, 2011). In the presence of the Ca2+-uniporter blocker ruthenium red, nemorosone induced mitochondrial swelling in a way sensitive to the classic mitochondrial permeability transition (MPT) inhibitor cyclosporine A. Unlike nemorosone, GA uncoupled mitochondria through a non-protonophoric mechanism (result not shown). In addition, mitochondrial swelling elicited by GA was not inhibited by cyclosporine A or EGTA and, therefore, it does not correspond to the MPT process (Zoratti and Szabò, 1995). Rather, the evidence that GA increased mitochondrial membrane fluidity suggests GSK1120212 that a direct

interaction with mitochondrial membrane, whose major structural lipids are cardiolipins, accounts for its permeabilizing action on the organelle. The evidence that isocitrate partly prevented GA-induced NADPH oxidation/depletion and

mitochondrial swelling in isolated mitochondria, as well as cell viability decrease, ATP depletion and ROS levels increase in HepG2 cells, suggests that NADPH oxidation/depletion is at least partly involved in the GA permeabilizing action on mitochondria and its consequence on cells. Isocitrate is the substrate of NADP+-dependent isocitrate dehydrogenase, a major Androgen Receptor Antagonist library NADPH source in mitochondria with a key role in cellular defense against ROS (Jo et al., 2001). In citosol, NADPH is provided primarily by the pentose phosphate pathway, including glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase. In this regard, the fact that HepG2 cells

incubated in medium with Plasmin low glucose levels were more sensitive to GA-induced death, mitochondrial membrane potential dissipation, ATP depletion and ROS levels increase reinforces the proposed GA toxicity mechanism. Low glucose impairs NADPH re-generation in citosol and may potentiate mitochondria-mediated cytotoxic actions. In conclusion, the present results suggest the following sequence of events for the GA action on mitochondria: 1) GA interaction with mitochondrial membrane increasing its fluidity and promoting its permeabilization; 2) mitochondrial membrane potential dissipation; 3) NAD(P)H oxidation/depletion due to inability of membrane potential-sensitive NADP+ transhydrogenase of sustaining its reduced state; 4) ROS accumulation inside mitochondria and cells; 5) additional mitochondrial membrane permeabilization due to ROS; and 6) ATP depletion. The evidence that Ca2+ efflux was only partially prevented by the Ca2+-uniporter blocker ruthenium red in isolated mitochondria and the inability of isocitrate to prevent mitochondrial membrane potential dissipation in HepG2 cells suggest that this latter is an early event associated to the GA action on mitochondria, which could ultimately, via energetic and oxidative stress implications, result in cell ATP depletion.

To prospectively evaluate the efficacy of RFA combined with ER in case of focal lesions, for BE with HGD/EC in 13 European centers with expertise in BE neoplasia. Patients with BE≤12 cm and HGD/EC on 2 separate endoscopies were included. Visible lesions (<2cm length; <50% circumference) were removed with ER, residual EC was excluded on 2 mapping endoscopies post-ER. Subsequent RFA was scheduled

every 3 months until clearance of BE was Selleck HSP inhibitor achieved, with max 5 RFA sessions allowed. Escape treatment was permitted for residual BE after RFA (max 2 APC sessions for islands <5mm, ER for islands >5mm or suspicious lesions). Follow-up (FU) endoscopy was scheduled at 3+9 mo after

last treatment and annually thereafter, with 4Q/2cm biopsies from ERK inhibitor mw neosquamous epithelium and gastric cardia. Endpoints: complete eradication of neoplasia (CE-neo) and intestinal metaplasia (CE-IM); durability of CE-neo/CE-IM. To ensure uniformity and protocol compliance, investigators were trained at the coordinating site and a study monitor attended all treatments and first FU on-site. Central pathology review of all ER/biopsies was performed at the coordinating site. 132 patients (107M, mean 65yrs, median BE C3M6) underwent en-bloc (n=63) or piecemeal ER (n=56); or no-ER (n=13). Worst ER histology: EC (n=78), HGD (n=31), LGD (n=7), no dysplasia (n=3). Worst grade post-ER/pre-RFA: HGD (n=36), LGD (n=45), no dysplasia (n=51). 124 pts Amine dehydrogenase completed the treatment phase, 8 discontinued due to unrelated causes. After a median of 3 (IQR 3-4) treatments, including ER (n=14) or APC (n=14), per intention-to-treat analysis (counting drop-outs as failures) CE-neo was reached in 122/132 (92%) and CE-IM in 115/132 (87%) pts. In a per-protocol analysis (censoring for drop-outs) CE-neo/CE-IM

