Structural Analysis of the Large Stokes Shift Red Fluorescent Protein tKeima.

The Keima family comprises large Stokes shift fluorescent proteins that are useful for dual-color fluorescence cross-correlation spectroscopy and multicolor imaging. The tKeima is a tetrameric large Stokes shift fluorescent protein and serves as the ancestor fluorescent protein for both dKeima and mKeima. The spectroscopic properties of tKeima have been previously reported; however, its structural basis and molecular properties have not yet been elucidated. In this study, we present the crystallographic results of the large Stokes shift fluorescent protein tKeima. The purified tKeima protein spontaneously crystallized after purification without further crystallization. The crystal structure of tKeima was determined at 3.0 Å resolution, revealing a ß-barrel fold containing the Gln-Tyr-Gly chromophores mainly with cis-conformation. The tetrameric interfaces of tKeima were stabilized by numerous hydrogen bonds and salt-bridge interactions. These key residues distinguish the substituted residues in dKeima and mKeima. The key structure-based residues involved in the tetramer formation of tKeima provide insights into the generation of a new type of monomeric mKeima. This structural analysis expands our knowledge of the Keima family and provides insights into its protein engineering.

Clinicoradiological Features of Alexia Without Agraphia.

Alexia without agraphia is a striking vascular syndrome of the acquired inability to read words just written down. This syndrome occurs after lesions in the splenium of the corpus callosum that disconnect the angular gyrus from the visual pathway. Most of the time, a lesion in the left occipital lobe is also present, and patients present with a visual field deficit. It is a classic neurological syndrome that is rarely seen. We present two cases of alexia without agraphia seen in our hospital the same week.

Experimental and Theoretical Rotational Diffusion Studies of 7DM4M1M1,8, N-2(1H)-one and 7A4T2H1B-2-one in Series of Alcohol Solvents: Stoke's-Einstein-Debye and Alavi-Waldeck Models.

Rotational diffusion studies of two solutes 7-(dimethylamino)-4-methoxy-1-methyl-1,8-naphthyridin-2(1H)-one (7DM4M1M1,8, N-2(1H)-one) and 7-amino-4-(trifluoromethyl)-2H-1-benzopyran-2-one (7A4T2H1B-2-one) having equal volumes but different chemical natures are studied in series of alcohol solvents at 303 K using steady-state methods. HOMO-LUMO, Electron density, Molecular electrostatic potential (MEP), etc., are obtained from computational calculations using Gaussian 09 software. Rotational reorientation times of 7DM4M1M1,8, N-2(1H)-one solute molecule is found to be less than 7A4T2H1B-2-one solute molecule indicates it rotates slowly in chosen solvents. Stoke's-Einstein-Debye (SED) model with stick boundary conditions for the 7A4T2H1B-2-one solute molecule is modeled to describe mechanical friction. Polar solutes along with mechanical friction also experience dielectric friction. Both these frictions being non-separable, the Alavi-Waldeck (AW) model is studied for dielectric friction contribution to the total friction solute experiences in solvents. AW model effectively explains the observed dielectric friction in alcohol solvents.

Male spondyloarthritis patients and those with longer disease duration have less severe disc degeneration: propensity score-matched comparison.

Objective: Using whole spine sagittal T2 MRI, we aimed to compare the severity and prevalence of disc degeneration (DD) in axial SpA patients vs the general population and to determine any association between spinal inflammation, structural changes, mobility and DD among SpA patients. Methods: Two prospectively collected cohorts of SpA patients (n = 411) and the general population (n = 2007) were recruited. Eventually, 967 participants from the populational cohort and 304 participants from the SpA cohort were analysed. Two hundred and nineteen matched pairs were generated by propensity score matching. Imaging parameters, including Pfirrmann grading, disc herniation, high-intensity zone, Schmorl's node, Modic change and anterior marrow change were studied and compared from C2/3 to L5/S1. DD was defined as Pfirrmann grade 4 or 5. Demographic factors, including age, sex and BMI, were collected. Multivariable linear regression was used to determine the association between spinal inflammation [Spondyloarthritis Research Consortium of Canada (SPARCC) spine MRI index], structural changes [modified Stoke Ankylosing Spondylitis Spinal Score (mSASSS)] and mobility (BASMI) with lumbar Pfirrmann score. Results: SpA patients had lower prevalence of DD (P < 0.001). The disease stage-stratified regression model showed that SPARCC spinal MRI index was associated with higher lumbar Pfirrmann scores in early disease (ß = 0.196, P = 0.044), whereas mSASSS was associated with lower lumbar Pfirrmann scores in later disease (ß = -0.138, P = 0.038). Males had higher mSASSS (P < 0.001) and lower odds of whole spine DD (odds ratio = 0.622, P = 0.028). Conclusion: SpA patients had lower DD severity than the general population. Males had higher mSASSSs, and increased mSASSS at later disease was associated with less severe DD.

