There are no Blue Flag beaches in Asia and the annual reports of

There are no Blue Flag beaches in Asia and the annual reports of the Environmental Protection Department of the Hong Kong SAR Government ( identify the measures it is taking to improve beach water quality. Hong Kong has a sub-tropical climate with most (80%) rain falling in the hot, wet, summers – an annual average of ∼220 cm. Although, during the summer months, typhoons have been known to dump that much rainfall and more in just a single day. Such rains cause problems with the drains, which overflow, and predictably, water quality standards fall – just at the time when people there, as here, are heading for PI3K inhibitor the beaches. At such times

in Hong Kong, as here too, beach water quality monitoring is halted – understandably. Great Britain’s 2012 summer weather has been similar to that typical of Hong Kong in that up to 27 June, total UK rainfall was >130 mm with the Meteorological Office confirming it had been the wettest April-June period in the United Kingdom in 100 years and the second wettest year since 1912. This was brought home to my town of Littlehampton on the night of 11 June when, because of torrential rain, the local Littlehampton Gazette

recorded that the basement apartments of 26 Victorian properties along South Terrace were flooded by storm water and raw sewage. Sewage? How could this be? The answer Ibrutinib cell line lies in the old sewerage system. About PRKACG eight years ago before he was sacked, the River Arun’s harbour master offered the occasional guided tour of his ‘patch’ and, in response to a question about the river sometimes smelling

of sewage, casually informed his group, including me, that sometimes, when the £53 million wastewater treatment scheme was overloaded, the excess was allowed to flow out along the old east–west Victorian sewerage pipe to be discharged into the River Arun exactly where it all used to be. On the night of the 11 June, this old sewer, now serving as a storm-water drain and, it transpires, an emergency sewer, failed – spectacularly. And, where did the emergency services pump this contaminated water from the basement apartments? Why into the Arun of course! In fact, moreover, Southern Water has a long history of ‘accidental’ discharges of raw sewage, one of the latest locally resulted in it being fined £5,000 on 11 June 2012 for allowing raw sewage to be discharged into a small tributary of the River Arun on 3 September 2009. As a postscript to the 5 July Sunday Times article discussed above, the author pointed out that ‘Water companies have allocated £1 billion in 2010–2015 to improve sewage overflows’ – such as has occurred in Littlehampton and the River Arun but at many other places too in England and Wales this summer. This equates to £250 million annually for the entire country.

Abnormalities in frontotemporal functional connectivity are also

Abnormalities in frontotemporal functional connectivity are also found in siblings of patients with schizophrenia Selleckchem Enzalutamide [16] and [17], but the heritability of functional connectivity determined from functional MRI (fMRI) is less well established, with one study estimating h2 at 0.42 [18]. It is known that ZNF804A is highly expressed in the brain and that the presence of A-allele

at rs1344704 creates a myelin transcription factor binding site [2] and [19]. The most comprehensive data on ZNF804A function come from neuroimaging and neuropsychology, collectively indicating that rs1344706 is associated with brain function. Esslinger et al. [20] reported reduced functional connectivity between the left and right dorsolateral prefrontal cortices and increased frontotemporal functional connectivity in carriers of the risk allele (A) during a working memory task, findings that were (partly)

replicated in two subsequent studies [16] and [21]. Importantly, Esslinger et al. [22] later showed that the reduced interhemispheric prefrontal connectivity was also apparent during selleck chemicals a facial emotion processing task and during rest, whereas the increased frontotemporal connectivity appeared specific to working memory processes both in the original study and in two replication studies [16] and [21].

This task-independent association of ZNF804A genotype on interhemispheric prefrontal functional connectivity prompted the hypothesis that these effects may be mediated Rutecarpine by effects on white matter integrity, especially in anterior interhemispheric connections. In contrast, the effects of ZNF804A on frontotemporal connectivity are less likely to be directly mediated by white matter structure since they have only been observed in the context of working memory tasks [21] and [22] and interact with task condition  [16]. In line with this hypothesis, Lencz et al. [23] showed that individuals homozygous for the ZNF804A risk allele (A) have reduced total white matter volumes compared to carriers of the nonrisk allele (C). However, total volumetric measures lack spatial specificity and are particularly susceptible to partial volume effects and segmentation difficulties. DT-MRI is more suited to the study of white matter, and FA is the most commonly used measure of white matter integrity in vivo. Surprisingly, using DT-MRI tractography, Voineskos et al. [19] did not detect any effects of ZNF804A on FA in the uncinate fasciculi, arcuate fasciculi, cingulum or corpus callosum of 62 healthy individuals, 39 C-carriers versus 23 A-homozygotes, aged between 18 and 59 years.

