Sewage and solid waste can be a valuable source of materials used directly or indirectly in manufacturing of usable products. These processes are associated with elimination of pollutants from liquid and

solid wastes. The best-known methods of waste management are production of biogas and composting. This

paper focuses on the possibility of obtaining biomass as a source of protein feed (whose value, in terms of the

composition of aminoacids and microelements, is comparable with conventional feed, e.g. soymeal, bonemeal

or fishmeal). Sewage components for bacterial, fungal, algal and vascular plants’ culture are characterized as

a source of protein feed. Methods of industrial scale production of enzymes, mainly proteases and lipases that

have broad applications in various industries, are discussed. Development perspectives of inexpensive methods

of usable products from waste production are showed. Interdisciplinary nature of presented issues, which requires cooperation of specialists in biology, chemistry and technology, is emphasized.

They are unfairly regarded as junk protein, but the truth is they have not been properly studied due to a lack of suitable tools. So how much do we really know about non-globular proteins?

Dr. Takao Ishikawa from the University of Warsaw talks about why perhaps not all scientists should aim to become professors, and explains what we can learn from yeast proteins.

Pseudomonas syringae pv. syringae (Pss) constitutes a diverse group of bacterial strains that cause canker of stone fruits, blight of cereals and red streak of sugarcane. The purpose of this study was to determine how diverse Iranian strains of Pss are when they come from different hosts. We compared a total of 32 Pss strains isolated from stone fruits, barley, wheat and sugarcane from different geographical regions of Iran based on their phenotypic and molecular properties. Strains showed some variation regarding carbon and nitrogen utilization. Pss strains were similar in their protein banding patterns. Additional bands were found in sugarcane strains. Most strains showed one indigenous plasmid DNA and a few had two and some none. The genes of syrB and syrD encoding syringomycin synthesis and secretion, respectively, were amplified using specific primers in polymerase chain reaction. Syringomycin, producing strains amplified two DNA fragments of 752 and 446 bp representing syrB and syrD genes, respectively. Primer specificity was shown for Pss using various genera. Based on the results of this study, it is suggested that Pss strains from different hosts and geographical regions show diversity in phenotypic and molecular characters. It is thought that phenotypic variation is due to adaptation to specific hosts and niches for survival and pathogenicity.

The measured rate of release of intercellular protein from yeast cells by ultrasonication was applied for evaluating the effects of sonication reactor geometry on cell disruption rate and for validation of the simulation method. Disintegration of two strains of Saccharomyces cerevisiae has been investigated experimentally using a batch sonication reactor equipped with a horn type sonicator and an ultrasonic processor operating at the ultrasound frequency of 20 kHz. The results have shown that the rate of release of protein is directly proportional to the frequency of the emitter surface and the square of the amplitude of oscillations and strongly depends on the sonication reactor geometry. The model based on the Helmholtz equation has been used to predict spatial distribution of acoustic pressure in the sonication reactor. Effects of suspension volume, horn tip position, vessel diameter and amplitude of ultrasound waves on the spatial distribution of pressure amplitude have been simulated. A strong correlation between the rate of protein release and the magnitude of acoustic pressure and its spatial distribution has been observed. This shows that modeling of acoustic pressure is useful for optimization of sonication reactor geometry.

SDS-PAGE electrophoresis was used to study the effect of NaCl on protein expression in two cultivars of tomato (Solanum lycopersicum L.): Edkawi (salt-tolerant) and Castle rock (salt-sensitive). Five-day-old seedlings were grown on MS agar media supplemented with 0, 50, 100, 150, 200 and 300 mM NaCl. Two days after treatment the seedlings were examined to determine the effect of salt on their growth and to relate that to protein banding variations. Gel analysis showed differences in at least 4 protein bands with molecular weights at 20, 25, 45 and 65 kDa. These proteins were induced in the 50 mM NaCl treatment in the salt-sensitive cultivar, then decreasing to undetectability at higher concentrations. In the salt-tolerant cultivar, most of the proteins exhibited a more or less steady expression pattern and maintained expression through the 200 mM NaCl treatment. All proteins gave weak or no expression signals at 300 mM NaCl, the treatment that proved lethal. Differentially expressed bands were identified using MALDI-TOF mass spectrometry. The putative function of each identified protein in relation to salt stress is discussed.

