Screening for pre-eclampsia with competing-risks model using placental growth factor measurement in blood samples collected before 11 weeks' gestation.

OBJECTIVES: To describe the distributional properties and assess the performance of placental growth factor (PlGF) measured in blood samples collected before 11 weeks' gestation in the prediction of pre-eclampsia (PE). METHODS: The study population consisted of pregnant women included in the Pre-eclampsia Screening in Denmark (PRESIDE) study with a PlGF measurement from the routine combined first-trimester screening (cFTS) blood sample collected at 8-14 weeks' gestation. PRESIDE was a prospective multicenter study investigating the predictive performance of the Fetal Medicine Foundation (FMF) first-trimester screening algorithm for PE in a Danish population. In the current study, serum concentration of PlGF in the cFTS blood samples was analyzed in batches between January and June 2021. RESULTS: A total of 8386 pregnant women were included. The incidence of PE was 0.7% at < 37 weeks' gestation and 3.0% at ≥ 37 weeks. In blood samples collected at 10 weeks' gestation, PlGF multiples of the median (MoM) were significantly lower in pregnancies with preterm PE < 37 weeks compared to unaffected pregnancies. However, PlGF MoM did not differ significantly between pregnancies with PE and unaffected pregnancies in samples collected before 10 weeks' gestation. CONCLUSIONS: The gestational-age range for PlGF sampling may be expanded from 11-14 to 10-14 weeks when assessing the risk for PE using the FMF first-trimester screening model. There is little evidence to support the use of PlGF in blood samples collected before 10 weeks' gestation. © 2023 The Authors. Ultrasound in Obstetrics & Gynecology published by John Wiley & Sons Ltd on behalf of International Society of Ultrasound in Obstetrics and Gynecology.

Dissociable and Paradoxical Roles of Rat Medial and Lateral Orbitofrontal Cortex in Visual Serial Reversal Learning.

Much evidence suggests that reversal learning is mediated by cortico-striatal circuitries with the orbitofrontal cortex (OFC) playing a prominent role. The OFC is a functionally heterogeneous region, but potential differential roles of lateral (lOFC) and medial (mOFC) portions in visual reversal learning have yet to be determined. We investigated the effects of pharmacological inactivation of mOFC and lOFC on a deterministic serial visual reversal learning task for rats. For reference, we also targeted other areas previously implicated in reversal learning: prelimbic (PrL) and infralimbic (IL) prefrontal cortex, and basolateral amygdala (BLA). Inactivating mOFC and lOFC produced opposite effects; lOFC impairing, and mOFC improving, performance in the early, perseverative phase specifically. Additionally, mOFC inactivation enhanced negative feedback sensitivity, while lOFC inactivation diminished feedback sensitivity in general. mOFC and lOFC inactivation also affected novel visual discrimination learning differently; lOFC inactivation paradoxically improved learning, and mOFC inactivation had no effect. We also observed dissociable roles of the OFC and the IL/PrL. Whereas the OFC inactivation affected only perseveration, IL/PrL inactivation improved learning overall. BLA inactivation did not affect perseveration, but improved the late phase of reversal learning. These results support opponent roles of the rodent mOFC and lOFC in deterministic visual reversal learning.

Contribution of the gut microbiota to the regulation of host metabolism and energy balance: a focus on the gut-liver axis.

This review presents mechanistic studies performed in vitro and in animal models, as well as data obtained in patients that contribute to a better understanding of the impact of nutrients interacting with the gut microbiota on metabolic and behavioural alterations linked to obesity. The gut microbiota composition and function are altered in several pathological conditions including obesity and related diseases i.e. non-alcoholic fatty liver diseases (NAFLD). The gut-liver axis is clearly influenced by alterations of the gut barrier that drives inflammation. In addition, recent papers propose that specific metabolites issued from the metabolic cooperation between the gut microbes and host enzymes, modulate inflammation and gene expression in the liver. This review illustrates how dietary intervention with prebiotics or probiotics influences host energy metabolism and inflammation. Indeed, intervention studies are currently underway in obese and NAFLD patients to unravel the relevance of the changes in gut microbiota composition in the management of metabolic and behavioural disorders by nutrients interacting with the gut microbiota. In conclusion, diet is among the main triggers of NAFLD and the gut microbiota is modified accordingly, underlining the importance of the concomitant study of the nutrients and microbial impact on liver health and metabolism, in order to propose innovative, clinically relevant, therapeutic approaches.