Impact of prior antiplatelet therapy on outcomes of acute ischemic stroke undergoing endovascular treatment: A systematic review and meta-analysis.

It is unclear how prior antiplatelet (APT) therapy affects outcomes of acute ischemic stroke (AIS) undergoing endovascular treatment. This review pooled data from the literature to compare outcomes of AIS between prior APT users vs non-users. PubMed, Embase, CENTRAL, and Scopus for studies were searched for studies comparing outcomes of AIS between APT users vs non-users up to 30th May 2023. Ten studies were included comparing 2648 APT users with 5076 non-users. Meta-analysis failed to demonstrate any statistically significant difference in symptomatic intracranial hemorrhage (sICH) but there was a tendency of higher mortality rates in prior APT users vs non-users. Although patients with prior APT therapy had significantly higher rates of successful recanalization as compared to patients with no prior APT treatment, meta-analysis showed significantly lower odds of functional independence amongst APT users vs non-users (OR: 0.77 95% CI: 0.68, 0.87 I2 = 22%). However, pooled analysis of adjusted data with fewer studies showed that there was no difference in sICH (OR: 1.04 95% CI: 0.78, 1.39 I2 = 0%), mortality (OR: 0.89 95% CI: 0.47, 1.68 I2 = 68%), successful recanalization (OR: 1.34 95% CI: 0.96, 1.88 I2 = 54%), and functional independence (OR: 0.96 95% CI: 0.81, 1.14 I2 = 0%) between APT users and non-users. Analysis of crude data indicates that prior APT therapy may improve successful recanalization without increasing sICH rates in AIS patients treated with endovascular therapy. However, there was an adverse effect of APT therapy on 3-month functional and survival outcomes. After adjustment of confounders, there was no difference in the odds of sICH, mortality, successful recanalization, and functional independence between APT users vs non-users.

A Young Female Newly Diagnosed With Takayasu's Arteritis Masquerading As Cerebrovascular Stroke.

The condition known as Takayasu's disease or Takayasu's arteritis is a type of vascular inflammation that affects the large and medium arteries. It can lead to a reduction in blood flow to various parts of the body, and it can cause severe complications. Patients with this disease may not have specific symptoms, which can lead to their diagnosis not being confirmed. Takayasu's disease is believed to be a probable cause of stroke in young patients. Although stroke is a common cause of morbidity, it is usually not an initial presentation in Takayasu's disease. In this study, a young female with left-sided hemiparesis was diagnosed with Takayasu's disease after a clinical and angiographic examination.

Association between Systemic Immune-Inflammation Index and Systemic Inflammation Response Index and Outcomes of Acute Ischemic Stroke: A Systematic Review and Meta-Analysis.

Systemic immune-inflammation index (SII) and systemic inflammation response index (SIRS) are being increasingly used to predict outcomes of various diseases. However, its utility for acute ischemic stroke (AIS) has not been established. Through this first systematic review and meta-analysis, we aimed to collate data on the prognostic ability of SII and SIRI for predicting functional outcomes and mortality after AIS. PubMed, CENTRAL, Scopus, Embase, and Web of Science were searched up to January 5, 2023, for studies reporting the association between SII or SIRI and outcomes of AIS. Adjusted data were pooled in a random-effects model. Meta-regression was conducted for variable cut-offs. Twelve studies were included. Pooled analysis of data showed that high SII was associated with poor functional outcomes after AIS (OR: 2.35 95% CI: 1.77, 3.10 I2 = 44% P < 0.00001). Meta-regression showed an increasing effect size with a higher cut-off of SII. Similarly, the meta-analysis demonstrated that AIS patients with high SIRI were at an increased risk of poor functional outcomes (OR: 1.69 95% CI: 1.08, 2.65 I2 = 78% P = 0.02). No association was noted with different cut-offs on meta-regression. Data on mortality were scarce but were suggestive of a higher risk of mortality with high SII and SIRI. SII and SIRI can be used to predict poor functional outcomes in AIS patients. Data on mortality are scarce to derive strong conclusions. Limited number of studies and variable cut-offs are important limitations that need to be overcome by future studies.

