology. Compared to manual microscopy, sediMAX 2 showed sensitivity and specificity of 100 % in the detection of intestinal parasites, as also recently demonstrated for sediMAX 1. However, all clinically important human protozoa were detected using only 15 images for each specimen, compared to 30 images required in sediMAX 1 analysis. Moreover, changing manually the focus, it is possible to carry out a discrimination between morphologically identical Entamoeba complex members, including the pathogenic E. histolytica and the non-pathogenic E. dispar, E. moshkovskii and E. Bangladeshi, from the non-pathogenic Entamoeba coli based on the number of nuclei present in the cells. This study presents sediMAX 2 as an automatic aid to traditional microscopy.     

Efficacy and Safety of Hizentra<Superscript>®</Superscript>, a New 20% Immunoglobulin Preparation for Subcutaneous Administration,in Pediatric Patients with Primary Immunodeficiency

Subcutaneous IgG treatment for primary immunodeficiencies (PI) is particularly well suited for children because it does not require venous access and is mostly free of systemic adverse events (AEs). In a prospective, open-label, multicenter, single-arm, Phase III study, 18 children and five adolescents with PI were switched from previous intravenous (IVIG) or subcutaneous (SCIG) IgG treatment to receive dose-equivalent, weekly subcutaneous infusions of Hizentra^® for 40 weeks. Mean IgG trough levels were maintained in patients previously on SCIG, or increased in those previously on IVIG, regardless of age. No serious bacterial infections were reported during the efficacy period of the study. The rates of non-serious infections were 4.77 (children) and 5.18 (adolescents) infections per patient per year. Related AEs were observed in seven children (38.9%) and two adolescents (40%). Three serious AEs and two AEs leading to discontinuation (all unrelated) were reported in children. Hizentra^® is an effective and well-tolerated treatment for pediatric patients.     

Effect of classic convulsants on Cl<Superscript>−</Superscript> conductance of the GABA<Subscript>A</Subscript> receptor complex in membranes of cerebral cortex cells at the early stage of kindling

Muscimol-stimulated Cl- conductance of synaptoneurosomes from the cerebral cortex of Wistar rats increased during the early stage of pharmacological kindling not inducing the seizure response in animals. Picrotoxin, bicuculline, and pentylenetetrazole potentiated inhibition of muscimol-dependent 36Cl- entry into synaptoneurosomes, which attested to increased sensitivity of the GABA(A) receptor/Cl- ionophore complex to classic convulsants.