2) Dry root weight

in the 0–20 cm soil layer peaked at 1

2). Dry root weight

in the 0–20 cm soil layer peaked at 14 d after pollination, and at 28 d for soils 20–40 cm and below. In the N0 treatment, dry root weight in the 0–20 cm layer peaked 14 d after pollination, but below 20 cm the dry root weight was reduced. Compared with N1, the N0 treatment showed a significant (P < 0.05) decrease in dry root weight at 0–20 cm soil depth, but there was a significant (P < 0.05) increase in the 70–100 cm layer. Changes in dry root weight in the 20–40 cm and 40–70 cm soil layers were not significantly different; however, the deep root ratio of N0 was significantly higher than that of N1. Root reductive activity is a comprehensive index that reflects root absorption function [13]. After pollination, root reductive activity in each soil layer changed as the plants matured (Fig. 3), exhibiting single-peak increases

before decreasing. Under N1, root reductive activities underwent significant increases in the 0–20 cm and 20–40 cm soil layers, with peaks exhibiting prolonged durations. Root reductive activity in the 70–100 cm layer under N0 showed a steady decrease compared with N1. Under both nitrogen levels, root reductive activity decreased in each layer of closely spaced plants, and the greatest difference between treatments was observed during the grain-filling Ceritinib purchase stage. At late grain filling, differences were not as evident. The effects of different plant spacing treatments on maize grain yield are influenced by interactions between aboveground and belowground resource competitions. Compared with competition for light aboveground, nutrient competition in roots includes more than 20 nutrient elements, which have

substantial differences in molecular weight, soil oxidation state and mobility, and there are more significant effects of nutrient competition in roots on the growth of plant [8]. Narrow spacing is chosen most often to increase photosynthetic capacity by increasing the interception of available solar radiation, resulting in improved maize yield [6]. However, some studies have Megestrol Acetate demonstrated that an increase in solar radiation does not increase but decrease maize production [23] and [24]. In this study, excluding interference due to aboveground competition for light, narrow spaced plants significantly decreased aboveground dry matter accumulation and grain yield by 8.4% and 5.0%, respectively. Aboveground dry weight and grain production are closely related to nitrogen accumulation, translocation and utilization. Above-ground nitrogen accumulation in the narrow plant spacing treatment was decreased by an average 12.8%.

Additionally, the authors thank Nicholas Walker for the English r

Additionally, the authors thank Nicholas Walker for the English review. “
“Coffee is the most consumed food product in the world. Roasting induces severe transformation on coffee’s chemical composition. Additionally, during storage, the roasted beans are susceptible to further chemical and physical changes that may greatly affect the quality and the acceptability of the brew.

Lipids are major coffee components and correspond to approximately 11–20 g/100 g of roasted Coffea arabica composition ( Oliveira, Franca, Mendonça, & Barros-Junior, 2006; Toci, Farah, & Trugo, 2006; Trugo, 2003). Furthermore, triacylglycerols (TAG) comprise the main lipid class in coffee and account for approximately Selleck PD0332991 8–17 g/100 g (75% of total coffee lipids) in freshly brewed coffee, whereas free fatty acids (FFA) account for 0.1–0.2 g/100 g (about 1% of total coffee lipids Metformin only) ( Trugo, 2003). Among the most important unsaturated fatty acids for coffee freshness are oleic (18:1n-9), linoleic (18:2n-6) and linolenic (18:3n-3) acids, which account, respectively, for approximately 0.6–1.1 g/100 g, 2.9–5.4 g/100 g and 0.08–0.15 g/100 g, representing 7%, 36% and 1% of TAG fraction