were achieved in 98% and 93%, respectively. Of 2 CE-neo failures, 1 was referred for surgery (T1bN0M0), 1 patient was treated endoscopically (off-protocol). Of 115 pts who reached CE-neo/CE-IM per-protocol, CE-neo was maintained in 112/115 (97%) pts during median 25 (IQR 18-34) mo FU since last treatment, with median 4 (2-5) FU endoscopies and 41 biopsies per patient. 3 pts with recurrent neoplasia (EC n=1; HGD n=2) were effectively re-treated by ER or APC. This is the largest prospective multicenter trial on RFA combined with ER for treatment of BE with HGD/EC. Our outcomes suggest that if performed by trained, expert endoscopists in carefully selected patients, this combined approach is highly effective, and appears durable in the majority (97%) of patients once complete eradication of neoplasia and IM is established. “
“Barrett’s esophagus (BE) is the pre-cursor lesion to esophageal adenocarcinoma (EAC).

At current injections double the strength of the rheobase (which were applied in a subset of cells), the mean latency to the first AP (the chronaxie) did not differ (median and interquartile values: wild-type, 9.5 (6.8, 9.5) ms, n = 29; Ts65Dn, 8.7 (6.9, 10.5) ms, n = 15; p = 0.310, Mann Whitney U test). Although the increased excitability of Ts65Dn GCs was not accompanied by changes in AP accommodation, it was associated with changes in AP waveform

(Fig. 3A). The average amplitude, measured between the overshoot and the afterhyperpolarization (Bean, 2007) for the first three APs evoked at or just above rheobase, was larger LDK378 mw by 4.4 mV in Ts65Dn cells (wild-type, 99.4 ± 1.4 mV, n = 33; Ts65Dn, 103.8 ± 1.1 mV, n = 20; p = 0.032, Student’s t-test). This was the result of a higher overshoot (by ~ 11%) without a change in afterhyperpolarization ( Fig. 3B). The larger APs in Ts65Dn GCs were also ~ 10% narrower (width at half amplitude: wild-type, PF-562271 714.9 ± 25.9 μs, n = 33; Ts65Dn, 643.5 ± 15.4 μs, n = 20; p = 0.045, Student’s t-test). It has

been shown previously that in wild-type GCs, membrane potential changes more slowly during the falling phase than the rising phase of the AP ( Brickley et al., 2007). Fig. 3C shows that this difference was maintained in Ts65Dn cells, indicating that the speeding of the APs was due to a proportionate increase in the maximum rates of rise and fall, of ~ 13% ( Fig. 3D). The finding that APs were faster in Ts65Dn cells, which have a longer membrane time constant because of their higher Cin and Rin, indicates that the speeding reflects changes in ion channel activity or distribution, which overcomes the slowing effect

of a longer membrane time constant on changes in membrane potential. It is known that there is a ~ 33% decrease in cerebellar volume and a 25–30% decrease in GC density in individuals with DS (Aylward et al., 1997, Baxter et al., 2000, Jernigan and Bellugi, 1990, Pinter et al., 2001 and Raz et al., 1995). We have found that in GCs of young adult Ts65Dn mice (P40–60), which replicate cerebellar changes in DS (20% shrinking of cerebellar volume, 14% narrowing of the granular layer, 24% drop in GC density) (Baxter et al., 2000 and Roper et al., 2006), the 4-Aminobutyrate aminotransferase electrical properties of the surviving GCs are not identical to those of GCs in wild-type mice. As the paucity of GCs in Ts65Dn mouse cerebellum and DS cerebellum stems from impaired division of precursor cells (Haydar and Reeves, 2011), changes in the electrical properties of Ts65Dn GCs could potentially be caused by arrested or slower development that results in immature electrophysiological characteristics. Wild-type GCs undergo marked changes in excitability, input resistance and AP waveform during postnatal development (Brickley et al., 2001 and Cathala et al.