Humidity is probably the most important abiotic factor influencing life cycles, distribution, survival, and population dynamics of stored product pests. Although most of these pests can complete their life cycles in any given relative humidity, their prolonged development time, as well as decreased emergence rate and fecundity, have been well documented in several previous studies. In the present study, we evaluated the changes in energetic substances (lipids, soluble carbohydrates, glycogen, and proteins) accumulated in different life stages of larvae and adults of Tribolium castaneum in response to different relative humidity levels (5, 12, 22, 30, 45, and 65%). The results showed that young larvae were more susceptible to low relative humidity levels and desiccation stress. Larvae tended to accumulate higher proportions of lipids during earlier stages while their energy content shifted towards proteins with an increase in their age. Adult beetles experienced a significant decrease in their protein content immediately after they initiated reproduction. The importance of these fluctuations in the biology of the red flour beetles was discussed in detail.

The aim of this study was to analyse and identify specific buffalo seminal plasma proteins (SPPs) responsible for sperm cryotolerance during low temperature storage. Computer Assisted Sperm Analysis (CASA) of the motility and viability of buffalo spermatozoa was performed before freezing and after thawing. Two sample groups were formed – ejaculates with high cryotol- erance (group A) and low cryotolerance (group B). CASA demonstrated that the initial progres- sive motility after thawing of the spermatozoa in group A is significantly higher than in group B (p<0.001). Group B showed a significant increase in the percentage of static and non-progressive spermatozoa at 240 min, when compared to group A (p<0.05). SPPs, proteins in the cryoprotec- tive medium (PM) and proteins in the mixture of PM and SP were separated by High Perfor- mance Liquid Chromatography (HPLC). Comparative analysis of the chromatographic profiles was performed to identify specific proteins related to sperm cryotolerance. SPPs profiles showed 5 distinct protein peaks in both groups, ranging from 500 kDa to 50 Da. Chromatograms of group A and group B showed quantitative and qualitative differences in protein content. Chromato- grams of proteins in PM showed 11 well-expressed peaks. HPLC analysis of the mixtures of SPPs from the two groups and PM visualized the formation of a new bio-complex structure expressed by a protein peak specific for group A (7.674 min, AU 1.50). This protein peak can be referred as a phenotypic trait for buffalo ejaculates with high sperm cryotolerance.

The aim of this study was to evaluate the effect of high doses of calcium bentonite on the blood parameters, anticoccidial activity and intestinal histology of broiler chickens. Three undred and sixty one-day old broilers were distributed into three treatments (T+VE, T-VE, TB )with three replicates. Amprolium was added to the feed of the positive control group, calcium bentonite powder was added to the TB group, and nothing was added to the feed of the T-VE group. Coccidiosis was induced on day 14, the birds were kept until day 49, measurements of the diffe- rent variables started from week 3, blood samples were collected via wing vein, and fecal oocysts were counted from the intestinal contents of each individual bird using the McMaster techni- que. A decrease in feed consumption, body weight gain and conversion ratio was noticed in the calcium bentonite group. Broilers in the calcium bentonite group (TB ) and negative control group (T-VE ) showed clinical signs of coccidiosis (blood in feces) and the number of oocysts in feces increased with time. Histopathological examinations of the affected caeca also demonstrated excessive tissue damage, hemorrhage, the presence of clusters of large schizonts and merozoites in the tissue, and coccidian oocysts in the lumen. Feed conversion was highest in the T+VE group.

Tight junction proteins are important for the maintenance and repair of the intestinal mucosal barrier. The present study investigated relationships among tight junction protein gene expres- sion, porcine epidemic diarrhea virus (PEDV) infection, and intestinal mucosal morphology in piglets. We compared the expression of six tight junction proteins (ZO-1, ZO-2, Occludin, Claudin-1, Claudin-4, and Claudin-5) between seven-day-old piglets infected with PEDV and normal piglets, as well as in PEDV-infected porcine intestinal epithelial cells (IPEC-J2). We also evaluated differences in mucosal morphology between PEDV-infected and normal piglets. The expression of six tight junction protein genes was lower in PEDV-infected piglets than in the normal animals. The expression of ZO-1, ZO-2, Occludin, and Claudin-4 in the intestine tissue was significantly lower (p<0.05) in PEDV-infected than in normal piglets. The expression of Claudin-5 in the jejunum was significantly lower in PEDV-infected piglets than in the normal animals (p<0.01). The expression of Claudin-1 and Claudin-5 genes in the ileum was signifi- cantly higher in PEDV-infected piglets than in normal piglets (p<0.01). Morphologically, the intestinal mucosa in PEDV-infected piglets exhibited clear pathological changes, including breakage and shedding of intestinal villi. In PEDV-infected IPEC-J2 cells, the mRNA expression of the six tight junction proteins showed a downward trend; in particular, the expression of the Occludin and Claudin-4 genes was significantly lower (p<0.01). These data suggest that the expression of these six tight junction proteins, especially Occludin and Claudin-4, plays an important role in maintaining the integrity of the intestinal mucosal barrier and resistance to PEDV infection in piglets.

