0001; Figure Figure1) 1) Of interest, all patients with no trans

0001; Figure Figure1).1). Of interest, all patients with no transfusion of RBC also did not receive any platelet concentrate, meaning total avoidance of allogeneic blood products in 29% of the patients in the fibrinogen-PCC group.Figure 1Platelet concentrate and red blood cell (RBC) transfusion in the emergency room and during surgery. *Platelet concentrate transfusion only reported for www.selleckchem.com/products/BIBW2992.html 371 of 601 patients from the trauma registry of the German Society for Trauma Surgery (TR-DGU). FFP, …For haemostatic therapy in the ER and during surgery, the FFP group received a median of 6 units of FFP (IQR: 4, 10; range: 2, 51), 6 units of RBC (IQR: 4, 11) and 0 units of platelet concentrate (IQR: 0, 2; range: 0, 8).

The patients in the fibrinogen-PCC group received a median of 6 g of fibrinogen concentrate (IQR: 3, 9; range: 0, 15) and 1,200 IU of PCC (IQR: 0, 2,400; range: 0, 6,600), while RBC median transfusion was 5.5 units (IQR: 0, 9.5) and platelet concentrate median transfusion was 0 units (IQR: 0, 0; range: 0, 2). The dosage of FFP, fibrinogen and PCC is described in Figure Figure2.2. During the first 48 hours after admission to the ICU, patients in the FFP group received median doses of 3 units of RBC (IQR: 1, 6; range: 0, 80; data reported for 424 patients) and 3 units of FFP (IQR: 0, 6; range: 0, 80; data reported for 405 patients). No information is available on platelet concentrate transfusion in this group during the stay on the ICU. For the fibrinogen-PCC group, a complete set of transfusion data was available.

During their stay at the ICU, these patients received a median dose of 2 units of RBC (IQR: 0, 3; range: 0, 11), platelet concentrate was transfused in 9% of the patients during this time (the dose was 1 or 2 units). The patients also received a median dose of 6 g of fibrinogen concentrate (IQR: 3, 10; range: 0, 22) and a median of 1,200 U of PCC (IQR: 0, 2,400; range: 0, 9,000).Figure 2Percentage of patients receiving the indicated amount of haemostatic agent (FFP, fibrinogen concentrate, PCC) in the emergency room and during surgery. Percentage of patients in brackets. FFP, fresh frozen plasma; PCC, prothrombin complex concentrate. …For haemostatic therapy in the ER and during surgery, the median ratio of FFP:RBC in the FFP group was 1 (IQR: 0.7, 1.3; minimum 0.1, maximum 6.5); Figure Figure33 shows the ratios for all patients in the group.

The same median value was observed in each of the four years Batimastat included in the analysis (2005 to 2008). In the fibrinogen-PCC group, the median ratio of fibrinogen concentrate:RBC for coagulation therapy in the ER and during surgery was 0.9 (IQR: 0.7, 1.2), and the ratio of PCC (in hundreds of units):RBC was 1.6 (IQR: 0, 3); Figure Figure33 shows the distributions of fibrinogen concentrate:RBC and PCC:RBC ratios.

Isl1 and CERN-922 immunoreactivities progressed in parallel follo

Isl1 and CERN-922 immunoreactivities progressed in parallel following a central-to-peripheral gradient, but Isl1 immunoreactivity extended to more peripheral regions (Figures 5(d)�C5(g)).Figure 5Morphological features and expression patterns of Isl1 and other cell differentiation markers in the St37/38 Xenopus laevis retina. (a) Toluidine blue-stained transverse both retinal resin sections showed that the plexiform layers were observed across the …At St40, the typical multilayered structure of the retina was clearly distinguishable. However, the GCL was many cells thick, and the plexiform layers were poorly developed (Figure 6(a)). A proliferative population of undifferentiated cells remained at the ciliary margin zone (CMZ), providing a source of new cells for the growing retina (Figure 6(a)).