The patient gave his consent to submit his images for publication purposes. “
“Podstawowym źródłem składników odżywczych powinna być właściwie zbilansowana

dieta. W sytuacji, gdy z różnych przyczyn codzienna dieta nie pokrywa zapotrzebowania na podstawowe składniki odżywcze, można rozważyć stosowanie suplementacji. Suplement diety to środek spożywczy, którego celem jest uzupełnienie normalnej diety, będący skoncentrowanym źródłem witamin lub składników mineralnych bądź innych substancji wykazujących efekt odżywczy lub inny fizjologiczny, pojedynczych lub złożonych, wprowadzany ABT 199 do obrotu w formie umożliwiającej dawkowanie, w postaci: kapsułek, tabletek, drażetek i w innych podobnych postaciach: saszetek z proszkiem, ampułek z płynem, butelek z kroplomierzem itp. [1]. Przez suplementację diety rozumiemy również jej wzbogacanie, tzn. dodawanie do środków spożywczych jednego lub kilku składników odżywczych, niezależnie od tego, czy naturalnie występują one w tym środku spożywczym, czy nie, w celu zapobiegania niedoborom lub korygowania niedoborów tych składników odżywczych w całych populacjach albo określonych grupach ludności (zgodnie z poniżej przywołanymi regulacjami). W Polsce suplementy diety to

produkty spożywcze podlegające następującym regulacjom prawnym: – Ustawa z dnia 25 sierpnia 2006 r. o Bezpieczeństwie Idelalisib in vitro Żywności why i Żywienia (Dz. U. Nr 171, poz. 1225) Niezbędnymi składnikami diety są kwasy tłuszczowe omega-3, z których kwas alfa-linolenowy (alphalinoleic acid, ALA) nie jest syntetyzowany przez organizm ludzki i uważany jest za prekursora pozostałych kwasów z tej rodziny, przede wszystkim długołańcuchowych wielonienasyconych kwasów tłuszczowych (long chain polyunsaturated fatty acids, LC-PUFA), w tym kwasów dokozaheksaenowego (docosahexaenoic acid,

DHA) i eikozapentaenowego (eicosapentaenoic acid, EPA). Podstawowym źródłem EPA i DHA są ryby morskie, olej rybi oraz owoce morza. Kwasy omega-3 posiadają właściwości przeciwzapalne, zapobiegają miażdżycy naczyń krwionośnych i dlatego znalazły zastosowanie w zapobieganiu chorób sercowo-naczyniowych, zespołu metabolicznego oraz w przewlekłych chorobach zapalnych [2, 3, 4, 5]. Zasadnicze znaczenie ma również zapewnienie właściwej podaży kwasów omega-3 w okresie ciąży, laktacji, a także w wieku rozwojowym. Szczególnie istotne w tym okresie jest zaopatrzenie organizmu rozwijającego się płodu i dziecka w kwas dokozaheksaenowy, który w dużych ilościach odkłada się w rozwijającym się ośrodkowym układzie nerwowym [6]. Polska należy do krajów szczególnie zagrożonych niedoborem kwasów tłuszczowych długołańcuchowych omega-3.

Sample recoveries were tested at two concentration levels (2 and 3 nM) mimicking the average field sample concentrations. Two subsequent analyses of the same water volume were performed. For the first, a known concentration was sampled while for the second the C59 wnt same water volume left from the first analysis was resampled. Recoveries (R %) shown in Table 4, varied from 90.3 to 100 % and were deemed very satisfactory. Repeatability was investigated also at concentration levels of 2 and 3 nM.

Four concentration replicates for each concentration level were analyzed and their relative standard deviations, shown in Table 4, ranged from 3.1 to 16 %. A trend of increasing RSD % with decreasing concentration was observed. The overall accuracy for the NTD method was estimated always better than 7.4 %. The estimation took into account a 5 % uncertainty for the concentration R428 in vitro measurement of each tracer as provided by the calibration gas bottle, a 2 % uncertainty for the measurement of the dilution volumes used for the preparation of the desired calibration concentration ranges, a 5 % uncertainty for the volume measurement of the 10 ml seawater sampling and a 1 % uncertainty for the measurement of the purging volume. As mentioned in the Experimental section, nine mesocosm enclosures with modified pCO2 concentrations were studied. Based on their CO2 concentration differences, the mesocosms were divided into three

