All of these events were monitored by an independent, unblinded Data and Safety Monitoring Board (DSMB) that met approximately twice a year during the course of the study. In addition, Bangladesh required additional monitoring by a local DSMB. The common protocol surveillance system was designed to capture severe GE occurring among participants upon presentation to medical facilities in the GSK1210151A study areas. Infants who underwent randomization were visited at least monthly to remind parents to bring their child to a clinic or hospital if they developed symptoms

of gastroenteritis [4] and [5]. GE was defined as three or more watery or looser-than-normal stools within a 24-h period and/or forceful vomiting [7]. Upon presentation to a medical facility, stool samples CCI-779 purchase were collected; history of symptoms of the current illness was collected through interview with the parent/guardian; and physical signs were documented by medical staff caring for the subject via direct observation. Data on ongoing symptoms and signs were collected throughout the course of the episode. These data were used to define severity using the 20-point modified Vesikari Clinical Scoring System

(VCSS) (“severe” was defined as a score of ≥11) [8], [10] and [11]. For this analysis, we also looked at a score of ≥15 and ≥19, indicating “very Ketanserin severe” or “extremely severe” GE. Rotavirus antigens in stool specimens were detected by enzyme immunoassay (EIA) [12]. Wild-type rotavirus was confirmed by reverse-transcriptase-polymerase-chain-reaction (RT-PCR) for identification of the VP6 genotype. Identification of rotavirus P and G genotypes was performed by RT-PCR as previously described [13]. EIA assays were conducted in the laboratory of Dr. Richard Ward at Children’s Hospital Medical Center, Cincinnati, OH; RT-PCR assays were conducted at Merck Research Laboratories. Statistical analysis. Efficacy was defined as 1–(Rvaccine/Rplacebo) × 100%, where R represented the incidence for the respective groups. It was assumed that the

number of cases in each group followed a Poisson distribution; the statistical analysis then conditioned on the total number of subjects with severe gastroenteritis from both treatment groups, such that the number of subjects with severe gastroenteritis in the vaccine group followed a binomial distribution. For subjects with multiple episodes, only the most severe episode (identified by the VCSS) was used for analysis. For efficacy calculations, we counted cases starting 2 weeks after receipt of third dose of vaccine (per-protocol definition). We also calculated efficacy by specific serotype of rotavirus according to the same methods. Exact inference was used, and follow-up time was accounted for in the calculations.

Recently, Shewell et al. demonstrated that deletion of the glycosylated immunodominant C-terminus of AniA produced a truncated protein that elicited antibodies that inhibited nitrite reductase activity [69]. Vaccine-mediated inhibition of AniA function may be an effective approach because the capacity to grow anaerobically is likely an important adaptation during infection of the genital tract where oxygen tension is reduced. This hypothesis is supported by the detection of AniA-specific antibodies from women with lower or upper genital tract

infections and one patient with DGI [70]. AniA is also required for mature biofilm formation, which may protect against innate defenses GS-7340 mouse [71]. The exciting development of group B meningococcal vaccines, which was a formidible challenge for many years, may provide a useful template for developing a gonorrhea vaccine [72], [73] and [74]. Some of these vaccines contain outer membrane vesicles (OMV) and some are genetically engineered to stabilize the expression

of phase variable antigens and increase the range of antigenic specificities. Detergent-treated OMVs or OMVs produced from LOS mutants have been used to diminish endotoxicity. Immunization and challenge studies with Gc OMV have not been reported; a Gc outer membrane protein preparation demonstrated protection in mice when delivered intranasally 3-Methyladenine mw with CT [54], but this approach was not successful in subsequent studies, possibly due to differences in the protein isolation methods used [35]. The Novartis 4CmenB vaccine consists of OMVs combined with the NadA protein and two fusion proteins, factor H-binding

protein (fHbp) and neisserial heparin binding antigen (NHBA) fused to two other conserved antigens [74]. None of the three proteins (fHBP, NHBA and NadA) in the 4CmenB vaccine [74] are predicted to be suitable vaccine targets for Gc [75]; however, gonorrhea research may benefit from the use of proteomics technology and, or genome mining, which have advanced much the development of vaccines for group B N. meningitidis. Immunization of the genital tract also challenges gonorrhea vaccine development, although we are encouraged by the success of the HPV vaccine. Most efforts to develop a vaccine against gonorrhea have focused on conventional parenteral immunization, which generates circulating, predominantly IgG antibodies, but is generally ineffective at inducing secretory (S) IgA at mucosal surfaces. However, the genital tract secretions of both males and females contain more IgG derived largely from the circulation than SIgA produced locally and transported through epithelial cells [57].