Influence of the corn form in the diet and length of fasting before slaughter on the performance of overfed mule ducks.

The aim of this trial was to study the effect of the corn form fed during the overfeeding period and of the length of the fasting (LF) before slaughter on the performance of overfed mule ducks. A total of 180 ducks were distributed into 9 groups of 20 birds each, according to a 3 × 3 factorial design that differed according to the LF (9, 12, or 15 h) and the ratio of whole corn/corn flour in the overfeeding diet (OD): constant ratio of 30/70 [C diet]; progressive change from 0/100 to 30/70 [P diet]; or constant ratio of 0/100 [F diet]). At the end of the overfeeding period, the birds were slaughtered to measure the weight and chemical composition of the fatty liver. No significant interaction between the OD and LF was observed. The fatty liver weight was higher (+8%; P = 0.031) in the ducks fed the P diet than those fed the F diet. The ones fed the C diet were intermediate (709 g). The fatty liver weight was heavier (+7%) in the ducks slaughtered after 9 h of fasting than in those slaughtered after 15 h of fasting, The weight of fatty liver of birds slaughtered after 12 h was intermediate (711 g; P < 0.05). The foie gras fat loss during cooking (20% vs. 27%; P = 0.003), DM (68.13% vs. 69.3%; P = 0.021), and ash content (0.40% vs. 0.36%; P = 0.010) were lower in the group of ducks slaughtered after 9 h of fasting than in the other 2 groups. In conclusion, the present results suggest that (i) incorporating whole corn in the OD at a constant or increasing rate up to 30% significantly improves the performance of overfed mule ducks, and (ii) the shorter the fasting time is, the greater the weight and the quality of the foie gras will be.

Plate failure by bending following tibial fracture stabilisation in 10 cats.

OBJECTIVE: To describe the clinical findings and management of tibial fractures in cats in which osteosynthesis failed due to plate bending. METHODS: Case records and radiographs of cat tibial fracture repairs from five referral centres were reviewed for signalment and to assess incidence of plate failure by bending. Cats that sustained plate bending following plate or plate-rod fixation were reviewed for fracture configuration, repair method, initial postoperative and postfailure tibial alignment, revision treatment and outcome. RESULTS: The incidence of plate bending in cat fractures managed with plate and plate-rod fixation in the four referral centres where the overall number could be established was 13% (8/60). In the 10 cats in which plates bent, initial fractures were generally oblique or spiral with mild comminution and located in the middle or distal third of the tibia. Mean time to implant failure was 24 days (range 2 to 56 days). Mean tibial valgus angle increased from 12·9° to 30·9° following bending of the plate. Short-term outcome following revision surgery using orthogonal plating or stacked medial plates was favourable with improvement in tibial valgus in all five fractures with follow-up data. CLINICAL SIGNIFICANCE: Plate bending following tibial fracture stabilisation in these 10 cats resulted in tibial valgus deformation. Consideration of plate and/or intramedullary rod selection and application should be given to avoid a plate strain environment that exceeds the yield point of the plate.

A kinetic study of the natural induction of liver steatosis in Greylag Landaise geese: the role of hyperphagia.

We have previously demonstrated that a sharp rise in feed intake (hyperphagia) and spontaneous liver steatosis could be experimentally induced in domestic Greylag geese by combining a short photoperiod and a sequence of feed restriction followed by ad libitum corn feeding during the fall and the winter. In this previous work, however, individual feed intake could not be recorded. The present study aimed at evaluating the relationship between level and pattern of hyperphagia and liver weight with an individual control of feed intake in individually housed (IH) geese, while comparing the performances with group housed (GH) geese. A total of 300 male geese of 19 weeks of age, were provided with corn ad libitum after an initial feed restriction period. From 21 to 23 weeks of age, the daylight duration was progressively reduced from 10 to 7 h and kept as such until the end of the experiment (week 36). In all, 30 GH and 30 IH geese were slaughtered at 19, 27, 30, 32 and 36 weeks of age. Feed intake was measured per group in GH geese and individually in IH geese. During the 1st week of corn feeding, the average feed intake rose up to 600 g/goose per day in GH geese but not in IH geese where the feed intake rose gradually from 300 to 400 g/day. The liver weight increased from 93 g (week 19) to 497 g (week 32; P<0.05) in GH birds. In IH birds, liver weights were, on average, much lower (ranging from 220 to 268 g) than in GH birds (P<0.05). In GH and IH bird, the variability in the individual response to corn feeding was very high (liver weight cv ranging from 63% to 83% depending on slaughter age). A close correlation between corn consumption and liver weight was evidenced in IH birds at each slaughter age (R 2 ranging from 0.62 to 0.79), except at 36 weeks of age where this correlation was weak (R 2=0.14). The variability in the extent of liver steatosis is very high and our results in IH birds clearly point out the major role of hyperphagia, mainly at the beginning of the ad libitum corn feeding period, on the development of spontaneous liver steatosis.