Mediation effect of stroke recurrence in the association between post-stroke interleukin-6 and functional disability.

AIM: Post-stroke inflammation increases the risk of functional disability through enlarged cerebral infarct size directly and follow-up stroke event indirectly. We aimed to use post-stroke proinflammatory cytokine interleukin-6 (IL-6) as a marker of inflammatory burden and quantify post-stroke inflammation's direct and indirect effect on functional disability. METHODS: We analyzed patients with acute ischemic stroke admitted to 169 hospitals in the Third China National Stroke Registry. Blood samples were collected within 24 h of admission. Stroke recurrence and functional outcome measured by the modified Rankin scale (mRS) were assessed via face-to-face interviews at 3 months. Functional disability was defined as an mRS score ≥2. Mediation analyses under the counterfactual framework were performed to examine the potential causal chain in which stroke recurrence may mediate the relationship between IL-6 and functional outcome. RESULTS: Among the 7053 analyzed patients, the median (interquartile range [IQR]) NIHSS score was 3 (1-5), and the median (IQR) level of IL-6 was 2.61 (1.60-4.73) pg/mL. Stroke recurrence was observed in 458 (6.5%) patients, and functional disability was seen in 1708 (24.2%) patients at the 90-day follow-up. Per stand deviation (4.26 pg/mL) increase in the concentration of IL-6 was associated with an increased risk of stroke recurrence (adjusted odds ratio [aOR], 1.19; 95% CI, 1.09-1.29) and disability (aOR, 1.22; 95% CI, 1.15-1.30) within 90 days. Mediation analyses revealed that 18.72% (95% CI, 9.26%-28.18%) of the relationship between IL-6 and functional disability was mediated by stroke recurrence. CONCLUSIONS: Stroke recurrence mediates less than 20% of the association between IL-6 and functional outcome at 90 days among patients with acute ischemic stroke. In addition to typical secondary prevention strategies for preventing stroke recurrence, more attention should be paid to novel anti-inflammatory therapy to improve functional outcomes directly.

Association of LIPC polymorphisms with stroke risk in the Chinese population.

Background: Stroke is a common cerebrovascular disease. The purpose of this study was to explore the association between LIPC single nucleotide polymorphisms (SNPs) and the risk of stroke in the Chinese population. Methods: This study recruited 710 stroke patients and 701 healthy controls. The four SNPs (rs690, rs6083, rs3829461, and rs6074) in LIPC were genotyped by the Agena MassARRAY. The correlation between LIPC polymorphisms and stroke risk was measured by odds ratio (OR) and 95% confidence interval (CI). In addition, multifactor dimensionality reduction (MDR) analysis was used to evaluate the impact of SNP-SNP interaction on stroke risk. Results: Overall analysis showed that rs690 was associated with an increased risk of stroke (T vs. G: OR = 1.19, 95% CI: 1.01-1.40, p = 0.041; additive: OR = 1.20, 95% CI: 1.01-1.42, p = 0.036). The stratified analysis revealed that rs690 was associated with an increased risk of stroke in subjects aged ≤ 64 years, male patients, and smokers, and rs6074 was associated with an increased risk of stroke in subjects aged > 64 years, male patients, drinkers, and non-smokers (p < 0.05). The results of the MDR analysis suggested the four-locus model as the most favorable model for assessing the risk of stroke. The analysis of clinical parameters of stroke patients showed that rs690 was correlated with platelet distribution width (PDW) (p = 0.014) and hematocrit levels (p = 0.004), and rs6074 was correlated with low-density lipoprotein cholesterol (LDL-C) level (p = 0.033). Furthermore, bioinformatics analysis results demonstrated that the expression levels of LIPC and its related genes (APOB, CETP, PNPLA2, and LMF1) were significantly different between the control and stroke groups (p < 0.05), and LIPC-related proteins were mainly related to lipid metabolism. Conclusion: This study indicated that rs690 and rs6074 in LIPC were significantly associated with increased risk of stroke in the Chinese population, possibly by regulating the levels of PDW, HCT, and LDL-C.

