Macaques vaccinated with live-attenuated SIV control replication of heterologous virus

An effective AIDS vaccine will need to protect against globally diverse isolates of HIV. To address this issue in macaques, we administered a live-attenuated simian immunodeficiency virus (SIV) vaccine and challenged with a highly pathogenic heterologous isolate. Vaccinees reduced viral replication by ∼2 logs between weeks 2–32 (P ≤ 0.049) postchallenge. Remarkably, vaccinees expressing MHC-I (MHC class I) alleles previously associated with viral control completely suppressed acute phase replication of the challenge virus, implicating CD8+ T cells in this control. Furthermore, transient depletion of peripheral CD8+ lymphocytes in four vaccinees during the chronic phase resulted in an increase in virus replication. In two of these animals, the recrudescent virus population contained only the vaccine strain and not the challenge virus. Alarmingly, however, we found evidence of recombinant viruses emerging in some of the vaccinated animals. This finding argues strongly against an attenuated virus vaccine as a solution to the AIDS epidemic. On a more positive note, our results suggest that MHC-I–restricted CD8+ T cells contribute to the protection induced by the live-attenuated SIV vaccine and demonstrate that vaccine-induced CD8+ T cell responses can control replication of heterologous challenge viruses

Glioblastoma-derived spheroid cultures as an experimental model for analysis of EGFR anomalies

Glioblastoma cell cultures in vitro are frequently used for investigations on the biology of tumors or new therapeutic approaches. Recent reports have emphasized the importance of cell culture type for maintenance of tumor original features. Nevertheless, the ability of GBM cells to preserve EGFR overdosage in vitro remains controversial. Our experimental approach was based on quantitative analysis of EGFR gene dosage in vitro both at DNA and mRNA level. Real-time PCR data were verified with a FISH method allowing for a distinction between EGFR amplification and polysomy 7. We demonstrated that EGFR amplification accompanied by EGFRwt overexpression was maintained in spheroids, but these phenomena were gradually lost in adherent culture. We noticed a rapid decrease of EGFR overdosage already at the initial stage of cell culture establishment. In contrast to EGFR amplification, the maintenance of polysomy 7 resulted in EGFR locus gain and stabilization even in long-term adherent culture in serum presence. Surprisingly, the EGFRwt expression pattern did not reflect the latter phenomenon and we observed no overexpression of the tested gene. Moreover, quantitative analysis demonstrated that expression of the truncated variant of receptor—EGFRvIII was preserved in GBM-derived spheroids at a level comparable to the initial tumor tissue. Our findings are especially important in the light of research using glioblastoma culture as the experimental model for testing novel EGFR-targeted therapeutics in vitro, with special emphasis on the most common mutated form of receptor—EGFRvIII

Immunological composition of human milk before and during subclinical and clinical mastitis

Mastitis, an inflammatory condition affecting more than 25% of breastfeeding women, is usually associated with reduced milk secretion, pain, and discomfort, which often leads to early cessation of breastfeeding. Although the etiology of mastitis is multifactorial, a pro-inflammatory state of the mammary gland might be a risk factor. However, changes in milk composition, and specifically in the milk immune profile, prior to and during mastitis have not been well described. To help close this research gap, we documented the immune profiles of milk produced by both breasts of 10 women experiencing clinical (CM) and 8 women experiencing subclinical (SCM) mastitis during the week of sign/symptom development as well as the week prior and compared them with milk produced by 14 healthy controls. CM was defined as having signs/symptoms of mastitis, whereas SCM was presumed if the participant did not have signs/symptoms of CM, but her milk had a somatic cell count >400,000 cell/mL and/or sodium-to-potassium (Na/K) ratio >1.0. Concentration of 36 immune factors (including immunoglobulins, cytokines, chemokines, and growth factors) was quantified via immunoassays. Milk produced by women who developed CM had distinct immune profiles the week prior to diagnosis, particularly elevated concentrations of pro-inflammatory cytokine IL-1β and regulatory cytokines IL-2, IL-4 and IL-10. In contrast, immune profiles in milk produced by women with SCM did not differ from that produced by healthy women or those with CM the week prior to mastitis onset. Once mastitis appeared, marked changes in milk’s immune profile were observed in both CM and SCM groups. CM was characterized by elevated concentrations of 27 compounds, including pro-inflammatory cytokines (IL-1β, IL-1ra, and TNFα) and chemokines (including IL-8, eotaxin, IP-10, MCP-1, MIP1α, and MIP1β), compared to healthy controls. Milk’s immune profile during SCM was intermediate, showing higher levels of IL-6, IFNγ, and MCP-1 compared to healthy controls, suggesting a milder, more controlled immune response compared to CM. Only milk produced by the mastitis-affected breast had altered immune profiles. Further research is needed to determine if these differences in milk’s immune profiles can be used to improve mastitis risk prediction prior to onset of symptoms

