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Alzheimer’s disease (AD) is an age-related neurodegenerative disorder associated with memory loss, but the AD-associated neuropathological changes begin years before memory impairments. Investigation of the early molecular abnormalities in AD might offer innovative opportunities to target memory impairment prior to onset. Decreased protein synthesis plays a fundamental role in AD, yet the consequences of this dysregulation for cellular function remain unknown. We hypothesize that alterations in the de novo proteome drive early metabolic alterations in the hippocampus that persist throughout AD progression. Using a combinatorial amino acid tagging approach to selectively label and enrich newly synthesized proteins, we found that the de novo proteome is disturbed in young APP/PS1 mice prior to symptom onset, affecting the synthesis of multiple components of the synaptic, lysosomal, and mitochondrial pathways. Furthermore, the synthesis of large clusters of ribosomal subunits were affected throughout development. Our data suggest that large-scale changes in protein synthesis could underlie cellular dysfunction in AD., Elder et al. investigated the de novo proteome in isolated hippocampal slices of young APP/PS1 Alzheimer’s model mice and revealed that large-scale changes in protein synthesis precede symptom onset. These results suggest that broad dysregulation of protein synthesis could contribute toward cellular dysfunction in Alzheimer’s disease.

Endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) has been shown to activate the eIF2α kinase PERK to directly regulate translation initiation. Tight control of PERK-eIF2α signaling has been shown to be necessary for normal long-lasting synaptic plasticity and cognitive function, including memory. In contrast, chronic activation of PERK-eIF2α signaling has been shown to contribute to pathophysiology, including memory impairments, associated with multiple neurological diseases, making this pathway an attractive therapeutic target. Herein, using multiple genetic approaches we show that selective deletion of the PERK in mouse midbrain dopaminergic (DA) neurons results in multiple cognitive and motor phenotypes. Conditional expression of phospho-mutant eIF2α in DA neurons recapitulated the phenotypes caused by deletion of PERK, consistent with a causal role of decreased eIF2α phosphorylation for these phenotypes. In addition, deletion of PERK in DA neurons resulted in altered de novo translation, as well as changes in axonal DA release and uptake in the striatum that mirror the pattern of motor changes observed. Taken together, our findings show that proper regulation of PERK-eIF2α signaling in DA neurons is required for normal cognitive and motor function in a non-pathological state, and also provide new insight concerning the onset of neuropsychiatric disorders that accompany UPR failure.

Background Development of a diverse T-cell receptor β (TRB) repertoire is associated with immune recovery following hematopoietic cell transplantation (HCT) for severe combined immunodeficiency (SCID). High-throughput sequencing of the TRB repertoire allows evaluation of clonotype dynamics during immune reconstitution. Objectives We investigated whether longitudinal analysis of the TRB repertoire would accurately describe T-cell receptor diversity and illustrate the quality of T-cell reconstitution following HCT or gene therapy for SCID. Methods We used high-throughput sequencing to study composition and diversity of the TRB repertoire in 27 infants with SCID at 3, 6, and 12 months and yearly posttreatment(s). Total RNA from peripheral blood was used as template to amplify TRB rearrangements. Results TRB sequence analysis showed poor diversity at 3 months, followed by significant improvement by 6 months after cellular therapies. Kinetics of development of TRB diversity were similar in patients with a range of underlying gene defects. However, in patients with RAG and DCLRE1C defects, HCT with no conditioning or immune suppression only resulted in lower diversity than did HCT with conditioning. HCT from a matched donor correlated with higher diversity than did HCT from a mismatched donor. Naive CD4+ T-cell count at 6 months post-HCT correlated with higher TRB diversity. A Shannon index of diversity of 5.2 or lower 3 months after HCT predicted a need for a second intervention. Conclusions TRB repertoire after hematopoietic cell therapies for SCID provides a quantitative and qualitative measure of diversity of T-cell reconstitution and permits early identification of patients who may require a second intervention.

Alzheimer’s disease (AD) is an age-related neurodegenerative disorder, but neuropathological changes in AD begin years before memory impairment. Investigation of the early molecular abnormalities in AD might offer innovative opportunities to target memory impairment prior to its onset. Decreased protein synthesis plays a fundamental role in AD, yet the consequences for cellular function remain unknown. We hypothesize that alterations in the de novo proteome drive early metabolic changes in the hippocampus that persist throughout AD progression. Using a combinatorial amino acid tagging approach to selectively label newly synthesized proteins, we found that the de novo proteome is disturbed in APP/PS1 mice prior to pathology development, affecting the synthesis of multiple components of synaptic, lysosomal and mitochondrial pathways. Furthermore, the synthesis of large clusters of ribosomal subunits were affected throughout neuropathology development in these mice. Our data suggest that large-scale changes in protein synthesis could underlie cellular dysfunction in AD.

