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Analogue of ampicillin, is a semisynthetic antibiotic with essentially the all patients who present agar (Biokar®) were prepared and sterilized according to the manufacturers’ instructions. Another drug and may not reflect the rates.

March – Please Check NICE guidance for all blood cells (erythrocytes) and the lining ongoing antibiotic use not directed against. That.

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The treatment duration of the study will be at least 5 days which may increase to 7 days, if Investigator feels the need of continuing the treatment at Day 5.

In such cases clinical assessment will be done again at Day 7.

Thus the efficacy evaluation will be performed at baseline (Day 0) prior to study drug administration and on Day 5 and/ or 7.

Safety laboratory evaluations will be performed at Screening Visit and will be repeated at end of study treatment i.e.

On Baseline, Day 2, Day 5 and/ or Day 7, Visual Analogue Scale (VAS) will be used to measure amount of pain and amoxicillin for ear infection in adults swelling that a subject will experience.

Zero would mean "No

pain

" and 10 would mean "Worst possible pain". The subject will be asked to choose the most appropriate score on VAS which best describes his/her pain status over last 24 hours.

While using VAS for swelling, zero would mean "No swelling" and 10 would mean "Maximum possible swelling". The Investigator will choose the most appropriate score on VAS which best describes subject's swelling. Sample for microbiological culture will be obtained at the discretion of Investigator while performing the surgical procedure.

When microbiological culture is indicated and feasible, two sets (each consisting of aerobic and anaerobic media) will be obtained. This sampling will be done in approximately 80 subjects randomly who will provide consent at Screening. The sample will be collected and dispatched in appropriate transport medium to the central laboratory for

culture

and antibiotic susceptibility.

Protocol waivers or exemptions will not be allowed. The total duration of each subject's participation will be 6-9 days including Screening

period

of 0-1 day (Day -1 to

Day

0) and Treatment period of 5-7 days (Day 0 to Day 5-7). The primary objective of the study is to compare clinical efficacy (cure and improvement) of both the treatment arms which uses VAS for assessment of pain and swelling. Since the clinical efficacy parameters are

subjective

, it is mandatory to keep the observer/Investigator blinded during the study assessment. In addition, designing blinded study is challenging owing to different dosage regimen and formulation of study drugs.

The

study design of double blind double dummy is not recommended in view of increased pill burden which in turn can have an impact on treatment compliance. The possibility of over-encapsulation is ruled out due to large pill size of amoxicillin + clavulanic acid.

Therefore to maintain blinding of Investigators, the study is designed as observer blind with Investigator remaining blinded throughout the study period. An unblinded study team member will be appointed for each site who will be involved in dispensing of the study drugs to subjects. The unblinded study team member will remain present during subject assessment for all study visits and will ensure that Investigator remains blinded to treatment assignment. Study treatment includes amoxicillin + clavulanic acid (875mg/125mg) two times daily or clindamycin (150mg) four times daily for 5-7 days both administered

orally

with meal. Augmentin™ (amoxicillin + clavulanic acid) from GSK and Dalacin-C (clindamycin) from Pfizer will be used as study treatments.

The contents of the label will be in accordance with all applicable regulatory requirements.

Patients can

receive

additional medical therapy such as analgesics or anti-inflammatory drugs. However,

opioid

analgesics and any other antibiotic apart from study drugs will not be permitted during entire study period. This study will be conducted in

accordance

with ICH GCP, all applicable subject privacy requirements, and the ethical principles that are outlined in the Declaration of Helsinki 2008, including IRB/ IEC review and approval of study protocol and any subsequent amendments, subject informed consent and Investigator reporting requirements. Maria Tereza Ferreira Duenhas Monreal I ; Paulo Camara Marques Pereira II ; Carlos Alberto de Magalhaes Lopes II. I Federal University of Mato Grosso do Sul (UFMS), Campo Grande, MS II State University of Sao

Paulo

(UNESP); Botucatu, SP, Brazil.

The intestinal tract harbors a huge diversity of metabolically-active aerobic and anaerobic bacteria that interact, forming a complex ecosystem.

This microbiota has an important role in human metabolism, nutrition, immunity, and protection against colonization by pathogenic microorganisms.

Several factors can influence the intestinal microbiota; these include age, diet, inflammatory and infectious processes, and the use of antimicrobials.

We investigated the influence of bacterial infection of the respiratory tract and of amoxicillin therapy on the normal intestinal microbiota of patients.

Bacterial infectious processes affecting the respiratory tract were found to influence the intestinal microbiota, significantly decreasing the number of colony-forming units (CFUs) of Bacteroides spp.

The use of amoxicillin also influenced the intestinal microbiota, significantly decreasing

the

CFU of Bifidobacterium spp.

Changes in the composition of the intestinal

microbiota

need to be observed, since a decrease in the normal microorganisms can pose a number of hazards for hosts, including decreased resistance to colonization. With proper follow-up, health-care teams can minimize such hazards by implementing suitable therapy- and diet-related measures, thus reducing the occurrence of detrimental effects on the gastrointestinal ecosystem.

Key Words: Intestinal microbiota, bifidobacterium, bacteroides, lactobacillus.

The gastrointestinal tract harbors a huge diversity of aerobic and anaerobic bacteria that interact in a complex ecosystem [1].

This microflora comprises 400 to 500 metabolically-active bacterial species, which have a pronounced impact on the host's intestinal function and health [2,3].

There is evidence that the dominant profile of

anaerobic

bacteria usually found in adults is established in the first four years of life [4].

Overall, intestinal bacteria can be grouped into species that have detrimental effects on the host and species that have beneficial effects.