% Yield: 69%, m.p: 182 °C, IR: (KBr in cm−1): 3337 (N–H str), 2944 (C–H str), 2130 (C–N str), 1661 (C O str), 826 (C–Cl str); 1H NMR: (DMSO d6): (δ, ppm) 2.65 (t, 2H, CH2), 2.49 (t, 2H, CH2), 2.21 (t, 2H, CH2), 3.5 (t, 2H, CH2), 3.6 (t, 2H, CH2), 7.6 (d, 1H, ArCH), 8.21 (d, 1H, ArCH), 8.67 (d, 1H, ArC); MS: (m/z: RA%): 443 (M+, 70%); 445 (M+2, 25%); Elemental analysis: Calculated for C18H17ClN8O2S; C, (48.59%); H, (3.85); N, (25.19%); R428 concentration found: C, (47.12%); H, (3.00%),

N, (25.16%). %Yield: 60%, m.p: 205 °C, IR: (KBr in cm−1): 3344 (N–H str), 2986 (C–H str), 2145 (C–N str), 1667 (C O str), 768 (C–Cl str); 1H NMR: (DMSOd6): (δ, ppm):δ 2.56 (t, 2H, CH2), 2.98 (t, 2H, CH2), 2.84 (t, 2H, CH2), 3.15 (t, 2H, CH2), 3.47 (t, 2H, CH2), 7.76 (d, 1H, ArCH), 8.59 (d, 1H, ArCH), 8.42 (d, 1H, ArCH); MS: (m/z: RA%): 443 (M+,60%); 445 (M+2,20%); Elemental analysis: Calculated for C18H17ClN8O2S; C, (48.59%), H, (3.85%), N, (25.19%); found: C, (48.37%), H, (3.56%), N, (25.12%). %Yield: 58%, m.p: 275 °C, IR: (KBr in cm1): 3319 (N–H str), 2978 (C–H str), 2190 (C–N str), 1619 (C O str); selleck chemicals llc 1H NMR: (DMSO d6): (δ, ppm) 2.32 (t, 2H, CH2), 2.32 (t, 2H, CH2), 2.66 (t, 2H, CH2), 3.1 (t, 2H, CH2), 3.79 (t, 2H, CH2), 7.79 (d, 1H, ArCH), 8.41 (d, 1H, ArCH), 8.76 (d, 1H, ArCH); MS: (m/z: RA%): 440 (M+,40%); Elemental analysis: Calculated for C19H20N8O3S; C, (51.81%), H, (4.58%), N, (25.44%); found: C, (51.77%), H, (3.54%), N, (25.32%). %Yield: 56%, m.p: 269 °C, IR: (KBr

in cm−1): 3496 (N–H str), 2998 (C–H str), 2306 (C–N str), 1686 (C O str); 1H NMR: (DMSO d6): (δ, ppm) 2.56 (t, 2H, CH2), 2.87 (t, 2H, CH2), 2.61 (t, 2H, CH) 3.23 (t, 2H, CH2), 3.81 (t, 2H, CH2), 7.68 (d, 1H, ArCH), 8.86 (d, 1H, ArCH), 8.19 (d, 1H, ArCH), M: (m/z: RA%): 440 (M+,70%); Elemental analysis: Calculated for C19H20N8O3S; C, (51.81%), H, (4.58%), N, (25.44%); found: C, (51.67%), H, (4.55%), N, (25.34%). %Yield: 60%, m.p: Dipeptidyl peptidase 260 °C, IR: (KBr in cm−1): 3412 (N–H str), 2918 (C–H str), 2394 (C–N str), 1619 (C O str); 1H NMR: (DMSO d6): (δ, ppm) 2.12 (t, 2H, CH2), 2.36 (t, 2H, CH2), 2.48 (t, 2H, CH2), 3.54 (t, 2H, CH2), 3.15 (t, 2H, CH2), 7.20 (d, 1H, ArCH), 8.40 (d, 1H, ArCH), 8.45 (d, 1H, ArCH); MS: (m/z: RA%): 440 (M+,60%); Elemental analysis: Calculated for C19H2N8O3S; C, (51.81%), H, (4.58%), N, (25.44%); found: C, (50.87%), H, (4.21%), N, (25.39%). % Yield: 62%, m.p: 265 °C, IR: (KBr in cm−1): 3417 (N–H str), 2935 (C–H str), 2305 (C–N str), 1624 (C O str), 1530 (N–O str); 1H NMR: (DMSO d6): (δ, ppm) 2.31 (t, 2H, CH2), 2.26 (t, 2H, CH2), 2.39 (t, 2H, CH2), 3.25 (t, 2H, CH2), 3.56 (t, 2H, CH2),7.