pCO2 groups. Mesocosms M2, M4, M6 (280, 280, 360 μatm pCO2, respectively) represent the low pCO2 group, mesocosms M1, M3, M8 (560, 840, 1120 μatm) the middle pCO2 group and mesocosms M5, M7, M9 (1400, 2000, 3000 μatm) the high pCO2 group. The applied low pCO2 values are characteristic of our present day environment, the middle ones represent the predicted atmospheric CO2 levels for 2100 and the third ones provide a more extreme future scenario ( Gattuso and Hansson, 2011). Seawater samples from all mesocosms were collected, purged and analyzed as described in the Experimental section. Throughout the experiment,

calibrations of one concentration level were performed against a working gas-mixture standard, routinely, every five sample measurements. The response factor of these standard analyses was used to calibrate the samples measured in between. selleck screening library On days where the ambient temperature remained stable within the day, the GC responded similarly to all calibration samples. On days with stronger ambient temperature differences the GC responses between the various calibrations were more diverse. Representative averages of the % variation of the calibration factor (% RSD) within a day were in the order of 21.5, 18.54 and 30 % for DMS, isoprene and the α-pinenes, respectively. At least one blank analysis was performed in each measurement sequence (day of analysis). Linearity of the system was confirmed regularly (five times) during the course of the experiment, over wide ranges of concentrations (1.3–9.3 nM for DMS, 1.5–10.4 nM for isoprene and 1.7–12.

No known deaminase acts on adenine in DNA, however, an adenosine deaminase (ADA) converts free dA to dI which can be further metabolized to Hx [7]. Hx can be salvaged to deoxyinosine monophosphate (dIMP) and subsequently converted to deoxyinosine triphosphate (dITP) by a currently poorly understood pathway [8]. dITP may also be produced by spontaneous deamination of dATP. DNA polymerases can use dITP as a substrate

during DNA replication and will most often insert dITP opposite a C (Figure 2a) [9 and 10]. Normally the intracellular dITP concentration is kept learn more low compared to the canonical deoxynucleotide triphosphates (dNTPs) [8]. The steady state level of inosine in DNA is 0.5–1 per 106 nucleotides in different mouse tissue, E. coli and S. cerevisiae [ 11, 12• and 13•]

comparable to the level of the more studied oxidation product of dG, 8-oxo-7,8-dihydro-2′-dG. In addition to its premutagenic properties, dI may lead to altered recognition sites for DNA binding proteins with consequences for example gene expression. To avoid such treats, cells harbor two main pathways for inosine repair: base excision repair (BER) and Endonuclease V. BER is the major pathway for repair of damaged DNA bases and proceeds through multiple steps requiring several enzymes [14]. The first step is initiated by DNA glycosylases recognizing and removing SD-208 solubility dmso damaged DNA bases such as alkylated, oxidized and deaminated bases. In most pro- and eukaryotic species the alkyl adenine DNA glycosylases (Escherichia coli AlkA; Saccharomyces cerevisiae MAG; mammalian Aag), remove Hx from genomic DNA ( Figure 2a) [ 15 and 16]. In Schizosaccharomyces pombe, thymine DNA glycosylase (Thp1)

rather than Mag1 appears to be the inosine-specific DNA glycosylase [ 17 and 18]. An alternative excision repair pathway for the removal of deaminated purine bases has been proposed, in which Endonuclease V (EndoV) initiates repair by cleavage of the second phosphodiester bond 3′ to inosine ( Figure 2 and Figure 3) [ 19]. Further, a small patch of DNA containing the lesion is removed by exonucleases or endonucleases and finally, DNA polymerase and DNA ligase completes repair by gap filling and ligation. Although this alternative C59 nmr excision repair mechanism has been reconstituted in vitro with recombinant proteins [ 20], in vivo data supporting this pathway is lacking. The molecular basis for recognition and cleavage of inosine-containing DNA by prokaryotic EndoV has been elucidated through structure determination of EndoV-DNA complexes [21]. EndoV has an αβα fold with a central 8-stranded β-sheet flanked on either side by α-helices (Figure 3b) — including a ribonuclease H-like motif shared with nucleases such as RNaseH [22 and 23], RuvC [24] and UvrC [25], as well as Piwi protein [26 and 27] and the Piwi subdomain of Argonaute [28 and 29], being part of the RNA-induced silencing complex (RISC).