Solvatochromism as a Novel Tool to Enumerate the Optical and Luminescence Properties of Plastic Waste Derived Carbon Nanodots and Their Activated Counterparts.

Herein, we have developed a one-pot methodology to synthesise three types of C-dots and their activated counterparts from three different types of waste plastic precursors such as poly-bags, cups and bottles. The optical studies have shown the significant change in the absorption edge in case of C-dots in comparison to their activated counterparts. The respective variation in the sizes is correlated with the change in electronic band gap values of formed particles. The changes in the luminescence behaviour are also correlated with transitions from the edge of the core of formed particles. The obtained variations in the Stokes shift values of C-dots, and their ACs were used to explore the types of surface states and their related transitions in particles. The mode of interaction between C-dots and their ACs was also determined using solvent-dependent fluorescence spectroscopy. This detailed investigation could provide significant insight on the emission behaviour and the potential usage of formed particles as an effective fluorescent probe in sensing applications.

Heart rate variability as a predictor of stroke course, functional outcome, and medical complications: A systematic review.

Background: Heart rate variability (HRV) is a non-invasive marker of autonomic nervous system function that is based on the analysis of length differences between subsequent RR intervals of the electrocardiogram. The aim of this systematic review was to assess the current knowledge gap in the utility of HRV parameters and their value as predictors of the acute stroke course. Methods: A systematic review was performed in accordance with the PRISMA guidelines. Relevant articles published between 1 January 2016 and 1 November 2022 available in the PubMed, Web of Science, Scopus, and Cochrane Library databases were obtained using a systematic search strategy. The following keywords were used to screen the publications: "heart rate variability" AND/OR "HRV" AND "stroke." The eligibility criteria that clearly identified and described outcomes and outlined restrictions on HRV measurement were pre-established by the authors. Articles assessing the relationship between HRV measured in the acute phase of stroke and at least one stroke outcome were considered. The observation period did not exceed 12 months. Studies that included patients with medical conditions influencing HRV with no established stroke etiology and non-human subjects were excluded from the analysis. To minimize the risk of bias, disagreements throughout the search and analysis were resolved by two independent supervisors. Results: Of the 1,305 records obtained from the systematic search based on keywords, 36 were included in the final review. These publications provided insight into the usability of linear and non-linear HRV analysis in predicting the course, complications, and mortality of stroke. Furthermore, some modern techniques, such as HRV biofeedback, for the improvement of cognition performance after a stroke are discussed. Discussion: The present study showed that HRV could be considered a promising biomarker of a stroke outcome and its complications. However, further research is needed to establish a methodology for appropriate quantification and interpretation of HRV-derived parameters.

Re-stooping after Corrective Osteotomy in Patients with Ankylosing Spondylitis.

Background: Corrective osteotomy is an effective surgery for correcting posture in patients with ankylosing spondylitis (AS). Despite satisfactory correction, some patients experience re-stooping during follow-up. However, there have been no studies on re-stooping in AS. We aimed to analyze the factors that affect re-stooping. Methods: Fifty patients (50 cases) who underwent thoracolumbar corrective osteotomy for AS from March 2006 to April 2018 were analyzed. We defined re-stooping as global kyphosis that recurs after corrective osteotomy. The patients were divided into two groups based on the ratio of correction loss: non-re-stooping group (N group) and re-stooping group (R group). We analyzed the demographic data and radiological parameters, such as modified Stoke Ankylosing Spondylitis Spine Score (mSASSS), sagittal vertical axis, and various angles. We also investigated the factors affecting re-stooping by analyzing the correlation between the ratio of correction loss and various factors. Results: A significant difference was seen in the change in the mSASSS from before surgery to the last follow-up between the N group (2.87 ± 3.08) and the R group (9.20 ± 5.44). In multivariate analysis, only the change in the mSASSS from before surgery to the last follow-up was significantly correlated with the ratio of correction loss. Conclusions: Thoracolumbar corrective osteotomy seems to provide high satisfaction among patients with AS but can lead to re-stooping during follow-up. The change in mSASSS was related with re-stooping in the current study. We recommend active rehabilitative exercises and appropriate medication depending on the patient's condition, which may help delay the postoperative progression of AS.