The influence of operating parameters on biogenic compounds removal in the laboratory sequencing batch reactors

Technologia sekwencyjnych biologicznych reaktorów, w której jako pierwszej wdrożono proces osadu czynnego na długi okres czasu została wyparta przez systemy przepływowe [1]. W ostatnich latach oczyszczanie ścieków systemem SBR zdobywa jednak coraz większe uznanie, szczególnie od momentu wprowadzania w poszczególnych krajach wymagań dotyczących intensywniejszego usuwania ze ścieków związków biogennych. Sekwencyjne reaktory mogą być już stosowane nie tylko do oczyszczania niewielkich ilości ścieków z terenów turystycznych czy zakładów przetwórczych, ale również pochodzących z większych miast (np. dla 19 tys. m3/d w Chełmnie). Stosując w jednym urządzeniu wszystkie podstawowe procesy oczyszczania ścieków, tj. napowietrzanie, sedymentację i dekantację (rys. 1), reaktor sekwencyjny (SBR) daje duże możliwości prowadzenia zintegrowanego nie tylko biologicznego utlenienia organicznych związków węgla i amonifikacji, ale także procesów nitryfikacji, denitryfikacji oraz zwiększonego wewnątrz komórkowego magazynowania ortofosforanów [2,3]. Praca reaktorów sekwencyjnych jest na tyle elastyczna, iż pozwala na dokonywanie szybkich zmian parametrów operacyjnych w sposób prosty, w zależności od ilości i składu dopływających ścieków. W laboratoryjnych badaniach procesu biologicznego oczyszczania ścieków, które często są badaniami rozpoznawczymi poprzedzającymi wprowadzanie nowych rozwiązań na skalę techniczną istotne jest określenie stabilnych warunków pracy układu, w celu zmniejszenia wpływu czynników zakłócających zdefiniowanie zachodzących procesów. Rektor sekwencyjny ze względu na łatwość obsługi oraz kontroli najważniejszych parametrów technologicznych jest bardzo często wykorzystywany w tych badaniach, jednakże uzyskanie stabilnej pracy osadu czynnego wymaga wielu tygodni adaptacji oraz optymalizacji parametrów operacyjnych. Przeprowadzone badania wpływu zmian współczynnika dekantacji oraz czasu trwania napełniania na efektywność usuwania związków azotu oraz fosforu ze ścieków syntetycznych nie stanowią nowości w szeroko poznanej i przebadanej problematyce technologii SBR, jednakże mogą ułatwić dochodzenie do wymaganej efektywności pracy reaktora. Dlatego też w oparciu o uzyskane wyniki badań sformułowano najistotniejsze wnioski: reaktor pracujący przy 33% i 67% dekantacji wykazywał tendencje do puchnięcia osadu ze wzrostem czasu fazy anaerobowej, najkorzystniejsze i stabilne właściwości sedymentacyjne osadu czynnego uzyskano dla f = 50%, skrócenie fazy beztlenowej zwiększyło intensywność nitryfikacji dla f = 50 i 60% dekantacji (wzrasta wiek aerobowy osadu), dla 33% zależność określona została odwrotnie, ze wzrostem wartości dekantacji oraz czasu trwania fazy anoksyczno-anaerobowej rosło stężenie ortofosforanów uwalnianych przez bakterie w warunkach beztlenowych, przy 30% dekantacji nie nastąpiło uruchomienie mechanizmu biologicznej defosfatacji, w reaktorze o wartości f = 50 i 67% ze wzrostem czasu fazy anoksyczno-anaerobowej od 1h do 3h malało stężenie ortofosforanów w ściekach oczyszczonych, największe usunięcie ortofosforanów uzyskano przy 50% dekantacji oraz 3-godzinnej fazie beztlenowego napełniania.The sequencing batch reactors (SBR) allow executing integrated process of carbon, nitrogen and phosphorus compounds removal on the basis of optimal planning of SBR duty cycle. Obtaining high effects of biological removal of biogenic compounds is practically only a matter of optimization the duration time of variable oxygen conditions: anaerobic, anoxic and aerobic, in which result the process of denitrification and dephosphatation happens on the level of efficiency required by the standards which determine the quality of treated sewage. The rich experience in the range of exploitation as well as designing of SBR, which results for example as the set of German guidelines ATV-M 210 P, makes easier resolving the majority of encountered difficulties in optimization of SBR work. These reactors however are often used under laboratory conditions to investigate sewage treatment in the process of activated sludge. For the sake of long culture, adaptation and optimization periods of SBR work, which can last even several months, large facilitation of conducting investigations may be experience in the range of influence of selected operating parameters on efficiency of the biological removal of nitrogen and phosphorus compounds from synthetic sewage, which make up the aim of this paper. In the paper author introduced possibilities of SBR work optimization in range of operating parameters: decantation coefficient f and duration of individual process phases - the anaerobic filling and aeration on possibilities of biological removal of biogenic compounds. The tests were conducted on laboratory SBR system stand working in 8 hour cycle in range of f = 33, 50 and 67% and filling phases from 1 to 3 hours for each decantation coefficient. The dynamics of course of nitrogen and phosphorus compounds changes for all variables was also determined in sequential process phases of the full SBR cycle. During laboratory investigations of biological sewage treatment, which often are diagnostic tests preceding application of new solutions on technical scale it is essential to determine stable conditions of system's work, to decrease the influence of factors disturbing defining of processes. SBR for the sake of operation and control of the most important technological parameters easiness is very often used in these investigations, but to obtain the stable work of activated sludge requires many weeks of adaptation and optimization of operating parameters. Conducted investigations on influence of decantation coefficient changes and filling duration on efficiency of nitrogen and phosphorus compounds removal from synthetic sewage are not new in widely known and tested problems of SBR technology, but they can facilitate reaching required efficiency of reactor work. Therefore basing on obtained results of investigations most essential conclusions are: the sludge index increases along with increase of anaerobic filling phase duration, particularly above 2 hours, reactor with 33% and 67% of decantation shows tendencies for sludge swelling by longer times of anaerobic phase than by 50% of decantation, the most stable on that score, shortening anaerobic phase enlarges intensity of nitrification for f = 50 and 60% of decantation (aerobic age sludge increases), for 33% dependence was determined inversely, along with increase of decantation level and the anaerobic phase time increases the quantity of orthophosphates freed by phosphorous bacteria, mechanism of biological dephosphatation does not activate by 30% of decantation, in systems with f = 50 and 67% along with increase of time of anaerobic phase decreases concentration of orthophosphates in treated sewage, the biggest removal of orthophosphates was obtained by 50% of decantation and 3 hours of anaerobic filling phase. Obtained results show, that the most optimal conditions for tested system with regard on parameters of the activated sludge and results of dephosphatation were gained for reactor with 50% of decantation and 3 hours of anaerobic filling phase. During conducted investigations integrated C, N and P was not obtained, but gained results allow to guide further investigations in the direction of laboratory SBR work cycle optimization