This is particularly important for tissues with high blood volume as this can make a particularly large contribution to the estimated concentration. In practice, pharmacokinetic modeling is used to relate the contrast agent concentration in the different compartments to underlying physiological

parameters. While such models have been applied to DCE-MRI data of tumors and multiple sclerosis [6], none has modeled exchange to and from the CSF, which may be necessary in more subtle disorders [14]. Statistical modeling has also been employed, but great care is required to ensure that parameters are adequately modeled between tissues. Further work is required to establish whether these complex models can be supported by the data generated from DCE-MRI studies of subtle BBB disorders. It may be that other contrast agents need to be investigated MAPK Inhibitor Library cell line with the aim of increasing the signal enhancement compared to that from gadodiamide, or scanner electronics and gain setting improved to increase the dynamic range of signal capture and reduce the influence of noise and signal discretization error. However, if the ultimate goal is to establish whether differences in concentration profiles are truly reflective of endothelial permeability in subtle disorders, then a quantitative assessment is required and these problems need to be overcome. ABT-199 mw DCE-MRI was performed on a group of mild stroke patients classified

into two groups using the Fazekas

white matter rating scale. No significant differences were found between patients with a high or low white matter rating, although there was a trend towards greater enhancement in patients with a higher degree of white matter abnormality. The effect of noise, scanner drift, intrinsic tissue parameters and imaging sequence parameters on the interpretation of the signal enhancement profiles was assessed. Background noise was found to be comparable in magnitude to the observed differences, while scanner drift had less influence except in the CSF where a progressive rise in signal was observed. Calculation of contrast agent concentration, correcting for systematic differences in intrinsic tissue parameters, noticeably altered the relationship between Ergoloid tissues when compared to signal enhancement measurements, although differences between patient groups remained insignificant. These results suggest that it may be inappropriate to draw conclusions about the amount of contrast agent present in a tissue, and hence it is likely BBB impairment, from signal enhancement data. Therefore, studies of subtle BBB abnormalities should establish the influence of noise, drift and intrinsic tissue parameters on their data before conclusions are drawn. If this is not done, systematic errors introduced by drift and intrinsic tissue parameters may be erroneously perceived as BBB differences between patients.

Thus, the current 16 PAHs measured is a good predictor of PAH presence in the sample, and is representative of the PAH list for which SQGs are generally available. However, this review could not assess whether this subset was a good predictor of overall PAH toxicity, which will be a function of the specific combination, levels and bioavailability of PAHs in a sample. A third workshop recommendation was that EC consider the examination of PCBs based upon the measurement of individual

congeners rather than aroclors, since these compounds provide a better platform of information INCB024360 chemical structure with which to evaluate toxicity and assess bioaccumulation potential. The current DaS aroclor-based LAL could not be used with the congener data reported in the study database for a critical assessment of this recommendation. However, a hypothetical conversion of this SQG to a congener-based one suggested that it was comparable in its outcomes to other DM PCB LALs. We found that most

congener-based PCB SQGs were based upon the ICES7 list of PCBs, and that the sum of these correlated well with total PCBs for a sample reported in the database, though this was biased by the fact that these congeners were by far the most reported ones. Gamma-secretase inhibitor EC will likely wish to give a high priority to the review of the basis for PCB evaluation in the DaS program, and provision of congener-based action levels and guidance similar to that used in other countries. A fourth workshop recommendation was that EC further considers the question of inclusion of a broader range of organic compounds in the assessment. Although the addition of extra pesticides on the CCME list had a minor impact on the conservatism of an assessment protocol when compared to the expansion of a metals list, the addition of the extra pesticides (lindane, aldrin

and HCB) and tTBT in the Consensus SQG list did result in a significant Tideglusib increase in conservatism. Thus, we found that the addition of some organic compounds for which SQGs were available significantly increased the proportion of samples assigned to Tier 2 or Tier 3, though the degree to which this happened depended upon the conservatism of the LAL and UAL levels. It should be noted that, due to the approach applied, this review focused only on organic contaminants for which SQGs were generally available, primarily a set of pesticides, but it is possible that a broader range of organic analytes (assuming data and SQGs were available) would provide similar results.