Comparison of endoscopic and open surgery in life-threatening large spontaneous supratentorial intracerebral hemorrhage: A propensity-matched analysis.

BACKGROUND: Conventionally, open surgery (OS), including standard craniotomy (SC) and decompressive craniectomy (DC) with hematoma evacuation, is adopted to treat life-threatening large spontaneous supratentorial intracerebral hemorrhage (ICH). Recently, endoscopic surgery (ES), a minimally invasive surgical treatment, has gained increased popularity. However, the safety and efficacy of ES for life-threatening large ICH is uncertain. AIM: The aim of this study was to evaluate the effectiveness and safety of ES for life-threatening large ICH and compare it with traditional OS. METHODS: We retrospectively analyzed the clinical and imaging data of consecutive supratentorial ICH patients with preoperative Glasgow Coma Scale (GCS) score ⩽ 8, who underwent ES or OS between May 2015 and October 2021. To minimize bias in case selection, propensity score matching was performed (ratio 1:2, caliper o.2). The primary outcome was a prognosis-based dichotomized (favorable or unfavorable) outcome of the 5-point Glasgow Outcome Scale (GOS) at 6 months. Favorable outcome was defined as a GOS score of 4 to 5 at 6 months. Sensitivity analysis was also performed to ensure the robustness of the findings. RESULTS: Of 695 patients who underwent surgical treatment for spontaneous ICH, 191 patients were identified to be eligible, with 58 patients in the ES group and 133 patients in the OS group. Propensity score matching improved covariate balance and generated a comparable cohort (53 ES and 106 OS) for all analyses. The ES group had a higher incidence of the primary outcome of favorable outcome at 6 months (ES 20/53 (37.7%) vs. OS 22/106 (20.8%); propensity score-matched relative risk (RR) (95% CI) = 1.74 (1.13-2.68); p = 0.013). Sensitivity analysis showed the result was stable. CONCLUSION: ES is a safe treatment for life-threatening large spontaneous supratentorial ICH patients and may achieve better outcomes than OS.

Protocol of End-PSCI trial: a multicenter, randomized controlled trial to evaluate the effects of DL-3-n-butylphthalide on delayed-onset post stroke cognitive impairment.

BACKGROUND: Delayed-onset post stroke cognitive impairment (PSCI) results from secondary neurodegeneration induced by stroke. Whereas targeted prevention or treatment strategies are still missing due to lack of evidences. This trial aims to evaluate the preventive effects of DL-3-n-butylphthalide (NBP) on delayed-onset PSCI. METHODS: Effects of NBP on Delayed-onset Post Stroke Cognitive Impairment (End-PSCI) is a prospective, parallel-group, open-label, multicenter, randomized controlled trial with blinded outcome assessment. Hospital patients with acute cerebral infarction (within 2 weeks of onset) will be randomized into either standard medical therapy group or standard medical therapy combined NBP treatment group (NBP 200 mg, three times per day for 24 weeks). The primary outcome is the difference of incidence of delayed-onset PSCI between two groups. The secondary outcomes include difference of white matter degeneration, cognitive scores and prevalence of early-onset PSCI between two groups. DISCUSSION: End-PSCI trial will provide evidences for NBP preventing delayed-onset PSCI. The secondary outcomes will also provide valuable insights into the pathogenesis of delayed-onset PSCI and mechanism of NBP's actions. TRIAL REGISTRATION: Trialsearch.who.int , ChiCTR2000032555, 2020/5/2, prospectively registered.

The potential risk factors of early-onset post-stroke depression from immuno-inflammatory perspective.