( Folstar, 1985; Lercker et al., 1996; Nikolova-Damyanova, Velikova, & Jham, 1998; Speer & Kolling-Speer, 2006). Lipids may contribute to loss of sensory quality during storage. TAG can be hydrolyzed either chemically or enzymatically to produce a mixture of diacylglycerols,

monoacylglycerols, FFA, and glycerols molecules (Folstar, 1985; Frankel, 2005). The rate at which these reactions occur depends mostly on factors related to environmental and technological aspects such Thalidomide as availability of oxygen and moisture, exposed surface area, temperature, as well as package material (Manzocco & Lagazio, 2009; Pérez-Martínez, Sopelana, Paz de Peña, & Cid, 2008; Speer & Kolling-Speer, 2006). Since during coffee roasting hydrolytic enzymes are thermally inactivated, moisture and temperature are the main factors that will rule hydrolysis reactions in roasted coffee. The presence of high moisture content in food storage systems reduces the contact between food and oxygen, which tends to cause a decrease in oxidation reactions, but promotes hydrolysis reactions. When moisture in the storage system is low, Entropy decreases in the system, which leads to a decrease in the kinetic energy of the molecules and thus in the rates of all types of reactions. However, when storage temperature is high, Entropy increases, accompanied by a raise in the rate of degradation reactions (Frankel, 2005, Chapter 11; Kim & Min, 2008).

5% reduction, respectively, Fig 1B) When mouse survival was eva

5% reduction, respectively, Fig. 1B). When mouse survival was evaluated, it was noted that the first animal died on the 9th day in EAT-inoculated group of mice. At the 10th day a 20% death toll was noted, and only 50% of the animals were alive on the 15th day. In the R-954-treated group of mice, only one animal died during the experimental period and this death occurred only after 14 days of consecutive treatment (Fig. 1C). On the 10th day after EAT cell inoculation into mice, a 12.6-fold increase of total blood cell count was observed (0.5 ± 0.2 × 107

cells in control group vs. 6.3 ± 2.1 × 107 cells in EAT-inoculated mice). This effect was accompanied by a proportional increase in total bone marrow cell count (0.12 ± 0.02 × 107 cells in selleck chemicals llc control

group vs. 1.3 ± 0.02 × 107 cells in EAT-inoculated mice) and on cells from ascitic lavage (0.8 ± 0.3 × 107 cells in control group vs. 11.7 ± 1.1 × 107 cells in EAT-inoculated mice). SCH772984 mw Vincristine (0.5 mg/kg, i.p.) a well known carcinostatic agent used for comparison purpose reduced total blood cell counts by 55.5% (6.3 ± 2.1 × 107 cells in EAT-inoculated mice vs. 2.8 ± 0.8 × 107 cells in vincristine-treated EAT inoculated mice), total bone marrow cell count by 76.9% (1.3 ± 0.02 × 107 cells in EAT-inoculated mice vs. 0.3 ± 0.2 × 107 cells in vincristine-treated EAT inoculated mice), and total cells in ascitic fluid by 71.8% (11.7 ± 1.1 × 107 cells in EAT-inoculated mice vs. 3.3 ± 1 × 107 cells in vincristine-treated EAT inoculated mice). Treatment of animals previously inoculated with EAT cells with B1 antagonist R-954 reduced total blood, bone marrow, and ascitic cell counts to values similar to vincristine-treated mice (3.2 ± 0.9; 0.5 ± 0.1; 4.8 ± 1.1 × 107 cells, respectively) (Fig. 2). In order to evaluate inflammatory mediator release after EAT inoculation, mice were sacrificed on the 10th day after tumor cell injection. Peritoneal ascitic fluid was collected and total protein, nitric oxide, PGE2, and TNFα were measured. In mice inoculated with EAT cells, marked increases

of the total proteins (from 13.8 ± 3.1 to 493.5 ± 33.8 mg/ml), O-methylated flavonoid nitric oxide (from 2.8 ± 3.3 to 76.9 ± 12.7 μM), PGE2 (from 28.4 ± 5.9 to 344.9 ± 45.8 pg/ml), and TNFα (from 31.7 ± 9.9 to 792.3 ± 113.4 U/ml) were noted when compared to the levels in the fluids of non-inoculated animals. Treatment of mice with R-954 reduced significantly the total protein extravasation (57.3%) as well as the production of nitric oxide (56%), PGE2 (82%) and TNFα (85.7%). The antitumoral drug vincristine also significantly reduced by 92% the protein extravasation, by 84.5% nitric oxide, by 94.7% PGE2, and by 92.2% TNFα levels (Table 1). When Ehrlich cells are inoculated intraperitoneally, the tumor process develops in an ascitic form but when the cells are inoculated subcutaneously, the tumor develops in a solid form.