The effect of products of adhesive wastewater oxidation with Fenton's reagent on biological treatment in SBR

The advanced oxidation process with Fenton’s reagent can be used in industrial wastewater treatment, especially for the degradation of not easily biodegradable contaminations. Chemical oxidation of industrial wastewater or other substances as a pre-treatment before biological purification is the most effective method of wastewater treatment. However the Fenton process requires low pH of wastewater and final neutralization with a big amount of iron-containing precipitate. Thes paper presents laboratory experiments dealing with the treatment of adhesive wastewater in an sequencing batch reactor following the Fenton process without final neutralization

Profile of orthophosphates concentration changes during sewage and sludge treatment

Wykorzystanie znajomości profilu zmian stężenia związków biogennych w oczyszczalniach ścieków stanowi istotny warunek ich projektowania. Wykorzystywanie zintegrowanych układów do usuwania ze ścieków związków azotu oraz fosforu wprowadziło znaczące zmiany w projektowaniu i eksploatacji tzw. gospodarki osadowej oczyszczalni. Dane statystyczne wskazują, że na 207 ankietowanych polskich oczyszczalni ścieków blisko 83% stosuje proces osadu czynnego i 80% z tych obiektów pracuje w systemie pełnego lub częściowego biologicznego usuwania związków biogennych - tzw. układy wysokoefektywnego usuwania fosforu (enhanced biological phosphorus removal) [1]. Uzyskując wyższą skuteczność oczyszczania ścieków, jednocześnie zwiększa się ilość osadów kierowanych do unieszkodliwiania oraz zmienia ich parametry jakościowe, wynikające m.in. z dużej akumulacji fosforu w komórkach bakterii. Podczas unieszkodliwiania osadów w warunkach beztlenowych (fermentacja metanowa), nawet 60% fosforu usuniętego ze ścieków może zostać wtórnie uwolniona podczas hydrolizy polifosforanów do cieczy osadowej. Stężenie fosforu wzrasta w bardzo szerokim zakresie, na co ma wpływ duża zmienność jakościowa osadów oraz problemy z utrzymaniem stabilności procesów ich unieszkodliwiania. Przykładowy zakres stężenia fosforu w oczyszczalni ścieków w Białymstoku wynosił od 20 do 435 mg Pog/L w cieczy osadowej z zagęszczaczy grawitacyjnych, natomiast z komór fermentacji od 18 do 590 mg Pog/dm3 [2÷3]. Ciecz osadowa ponadto charakteryzuje się wysoką zawartością związków organicznych, zawiesin ogólnych trudno opadających i koloidalnych, gazów rozpuszczonych (metan, amoniak, CO2, H2S), metali ciężkich i tłuszczów oraz amoniaku, którego stężenie może osiągnąć nawet kilkaset gramów na metr sześcienny. W cieczy pofermentacyjnej stwierdza się ponadto duże stężenia lotnych kwasów tłuszczowych (LKT), węglanów oraz kationów potasu i magnezu [4÷5]. Rosnąca ilość uwalnianego fosforu w cieczy osadowej, przy równie dużym stężeniu azotu amonowego oraz udziale jonów magnezu stanowi także źródło wytrącającego się twardego osadu fosforanu amonowo-magnezowego MgNH4PO4ź6H2O, czyli struwitu [6]. Powszechnym sposobem oczyszczania cieczy osadowej, powstającej podczas przeróbki osadów jest jej zawracanie do oczyszczalni ścieków. Pod względem ilościowym wody osadowe stanowią jedynie kilkuprocentowy udział w ogólnym bilansie ilościowym ścieków. Jednakże ładunek zanieczyszczeń, szczególnie związków biogennych, jaki jest w ten sposób zawracany może dochodzić nawet do kilkudziesięciu procent ogólnego ładunku dopływającego do oczyszczalni. Nieregularne, okresowe oraz zmienne jakościowo dopływy, wynikające z cykliczności oraz różnej efektywności pracy urządzeń gospodarki osadowej mogą stanowić problem dla stabilności pracy oczyszczalni oraz jej eksploatacji. Szczególnie ujawnia się to przy braku monitoringu związków fosforu podczas poszczególnych faz oczyszczania ścieków i przeróbki osadów [7]. W prezentowanym artykule autorzy przedstawili profil zmian stężenia