For patients receiving haploidentical donor (haplo) hematopoietic cell transplant (HCT), depletion of TCRαβ T cells from the haplo-graft allows for excellent outcomes and low rates of graft-versus-host disease (GVHD), albeit with a significant delay in immune reconstitution (IR). The adoptive transfer of donor T-cells has been used to optimize IR but increases the risk of GVHD. CD45RA-depleted (memory) T-cells are associated with low rates of alloreactivity and thereby GVHD but retain specificity for leukemic and viral antigens. We implemented a prospective trial utilizing escalating doses of CD45RA-depeleted T-cell as addback following TCRαβ/CD19-depleted haploHCT to improve IR. Patients with acute lymphoblastic leukemia (ALL) also received prophylactic Blinatumomab (Blina) following infusion of CD45RA-depeleted T-cell to overcome the risk immune escape secondary to HLA-loss and relapse (NCT03849651). Between 2019 to 2020, 30 pediatric patients (9 males, 21 females) with high-risk acute leukemia were enrolled. Median age at HCT was 8.7 years (range 0.9-18.8). Nineteen patients had ALL, 11 patients had AML. Ten patients were in CR1, 13 in CR2 and 7 in CR3/>. Five patients received prior CD19-CAR T-cell therapy. The donors used were mothers (n=15), fathers (n=13), or sibling/others (n=2). All patients received a reduced intensity preparative regimen consisting of Fludarabine, Melphalan, Cyclophosphamide and Thiotepa. ATG was given on days -5, -4 and -3. Mobilized peripheral blood graft were infused on day 0 with a median number of CD34+ cells, γδ+ T-cells, αβ+ T cells, and B cells of 14.8, 22.7, 0, and 0.09x 10 6/kg, respectively. No GVHD prophylaxis was used post-HCT. Two weeks following engraftment, patients received CD45RA-depleted T-cell addback in 3 escalating doses (DL1: 1x10 5cells/kg, DL2: 1x10 6 cells/kg, DL3:1x10 7 cells/kg). All 30 patients engrafted with a median time for neutrophil and platelet engraftment of 10 (range: 9-11) and 15 (range: 13-20) days, respectively. Infusion of escalating doses of CD45RA-depleted T-cells was well tolerated. One month post infusion, there was a significant increase in the median number of CD3 T-cells, including CD8 and CD45RO+ T-cell subsets (**p In this interim analysis, addback of CD45RA- depleted T-cells following TCRαβ/CD19-depleted haplo HCT was safe and led to enhanced functional immune reconstitution. Prophylactic infusion of Blina is well tolerated and its use post-transplant warrants further investigation. Analyses into the effect of ATG on immune reconstitution are underway. Figure 1 Figure 1. Disclosures Sharma: CRISPR Therapeutics: Other, Research Funding; Novartis: Other: Salary support paid to institution; Vertex Pharmaceuticals/CRISPR Therapeutics: Other: Salary support paid to institution; Spotlight Therapeutics: Consultancy; Medexus Inc: Consultancy; Vindico Medical Education: Honoraria. Gottschalk: Catamaran Bio: Consultancy; Immatics: Membership on an entity's Board of Directors or advisory committees; Other: Other: patents and patent applications in the field of cancer cell and gene therapy ; Novartis: Consultancy; Tidal: Consultancy; Tessa Therapeutics: Consultancy. Triplett: Miltenyi: Other: Travel, meeting registration.

To survive in a dynamic environment, animals need to identify and appropriately respond to stimuli that signal danger1. Survival also depends on suppressing the threat-response during a stimulus that predicts the absence of threat (safety)2-5. An understanding of the biological substrates of emotional memories during a task in which animals learn to flexibly execute defensive responses to a threat-predictive cue and a safety cue is critical for developing treatments for memory disorders such as post-traumatic stress disorder5. The centrolateral amygdala is an important node in the neuronal circuit that mediates defensive responses6-9, and a key brain area for processing and storing threat memories. Here we applied intersectional chemogenetic strategies to inhibitory neurons in the centrolateral amygdala of mice to block cell-type-specific translation programs that are sensitive to depletion of eukaryotic initiation factor 4E (eIF4E) and phosphorylation of eukaryotic initiation factor 2α (p-eIF2α). We show that de novo translation in somatostatin-expressing inhibitory neurons in the centrolateral amygdala is necessary for the long-term storage of conditioned-threat responses, whereas de novo translation in protein kinase Cδ-expressing inhibitory neurons in the centrolateral amygdala is necessary for the inhibition of a conditioned response to a safety cue. Our results provide insight into the role of de novo protein synthesis in distinct inhibitory neuron populations in the centrolateral amygdala during the consolidation of long-term memories.

The purpose of this study was to analyse the influence of water salinity on the incubation of eggs and growth of the larvae of sichel Pelecus cultratus (L.) under controlled conditions. The following water salinity was considered: 3, 6, 9, 12, 15 and 0 ppt for the control. The eggs incubation time depended on the water salinity, lasting from 56.3°D [9 ppt] to 107.7°D [0 ppt]. The higher water salinity, shorter the incubation time. The highest survivability of larvae was obtained in water of the salinity equal 3 ppt and in freshwater, where respectively 87% and 84% of the larvae survived. No hatched larvae were obtained in water of the salinity of 12 and 15 ppt, as all embryos died during the experiment. Larvae were reared up for 21 days. The final body weight, depending on the water salinity, reached from 43.5 mg [9 ppt] to 74.6 mg [3 ppt], at the respective body lengths of 19.3 mm and 23.2 mm. The highest body gains and lengths of larvae were observed in water of the salinity of 3 ppt. The lowest survivability of sichel larvae occurred in water of the salinity 9 ppt, where it was only 53.2%. The survivability of larvae in freshwater and water of the salinity of 3 ppt was quite similar: 96.2% and 95.6% respectively. Water salinity degree of 12 and 15 ppt appeared to be lethal for both embryos and larvae of the sichel.

Whether fragile X mental retardation protein (FMRP) target mRNAs and neuronal activity contributing to elevated basal neuronal protein synthesis in fragile X syndrome (FXS) is unclear. Our proteomic experiments reveal that the de novo translational profile in FXS model mice is altered at steady state and in response to metabotropic glutamate receptor (mGluR) stimulation, but the proteins expressed differ under these conditions. Several altered proteins, including Hexokinase 1 and Ras, also are expressed in the blood of FXS model mice and pharmacological treatments previously reported to ameliorate phenotypes modify their abundance in blood. In addition, plasma levels of Hexokinase 1 and Ras differ between FXS patients and healthy volunteers. Our data suggest that brain-based de novo proteomics in FXS model mice can be used to find altered expression of proteins in blood that could serve as disease-state biomarkers in individuals with FXS., Elevated protein synthesis, and dysregulated mGluR signalling, are documented in fragile X syndrome (FXS) Here the authors use proteomic analysis in a mouse model of FXS, and following mGluR5 stimulation, to identify potential biomarkers for the disease.