The filtrate was used for the preliminary phytochemical analysis. The tests were performed according to methods described by Khandelwal (1998) and Kokate (2007). 12 and 13 TLC for various phytoconstituents was carried out as per methods described by Wagner and Bladt (1996).14 Albino Wistar rats, 8–12 weeks old, weighing in range of 120–180 g, was procured from Haffkine Institute, Parel. The animals were accommodated selleck in groups of five in polypropylene cages with stainless steel grill

top and a bedding of clean paddy husk was provided. The animals were maintained in air conditioned room with controlled temperature maintained in the range of 22–25 °C and alternating 12 h periods of light and dark cycle. The relative humidity was close to 60%. The animals were acclimatized to standard laboratory conditions prior to experimentation. The guidelines issued by Institutional Animal Ethics Committee of Ramnarain Ruia College, Mumbai, India with CPC SEA registration selleck chemicals no. CPC SEA/315, regarding the maintenance and dissection of small animals were strictly followed. Rats were administered a dose of 2000 mg/kg body weight for 14 days and were then examined for any signs of behavioural changes and mortality. All experiments were performed on female Albino Wistar rats (200–250 g)

obtained from the Haffkine Institute, Parel, Mumbai, Maharashtra, India. The animals were accommodated in groups of six in polypropylene

cages with stainless steel grill top and a bedding of clean paddy husk. Animals were maintained under a constant 12-h period of light and dark cycle and an environmental temperature of 22–25 °C. The Dichloromethane dehalogenase animals were acclimatized for 15 days before being used for the experiments. The guidelines issued by Institutional Animal Ethics Committee of Ramnarain Ruia College, Mumbai, India with CPC SEA registration no. CPC SEA/315, regarding the maintenance and dissection of small animals were strictly followed. The animals were fed on the standard pellet diet (Amrut Feed, Pune) and water was given ad libitum. The overnight fasted rats were made diabetic with streptozotocin (STZ) (Sigma, St Louis, MO; 60 mg/kg; intraperitoneally). The STZ was prepared freshly by dissolving it in Na-citrate buffer (0.01 M, pH 4.5) and maintained on ice prior to use; the injection volume was 0.2 ml. Diabetes was confirmed in the rats by measuring the fasting blood glucose concentration after 72 h of STZ administration. The rats with glucose level above 300 mg/dl were considered to be diabetic and were used in the experiment. Animals had free access to food and water after the STZ injection.

This can cause a bias toward the null, diluting an existing risk see more because of inclusion of cases that were not exposed during embryogenesis. However, in August of 2013, Andersen et al9 from Denmark presented a second study using the same Danish registries covering more years (1997-2010) and more pregnant women (897,018 vs 608, 835). In contrast to Pasternak et al,8 Andersen’s study detected a 2-fold increased risk of cardiac malformations with ondansetron (odds ratio [OR], 2.0; 95% confidence interval [CI], 1.3–3.1),

leading to an overall 30% increased risk of major congenital malformations. To rule out confounding by indication, Andersen et al9 also examined metoclopramide taken for morning sickness, detecting no increase in teratogenic risk. The fact that the same large registry can be investigated to yield such opposing results is concerning. There

is an exponential rise in use of prescription database linkage to birth registries. None of these were designed specifically to address fetal drug safety, and there may be flaws in the quality and completeness of the available data. Of potential importance, a recent large case control study by the Sloan epidemiology unit and the Centers of Disease Control and Prevention, has reported a 2-fold increased risk for cleft palate associated with ondansetron taken for NVP Regorafenib in the first trimester of pregnancy