Background: Mounting evidence strongly uncovered that peripheral immuno-inflammatory response induced by acute stroke is associated with the appearance of post-stroke depression (PSD), but the mechanism remains unclear. Methods: 103 stroke patients were assessed at 2 weeks after onset using Diagnostic and Statistical Manual of Mental Disorders, 5th edition and then divided into PSD and non-PSD groups. Polymorphisms of inflammatory molecules (interleukin [IL]-1ß, IL-6, IL-10, IL-18, tumor necrosis factor-α [TNF-α], interferon-γ [IFN-γ] and C-reactive protein [CRP]), complete blood count parameters, splenic attenuation (SA) and splenic volume (SV) on unenhanced chest computed tomography, demographic and other clinical characteristics were obtained. Binary logistic regression model was used to analyze the associations between inflammation-related factors and the occurrence of PSD at 2 weeks after stroke. Results: 49 patients were diagnosed with PSD at 2 weeks after onset (early-onset PSD). The C/T genotypes of CRP rs2794520 and rs1205 were less in PSD group than non-PSD group (both adjusted odds ratio = 3.364; 95%CI: 1.039-10.898; p = 0.043). For CRP rs3091244, the frequency of G allele was higher (80.61% vs. 13.89%) while the frequency of A allele was lower (6.12% vs. 71.30%) in PSD patients than non-PSD patients (χ2 = 104.380; p<0.001). SA of PSD patients was lower than that of non-PSD patients in the presence of CRP rs2794520 C/T genotype and rs1205 C/T genotype (both t = 2.122; p = 0.039). Peripheral monocyte count was less in PSD group than non-PSD group (adjusted odds ratio = 0.057; 95%CI: 0.005-0.686; p = 0.024). Conclusions: CRP polymorphisms, SA based on CRP genotype, and peripheral monocytes are associated with the risk of early-onset PSD, suggesting peripheral immuno-inflammatory activities elicited by stroke in its aetiology.

Medications adherence and associated factors among patients with stroke in the Kingdom of Saudi Arabia

Background: Stroke is one of the most significant neurological problems around the world, and is considered a leading cause of death. Due to polypharmacy and multimorbidity, stroke patients are susceptible to have lower levels of adherence to their medications and self-care activities. Methods: Patients who have suffered a stroke and had recently been admitted to public hospital were approached for recruitment. Patients’ adherence to their medications was examined using a validated questionnaire during an interview between the principal investigator and the patients, where patients’ adherence to their selfcare activities was assessed using a developed, validated and previously published questionnaire as well. Reasons for lack of adherence was explored from the patients. Verification of patient’s details and medications was done via the patient’s hospital file. Results: The mean age of the participants (n=173) was 53.21 (SD= 8.61) years. Assessing patients’ adherence to medications showed that more than half of them stated that they sometimes/often forgot to take their medication/s, while 41.0% sometimes/often stopped their medication/s from time to time. The mean adherence to medications score (out of 28) was 18.39 (SD=2.1), with 83.8% having a low adherence level. It is found that patients who did not take their medications were due to forgetfulness (46.8%) and complications from taking the medications (20.2%). Better adherence was associated with higher educational level, higher number of medical conditions, and higher frequency of glucose monitoring. Adherence to self-care activities showed that majority of patients performed correct self-care activities three times a week. (AU)

Acute Ischemic Stroke Presentation Masked by Falsely Localizing Motor Seizure: A Clinical Case Series.

In this case series, we describe a novel observation in which 4 patients with acute ischemic stroke secondary to large vessel occlusion and no history of seizure present with focal seizure activity localizable to a chronic, contralateral infarct. The explanation for this phenomenon is unknown but may be due to a combination of effects involving disrupted interhemispheric inhibitory connections and epileptogenic changes involving chronically infarcted tissue.

Acute Thalamic Ischemic Stroke in an Older Patient Newly Vaccinated with COVID-19 Vaccine Based on Adenoviral Vectors.

Introduction: Recent reports have shown several cases of cerebrovascular events after vaccination against COVID-19. The effects have been described mainly in women within the first two weeks of receiving the vaccine. Clinical Case: We describe here the first Colombian case of a cerebrovascular event after vaccination against COVID-19 in a 67-year-old woman with a vascular history. Four days after application of the messenger ribonucleic acid (mRNA) vaccine, she exhibited deviation of the labial commissure, ipsilateral ptosis, and limitation of march with lateralization. The event was associated with a subacute ischemic event in the right thalamus in parasagittal situation, changes in chronic ischemic microangiopathy of small vessels, and vascular crossing in the right cerebellar angle, without other alternative causes. Conclusion: The development and rapid use of vaccines has allowed the hospitalization and mortality statistics associated with COVID-19 to be reduced, but at the same time, it has generated concern about the potential side effects, generating controversy among the general population, especially in individuals with cardiovascular diseases. In our case, we provided evidence for the discussion of potential cerebrovascular events related to the application of vaccines in older people with a history of cerebrovascular diseases. This was done in order to analyze and control in subsequent studies the modulation of medical history on the likely effects of vaccination. However, despite the unavoidable side effects, the benefits of vaccination are superior.