Prevalence and heritability of symptomatic syringomyelia in Cavalier King Charles Spaniels and long-term outcome in symptomatic and asymptomatic littermates.

BACKGROUND: Syringomyelia (SM) is common in the Cavalier King Charles Spaniel (CKCS). Dogs with syringes express clinical signs or might be clinically silent. OBJECTIVES: To investigate the prevalence and heritability of symptomatic SM, the association between clinical signs and magnetic resonance imaging (MRI) findings, and long-term outcome. ANIMALS: All CKCS registered in the Danish Kennel Club in 2001 (n = 240). METHODS: A cross-sectional questionnaire-based prevalence study validated by telephone interviews and clinically investigated clinical signs of SM. Dogs were 6 years at the time of investigation. A prospective observational litter study including clinical investigations, MRI and 5-year follow-up of symptomatic and asymptomatic siblings. Heritability was estimated based on the scale of liability in the study population and litter cohort. RESULTS: The cross-sectional study estimated a prevalence of symptomatic SM at 15.4% in the population. Thirteen symptomatic and 9 asymptomatic siblings participated in the litter study. Spinal cord syringes were confirmed in 21 of 22 littermates (95%). Syrinx diameter and mean syrinx : spinal cord ratio were significantly correlated with clinical signs (P < .01). Estimated heritability of symptomatic SM was 0.81. Symptomatic SM motivated euthanasia in 20%. Dogs with syringes, which expressed no clinical signs at the age of 6, remained asymptomatic in 14/15 cases (93%). CONCLUSIONS AND CLINICAL IMPORTANCE: The prevalence of symptomatic SM is high and genetics have a high impact on clinical disease expression. Further investigations of factors influencing the outbreak threshold of clinical signs of SM are desirable.

Impact of feed restriction on health, digestion and faecal microbiota of growing pigs housed in good or poor hygiene conditions.

Feed restriction could be a relevant strategy to preserve gut health, reduce systemic inflammatory response and finally limit antibiotic use. This study assessed the effect of feed restriction on growing pigs submitted to a moderate inflammatory challenge induced by the degradation of the environmental hygiene that is known to alter growth rate. The experiment was run on 80 pigs selected at 7 weeks of age according to a 2×2 factorial design: two feeding levels, ad libitum (AL) and feed restricted (FR) at 60% of AL, and two conditions of environmental hygiene, clean and dirty. Pigs were housed individually throughout the experiment. From 61 to 68 days of age (day 0 to 7), pigs were housed in a post weaning unit and feed restriction was applied to half of the pigs from day 0 to day 29. At 68 days of age (day 7 of the experiment), pigs were transferred in a growing unit where half of FR and half of AL pigs were housed in a dirty environment (poor hygiene) and the other half in a clean environment (good hygiene) until day 42. Growth performance was recorded weekly. Blood and faeces samples were collected to measure indicators of inflammation, nutrient digestibility and microbiota composition. Faecal consistency was monitored daily to detect diarrhoeas. Feed restriction decreased daily weight gain (-35% to -50%, P<0.001), increased the feed conversion ratio (+15%, P<0.001) and CP digestibility (+3%, P<0.05) and reduced the occurrence of diarrhoeas irrespective of hygiene conditions. Poor hygiene conditions decreased growth performance (-20%, P<0.05) and total tract digestibility of all nutrients (P<0.001). Haptoglobin (+50%) concentrations and lymphocyte (+10%) and granulocyte (+40%) numbers were higher in poor hygiene conditions (P<0.05), confirming that the model was effective to induce a systemic inflammatory response. Both feed restriction and hygiene modified the profile of the faecal microbiota. In this study, feed restriction did not reduce the systemic inflammatory response caused by poor hygiene conditions despite the limitation of the occurrence of digestive disorders. However, our study opens discussions regarding the impact of hygiene and feed restriction on gut microbial communities and digestive health.