Aberrant neuronal translation is implicated in the etiology of numerous brain disorders. Although mTORC1-p70 ribosomal S6 kinase 1 (S6K1) signaling is critical for translational control, pharmacological manipulation in vivo has targeted exclusively mTORC1 due to the paucity of specific inhibitors to S6K1. However, small molecule inhibitors of S6K1 could potentially ameliorate pathological phenotypes of diseases, which are based on aberrant translation and protein expression. One such condition is fragile X syndrome (FXS), which is considered to be caused by exaggerated neuronal translation and is the most frequent heritable cause of autism spectrum disorder (ASD). To date, potential therapeutic interventions in FXS have focused largely on targets upstream of translational control to normalize FXS-related phenotypes. Here we test the ability of two S6K1 inhibitors, PF-4708671 and FS-115, to normalize translational homeostasis and other phenotypes exhibited by FXS model mice. We found that although the pharmacokinetic profiles of the two S6K1 inhibitors differed, they overlapped in reversing multiple disease-associated phenotypes in FXS model mice including exaggerated protein synthesis, inappropriate social behavior, behavioral inflexibility, altered dendritic spine morphology, and macroorchidism. In contrast, the two inhibitors differed in their ability to rescue stereotypic marble-burying behavior and weight gain. These findings provide an initial pharmacological characterization of the impact of S6K1 inhibitors in vivo for FXS, and have therapeutic implications for other neuropsychiatric conditions involving aberrant mTORC1-S6K1 signaling.

The aim of this study was to characterize changes in maximal oxygen uptake over several years and to elaborate current reference values of this index based on determinations carried out in large and representative groups of top Polish rowers. For this study 81 female and 159 male rowers from the sub-junior to senior categories were recruited from the Polish National Team and its direct backup. All the subjects performed an incremental exercise test on a rowing ergometer. During the test maximal oxygen uptake was measured with the BxB method. The calculated reference values for elite Polish junior and U23 rowers allowed to evaluate the athletes’ fitness level against the respective reference group and may aid the coach in controlling the training process. Mean values of VO2max achieved by members of the top Polish rowing crews who over the last five years competed in the Olympic Games or World Championships were also presented. The results of the research on the “trainability” of the maximal oxygen uptake may lead to a conclusion that the growth rate of the index is larger in case of high-level athletes and that the index (in absolute values) increases significantly between the age of 19-22 years (U23 category).

This study analyzed the influence of temperature on the rearing of sichel Pelecus cultratus larvae under controlled conditions. Five constant temperature regimes were tested: 14 °C, 18 °C, 22 °C, 26 °C and 30 °C. The larvae were reared for 30 days in aquaria with 50 dm3 working capacities. The larvae density was 40 individuals/dm3 and the fish larvae were fed live artemia nauplii. The assessment of the effect of thermal conditions during the rearing period on fish was based on the final size of fish, increment in the total length, relative growth ratio, survivability and condition (the Fulton index). At the end of the 30-day rearing period, it was determined that the largest larvae had been obtained in the 30 °C water (average weight 374.2 mg with average length 43.1 mm). The smallest larvae grew in the coldest water (14 °C), where their average weight was 32.2 mg with an average length of 19.4 mm. No negative effect of temperature was observed on the survivability rate of reared larvae, which oscillated around 91 ± 5 %. Sichel larvae are characterized by high tolerance and high survivability when reared in high-temperature water.

The common bream, Abramis brama (L.) in Kortowskie Lake (north-eastern Poland) usually spawn when the water temperature during the late spring rises to 20 °C. The optimal water temperature for embryonic development of this population is 21 °C. However, in some cases, the temperatures of the spawning grounds during bream spawning are much higher, becoming semi-lethal or even lethal for bream embryos. The aim of the study was to investigate the natural and artificial spawning effectiveness of bream during a 4-year study regarding the reproductive effectiveness of the wild bream population in relation to thermal fluctuation (optimal and semi-lethal) of their ecosystem during the natural spawning season. During four following seasons, the bream spawners, as well as the eggs, were collected from the spawning grounds and incubated at optimal (21 °C) and semi-lethal (23 °C) temperatures. The spawning of mature bream was artificially induced and the obtained eggs were also incubated at the same temperatures. The period of observed spawners on the spawning grounds was different in different years: the longest time of bream spawning was in the second year of the study and the shortest was during the third year of the study. In the last year, a decrease in water temperature caused a break in the spawning. Thermal changes in water temperature during the spawning season may have caused a high mortality level of bream embryos and decreased the recruitment of the next species generation.

Feeding and food selection of burbot (Lota lota L.) larvae reared in illuminated cages were studied. The experiment was carried out in mesotrophic Lake Maroz, in north-eastern Poland, for 6 weeks in two successive years. The initial stocking density was 1,250 larvae (20 DPH) per cage. Food selection according to the zooplankton groups (Rotifera, Cladocera and Copepoda) and length classes was expressed by the Strauss linear selectivity index (L). Zooplankton species composition in the lake was similar in the two seasons of the study and organisms shorter than 0.5 mm prevailed in the plankton. The mean number of prey found in burbot alimentary tracts increased from about 40 up to over 200 during the course of the study. A very large inter-individual variation in the amount of food organisms consumed by fish was noted. Analysis of the values of the Strauss food selectivity index shows that at the beginning of the first year of the experiment, burbot larvae preferred copepods, most numerous in the environment at that time; later, fish tended to select cladocerans. In the second year of the study, fish more often ate copepods, irrespective of their quantities in the environment. During the whole study, reared burbot larvae did not eat rotifers, even when they were numerous in cages. Similarly to the rotifers, the smallest planktonic organisms, measuring up to 0.5 mm in length, were typically neglected by fish, while the 0.6–1.0 mm group was most frequently selected. There were also considerable individual differences between particular burbot specimens in their food preferences.