At this stage, the expression patterns for Isl1 (Figures 6(b), 6(e), and 6(f)), SV2 (Figure 6(c)), and CERN-922 (Figures 6(d) and 6(f)) followed a central-to-peripheral gradient. The number of Isl1 immunoreactive elements and the immunostaining intensity increased markedly in the INL and began to resemble those observed in the St53 retina (Figure 2). The patterns of distribution of Isl1 positive cells during X. laevis retinal development are summarized in Figure 7.Figure 6Morphological features and expression patterns of Isl1 and other cell differentiation markers in the St40 Xenopus laevis retina. (a) Toluidine blue-stained transverse retinal resin sections revealed that the typical multilayered structure was clearly …Figure 7Schematic drawings of the Isl1 positive cells in the developing retina of Xenopus laevis.

Black nuclei represent Isl1-immunostained cells. (a) Isl1 immunoreactivity during St29�C31 was restricted to nuclei located near the vitreal surface of the …4. DiscussionRecent studies conducted in our laboratory confirmed the presence of the LIM-domain transcription factor Isl1 in differentiating and mature ganglion, amacrine, bipolar, and horizontal cells in the retina of mammals [54], birds [18], reptiles [12], and fish [9�C11]. In the present study, we extend our observations to the retina of the anuran Xenopus laevis, an amphibian model extensively used in studies of retinal cell organization [44, 55�C57]. We first established the precise cell types that showed immunoreactivity for Isl1 in the differentiated retina, and subsequently analyzed the Isl1 immunoreactivity in the corresponding cells in the retina throughout development.

The Isl1 spatiotemporal expression pattern spans the differentiation of four neuronal classes��ganglion, amacrine, bipolar, and horizontal cells��in the developing X. laevis retina.4.1. Isl1 Expression in the Nonlayered Retina of X. laevis Dacomitinib Between St29 and 30, the first detectable Isl1 expression, and St 35-36, the plexiform layers were not recognizable in the developing X.

However, because usefulness of ECLS in this setting remains debat

However, because usefulness of ECLS in this setting remains debatable [20], we believe this study adds important information neither about ECLS as a rescue therapy in patients with drug-induced cardiac arrest and refractory shock.ConclusionsBased on our experience, we consider ECLS as a last resort, efficient, and relatively safe therapeutic option in critically ill poisoned patients (i.e. cardiac arrest and refractory shock) who do not respond to conventional therapies, providing the cardiac surgeon with the means to rapidly intervene and control ECLS-related complications. However, because there is insufficient evidence concerning the use of ECLS as a treatment for severe cardiac impairment due to poisoning, further studies are needed to clarify criteria for unresponsiveness to conventional treatment and the indications of ECLS in this setting.

Key messages? Cardiovascular failure is the leading cause of death following a cardiotoxic drug overdose.? This report supports the hypothesis that ECLS may be considered as a last resort, efficient, and relatively safe therapeutic option in critically ill poisoned patients (i.e. cardiac arrest and refractory shock) who do not respond to conventional therapies.AbbreviationsACT: activated clotting time; CPC: cerebral performance class; ECLS: extra corporeal life support; MSA: membrane stabilizing activity; SAPS: Simplified Acute Physiology Score; SOFA: Sequential Organ Failure Assessment.Competing interestsThe authors declare that they have no competing interests.Authors’ contributionsCD and MB initiated the study, and the design.

CD and PC were involved in the interpretation of the results. CD wrote the manuscript, and PC helped to draft the manuscript. PL, CI, MT, OL, MB, CQ, MM, and PC contributed to the conception of the study and revision of the manuscript. All authors read and approved the final manuscript.Authors’ informationThe work has been presented in part at the annual congress of the Soci��t�� de GSK-3 R��animation de Langue Fran?aise (SRLF) held in January 2008, Paris, France.NotesSee related commentary by Ashrafian and Athanasiou, http://ccforum.com/content/13/5/187AcknowledgementsWe are indebted to Dr Jean-Jacques Parienti from Department of Biostatistics and Clinical Research, Caen University Hospital, France, for his critical review of the manuscript. We thank Madame Val��rie Valfong for her contribution to polishing the manuscript.
Sepsis affects 750,000 patients per year in the USA, killing 250,000 of these people. In the UK severe sepsis has a mortality rate of 45% [1-3] and despite putative therapeutic options including early goal-directed therapy [4] and activated protein C [5], outcome in septic patients has not vastly improved.