ortofosforanów podczas oczyszczania ścieków oraz unieszkodliwiania osadów ściekowych miejskiej oczyszczalni "Jamno", ze szczególnym uwzględnieniem wód osadowych.Knowledge of biogenic compounds concentrations change profile in sewage treatment plants utilization makes up the essential condition of the optimum work of individual stages of treatment. This is caused by application of integrated arrangements for nitrogen and phosphorus compounds removal from sewage which introduced significant changes in sludge management of plant. Statistical data show that near 83% of 207 questioned Polish sewage treatment plants applies the process of activated sludge and 80% from these objects works in the system of full or partial biological removal of biogenic compounds - so-called arrangements of highly efficient removal of phosphorus (enhanced biological phosphorus removal). This paper presents profile of orthophosphates concentration change in sewage and sludge treatment parts of sewage treatment plant "Jamno". This plant consists of three integral parts: - mechanical with installation for preliminary chemical precipitation, - biological with installation of final precipitation, - and sewage sludge processing with installation of chemical conditioning. Following media are emitted during processes of sewage sludge processing: - sedimentation water from thickeners, - eluate from centrifuges, - sedimentation water from chemical reactor Sewage from Koszalin are treated in mechanical and multistage advanced biological system with the chemical preliminary and final precipitation with PIX coagulant. Orthophosphates removal efficiency reached 98÷99%. But large part of orthophosphates were cumulated in the bacteria cells and at thickening, anaerobic digestion and dewatering of biological sludge phosphorus is released from bacteria to sedimentation waters. The return water was only 1.4% of total volume of inflowing sewage to treatment plant "Jamno". Its quality may be substantial load for treatment process because it was characterized by variability of properties and amount, high concentration of nutrients (80% of orthophosphates was in suspended solids) and irregularity of inflow. Return load of orthophosphates share in the total load was 40% and reduction this value may only by achieved by replacement of centrifuges and better operation of gravity thickener. In such case decrease of coagulant consumption is possible to achiev

Study on ammonia nitrogen removal efficiency on selected sorbents

In this paper the authors present the impact study on the electrokinetic potential of ammonium nitrogen removal capabilities using various sorption materials. The study included an analysis of the electrokinetic potential zeta of the selected sorbents and their suitability to determine the removal efficiency of ammonium ions from the model solution and a liquid sludge from the sludge digestion process. Used materials were both natural and modified in the form of dust: activated carbon, natural zeolite, bentonite, sepiolite and ferric sludge from water treatment. Tested materials had a negative electrokinetic potential. The lowest potential of activated carbon had a value of zeta = -20.4 mV and the highest value zeta = -12.1 mV obtained for sepiolite. The results showed a clear dependence of ammonia nitrogen removal on the value of the electrokinetic potential. Sorbents with the most negative zeta potential values had a highest removal efficiency of ammonium ions (carbon, zeolite). The maximum value of the individual removal of ammonium ions was 10 mg N-NH4/g of activated carbon, which is characterized by the most negative value of the electrokinetic potential zeta. The results from this study suggest that, the usefulness of knowledge of the electrokinetic potential, as one of the key parameters of evaluating the physicochemical properties of sorbents