(OR, 2.37; 95% CI, 1.28–4.76).10 The maternal safety of ondansetron has been challenged in June 2012, when the FDA issued a warning of possible serious cardiac output (QT) prolongation and Torsade the Pointe among people receiving ondansetron. 11 As a result, the FDA requires strict workup of patients receiving ondansetron, to rule out long QT, electrolyte imbalance, congestive heart failure or taking concomitant medications that prolong the QT interval. 12 Because this drug is not approved by the FDA for pregnant women, the FDA did not specifically address precautions in pregnancy. However, in the context of NVP, women with severe NVP often exhibit electrolyte abnormalities (hypokalenia or hypomagnesemia). Idoxuridine Presently, counseling of women who receive ondansetron for morning sickness suggests that these FDA precautions are not being followed. Serotonin syndrome is a life-threatening disorder of excessive serotonergic activity, typically occurring when 2 or more serotonin-modifying agents are used simultaneously, although it may also occur with a single agent.12 From Jan. 1, 1998, to Dec. 30, 2002, Health Canada received 53 reports of suspected serotonin syndrome, most often reported with the use of selective serotonin reuptake inhibitors (SSRIs), monoamine oxidase inhibitors and selective serotonin- norepinephrine reuptake inhibitors.

Where comparison was possible, the results of the current study where relatively high: 4–12% higher than those of De Smet et al (2001) who allowed only one attempt with each hand, and 8–14% higher than those of Molenaar et al (2010) where three attempts were allowed.

The study by Butterfield et al (2009) reported 4% lower to 6% higher scores. Besides differences in methods, the higher results may be a consequence of the ongoing trend in the Netherlands, ie, height is still increasing over the decades (Fredriks et al 2000). This is supported by data from Statistics Netherlands (Frenken 2007). Another factor that must be taken into consideration is that the Dutch population, and in particular those in the three most northern provinces, is known to be relatively tall (Frenken 2007). Besides including a large number of children, a relatively large Protein Tyrosine Kinase inhibitor geographical area was covered and both rural and urban schools were included to Talazoparib chemical structure ensure a broad diversity and heterogeneity of participants. A vast number of different instruments are available to measure grip strength. The Jamar hand dynamometer was selected because most normative studies have used this device and therefore it allows data to be compared with other (and future) studies (Innes 1999, Roberts et al

2011). Moreover, besides having a high test-retest and inter-investigator reliability, it also has high reproducibility when used by children (Lindstrom-Hazel et al 2009, Mathiowetz et al 1984,

Roberts et al 2011, Van den Beld et al 2006). To ensure all children were measured in the same manner, and again to follow standardised methods, participants were measured according to the ASHT protocol (Innes 1999, Roberts et al 2011). However, we implemented three exceptions. First, for the 4 and 5 year olds, the handle of the device was Edoxaban set to the first setting, which is considered to be less accurate than the second (Bechtol 1954, Boadella et al 2005, Firrell and Crain 1996, Hamilton et al 1994). These findings result from studies that focus on adults, and young children obviously have smaller hands. Therefore the distance to the handle of the device (3.8 cm) is relatively large compared to their average hand size (Bear-Lehman et al 2002). In practice, they could not reach the second setting adequately, and the first setting has also been used for adults with small hands (Ruiz-Ruiz et al 2002). Second, it is preferred to use the mean of three attempts (MacDermid et al 1994, Mathiowetz et al 1984). However, other studies showed that scoring fewer attempts, taking fewer attempts into consideration, or even using the maximum attempt, does not lead to significant differences compared with the mean of three attempts (Coldham et al 2006, Crosby and Wehbé 1994, Haidar et al 2004). Additionally, although fatigue does not seem to influence grip strength measurement in adults, we could not find any studies regarding this matter in children.

Vaccines recommended in the categories 1, 2, and 3 are also assessed to determine the public health interest of their integration into the Health Care Benefits Ordinance (Article 12) (vaccines targeting travelers are not considered). Such a request for integration would then be evaluated by appropriate independent commissions (see below). The commission obtains technical data and expertise for deliberation from a variety of sources, including official commission members, national reference centers such as the national influenza center or the influenza working

group, Navitoclax mw and invited national ad hoc experts. Use is made of WHO position papers, as well as national position statements and information found on websites, such as the European Centre for Disease Surveillance and Control (ECDC) and the U.S. Centers for Disease Control and Prevention (CDC). Recommendations from other NITAGs such as the U.S. Advisory Committee on Immunization Practices are taken into account. Working groups set up by the commission are a preferred source of information and expertise (Table 2), some of which are permanent, while others are set up for a specific period of time. They provide a foundation for decisions in adherence with the analytical framework (see above). Membership in a working group is voluntary and is decided upon by the commission members; any commission member