Two recent studies reporting adverse effects

Two recent studies reporting adverse effects selleck chemical Axitinib of catecholamine vasopressors on organ function [23] and mortality [24] in septic shock support our results. Furthermore, the findings of the present analysis are in line with earlier data suggesting harmful effects of excess catecholamine exposition in general critically ill patient populations. For example, Boldt and colleagues showed that circulating plasma levels of catecholamines were higher in non-surviving when compared with surviving surgical intensive care unit patients [25]. A randomized trial that investigated the outcome effects of supranormal oxygen delivery in a diverse group of critically ill patients reported higher in-hospital mortality in patients receiving liberal catecholamine therapy than in control patients exposed to standard care [26].

Important limitations must be considered when interpreting the results of this study. First, and probably most importantly, mean values of punctually instead of continuously recorded MAPs were analysed. Thus, the true course of MAP may have been under- or over-estimated. Additionally, it is possible that some patients changed between MAP quartiles during the shock period but were eventually grouped into one quartile based on their average MAP. Second, as the original study was performed in the late 1990s the definition of some disease-related events does not correspond to current recommendations. This is particularly relevant for the definition of renal failure [27] and disseminated intravascular coagulation [28], which has recently been newly defined based on international consensus.

Furthermore, the occurrence of disease-related events was documented during the intensive care unit stay after study randomization. Although more than half of non-surviving study patients did not achieve shock resolution and developed disease-related events during the evaluated shock period, it is possible that some disease-related events occurred either after shock resolution or during a renewed shock episode during which MAP and the mean vasopressor load were not evaluated. When drawing clinical conclusions from our results caution is warranted because the MAP quartiles analysed were retrospectively defined and can not be considered as treatment goals. Finally, it must be considered that this post hoc analysis was performed in an uncontrolled patient cohort, and its results must not be considered to have the same validity as those of a randomized, controlled trial.

ConclusionsMAP GSK-3 levels of 70 mmHg or higher do not appear to be associated with improved survival in septic shock. However, augmenting vasopressor dosages to elevate MAP to more than 70 mmHg may increase mortality. Future trials are needed to identify the lowest acceptable MAP level to ensure tissue perfusion and avoid unnecessary high catecholamine infusions.

Ho and colleagues predicted 1 to 1 5 units FFP to each RBC to pre

Ho and colleagues predicted 1 to 1.5 units FFP to each RBC to prevent dilutional coagulopathy in mathematical models [55].Since 2007, growing numbers of retrospective Brefeldin A clinical military and civilian papers have studied early formula-driven haemostatic resuscitation with different FFP:RBC ratios (mostly near 1:1) and mortality [11-20,56,57]. Overall, these studies demonstrate a significant association between higher ratios and lower mortality in massive traumatic bleedings, with absolute mortality reductions ranging between 15 and 62% [11-20]. These figures surpass any predictions of potentially preventable deaths in trauma [47]. While the survival advantage of early and aggressive FFP transfusion in early formula-driven resuscitation cannot be ignored, the evidence behind it has limitations that are discussed next.

Survival advantageBorgman and colleagues reviewed 246 massively transfused (��10 units RBC/24 hours) combatants and analysed mortality at three different FFP:RBC ratios (1:8, 1:2.5 and 1:1.4) [11]. A 55% absolute reduction in mortality occurred between the highest and lowest ratios. While mortality reduction was impressive, patients with a higher FFP:RBC ratio (1:1.4) had a longer median time to death (38 hours) than those with a lower ratio (2 hours). These data suggest that lower ratio patients may not have lived long enough to receive FFP. Another study by the same group on civilian trauma patients reported a similarly impressive survival advantage for higher ratios than lower ratios, but also a markedly dissimilar time to death (35 hours versus 4 hours) [58].

Both studies disclose survivorship bias, where arguably patients had to survive long enough to receive FFP, thus questioning their conclusions.Addressing survivorship biasTwo studies specifically addressed the survivorship bias in high-FFP:RBC studies. Scalea and colleagues used stepwise logistic regression analysis on 806 patients, demonstrating no survival benefit for higher ratios when early deaths were excluded [56]. This study has its own limitations, however, including a failure to report the time to intensive care unit admission, an inability to include major factors (acidosis and coagulation) in the statistical model and a surprisingly low mortality (6%) for massive transfusions. Snyder and colleagues also attemp-ted to correct for survivorship bias in another study where mortality in high (>1:2) and low (<1:2) ratios was compared in regression models [57]. Using the FFP:PRBC ratio as a fixed value at 24 hours, as in many studies on this topic, the high ratio resulted in better survival. This survival advantage was lost, however, when the ratio was treated as a time-dependent Brefeldin_A variable (relative risk = 0.84, 95% confidence interval = 0.47 to 1.5).