The objectives of this paper were (1) to simulate flow velocity a

The objectives of this paper were (1) to simulate flow velocity and surface wave fields in the Suur Strait and to validate these with in situ observations; (2) using simulation results, to estimate the proportion of surface waves in the

flow field and water exchange through the Suur Strait; and (3) using observation data and model simulations, to estimate wave-induced and current-induced shear velocities. This paper is structured as follows. In section 2, the field data, circulation model and wave model are briefly described, and the wave and current shear velocities are calculated. In section 3, the model results are presented, discussed and compared with the measurements. The conclusions are drawn in section 4. Current velocity and wave measurements CDK inhibitor in the Suur Strait were performed in November–December 2008. A buoy station equipped with a Sensordata current meter SD-6000 and a pressure sensor was deployed on 13 November near Virtsu (58°34.95′N; 23°29.30′E, Figure 1c). The water depth at the location of the buoy station was 9 m. The current meter was at a depth of 3.5 m and the wave gauge at 2.5 m. The current speed and direction recording interval was 5 min, that of the wave gauge 0.25 s. Current measurements lasted until 4 December and wave measurements AZD6244 until 6 December. The method for reconstructing surface elevation spectra from sub-surface pressure recordings is described in detail by

Alari et al. (2008). Wind speed and direction were recorded with the Väisälä Weather Transmitter WXT520

installed at a height of 30 m at the Kessulaid weather station (Figure 1c). It recorded wind data at 5 min intervals from 21 November to 13 December. We used a height correction factor of 0.91 to reduce the recorded wind speed to the reference height of 10 m (Launiainen & Laurila 1984). We used a two-dimensional circulation model based on the hydrodynamic equations for a shallow sea. The model had been applied earlier to different regions of the Estonian coastal sea (Sipelgas et al. 2006). The model consists of vertically integrated motion equations equation(1) ∂u∂t+u∂u∂x+v∂u∂y−fv=−g∂η∂x+Fwxh−Fbxh+Fwavexh+Gx,∂v∂t+u∂v∂x+v∂v∂y+fu=−g∂η∂y+Fwyh−Fbyh+Fwaveyh+Gy Sulfite dehydrogenase and a continuity equation equation(2) ∂η∂t+∂uh∂x+∂vh∂y=0, where (u, v) are the vertically averaged velocities in the water column in the Cartesian coordinates, (Fxw, Fyw) are the kinematic wind stresses, (Fxb, Fyb) are the bottom friction stresses, (Fxwave, Fywave) are the wave-induced forces, (Gx, Gy) are the horizontal turbulent viscosities in the (x, y) directions, f is the Coriolis parameter, g is the acceleration due to gravity, η is the sea surface elevation (deviation from the equilibrium depth) and h(x, y) is the depth. In order to take into account the wave-induced currents, a wave-induced force per unit surface area is added to the kinematic wind stress in both the x and the y directions.

From another perspective, an established state of cross-modal sen

From another perspective, an established state of cross-modal sensory adaptation may in fact impair the ability of an implant to elicit simple phosphenes at all, in favor of phantom perceptions related to extra-visual sensory cortices,

e.g. touch Selumetinib clinical trial (Kupers et al., 2006, Merabet et al., 2007 and Ptito et al., 2008a). More research needs to be done to determine the impact of neuroplasticity on the likely performance of an implant in the long term, or conversely any negative influence of the implant on the recipients׳ adaptations to blindness. The implantation of penetrating cortical electrode arrays is a major neurosurgical procedure. As with any surgery involving the opening of the skull and intradural space, there is a demonstrable risk of acute and longer-term complications resulting in a poor surgical and clinical outcome. These risks include but are not limited to postoperative hemorrhage, swelling, tissue infarction, infection, seizures and neurological deficits, each of which may delay or preclude progressing

to the implant testing stage (discussed in more detail in Section 6.2). A key component to minimizing these risks will be the selection of candidate recipients in whom the burden of comorbidities known to negatively impact on neurosurgical outcomes is acceptably low. For example, the risk of postoperative bleeding is increased by hypertension, diabetes and coagulopathy buy Linsitinib (Seifman et al., 2011). Poor nutritional status due to advanced age, malignancy or obesity may increase the risk of infection (Walcott et al., 2012), and preoperative screening for MRSA colonization may be helpful in avoiding a complicated postoperative course in the event of infection (Harrop et al., 2012). Patients with a history of epilepsy are innately at greater risk of suffering postoperative seizures, and should be excluded (Weiss and Post,

2011). The complexity and experimental status of the implant procedure and rehabilitation Enzalutamide price process dictates that obtaining informed consent, for which a detailed discussion of surgical risks is required, will need to be undertaken carefully to ensure a thorough understanding by potential recipients. The mental health and capacity of a potential recipient is therefore of paramount importance in this context, as it may potentially impact on the treating physician׳s ability to ensure that a truly informed consent can be obtained (Lane et al., 2012 and Merabet et al., 2007). Moreover, it may impact on the perception of the soundness of potential recipients׳ motivation to participate, or the likelihood of effective rehabilitation (Dagnelie, 2008 and Merabet et al., 2007). Lane et al.