Complications of lateral plate fixation compared with tension band wiring and pin or lag screw fixation for calcaneoquartal arthrodesis. Treatment of proximal intertarsal subluxation occurring secondary to non-traumatic plantar tarsal ligament disruption in dogs.

OBJECTIVE: To compare complication rates and the outcomes of these complications after lateral plate fixation with figure-of-eight tension-band-wire and pin or lag screw fixation for arthrodesis of the calcaneoquartal joint, following non-traumatic disruption of the plantar tarsal ligament in dogs. METHODS: Data were collected retrospectively from five UK referral centres. Diplomate specialists and their residents performed all procedures. Referring veterinarians were contacted for long-term follow-up. RESULTS: Seventy-four procedures were undertaken in 61 dogs. There were 58 arthrodeses in the lateral plate group (Plate), nine in the pin and tension-band-wire group (Pin), and seven in the lag screw and tension-band wire-group (Screw). Compared to Plate (17%), further surgical intervention was required more frequently following Pin (56%, OR = 3.2) or Screw (43%, OR = 2.5) fixation. Clinical failure of arthrodesis occurred less frequently with Plate (5%) compared with Screw (43%, OR = 8.6) and Pin fixation (22%, OR = 4.4). Cases managed with external coaptation postoperatively were more likely to suffer from postoperative complications (OR = 2.2). CLINICAL SIGNIFICANCE: Lateral plating was associated with fewer postoperative complications than pin and tension-band-wire fixation for arthrodesis of the calcaneoquartal joint in dogs with non-traumatic disruption of the plantar tarsal ligament.

COX-2 expression in canine anal sac adenocarcinomas and in non-neoplastic canine anal sacs.

Anal sac adenocarcinoma (ASAC) is a clinically significant canine neoplasm characterized by early lymphatic invasion. Up-regulation of cyclooxygenase isoform 2 (COX-2) has been confirmed in several animal and human neoplastic tissues. The aim of the current study was primarily to evaluate COX-2 expression in canine ASAC and compare it to COX-2 expression in non-neoplastic canine anal sac tissue using immunohistochemistry with scoring for percentage positivity and intensity. Twenty-five ASAC samples and 22 normal anal sacs were available for evaluation. All canine ASAC samples and the normal anal sac tissues stained positively for COX-2. However, while normal anal sac tissue showed strong staining of the ductal epithelial cells, ASAC samples showed staining of the neoplastic glandular epithelial cells, with varying percentage positivity and intensity between ASAC samples. COX-2 immunoreactivity of ASAC samples was of low intensity in 52% and high in 12% of the cases; the remaining samples were of intermediate intensity. Seventy-six per cent of the ASAC had over 50% of the neoplastic glandular cells staining positive. These results confirm that COX-2 is expressed in the neoplastic glandular epithelial cells in canine ASAC and suggest a potential role for COX-2 inhibitors in the management of ASAC. Furthermore, the results indicate that COX-2 is expressed in ductal epithelial cells of the normal anal sac.

Long bone fracture as a complication following external skeletal fixation: 11 cases.

OBJECTIVES: To report on a series of dogs and cats with long bone fractures that occurred as a direct consequence of linear external skeletal fixation (ESF) application. METHODS: Retrospective study. Data from the medical records and radiographs of canine (n = 4) and feline (n = 7) cases were collected from three referral and three first opinion practices in the UK (1999 to 2011). RESULTS: Long bone fractures occurred following the application of linear ESF either while the ESF was in situ or after removal. All fractures occurred through either a pin tract or an empty drill hole. Pins associated with ESF-related fracture tended to be in the higher end of the recommended size range. The majority of cases had additional complicating factors such as multiple injuries, revision surgery, poor owner compliance with postoperative exercise restriction and the presence of empty drill holes. CLINICAL SIGNIFICANCE: In cases with features that could complicate outcome, careful attention should be paid to recommendations for ESF application. Leaving empty drill holes is suboptimal. The retrospective nature of the study, low numbers of, and diversity amongst, cases should be taken into consideration when interpreting the results from this study.

Fatigue, mood and quality of life improve in MS patients after progressive resistance training.