The aim of the present study was to analyse the occurrence of macroscopically visi- ble ovary alterations in 2 populations of northern pike Esox lucius L. originating from lakes in the Mazurian Lake District (NE Poland). The alterations were characterised by ovary tissue that was morphologically malformed, in part or in whole, and contained immature oocytes, i.e. trophoplas- tic or previtellogenic oocytes instead of vitellogenic oocytes. These alterations were found only in the ovaries, and no morphological alterations of the testes were noted. Macroscopic and histolog- ical analyses were carried out in order to classify the observed alterations in the ovaries. Three types of alterations were identified in which morphological malformations as well as histological investigation of the ovaries were considered. An analysis of the size and age of the fish in relation to the occurrence of alterations as well as of the macroscopic and histological nature of the alter- ation types was made. The data obtained revealed no lake or age dependency of the observed alterations. Based on the results obtained, we suggest that the presence of endocrine disruptors in the environment or/and genetic factors could be responsible for these kinds of gonad anomalies. However, our results did not allow us to determine the aetiology of the alterations.

Caffeine and melatonin have been shown to protect the Swedish mutant amyloid precursor protein (APP(sw)) transgenic mouse model of Alzheimer's disease from cognitive dysfunction. But their mechanisms of action remain incompletely understood. These Alzheimer's mice have extensive mitochondrial dysfunction, which likely contributes to their cognitive decline. To further explore the mechanism through which caffeine and melatonin protect cognitive function in these mice, we monitored the function of isolated mitochondria from APP(sw) mice treated with caffeine, melatonin, or both in their drinking water for one month. Melatonin treatment yielded a near complete restoration of mitochondrial function in assays of respiratory rate, membrane potential, reactive oxygen species production, and ATP levels. Caffeine treatment by itself yielded a small increase in mitochondrial function. However, caffeine largely blocked the large enhancement of mitochondrial function provided by melatonin. Studies with N2a neuroblastoma cells stably expressing APP(sw) showed that specific inhibition of cAMP-dependent phosphodiesterase (PDE) 4 or cGMP-dependent PDE5 also blocked melatonin protection of mitochondrial function, but A(2a) and A₁ adenosine receptor antagonists were without effect. Melatonin or caffeine at the concentrations used to modulate mitochondrial function in the cells had no effect on cAMP-dependent PDE activity or cellular cAMP or cGMP levels. Therefore, caffeine and increased cyclic nucleotide levels likely block melatonin signaling to mitochondria by independent mechanisms that do not involve adenosine receptor antagonism. The results of this study indicate that melatonin restores mitochondrial function much more potently than caffeine in APP(sw) transgenic mouse and cell models of Alzheimer's disease.

Alzheimer's disease (AD) affects millions of people world-wide and new effective and safe therapies are needed. Cotinine, the main metabolite of nicotine, has a long half-life and does not have cardiovascular or addictive side effects in humans. We studied the effect of cotinine on amyloid-β (Aβ) aggregation as well as addressed its impact on working and reference memories. Cotinine reduced Aβ deposition, improved working and reference memories, and inhibited Aβ oligomerization in the brains of transgenic (Tg) 6799 AD mice. In vitro studies confirmed the inhibitory effect of cotinine on Aβ1-42 aggregation. Cotinine stimulated Akt signaling, including the inhibition of glycogen synthase kinase 3β (GSK3β), which promotes neuronal survival and the synaptic plasticity processes underlying learning and memory in the hippocampus and cortex of wild type and Tg6799 AD mice. Simulation of the cotinine-Aβ1-42 complex using molecular dynamics showed that cotinine may interact with key histidine residues of Aβ1-42, altering its structure and inhibiting its aggregation. The good safety profile in humans and its beneficial effects suggest that cotinine may be an excellent therapeutic candidate for the treatment of AD.

Aims: The athlete’s heart is a widely discussed topic regarding the adaptation of the left ventricle (LV) to regular training. The data on the morphology and—even more—the function of the right ventricle (RV) are less well studied. The aim of the study was to assess the influence of prolonged exertion on morphology and function of the RV. Methods and Results: We examined 38 elite athletes, members of the Polish Olympic Team and a control group of 41 age and sex-matched healthy volunteers. Specifically, we assessed the details of RV size and function including: RV enlargement, transtricuspid systolic gradient, and dilatation of main pulmonary artery (PA) as compared with the values derived from the control group. There was no significant difference in the function of the RV assessed using tissue Doppler echocardiography (TDE) between the athletes and controls (S’: 15.0 cm/sec vs. 14.0 cm/ sec; E’: 15.8 cm/sec vs. 15.7 cm/sec; A’: 9.9 cm/sec vs. 10.4 cm/sec), but the athletes had a higher transtricuspid systolic gradient (23.6 mm Hg vs. 19.0 mm Hg, P = 0.004). There were no significant differences in TDE velocities in athletes with dilated RV or PA. However, those with elevated tricuspid regurgitation velocity had lower systolic velocities of the tricuspid annulus then the rest (S’: 12.3 cm/ sec vs. 15.5 cm/sec, P = 0.01). Conclusions: RV enlargement in professional athletes is not connected with deterioration of diastolic or systolic RV function. Athletes with elevated pulmonary systolic pressure at rest, however, present with lower longitudinal systolic velocities of RV assessed using TDE. (Echocardiography 2011;28:753-760)