Traumatic brain injury, malignant stroke, tumor, diffuse hypoxic–

Traumatic brain injury, malignant stroke, tumor, diffuse hypoxic–ischemic brain damage are supposed to be the main causes of BD. All these factors affect the brain and lead to brain edema and swelling, intracranial pressure increase, gradual reduction of cerebral perfusion pressure, decrease and termination of intracranial blood flow and necrosis of brain parenchyma up to 2nd cervical segment [1], [2] and [3]. According to the Russian Metformin in vitro National Guidelines of BD there are Diagnostic criteria for clinical diagnosis of BD [4]: 1. Defined cause irreversible deep coma. In general, these criteria correspond to neurologic criteria for the diagnosis

of brain death of American Academy of Neurology [2] and [5]. The following two confirmatory tests are approved for BD diagnosis in Russia: 1. Electroencephalography (EEG) – reveals no electrical activity of brain in BD patients. Angiography is believed to reduce the observational period only and does not substitute to any clinical criteria of BD. According to the Russian National Guidelines on Diagnostics of Brain Death, ultrasound confirmatory tests are being Crizotinib investigated and can not be recommended for BD diagnosis, at the same time, all over the world ultrasound tests are the 3rd in order of sensitivity and frequency for BD diagnostics [6] and [7]. Transcranial Doppler (TCD) is notably desirable in patients

in whom specific components of clinical testing cannot PTK6 be reliably performed or evaluated such as barbiturate brain protection, hypothermia or face trauma [8], [9] and [10]. Our

department has gained experience in ultrasonography in clinical and confirmatory tests, 438 cases of BD were diagnosed since January 1995 to December 2010 [11]. The diagnosis of BD was confirmed by TCD and EEG. Color duplex sonography (CDS) was started to be performed in 2009. We initiated a prospective observational study of the extra- and intracranial artery CDS in BD diagnostics in 2009. 20 patients with BD have been enrolled in the study up to December 2010. The study was approved by Local Ethic Committee of Moscow State University for Medicine and Dentistry in 2008. The aim of the study was – to investigate whether CDS of both extra- and intracranial arteries increases sensitivity of the test in patients with BD compared with CDS of intracranial arteries alone; The study was started in Moscow hospital intensive care units in 2009 and has still been going on. 20 patients with BD due to traumatic brain injury and intracranial hemorrhage were included in the study and underwent a sonographic study which included color duplex sonography (CDS) of extracranial and intracranial arteries. BD was diagnosed according to the Russian National Guidelines of BD. The average age of patients was 25 ± 5.4 years. The average time from ICU admission to BD development was 27 ± 6.5 h. The diagnosis of traumatic brain injury and intracranial hemorrhage was detected by computer tomography at the admission.

Currently, if a research or clinical study requires both PET and

Currently, if a research or clinical study requires both PET and MRI data, the patient must endure two exams in confining scanners, which is problematic for patients who suffer

from even mild claustrophobia. This duplication not only increases the discomfort (both physical and psychological) the patient must endure, but also effectively doubles the chances of motion during one or both scans with the subsequent need to rescan particular sequences or even the entire study. In light of the discussion in the previous section if, as some studies suggest, there is an added diagnostic benefit to combing PET and MRI, then it is of great import to minimize the difficulties associated with acquiring both data sets. The problem of patient anxiety and discomfort is a well-known phenomenon extending back (at least) AG-014699 in vivo to the first few years after the widespread introduction of clinical MRI [86], [87], [88] and [89]. A review of the topic shows that as many as 37%