Fatigue occurs in the majority of multiple sclerosis patients and therapeutic possibilities are few. Fatigue, mood and quality of life were studied in patients with multiple sclerosis following progressive resistance training leading to improvement of muscular strength and functional capacity. Fatigue (Fatigue Severity Scale, FSS), mood (Major Depression Inventory, MDI) and quality of life (physical and mental component scores, PCS and MCS, of SF36) were scored at start, end and follow-up of a randomized controlled clinical trial of 12 weeks of progressive resistance training in moderately disabled (Expanded Disability Status Scale, EDSS: 3-5.5) multiple sclerosis patients including a Control group (n = 15) and an Exercise group (n = 16). Fatigue (FSS > 4) was present in all patients. Scores of FSS, MDI, PCS-SF36 and MCS-SF36 were comparable at start of study in the two groups. Fatigue improved during exercise by -0.6 (95% confidence interval (CI) -1.4 to 0.4) a.u. vs. 0.1 (95% CI -0.4 to 0.6) a.u. in controls (p = 0.04), mood improved by -2.4 (95% CI -4.1 to 0.7) a.u. vs. 1.1 (-1.2 to 3.4) a.u. in controls (p = 0.01) and quality of life (PCS-SF36) improved by 3.5 (95% CI 1.4-5.7) a.u. vs. -1.0 (95% CI -3.4-1.4) a.u. in controls (p = 0.01). The beneficial effect of progressive resistance training on all scores was maintained at follow-up after further 12 weeks. Fatigue, mood and quality of life all improved following progressive resistance training, the beneficial effect being maintained for at least 12 weeks after end of intervention.

Resistance training improves muscle strength and functional capacity in multiple sclerosis.

OBJECTIVE: To test the hypothesis that lower extremity progressive resistance training (PRT) can improve muscle strength and functional capacity in patients with multiple sclerosis (MS) and to evaluate whether the improvements are maintained after the trial. METHODS: The present study was a 2-arm, 12-week, randomized controlled trial including a poststudy follow-up period of 12 weeks. Thirty-eight moderately impaired patients with MS were randomized to a PRT exercise group (n = 19) or a control group (n = 19). The exercise group completed a biweekly 12-week lower extremity PRT program and was afterward encouraged to continue training. After the trial, the control group completed the PRT intervention. Both groups were tested before and after 12 weeks of the trial and at 24 weeks (follow-up), where isometric muscle strength of the knee extensors (KE MVC) and functional capacity (FS; combined score of 4 tests) were evaluated. RESULTS: KE MVC and FS improved after 12 weeks of PRT in the exercise group (KE MVC: 15.7% [95% confidence interval 4.3-27.0], FS: 21.5% [95% confidence interval 17.0-26.1]; p < 0.05), and the improvements were better than in the control group (p < 0.05). The improvements of KE and FS in the exercise group persisted at follow-up after 24 weeks. Also, the exercise effects were reproduced in the control group during the 12-week posttrial PRT period. CONCLUSIONS: Twelve weeks of intense progressive resistance training of the lower extremities leads to improvements of muscle strength and functional capacity in patients with multiple sclerosis, the effects persisting after 12 weeks of self-guided physical activity. LEVEL OF EVIDENCE: The present study provides level III evidence supporting the hypothesis that lower extremity progressive resistance training can improve muscle strength and functional capacity in patients with multiple sclerosis.

Characterization of human mucin (MUC15) and identification of ovine and caprine orthologs.

The glycoprotein MUC15 (mucin 15) was initially isolated from the bovine milk fat globule membrane. The present work demonstrates the existence of immunologically similar proteins ( approximately 130 kDa) in ovine, caprine, porcine, and buffalo milk samples. Purification and N-terminal amino acid sequencing confirmed the presence of ovine and caprine MUC15 orthologs in milk fat globule membranes. Expression of MUC15 in human milk was demonstrated by immunostaining ( approximately 150 kDa) as well as by mass spectrometry. Screening of a human multiple tissue expression array showed abundant MUC15 gene expression in placenta, salivary gland, thyroid gland, trachea, esophagus, kidney, testis, and the leukemia K-562 cell line. Furthermore, moderate expression was seen in the pancreas, adult and fetal lung, fetal kidney, lymph node, adult and fetal thymus, and parietal lobe. Structural motifs for interactions (epidermal growth factor receptor and Src homology 2 domains) are identified in the intracellular region. Implication of the mucin in signal transduction and the potential physiological function of MUC15 are discussed.