An adequate learning curve is important in order to minimize comp

An adequate learning curve is important in order to minimize complications. The surgeon should also be confident about the instrumentation to reduce the duration of surgery and radiation exposure. The major complications primarily occur in the immediate postoperative period and can be related both to the implant and to the surgical procedure. The correct surgical indication selleck chemical Dasatinib remains mandatory. Patients should be informed about the potential complications and the possible need for instrumentation removal.
Morbid obesity is one of the major health problems of the 21st century. Formally recognized by the WHO as a global epidemic in 1997, it was estimated that in 2008, 1.5 billion adults, 20 and older, were overweight. Of these, over 200 million men and nearly 300 million women were obese, with higher rates among women than men.

The rate of obesity also increases with age at least up to 50 or 60 years old and severe obesity in the United States, Australia, and Canada is increasing faster than the overall rate of obesity. Once considered a problem only of high-income countries, obesity rates are rising worldwide and affecting both the developed and developing world. These increases have been felt most dramatically in urban settings [1]. Concurrently research on factors regulating obesity as well as possible treatments has been ongoing, with bariatric surgery making the greatest leaps and providing the means for better understanding of the metabolic and endocrine parameters involved in weight gain and weight loss [2, 3].

As the most effective means for excess weight loss available, bariatric surgery has been growing continuously, with more and more patients opting for surgical treatment of their condition, new operations and techniques being developed, and new instruments being produced. The advantages of minimally invasive surgery have been instrumental for this growth [4]. Many operations have been devised, with the Roux en Y gastric bypass being the most effective as far as excess weight loss is concerned, and sleeve gastrectomy being preferred by a growing number of surgeons due to its simplicity, effectiveness, and low rate of complications. In 2006, a new technique was presented, initially named total vertical gastric plication, better known today as laparoscopic greater curvature plication (Evidence Level III) [5].

Developed in Iran by Dr Talebpour as a cheap alternative to Laparoscopic Sleeve Gastrectomy, it appears to be gaining ground as its theoretical advantages of technical simplicity and low complication rate are of major importance Anacetrapib to the growing industry that Bariatric Surgery has become, as well as to the industry of Bariatric Tourism. 2. Aim Laparoscopic Greater Curvature Plication (LGCP) or Gastric Plication is a relatively new technique. Gastric Plication was initially proposed by Wilkinson and Peloso [6] in 1981 and introduced in 2006 by Dr Talebpour in Iran [5].

In 2010, another study performed by O’Malley Jr and Weinstein as

In 2010, another study performed by O’Malley Jr. and Weinstein assessed the outcomes of 10 patients undergoing parapharyngeal space resection using the TORS approach. The surgery was performed in 9 of the 10 patients, with acceptable operative time and blood loss, and no significant complications such as hemorrhage, infection, trismus or tumor spillage. One patient was converted to an Enzalutamide pancreatic cancer open transcervical approach due to difficulties found during resection and to avoid the risk of tumor spillage. In 7 patients that had resection of a parapharyngeal space pleomorphic adenoma, local control was obtained in all 7 patients, although tumor spillage was reported in one patient. The TORS approach was found to offer reduced complication rates when compared to the transcervical approach [79, 80].

Another approach to the infratemporal fossa was developed by McCool et al. [81], in which 6 complete and 2 partial resections were performed using a suprahyoid port, while the other arms were placed transorally. In another report, Hanna et al. [82] obtained excellent access to the anterior and central skull base in cadavers, including the cribriform plate, fovea ethmoidalis, medial orbits, planum sphenoidale, nasopharynx, pterygopalatine fossa, and clivus. In addition, sella turcica and suprasellar and parasellar access was achieved using the robotic arms. However, there is a continuing need for further development of appropriate instruments, in terms of size, flexibility, and function. 7.6.