(range: 4%–-37%) of patients undergoing MRI had an anxiety-related reaction to the procedure [90] and [91]. In one study, which found that approximately 14% of MRI patients required some form of sedation to tolerate a standard-of-care MRI, the use of sedation was actually more common in patients who had already had previous MRI exams, indicating that familiarity with the procedure may not reduce stress related to the procedure [92]. The problem of anxiety and discomfort Branched chain aminotransferase during imaging is not unique to MRI, as similar issues arise for PET examinations. click here It has been noted that a patient that is stressed and fidgeting can have elevated FDG uptake in skeletal muscle, which may adversely affect tumor-to-muscle ratio measurements [93]. Additionally, there is a well-known anxiety-induced increase in FDG uptake in brown fat that has been linked to false-positive

interpretations in 2%–4% of all studies, as well as false-negative interpretations due to brown fat uptake masking lesion detectability [94], [95] and [96]. The problem is often exacerbated in pediatric patients where stress-induced muscle tension, crying and the associated coughing can yield increased muscle FDG uptake [97]; these issues are well known amongst technologists, and efforts have been made to address the particular issues surrounding pediatric PET studies [98]. A final, extremely practical, point to note is that a combined PET–MRI exam would preclude the patient from having to endure the (sometimes lengthy) periods in multiple waiting rooms waiting for their scans. As many of these patients are missing work and/or traveling from far distances to undergo their testing, a combined exam would undoubtedly enhance their experience and make it more tolerable. For the cancer patient who already may not have a great deal of strength to attend these imaging tests, eliminating one set of waiting rooms and preps would be greatly appreciated.

1) 1 cm3 of the upper 0–5 cm section of each core was removed an

1). 1 cm3 of the upper 0–5 cm section of each core was removed and stored frozen into 15 ml centrifuges tubes until the later analysis. ELISA and protein phosphatase 1 inhibition assay (PPIA 1) are the most sensitive methods widely used for determination of microcystin (Adamovsky et al., 2007, Amorim and Vasconcelos, 1999, Babica et al., 2006, Kankaanpaa et al., 2007, Msagati et al., 2006, Nicholson et al., 2007, Sipia et al.,

2006 and Yu et al., 2002). ELISA is often advised for the analyses of cyanobacterial toxins when their concentrations are lower than high-performance liquid chromatography (HPLC) detection limit (Mazur-Marzec et al., 2006). However, occasionally ELISA can give false positive results, therefore to confirm the occurrence of microcystin in samples PPIA was additionally employed. Mussels and sediment samples were lyophilized (TEGA, Germany) and then extracted using 30 ml PI3K inhibitor of pure methanol per 1 g mussel and sediment dry weight. Extracts were disrupted by sonication (5 min) and then centrifuged for 15 min at 10,000 rpm 20 °C. The solvents were removed by rotary evaporation and the residue was re-dissolved in 1 ml of MiliQ water. After that samples were vortexed for 1 min and then centrifuged

for 15 min at 12,000 rpm 20 °C. Later on, the samples Androgen Receptor Antagonist screening library were subjected to solid phase extraction on Sep-Pak Vac C18 cartridges (200 mg, Waters, Massachusetts, USA). Chlorophyll a was extracted by adding 80% ethanol to sediment samples ( Jespersen and Christoffersen, 1987). After 24 h samples were centrifuged and obtained supernatant analyzed spectrophotometrically according to Lorenzen (1967).

Extracts of mussels and sediments were diluted in MilliQ water (10–5,000 times) and analyzed by enzyme-linked immunosorbent assay (ELISA). The ELISA test was performed using 3-mercaptopyruvate sulfurtransferase the EnviroGuard kit (Strategic Diagnostics, Newark, DE, USA), according to the manufacturers’ instructions. The same extracts were also analyzed by colorimetric protein phosphatase 1 inhibition assay (PPIA). The PPIA was carried out on a 96-well microplate according to the method described by Rapala et al. (2002). Bovine serum albumin (BSA) was obtained from Sigma–Aldrich (St. Louis, MO, USA). Dithiothreitol (DTT), MgCl2·6H2O, MnCl2·4H2O, Na2SO4, p-nitrophenyl phosphate (p-NNP – the substrate), tris-(hydroxymethyl)-aminomethane (Tris) were of analytical grade. The substrate and enzyme buffers were prepared immediately before the test. Catalytic subunits (2.5 U) of commercially available enzyme (PP1; New England Biolabs, USA) were diluted in 1.5 ml of the enzyme buffer. Subsequently 10 μl of standard solutions or sample were added to the well and mixed with 10 μl of PP1 in buffer. After 5 min incubation, 200 μl of p-NPP in buffer solution was added to each well. The content of the wells was mixed by swirling the plate sideways. After 2-h incubation at 37 °C, the absorbance of the solutions was measured.