Mitochondrial dysfunction is a hallmark of Alzheimer's disease (AD) and is observed in mutant amyloid precursor protein (APP) transgenic mouse models of familial AD. Melatonin is a potent antioxidant, can prevent toxic aggregation of Alzheimer's beta-amyloid (Aβ) peptide and, when taken long term, can protect against cognitive deficits in APP transgenic mice. To study the effects of melatonin on brain mitochondrial function in an AD model, APP/PS1 transgenic mice were treated for 1 month with melatonin. Analysis of isolated brain mitochondria from mice indicated that melatonin treatment decreased mitochondrial Aβ levels by two- to fourfold in different brain regions. This was accompanied by a near complete restoration of mitochondrial respiratory rates, membrane potential, and ATP levels in isolated mitochondria from the hippocampus, cortex, or striatum. When isolated mitochondria from untreated young mice were given melatonin, a slight increase in respiratory rate was observed. No such effect was observed in mitochondria from aged mice. In APP-expressing neuroblastoma cells in culture, mitochondrial function was restored by melatonin or by the structurally related compounds indole-3-propionic acid or N(1)-acetyl-N(2)-formyl-5-methoxykynuramine. This restoration was partially blocked by melatonin receptor antagonists indicating melatonin receptor signaling is required for the full effect. Therefore, treatments that stimulate melatonin receptor signaling may be beneficial for restoring mitochondrial function in AD, and preservation of mitochondrial function may an important mechanism by which long term melatonin treatment delays cognitive dysfunction in AD mice.

This paper focuses on the qualitative and quantitative characterization of zooplankton in a section of the Nida (the Upper Wkra) River subjected to revitalization treatments. At four established sites, biological samples were taken for analyses. In total, 44 taxa of zooplankton were determiend in the collected material. The upper section of the Wkra River, along which two sites were set up, was in general more varied in species composition, and the structure of biocenosis was shaped by rotifers and protozoa (mainly ameba species) in nearly equal halves. At the other two sampling sites, localized below, along a further section of the watercourse, the structure of zooplankton was poorer. It was colonized by larger numbers of individuals belonging to a few species of rotifers, among which Keratella cochlearis var. tecta, Trichocerca similis and T. pusilla are the species indicating a raised level of the trophy of the river water.

A nine-year-old female with refractory Acute Myelomonocytic Leukemia (AML) was referred to our institute for Hematopoietic Cell Transplant (HCT) in 2015. While preparing for transplant, this patient became severely refractory to platelet transfusion. HLA antibody identification using single antigen bead technology revealed that she was highly sensitized to both HLA class I and class II antigens and had donor specific antibodies (DSA) to both parents (antibodies against HLA-B35 specific for her father; antibodies against A36 and DQ2 specific for her mother). Flow cytometric T cell allo-crossmatch of patient serum was positive with both parents. Unfortunately, this patient had no full siblings and an unfavorable matched unrelated donor search. The patient had few options other than haploidentical HCT from one of her parents and was therefore given a desensitization regimen of rituximab and plasmapheresis. Following two cycles of desensitization, overall class I PRA remained high (91%), but the patient’s DSA against her father (B35) became negative. The patient’s maternal DSA to DQ2 persisted. Patient/father T cell crossmatch after the desensitization treatment was diminished to borderline. The patient proceeded to HCT using her father as the donor, and was supported with HLA compatible platelets in the post-transplant period. She had neutrophil and platelet engraftment by days +13 and +19 respectively, following HCT. HLA antibody testing on the patient’s serum nine months post-transplant revealed greatly reduced antibody positivity with only two positive class I specificities and no antibodies to class II. This patient continues to remain in remission nearly three years post-transplant. This case study highlights the value of desensitization regimens for patients with donor specific antibodies and few donor options for HCT.

Experiments were conducted to determine full range of tolerated and optimal water condition for eggs incubation of three species from genus Leuciscus i.e: dace Leuciscus leuciscus (L.), ide Leuciscus idus (L.), and chub Leuciscus cephalus (L.). Spawners were caught from rivers of the northern (Pasleka River drainage) and central (Pilica River drainage) parts of Poland. Fertilized eggs were incubated under controlled conditions at ten different constant temperatures ranging from 4.5 to 29.0 o C. The optimal temperature ranges for the incubation of dace, ide and chub eggs were 7.5 to 12.3oC; 15.7oC and 19.0 to 23.0oC, respectively (considering hatching percentage, incidence of abnormalities and size of hatched larvae), which are close to the water temperature during spawning season. This study also reveals that embryos of studied species can adapt to increasing water temperature due to global warming up to 23.0 o C (dace and ide) and 27.5 o C (chub). Besides, some inter-population differences in the response to temperature were observed.

Artificial reproduction of asp under controlled conditions was investigated using two different spawning agents based on luteinizing hormone releasing hormone (LHRH) analogues and dopamine antagonists (Ovopel and Ovaprim), during two successive spawning seasons. Fish in the Ovopel and Ovaprim treatment groups were treated with a dose equivalent to 2 pellets and 0.5 mL per kg of body weight respectively. Out-of-season spawning and reproduction after a sudden increase in water temperature were also examined. No control fish ovulated although in both years, fish treated with Ovaprim showed a higher percentage of ovulation (75–88%) and embryo survival to the eyed-egg-stage (69–78%) than those treated with Ovopel (ovulation 25–38%; embryo survival 36–47%). The latency time was shorter in the groups where Ovopel was applied (40–42 h) than in Ovaprim groups (42–48 h). Similar results were obtained in out-off-season spawning. After a sudden increase in water temperature, females treated with Ovopel did not ovulate. This indicates that Ovaprim might be successfully used for artificial reproduction of asp by applying the induction protocol and the conditions reported in the present study.

This paper presents and describes a practical application of an experimental system for spawn incubation and fish larvae rearing under laboratory conditions in a closed water circulation. The experiment determined the effectiveness of water filtration in the system during the rearing of larvae of selected ornamental fish and native cyprinids. The results have shown full usability and diversity of application of the system for experimental purposes. The results obtained during experimental rearings with the system can be successfully used in practice.