Pediatric Surgery Although there are studies of robotic surgery thyroidectomy in children [47, 48], which we have discussed previously, studies of robotic surgery in the pediatric population are sparse. To date, the only pediatric case series is that described by Rahbar et al. [83] in 2007 at Children’s Hospital Boston. In this study, 4 pediatric cadaver larynxes were used to assess precision and tissue handling using a robotic-system. 5 living patients were enrolled to undergo a laryngeal cleft repair. Equipment size was the main limiting factor for these procedures, resulting in limited transoral access in 3 of 5 the patients. The other 2 patients, who had type 1 and type 2 laryngeal clefts, had successful surgical repairs using the robotic system. 8. Conclusion The trend towards the use of minimally invasive surgery has had an impact on the way new technology is thought of, developed, and incorporated into clinical practice.

Robotic surgery is continuing to advance, and is overcoming its limitations. It is improving the outcomes, such as reducing hospital stays and infection rates, and allowing for better cosmetic results. However, surgical robots were developed to perform procedures in spacious cavities, such as the abdomen, and thus, the instruments are over sized to perform many of Dacomitinib the otolaryngology and head and neck procedures.

2 1 Data Analysis SPSS version 11 0 was used for statistical ana

2.1. Data Analysis SPSS version 11.0 was used for statistical analysis. The prevalence of underweight, stunting, and wasting were calculated for the different age groups and both sexes selleck screening library and the severity classified based on z scores. Differences in the proportions of wasting, stunting, and underweight among boys and girls and at various ages were tested with the chi-square test. Correlation between CD4% and growth indices was obtained using Pearson’s correlation coefficient. Receiver Operating Characteristic curves were constructed to assess the relationship between HAZ and WAZ with CD4% and to determine the cutoff which would predict immune deficiency with optimal sensitivity and specificity. 3. Results A total of two hundred and thirty one antiretroviral-na?ve HIV-infected children were enrolled during the period under study.

The average age of the children at presentation was approximately 71 months with 17% under 3 years of age. 42% were boys and majority of the children were in WHO clinical stage 3. The mean CD4 percentage was 17.7 �� 10 (SD)% and the average BMI was 14.2 �� 2 (Table 1). Table 1 Demographic profile of the study population. In this cohort, the prevalence of underweight (WAZ < ?2) was 63%, stunting (HAZ < ?2) 58%, and wasting (WHZ < ?2) 16%, respectively, (Table 2). While a higher proportion of boys were underweight compared to girls, the rates of stunting and wasting were similar. Figure 1 shows the proportion of underweight and stunted children in the various age categories.

Children below 3 years of age and above 10 years of age were at a significantly higher risk of severe underweight when compared with children between the ages 3�C10 years but the proportion of stunted children was similar across all age groups. Severe stunting was found in >40% of children at all age groups. The proportion of children with normal nutritional status (WAZ and HAZ > ?1) tended to decrease with increasing age. Table 3 shows the mean CD4% and CD4 cell count of all children as well as in the subsets with either underweight or stunting, with a clear trend towards lower cell count and percentage among children above the age of 10 years in all three groups. Figure 1 Proportion of children with underweight (WAZ < ?2 SD) and stunting (HAZ < ?2 SD) in different age groups. *P < .05 versus 3�C5 and 5�C10 years age group.

Table 2 Gender wise prevalence of malnutrition among HIV-infected Children. Table 3 Mean CD4% and CD4 cell count of children in different age groups as well as in those with underweight and stunting. We examined the relationship between CD4 percentage and the various growth parameters. CD4% was available for 194 children with 41% of them showing severe immunodeficiency (CD4 <15%) at the time of initial presentation. Of these children with CD4 <15%, 71% were stunted (HAZ < ?2) Batimastat and 76% were underweight (WAZ < ?2).

However, they later abandoned this technique, given the lack of d

However, they later abandoned this technique, given the lack of difference compared with their ��standard�� approaches. Recently in 2012, kinase inhibitor Tofacitinib Gao et al. reported successful robotically assisted mitral valve replacement with excellent results [33]. Figure 2 Heart Lung machine with peripheral cannulation via the femoral vessels. Figure 3 Direct transthoracic aortic clamping. 3. Results During the past 16 years, cardiac surgeons worldwide have reported their MIMVS data with promising results. The majority of these results suggest that MIMVS provide excellent, safe, and familiar exposure of the mitral valve with results comparable to those with conventional approaches. Unfortunately we lack data from large prospective randomized control series comparing the results of minimally invasive versus the conventional sternotomy technique.