Table 3 shows the rate of hydrolyzes of angiotensin I, dynorphin1-13, neurotensin1-13 and bradykinin, by the B. jararaca venom. In this set of putative substrates, only bradykinin was not hydrolyzed by the BjV and a good cleavage of angiotensin I was observed. Dynorphin1-13 was also well hydrolyzed by the B. jararaca crude venom, followed by the neurotensin1-13 degradation. Table 3 also shows the cleavage points determined in angiotensin I and dynorphin1-13. As can be observed, Z-VAD-FMK in vivo angiotensin I presents one cleavage point between the residues Tyr–Ile, that was totally blocked

by PMSF and not affected by EDTA or 1,10-phenantroline. Moreover, the commercial serum produced www.selleckchem.com/products/Thiazovivin.html by the Butantan Institute was able to reduce only 44% of the hydrolysis of angiotensin I by BjV. Dynorphin1-13 presents two scissile bonds, between the residues Arg–Arg and Lys–Leu, that were principally blocked by PMSF (88%) and partially blocked by EDTA (28%), and 1,10-phenantroline (6%). Table 3 shows that the antibothropic serum was able to block 48% of the hydrolytic activity of

the venom on dynorphin A cleavage. Since the observation of angiotensin I cleavage is mainly due by serine peptidases and partially blocked by the antivenom, we decided to test the other four bothropic venoms used to make the immunization pool. The results obtained with the venoms used to compose the immunization pool, again showed the presence of a chymotrypsin-like activity in these venoms, although with distinct specific activities (Table 4). The cleavage points were unique between Tyr–Ile bonds (data not shown) and were determined by the internal standardization of the HPLC conditions using the BjV. The blocked effect of the antibothropic

serum was different for each old venom, showing variations in their composition. The angiotensin-I hydrolyzes by the venom from B. jararacussu and B. jararaca were only partially blocked by the commercial antivenom ( Table 4). In contrast, angiotensin I degradation was fully inhibited by using the antivenom when the venoms from B. moojeni and B. neuwiedi were used. Although it was proposed that B. jararaca and Bothrops neuwied should be included in the genus Bothropoides, and B. alternatus into genus Rhinocerophis, there is no clear consensus about the systematics of this group ( SBH, 2007). Since human envenomations involving these species are treated with the antibothropic serum, this study still considers these snake venoms as belonging to the genus Bothrops. The objective of the present study was to analyze the ability of the antivenom produced by the Butantan Institute, São Paulo, Brazil, to neutralize B. jararaca major venom toxins. A set of FRET peptides (Free Ressonance Energy Transfer) was studied using the BjV and site-directed inhibitors PMSF, EDTA and 1,10-phenanthroline.

This assumption also yields marginally conservative values of the deficit-volume which is a desirable feature in the design of water resources systems towards ameliorating the drought conditions. It is worthy to mention that Millan and Yevjevich (1971) developed the regression equations for predicting E(LT) and E(MT) which were also tested for the annual and monthly hydrological droughts using Canadian

river flows. These relationships were found reasonably ERK inhibitor library reliable although at times they tended to under predict in the range of 3–10%. As a note in the context of analysis of monthly droughts, it is prudent to mention that the values of ρ1 in the SHI sequences were low suggesting a weak dependence structure. Therefore, the first order Markov chain model (Markov chain-1, Eq. (8)) was tried to estimate E(LT). It was noted that the predictions of LT tended to be almost the same as predicted by the extreme number theorem. However, at times the predictions by the extreme

number theorem tended to be marginally higher than the Markov chain-1 model and also be nearer to the observed counterparts. This observation vindicates the applicability of the extreme number theorem on monthly as well as annual basis. In fact as the name reads “theorem of extremes of random numbers of random variables” essentially is meant for random sequences, which is evidenced by the results in the present case (annual flows). It has the capability to perform reasonably well in the presence of weak dependence structure and for this reason MS 275 it performed satisfactorily even in monthly streamflow series. It was also observed that when the degree of the first order dependence is remarkable (i.e. ρ1 being above 0.5) then the extreme number theorem breaks down and recourse to the Markov chain models,

among others becomes a necessity. The weekly SHI sequences of rivers with negligible lake effects such as those in Atlantic Canada tended to follow AR-1 process, therefore the extreme number theorem based relationships (Eqs. (1), (2), (3), (4) and (5)) were attempted to model E(LT). In general, such a model resulted in consistent under prediction. As noted earlier, the weekly SHI sequences PI-1840 of rivers riddled with significant lake storages tend to obey AR-2 process or even higher order dependence processes ( Table 2). For such rivers, the extreme number theorem does not hold because of a lack of accountability for the second order dependence. Therefore, a second order Markov chain model (Eq. (7)) was envisaged in which the parameters were computed using the counting method ( Sen, 1990 and Sharma and Panu, 2010). The best estimates of the first order probabilities were obtained using the non-standardized weekly flow series ( Table 2).