Improved parameter estimation for systems with an experimentally located Hopf bifurcation.

When performing system identification, we have two sources of information: experimental data and prior knowledge. Many cell-biological systems are oscillating, and sometimes we know an input where the system reaches a Hopf bifurcation. This is the case, for example, for glycolysis in yeast cells and for the Belousov-Zhabotinsky reaction, and for both of these systems there exist significant numbers of quenching data, ideal for system identification. We present a method that includes prior knowledge of the location of a Hopf bifurcation in estimation based on time-series. The main contribution is a reformulation of the prior knowledge into the standard formulation of a constrained optimisation problem. This formulation allows for any of the standard methods to be applied, including all the theories regarding the method's properties. The reformulation is carried out through an over-parametrisation of the original problem. The over-parametrisation allows for extra constraints to be formed, and the net effect is a reduction of the search space. A method that can solve the new formulation of the problem is presented, and the advantage of adding the prior knowledge is demonstrated on the Brusselator.

Biomechanical analysis of the effect of changing patient-handling technique.

The objective of the study was to assess the changes in the mechanical load on the low-back when shifting from a self-chosen to a recommended patient-handling technique. Nine female health care workers without formal education in patient-handling carried out 8 different tasks involving moving, turning and lifting situations. By means of a dynamic 3D biomechanical model of the lower part of the body, peak torque, compression and shear forces at the L4/L5 joint were compared using the two different patient-handling techniques. In 5 of the 8 tasks, a significant reduction was observed in spinal loading. Application of the recommended technique decreased the compression value significantly for all tasks with a mean value above 3000 N. For the two tasks with the highest compression values when using the self-chosen technique (4223, 4446 N), the loading was reduced with 36% and 25%, respectively. If the principles behind the recommended technique are implemented and maintained, a decrease in the risk of low-back disorders during patient-handling should thus be expected.

Intra-abdominal pressure increases during exhausting back extension in humans.

The aim of this study was to investigate in humans the effect of maintained spinal load on the intra-abdominal pressure (IAP), low back kinematics and trunk muscle activity. This study consisted of two endurance tests for the low back muscles performed 3 weeks apart. Nine healthy subjects participated in the study. In the first test (upright-test), the subject had to pull with the back muscles at a constant force in an upright position, and in the second test (incline-test), the subject had to resist a constant forward pulling force while standing with a 45 degrees inclination of the back in relation to vertical. The IAP, rate of perceived exertion and electromyogram (EMG) from the erector spinae, iliocostalis lumborum, rectus abdominus and the external and internal oblique muscles were measured using surface electrodes. There was no significant difference in endurance time between the two tests. Both tests showed a significant increase in EMG amplitude with time for all muscles except the erector spinae muscle. A decrease in the median frequency for the erector spinae muscle was found indicating fatigue, and since no increase in EMG amplitude was found a decrease in force output from the muscle must be assumed. The IAP increased significantly during both tests. There was a considerable variation between the subjects in the size of this development, but the pattern was the same for all subjects. In conclusion it was found that trunk extension until exhaustion initiates an increase in the activity of the abdominal muscles and an increase in the IAP as the low back muscles become fatigued.

The importance of structural transitions of the switch II region for the functions of elongation factor Tu on the ribosome.

Elongation factor Tu (EF-Tu) undergoes a large conformational transition when switching from the GTP to GDP forms. Structural changes in the switch I and II regions in the G domain are particularly important for this rearrangement. In the switch II region, helix alpha2 is flanked by two glycine residues: Gly(83) in the consensus element DXXG at the N terminus and Gly(94) at the C terminus. The role of helix alpha2 was studied by pre-steady-state kinetic experiments using Escherichia coli EF-Tu mutants where either Gly(83), Gly(94), or both were replaced with alanine. The G83A mutation slows down the association of the ternary complex EF-Tu.GTP.aminoacyl-tRNA with the ribosome and abolishes the ribosome-induced GTPase activity of EF-Tu. The G94A mutation strongly impairs the conformational change of EF-Tu from the GTP- to the GDP-bound form and decelerates the dissociation of EF-Tu.GDP from the ribosome. The behavior of the double mutant is dominated by the G83A mutation. The results directly relate structural transitions in the switch II region to specific functions of EF-Tu on the ribosome.