can chair and participate in a working group. External experts can be invited to join as well. People from the pharmaceutical Carnitine palmitoyltransferase II industry may selleck compound be consulted but they cannot participate in a working group. The working group creates a basic document that functions as a strategic pre-position statement. It is then circulated among the membership of the commission. Members can ask questions and give feedback, after which the document is presented in a plenary meeting. The Secretariat verifies the references

used, as well as independence of the work. In making its assessments, the commission considers the following vaccine-preventable outcomes, which are ranked in order of descending importance: mortality, hospitalizations, overall morbidity, epidemic potential, and equity and disability-adjusted life years (DALYs) or quality-adjusted life years (QALYs) lost. Disease burden is an evaluated criterion for each vaccine, but there are no predefined limits on criteria. The criteria are ad hoc, and are made according to the disease and on the synthesis of all available data. A vaccine is recommended only if its benefits, in terms of morbidity and mortality (diseases and their complications), are significantly greater than the risk of it causing adverse effects. Recommendations are usually decided upon by open vote, but occasionally a secret vote may be held. If experts do not agree on issues, they are resolved on a case-by-case basis.

1, 2 and 21 Different clinical subtypes of drusen have been described in AMD, but all drusen seem to be indistinguishable in location, composition, and substructure.5 “Basal laminar drusen,” also termed “cuticular drusen,” refers to an early-onset drusen phenotype with innumerable small (25

to 75 μm) hard drusen.22 and 23 This subtype of AMD is more easily visualized angiographically than biomicroscopically, with a typical “stars-in-the-sky” PD98059 appearance during the early arteriovenous phase of fluorescein angiography (Figure 1).24 In later stages, the number of drusen often increases, with clustered groups of drusen scattered throughout the retina.22 In general, color fundus photographs are used to evaluate the morphology of drusen over time. However, color images do not provide detailed information about the changing morphology

of small drusen.25, 26 and 27 PR-171 mw The introduction of spectral-domain optical coherence tomography (SD-OCT) has enabled an improved in vivo visualization of drusen morphology.28 SD-OCT is able to acquire 3-dimensional images of the retina with high speed and high resolution. Subsequently, studies of the fine details of small drusen and adjacent retinal structures become possible.28 and 29 After we observed occasional changes of drusen morphology in routinely followed eyes with basal laminar drusen, we decided to longitudinally investigate the appearance Adenosine of small hard drusen in eyes with this phenotype. The focus of our investigation was to determine whether morphologic parameters may be predictive for processes of progression or regression of small hard drusen in basal laminar drusen affected eyes. A total of 10 subjects who met the diagnostic criteria of basal laminar drusen were retrieved from the European Genetic Database (EUGENDA, www.eugenda.org), a large multicenter database for clinical and molecular analysis of AMD and different early-onset drusen phenotypes.

For inclusion in the study, subjects had basal laminar drusen of the posterior pole and ocular media allowing adequate SD-OCT scanning, defined by a maximum score of NO3/NC2/C1/P1 according the Lens Opacities Classification System III.30 Study participants had to be able to fixate for at least 1 minute per eye to allow adequate SD-OCT scanning. The basal laminar drusen phenotype was defined as a symmetrically distributed pattern between both eyes of at least 50 scattered, uniformly sized, small (25 μm to 75 μm), hyperfluorescent drusen on fluorescein angiography in each eye, of which at least 20 drusen are located outside the Wisconsin age-related maculopathy grading template.31 Eyes with choroidal neovascularization (CNV), a large area of central geographic atrophy (>1500 μm), and retinal abnormalities other than AMD-related were excluded from the study.