We therefore have to rely on retrospective analysed registry data (mostly single centre). 4. Mortality Reviewing all comparative MIMVS studies evaluating mortality, no study has shown a significant difference between minimally invasive and conventional approaches [34�C40]. In 2003, Greelish et al. [20] reported the first long-term results (5-year followup) of mini-VS, indicating a freedom from mitral regurgitation and reoperation >90%. In their early port access cases, Mohr et al. [23] reported a high mortality rate (9.8%) for mini-MVS, partially procedure related, with 2 of 51 patients experiencing an aortic dissection [23]. After discontinuing the port access technique and modification and simplification of the surgical procedure, the mortality decreased to an in-hospital mortality rate of 3.

9% [41]. The Leipzig long-term results revealed an actuarial survival rate of 83% at 6.8 years [42]. When excluding the initial 200 patients in whom an endoclamp was used, the overall results are even more impressive [42]. In 2002, Mohr’s group (Onnasch et al. [36]) reported their 5-year experience performing mini-MVS in 449 patients, with a mean survival rate of 96.3% at 2-year followup. The East Carolina University group reported a combined series with Hargrove consisting of 1178 successful video-assisted mitral valve operations between 1996 and 2008 [43]. The operative mortality rates for mitral valve repair and replacement for this two center series were 2.1% and 4.6%, respectively, but only 0.2% for isolated primary mitral valve repair.

A recent meta-analysis by Modi et al. [44] identified ten papers published between 1998 and 2005 which were suitable for analysis. This study included 1358 minimally invasive patients and 1469 sternotomy patients. Although cross-clamp and cardiopulmonary bypass time were longer in the minimally invasive group, there were no difference Anacetrapib in mortality, stroke, reoperation for bleeding, new onset atrial fibrillation, or duration of ICU stay or hospital stay [44]. In a more recent study, Stevens et al.

Methods DNA constructs Constitutively active Notch constructs wer

Methods DNA constructs Constitutively active Notch constructs were made with cDNA encoding either membrane tethered, Drosophila selleck chem Notch, constitutively activated by the removal of the extracellular domain, or the soluble intracellular domain. These truncated Notch constructs were cloned into the pIZ V5 His expression vector producing non tagged proteins. The control expression plasmid was constructed by cloning firefly luciferase into the same pIZ V5 His expression vector. The luciferase reporter construct con tains a 1. 4 kb tandem duplication of E m3 upstream regulatory sequences, cloned into a pGL2 Basic vector, as described. Genome wide RNAi method A total of 23,560 dsRNAs, made available from the Dro sophila RNAi Screening Center at Harvard Medical School, were screened by the following method, Kc167 cells were washed three times and resuspended in serum free Sangs M3 medium at a concentration of 5 �� 105 cells ml.

Using a robotic liquid handler, 104 cells were uniformly dispensed into the wells of 384 well polypropylene plates containing dsRNA and incubated for 45 min at room temperature. An equal volume of M3 medium containing 10% fetal bovine serum was added and incubated for four days. On day four, the RNAi treated cells were diluted with 100 ul of medium, mixed and 20 ul were dispensed into the wells of six new 384 well plates, pre aliquoted with 20 ul of transfection mix. The six plates contained the three different transfection mixes, each in duplicate. Transfection mixes were prepared with Effectene Trans fection Reagent, following the manufacturers guidelines.

Luciferase activity was measured 24 h post transfection using the Steady Glo Luciferase Assay Sys tem. This method requires only two plasmids to be transfected at one time and gave acceptable signal to noise ratios for high throughput screening in 384 well plate format. Whereas, the conven tional renilla dual glo assay was not robust enough to scale to 384 well format with this Notch reporter sys tem using an endogenous target. In contrast to path ways with soluble ligands, the reporter and constitutively active Notch constructs are required to transfect the same cell to activate transcription. With the renilla dual glo system, adding the control con struct required the co transfection of three individual plasmids and this reduced the signal to noise ratio to insufficient levels.

Data analysis Duplicate measurements for each of the three signals were averaged, Notch specific E m3 promoter in the presence of activated Notch, the E m3 promoter alone and the unrelated viral promoter OplE2. The Necn m3 luc signal was Dacomitinib normalized two different ways, by either the m3 luc or con luc signals. The z scores of the log2 ratios were calculated by using the standard deviation and mean of the measurements that corresponded to the 96 wells of the original dsRNA stock plates.