The aim of this study was to define an influence of water temperature on successful reproduction of burbot under the hatchery conditions. Research was conducted during four successive spawning seasons where wild spawners were used. In the first three years of study fish were kept in three different (natural) thermal conditions. In the fourth year it was confirmed that the highest efficacy of synchronization of the spawning could be reached only under controlled thermal regimes. That year one group of spawners was kept at 6 o C before spawning and then a sudden decrease of the

The aim of the study was to compare the female dace Leuciscus leuciscus (L.) and ide Leuciscus idus (L.) hormonal stimulation effectiveness with two preparations (Ovopel and Ovaprim) separately and in combination. The experiment was conducted under controlled conditions. During the experiment the percentage of ovulating females, ovulation time and survival of embryos to the eyed stage were recorded. Stimulation with Ovopel resulted in shorter ovulation appearance time in females of both species (48–50 h for dace and 36–38 h for ide) while stimulation with Ovaprim in a higher percentage of live embryos in the eyed stage (almost 80 and 60% for dace and ide respectively). The highest hormonal stimulation effectiveness was recorded in the group where a combination of those two hormonal substances was applied as shortening of the time for achievement of spawning readiness (48 and 36 hours for dace and ide respectively), a significant ovulation synchronization and the highest embryos survival rates (almost 80 and 70% for dace and ide respectively) were recorded.

The influence of dose and time of eggs exposure in tannin solution on tench embryonic development, survival of embryos and percentage of larvae hatched were studied. The eggs obtained under controlled conditions from 11 tench females were fertilized with semen from 7 males and then rinsed for 1 h in Woynarovich solution (40 g urea, 30 g NaCl per 10 dm3 of water). Random samples (50 cm3) of the fertilised eggs were placed into three tannin solutions 0.05, 0.1 or 0.15% for 30, 60 or 90 s, then they were transferred to Weiss jars where incubation at 25°C took place. Tannin solution of 0.05% for 30–90 s or 0.1% for 30 s allowed effective unsticking of eggs and resulted in a very high per cent of hatched larvae. Extending the eggs rinsing time to 60 s in 0.10 and 0.15% solutions resulted in a high mortality of embryos during hatching because of hardened egg capsule.

Wild living spawners of dace Leuciscus leuciscus (L.), ide L. idus (L.) and chub L. cephalus (L.), were obtained from rivers of central (the Pisa River basin) and northern (the Pasleka River basin) Poland and next transported to a hatchery for artificial reproduction. The obtained eggs were dry fertilized and next incubated in water as different constant temperatures ranging from 4.5 to 29.0 o C. The hatched embryos were kept at the same temperatures until the moment of complete yolk sac resorption and later fed ad libitum on live nauplial Artemia sp. stages. The power function was applied for determination of the relation between the temperature and time until and achievement of each of the thirteen characteristic stages of embryonic development. It was shown that the pace of embryonic development increased with the temperature increase. In the majority of cases, at identical thermal conditions, both the incubation time and the pace of embryonic development were much less diversified among individual of the same species originating from different population than between the individual species. The obtained data finds application in optimization of early raring process and as a consequence will be an important tool in protection of endangered species.

Research into arti¢cial reproduction of chub was stu-died during three subsequent reproductive seasons.Chubreproductionwasconductedintwodiierentvar-iations. In the ¢rst experiment diierent hormonalagentsweregivento¢sh,suchasCPE,hCG,oranana-logue of GnRH with dopamine antagonist (ovopel). Inthe second one, the optimum moment of applying ofarti¢cial reproduction was sought and the ¢sh wereonlyobtained an analogue of GnRH in diierent peri-ods.Theresultsobtainedintwo¢rstseasonsindicatedthat ovopel is the mosteiective agent for the arti¢cialreproduction of chub. Consequently, it was decidedthat the ¢sh would obtain only this agent in the ¢nalyear of research. In last year (experiment II), chubspawnersweretakentothehatcheryundercontrolledconditions every few weeks from February to thebeginningofJune.Periodically,thematurityofoocyteswas checked. As soon as the oocytes had achievedstage 2^3 of maturity, ¢sh were obtained ovopel. Itwas noted that there was no problem with collectingsemen. Obtaining oocytes was much more di⁄cult.Most frequently, they were not obtained at all orobtained in very small amounts. It was also observedthat the administration of hormones caused a veryquickmaturationofgonadsandtheirover-maturation.Keywords: chub Leuciscus cephalus,arti¢cialspawning, wildpopulation, gametesqualityIntroductionUnder hatchery conditions, ¢sh reproduction isconducted primarilyduetothe use of hormonalaids(Kucharczyk 2002). Until quite recently, injections ofa suspension of homogenized pituitary glands werethe only way of inducing ovulation in females andspermiationinmales.Nowadays,manydiierenthor-monal agents are available, which are slowly repla-cing the pituitary gland suspension (Brzuska A Brzuska 2000; Kucharczyk, Kujawa,Mamcarz, TargonŁska, Krejszei & Wyszomirska2007).Stimulationofpubescenceis possibleat diier-ent levels: hypothalamus, stipule and gonads. At thelevel of the hypothalamus, stimulation is executedmostly by a change of environmental parameters,mainly temperatureand daylighttime, whereas hor-monal agents are used in stimulationat other levels:pituitarygland^naturalorsyntheticgonadoliberins;gonads^ ¢shorhumangonadotropins (Yaron1995).To develop a reproduction biotechnology, varioushormonal agents are checked. The agents and thedoses in which they will be used depend onthe spe-ciesandthetypeofspawn.Onlyafewpapersonarti-¢cial reproduction of the chub Leuciscus cephaluisL.have been published before the beginning of thisstudy (Kucharczyk 2002). The following hormonalagents have been used for hormonal stimulation:hCG (human chorionic gonadotropin) and CPE (ex-tract of carp pituitary glands) (Kucharczyk, Kujawa,Mamcarz, Skrzypczak& Wyszomirska1998).Chubhasnogreatereconomicimportance,butitisvalued in recreational ¢shery. As a reophilic species,it is also important as a live component of environ-ment (Bzachuta 1998; Penczak, Kruk & Koszalin ski1998). It is one of the most widely distributed fresh-water ¢sh in Europe, although in recent years it hasdisappeared from manyhabitats and has become an