If task difficulty is used as the indicator for balance exercise intensity, exercise prescription across broad populations cannot be monitored or graded to ensure training effects for individual patients. If all patients had the same balance capacity at the beginning of a program, then a linear progression in task difficulty through a program may represent an increase in balance exercise intensity for individuals from session to session. Apart from the fact that no group of participants

is ever homogeneous, one would still be left with this dilemma regarding the level at which the exercise intensity was pitched through the program. It would be unclear whether all participants started the balance exercises at a low intensity and stayed low, or started at a moderate intensity and practised high intensity exercises by the end of the intervention. One program selleck kinase inhibitor that explicitly presented a rubric to guide balance exercise intensity prescription was identified (Littbrand et al 2006a). This HIFE program includes a table (p. 8) that defines low, medium, and high intensity exercise prescriptions. For the strength training exercises, the repetition maximum principle is used. For balance exercise a three-point scale ranging from ‘no challenge’

to ‘fully challenged’ postural stability is used. The authors provide a definition for full challenge of postural stability as ‘balance exercises else performed near the limits of maintaining postural Selleckchem Everolimus stability’ (Littbrand et al 2006a p. 8) This attempt

at standardisation carries some face validity given that repetitive work at the limits of stability is likely to represent an overload, however the ordinal scaling limits the usefulness of this rating of balance exercise intensity. If the level of balance exercise intensity cannot be measured in a reliable and valid way then questions of how hard we need to challenge balance in order to induce improvements in balance cannot be answered. This issue is of particular relevance for the development and implementation of home exercise or unsupervised programs, as it has been found that clinicians often prescribe programs of lower challenge in the home environment compared to supervised situations (Haas et al 2012). While still ordinal in nature, another rating scale that may inform a future measure of balance exercise intensity is the Borg scale. Studies in this review that utilised the Borg scale, also known as the rating of perceived exertion scale, reported the intensity of interventions of mixed exercise types, attributing the rating to the program in its entirety (Means et al 2005, Nelson et al 2004, Pereira et al 2008).

3 and 10 culture volume per day) at days 3 and 4. Prior to virus infection, using the same bioreactor vessel used for Vero cell culture, the media feed was stopped and pH, DO and temperature settings were adjusted to 7.4, 25% and 32.5 °C, respectively. Media was not refreshed but glucose and glutamine

were fed when concentrations were below 5 mM and 0.5 mM, respectively. Cells were infected with poliovirus with an MOI (multiplicity of infection) of 0.01. Virus cultivation was considered finished when 100% CPE (cytopathic effect) was observed microscopically. Cells were counted daily using a Nucleocounter NC-100 (Chemometec). Cell culture metabolites such as glucose, lactate, glutamine, glutamate and ammonia were monitored using a Bioprofile 100 Plus (Nova Biomedical Waltham, MA). Poliovirus was quantified with a virus titer Metformin in vivo assay as described previously [10]. The amount of d-antigen was assessed using a d-antigen ELISA [11]. Vero cell cultures were performed

in four different cultivation modes, batch, semi-batch, perfusion and recirculation. Batch cultivations were performed to obtain a reference growth curve for later comparison with the more sophisticated culture methods where either media is refreshed (semi-batch and perfusion) or circulated (recirculation). After 3–4 days of cultivation, a cell density at 1.0 × 106 cells mL−1 was reached in batch cultivation with an average growth rate of 0.036 h−1 during exponential growth and a growth rate of 0.022 h−1 at the moment of virus infection on day 4 (Fig. 1; Table 1). At this point cells are present very as a monolayer on the microcarriers (Fig. 2). Applying a daily partial check details medium renewal in a semi-batch mode allowed cell growth to continue and after 2 additional days of culture (6 days in total) a cell density of 1.8 × 106 cells mL−1 was obtained. Here comparable growth rates to batch cultivation were observed. The growth rate declined during the feed phase from

0.034 h−1 at day 3 to 0.006 h−1 at day 6. Using a perfusion mode, where medium renewal is continuous, cell growth could be prolonged to yield a cell density of 2.7 × 106 cells mL−1 in 7 days. The growth rates of the Vero cells were lower during the feed phase compared to the growth rates observed in semi-batch cultivations and decreased from 0.018 h−1 at day 3 to 0.005 h−1 at day 7. Cells were present in a multilayer on the microcarriers at these cell concentrations (Fig. 2). In the so-called recirculation method [9] cells were retained in the bioreactor while medium from an external container was circulated. When starting with an inoculation density of 0.6 × 106 cells mL−1 a monolayer was already formed after one day of cultivation, and cells started to grow in a multilayer rapidly. Cell concentrations of 5.0 × 106 cells mL−1 were found after a culture time of 4 days, while growth rates decreased linearly during the feed phase from 0.025 h−1 at day 2 to 0.0004 h−1 at day 4.