The aim of this study was to identify the proteoforms of albumin and kallikrein in stallion seminal plasma (SP), and to determine their correlations with sperm motility parameters. The experimental material consisted of ejaculates from 8 stallions, which were collected during the breeding and non-breeding seasons (BS and NBS, respectively). SP proteins were identified by 2-D PAGE and mass spectrometry (MALDI TOT/TOF MS). Sperm motility parameters were analyzed using the CASA system. Protein expression (integrated optical density-IOD) of albumin proteoforms 1 (ALB 1) and 2 (ALB 2) and kallikrein proteoforms 1 (KAL 1) and 2 (KAL 2) was correlated (p<0.05) with sperm motility parameters (total motility and progressive motility) during the BS. No significant correlations were found between the expression of albumin or kallikrein and sperm motility parameters during the NBS. The presence of correlations between the expression of ALB 1, ALB 2, KAL 1, KAL 2 and selected sperm motility parameters could suggest that the analyzed components of the SP belong to the group of fertility-associated pro- teins (FAPs).

Therefore, the aim of the present study was to evaluate the possible effect of bilberry fruit (Vaccinium myrtillus L.) supplement in a daily diet on the cognitive behaviour of the rats and the expression of paravalbumin (PV) in populations of hippocampal neurons. It has been postulated that the antioxidants present in bilberry fruit may act as neuroprotective factors playing also a significant role as memory enhancements. Forty Wistar rats with a similar average body weight (460 ± 0.4 g) were divided into four groups (n=10 per group). The control group received standard feed (210 g/week), whereas animals of experimental groups received standard feed supplemented with bilberry (per os) at consumed doses of 2 g (group I), 5 g (group II), and 10 g/kg b.w./ /day (group III). After three months of feeding with bilberry, the modified elevated plus-maze test (mEPM) was performed. After 32 weeks of feeding, brains were collected and PV-immunoreactive (ir) neurons were immunohistochemically visualized. In the modified elevated plus-maze test, transfer latency examined 2 h and 24 h after the acquisition session was significantly shorter (p<0.05) in the group II in comparison with the control group. In CA1 and CA2/CA3 hippocampal fields as well as dentate gyrus of all experimental groups, a significant (p<0.05) decrease in number of PV-ir neurons were found. In relation to the control group, the mean subpopulation of PV-ir neurons found in groups II and III were significantly reduced. The subpopulations of PV-ir neurons found in DG of all experimental groups were significantly reduced in comparison to the control. In conclusion the in the present paper we demonstrated a relationship between the diet rich in a bilberry fruit and process of memory as well as numbers of calcium- binding protein-expressing hippocampal neurons. Our results may be source of basic knowledge for further research aiming at neuroprotective role of the bilberry fruit.

In this study, we examined whether and to what extent oxidative stress is induced in seedlings of two winter triticale (Triticosecale Wittm.) varieties (susceptible Tornado and resistant Witon) in response to infestation by the cereal grain aphid (Sitobion avenae L.) and bird-cherry-oat aphid (Rhopalosiphum padi L.). We compared the level of hydrogen peroxide (H2O2) and lipid peroxidation products as well as markers of protein damage (protein-bound thiol and carbonyl groups). The studied parameters were measured at 6, 24, 48 and 96 h post-initial aphid infestation compared to the non-infested control seedlings. Our studies indicated that the cereal aphid feeding evoked oxidative stress in the triticale seedlings. Cereal aphid feeding increased the H2O2 level in triticale tissues, with maximum levels observed at 24 and 48 h post-infestation. Triticale infestation with aphids also increased lipid peroxidation products in triticale seedlings, with the maximal levels at 48 or 96 h post-infestation. Further, there was a reduction in protein thiol content and an increase in protein carbonyl content in the triticale seedlings after infestation with female aphids. Stronger triticale macromolecule damages were evoked by the oligophagous aphid R. padi. There was a more substantial protein thiol content reduction in the resistant Witon cultivar and higher accumulation of protein-bound carbonyls in the tissues of the susceptible Tornado cultivar. The changes were proportional to the aphid population and the time of aphid attack. These findings indicate that the defensive strategies against cereal aphid (S. avenae and R. padi) infestation were stimulated in triticale Tornado and Witon seedlings. Our results explain some aspects and broaden the current knowledge of regulatory mechanisms in plant-aphid interactions.

Professor Agnieszka Chacińska from the International Institute of Molecular and Cell Biology talks about her research on mitochondrial proteins and their association with neurodegenerative diseases and metabolic disorders.