In this work were shown some crucial moments in early Troheus moorii development (for example: begin of exogenous feeding period, end of endogenous feeding period, time of finish fins development and others). Fertilized eggs, obtained after natural spawn from females of Tropheus species kept in tank, were incubated in laboratory conditions. Hatched embryos were reared in incubation tank, in constance temperature, until resorption of the yolk sac and finish fins development. Embryos were fed ad libitum with live Artemia nauplii. In Tropheus, the fins became fully developed and the yolk sac supplies depleted on the same day, 25 th since the mating. Since this species keeps its offspring in the mouth for 30–35 days (YANAGISAWA ,S ATO 1990), the young stay in they shelter for another one to two weeks. When they are let out for the first time, they are in an advanced stage of their development, mature enough to live like the adults of their species. Since their yolk sack supplies are depleted before the termination of parental care period, it is obvious that they must be intrabucally fed in this time. It is confirmed by the fact that in aquarium conditions incubating females pick the food particles as the non-incubating females do. The food had been found on a regular basis in incubating females mouths when acquiring eggs and embryos for observations. Since the young Tropheus already can ingest exogenous food while being still in mothers mouth, it is on purpose to feed crushed food to the incubating females in aquarium conditions.

Catfish frays were distribute on three groups: A – fish fed artificial diet, B – fish fed combined diet, C – fish fed natural food, during three weeks stocking under controlled condition. Two variants of density were applied in each experimental group – 7.5 (1) and 15 (2) ind dm -3

is among the most popular species of thatfamily in aquarium breeding. In the natural environment they are foundmainly in tropical waters of South America although they are also highlynumerous outside their natural geographic location being objects of interestin high demand among aquarium keepers worldwide. Their high popularityresults from both attractive coloring and ease of breading. Buenos Aires tetraisa small fish reaching7–9cm inlength. In thenatural environmentitis foundmost numerously in the basin of the La Plata. The species was imported toEurope during early 1920’s. It is relatively fecund for its size. During onespawning act the female lays even more than 2000 of eggs (K

Studies on the influence of individual spermatozoa motility parameters (% of motility, motility time) on survival of embryos to the eyed-egg-stage were conducted on four freshwater fish species: burbot, ide, Eurasian perch and goldfish. Semen from individual males was collected to separate syringes and next the spermatozoa motility parameters were determined. Samples of oocytes originated from one female from each of the species covered. Each sample of eggs was fertilized with semen originating from a different male. Additionally an attempt was made to fertilize the oocytes with the pooled semen collected from many males, which formed as the control. At the eyed-egg-stage the numbers of live and dead embryos were counted. Aiming at determining the influence of the examined spermatozoa motility parameters single and multiple regression analysis was conducted. It was established that the highest embryos average survival values obtained after fertilization of spawn samples with semen originating from individual males did not differ from the results obtained for the controls that were fertilized with semen from pooled sample. In all cases investigated significant relationships between the spermatozoa motility parameters and the survival of embryos were recorded.

This paper describes the resistance of european catfish (Silurus glanis L.) larvae to temporary lack of food. Then PNR (point of no return) for european catfish larvae kept in water at 15, 20 and 25 o C( ±0.5 o C) was determined. As a result of the conducted experiments it was observed that european catfish larvae reach the PNR50 in the water at 15, 20 and 25 o C after 11.0, 9.2 and 4.4 days respectively. The conducted experiment confirmed that water temperature plays the key role in determining the period of resistance to lack of food. The observed, at least one week long period of european catfish larvae kept in water at 20 o C resistance to shortage of food allows freedom

Chromosomes of Siamese fighting fish Betta splendens were examined by Giemsa, CMA3 and Ag-NOR. The karyotype consisted of 6 pairs of submetacentric, 7 pairs of subtelocentric and 8 pairs of acrocentric chromosomes (2n=42, NF=54). NORs were observed at two submetacentric pairs, nos. 1 and 6, at telomeric regions of the short arms. The Betta’s karyotype seems to have a derived status when compared to most species of the order Perciformes.

Induction of gynogenetic development of ide using semen of dace was performed. The results obtained during genetic inactivation of dace sperms showed that the most optimal time of UV irradiation dose was from 5 to 10 minutes with the optimum in minute 9 (3456 J m -2 ). The ide oocytes were next fertilized with genetically inactivated semen of dace and subjected several minutes later to environmental shock aiming at retaining the second polar body inside the oocyte. The obtained results were analyzed using a color marker. The highest percentage of gynogenetic ide breed (exceeding 20%) was obtained when fertilized oocytes were subjected to environmental shock lasting for 3 minutes during the 14th minute after fertilization.

Fourteen easily recognizable developmental stages (DS) for larval and juvenile asp Aspius aspius (L.) were described, beginning from hatching until the time of complete squamation of juveniles. Rearing temperature ranged between 14.0 and 18.7 o C (mean 17.2 o C). The proposed system of developmental stages for asp can be used both in laboratory studies and field work in order to follow the course and rate of asp early development.

The manuscript presents a comparative analysis of slaughter yield, sensory traits and proximate and fatty acid composition of a little-known rapfen ( Aspius aspius L) with bream ( Abramis brama L) and pike ( Esox lucius L). The experimental fish of each species were obtained under natural conditions between July and August. The rapfen was characterized by the highest slaughter yield of gutted and deheaded carcass (74.57%). The slaughter yields of fillet with skin (59.44%) and deskinned fillet (51.26%) were similar to those reported for pike, but higher than those of bream. Aroma and consistency of evaporated meat carcass obtained similar scores, whereas differences were demonstrated between flavour, colour and juiciness of rapfen and bream, and between colour and juiciness of rapfen and pike. The fat contents of rapfen and bream carcasses were similar (2.52% and 3.63%, respectively) and higher than that of pike (0.64%). The contents of protein (18.00–18.83%) and ash (0.99–1.01%) were alike in the three species examined. The concentrations of polyunsaturated fatty acids (PUFA) in 100 g carcasses of rapfen and bream were similar, and higher than in pike. The carcasses of the fish species examined differed in terms of the concentrations of monounsaturated fatty acids (MUFA). The total concentrations of n-6 fatty acids were different in rapfen, bream and pike, whereas concentrations of n-3 acids were higher than those reported for pike. Rapfen carcass had the highest concentration of docosahexaneoic acid (DHA; 22:6 n-3).

Commercially exploited tench populations in 755 lakes in the Northeastern part of Poland were observed on the basis of the fishing data from 1951 to 1994. The observations were analyzed in five size categories of water reservoirs, whose total area reached over 123,900 ha. The analyses were performed according to the parameter of a commercial fish catch per area unit, taking into consideration the number of months per year when the lakes were exploited. Statistically significant differences were determined in the mean value of the tench catch parameter between the different size categories of lakes. Similar time-related differences were observed. In the lakes >100 ha the relative tench catch gradually declined in the successive time intervals. Tench was the rarest in catches from lakes

Commercially exploited tench, Tinca tinca, (L.) populations in 480 lakes in the northeastern Poland were analyzed on the basis of the fishery data from 1949 to 1994. The long-term changes in tench catch were studied in four categories of lakes with respective proportions of littoral zones ( 30%) to 0.93 kg ha−1 month−1 in lakes with the largest littoral ( 60%). Mean stocking rates of tench (number of 1-year-old fish per ha of littoral) for 1949–1994 were not significantly different in lakes of studied categories. However, lakes with proportionally less littoral zone had reduced stocking efficiency estimated as number of stocked fish per ha of littoral to require 1 kg ha−1 month−1 of tench.

Reciprocal hybrids between tench Tinca tinca (L.) and carp Cyprinus carpio L., and tench and bream Abramis brama (L.) were produced artificially. The survival of all these hybrids during embryogenesis was quite high. The highest survival rate (over 60%) at the eyed stage was observed for tench and bream hybrids (both sex combinations). The hatching rates of these hybrids were also over 60%. The number of larvae with some abnormalities (i.e. deformed body) was low. In contrast, the hatching rates of tench and carp hybrids were very low (0.2%). From over 1000 fertilized eggs, only three specimens started swimming, and only one specimen survived to juvenile stage. Embryos of hybrids and their parental species differ in morphological features. These differences were also visible in the juveniles. Body parameters of juvenile hybrids produced from three species had intermediate values in comparison to parental fish.

BACKGROUND T-cell depletion (TCD) effectively reduces severe graft-versus-host disease in recipients of HLA-mismatched allografts. However, TCD is associated with delayed immune recovery and increased infections. We hypothesized that specific depletion of CD45RA+ naive T cells, rather than broad depletion of CD3+ T cells, can preserve memory-immunity in the allografts and confer protection against important viral infections in the early post-transplant period. METHODS Sixty-seven patients who received TCD haploidentical donor transplantation for hematologic malignancy on 3 consecutive trials were analyzed. RESULTS Patients receiving CD45RA-depleted donor grafts had 2000-fold more donor T cells infused, significantly higher T-cell counts at Day +30 post transplant (550/μL vs 10/μL; P

This study presents artificial induction using tench eggs, Tinca tinca (L.), of androgenetic origin. The oocytes taken from common bream, Abramis brama (L.) and common carp, Cyprinus carpio L. were genetically inactivated using UV irradiation and then inseminated using tench spermatozoa. Androgenetic origin (haploid or diploid embryos) was checked using a recessive colour (blond) and morphological markers. The percentage of hatched embryos in all experimental groups was much lower than in the control groups. All haploid embryos showed morphological abnormalities, which were recorded as haploid syndrome (stunted body, poorly formed retina, etc.). The optimal dose of UV irradiation of common bream and common carp eggs was 3456 J m–2. At this dose, almost 100% of haploid embryos were produced at a hatching rate of over 6%. Lower UV-ray doses affected abnormal embryo development. The highest yield of tench androgenesis (about 2%) was noted when eggs were exposed to thermal shock 30 min after egg activation.

In this paper we present a system with controlled temperature and photoperiod for keeping breeders before their artificial spawning and give a general outline of its use. The advantages of this system were: utility, small area in which it was placed, wide range of temperature and photoperiod, possibility of worked tanks together or alone and in open, closed or semi-closed systems, cheap to construct and with low running costs. Additionally, this unit was tested for artificial spawning for several wild and domestic European cyprinid and percid fish species, as well as burbot, not uncommonly in different periods than the time of their natural spawning.

Induced spawning in perch, Perca fluviatilis L., was studied using follicle stimulating hormone and lutenizing hormone (FSH + LH) with the addition of pimozide or metoclopramide. The doses administered to fish were: 75 IU kg−1 of FSH + LH with 5 or 10mgkg−1 of pimozide (P) or metoclopramide (M) in females, and 25 IU kg−1 of FSH + LH with 2.5 or 5 mg kg−1 of P or M in males. Hormonal injections in males (except the injection of FSH + LH with 10 mg kg−1 of M) did not influence the quantity of milt obtained. All females treated in this experiment had oocytes at the same stage of nucleus migration, so the time of ovulation was very synchronous, i.e. 16-24 h after injection. The relationship between the quality of the eggs, expressed as a survival to eyed-egg stage, and latency was inverse. Spawning success, expressed as a spawning effectiveness coefficient (Se), was highest in the fish that had been treated with FSH + LH with a higher dose of M.