The detrimental effects include diarrhea, infections, liver damage, carcinogenesis, and intestinal putrefaction.

Inhibition of harmful bacteria (a mechanism known as 'resistance to colonization') [5], stimulation of the immune system, improvements in the digestion and absorption of essential nutrients, and vitamin synthesis are examples of the protective effects brought about by the intestinal microbiota [6]. The normal microbiota acts as a barrier, preventing colonization by potentially pathogenic microorganisms and an overgrowth of microorganisms that are already present, such as yeasts, which can cause systemic infections in immunodepressed patients, and Clostridium difficile , which can be a cause of diarrhea and pseudomembranous colitis [7,8]. Microorganisms of the genera Bifidobacterium and Lactobacillus perform a variety of functions important for the host's

health

. Whereas microorganisms of the genus Bacteroides have beneficial as well as detrimental effects [9], those of the genus Lactobacillus contribute to sustaining resistance to colonization, possibly by producing acetic and lactic acids, which lower intestinal pH, thus preventing overgrowth of many potentially pathogenic microorganisms, whose spread is curbed by intestinal acidity. also produce hydrogen peroxide, which prevents the development of yeasts ( Candida albicans ) [10]. In addition, they can stimulate cells of the immune

system

, inducing the production of IL-12 by mononuclear cells of the peripheral blood [11]. They are also capable of converting cholesterol into coprostanol, which, being less soluble, can be excreted; this mechanism precludes its absorption and consequential increase in plasma [12].

and Bifidobacterium spp., when administered in food items known as probiotics [13-15], can survive the transit through the gastrointestinal tract and temporarily settle in the intestine [6], with a number of desirable effects on the immune system, such as stimulation of phagocytic function [16] and control of the balance of pro- and antiinflammatory cytokines [15].

Bifidobacteria constitute a numerically-important group that is capable of a wide

variety

of biological activities important for host health.

One of these activities is an inhibitory effect against other species, often preventing colonization by invasive pathogens [17,18].

It has been suggested that the inhibitory mechanism is related to the production of acetic and lactic acids and other wide-spectrum antimicrobial compounds [19]. In fact, microorganisms of the genus Bacteroides are nutritionally versatile, being able to use a wide range of carbon sources.

They are responsible for most of the digestion of polysaccharides that takes place in the large intestine [20,21].

play an important function in the mechanism of resistance to colonization by C.

difficile [22]; they are found in large numbers in the large intestine [23].

Although the intestinal microflora remains relatively stable throughout life [8], factors such as disease and certain drugs can affect this balance [17]. This ecosystem can also be influenced by diet, geographical location, and gastrointestinal surgery [24].

Over the past years, studies have revealed the importance and participation of the intestinal microbiota in pathological processes, such as rheumatoid arthritis and atopic diseases [25-30]. Infections in the respiratory tract have also attracted the attention of investigators, as these infections are

commonly

seen in general clinical practice.

Although antibiotics are routinely prescribed to treat such episodes, one result of antimicrobial therapy may be a reduction in the number of microorganisms that normally live in the gastrointestinal tract, which allows for an overgrowth of bacterial species that are already present and consequent colonization by potentially-pathogenic microorganisms [31].

Amoxicillin (aminopenicillin) is the drug of first choice for the treatment of respiratory tract infections.

This beta-lactam antibiotic with bactericidal action is widely prescribed in clinical practice, particularly when a patient's airways are compromised [32]. Knowledge on the influence of infectious processes and antimicrobial agents on the gastrointestinal ecosystem is highly important clinically, since this microbiota has functions that affect host health.

We investigated the influence of respiratory tract infections and of amoxicillin therapy on the normal intestinal microbiota of patients.

A prospective study was carried out on 42 individuals distributed into two Groups: Group 1 (G1) : 22 patients with bacterial infections of the respiratory tract (sinusitis, pneumonia), of both sexes, 19 to 50 years old, seen at the emergency department of the Hospital das Clinicas of the School of Medicine of the Universidade Estadual Paulista (UNESP) in Botucatu, SP, Brazil, from July to December 2002. Control Group (CG) : 20 blood donors of both sexes, 18 to 50 years old, screened at the blood center of the same institution over the same period.

Criteria for inclusion : patients of both sexes, aged 18 years and over, with

epidemiological

, clinical, and imaging diagnosis of acute bacterial infectious diseases of the respiratory tract.

Criteria for exclusion : pregnancy; lactation; women with hormonal disorders; individuals with other underlying diseases; use of medication, particularly antimicrobials, within the

past

30 days.

The diagnoses of infection with bacterial microorganisms were based on clinical and epidemiological data and on nonspecific supplementary exams (

complete

blood counts, chest and/or sinus radiographs).

Blood counts revealing leukocytosis and radiographic tests showing sinus opacity (sinusitis cases) and/or revealing lung condensation (pneumonia cases) were considered indicative of bacterial infection.

The nutritional assessment took into account dietary aspects and anthropometric measurements (weight and height).

Three-day intake records were used for food intake assessment.

The calculations of total calories, carbohydrates, proteins, lipids, and other specific nutrients ingested were performed with the program Virtual Nutri [33]. In each group, the nutritional classification was based on body mass index (BMI) [34].

The intestinal bacterial microbiota was assessed with the method proposed by Sutter et al. Stool samples were collected from both groups in sterile containers with Transbac transport medium (Probac).

The interval between sample collection and laboratory handling did not exceed 1 h [36,37]. Three stool samples were collected from patients in Group 1: before treatment (time point T 1 ), at the end of treatment (time point T 2 ), and 30 days after treatment (time point T 3 ).

Only one sample was collected from each individual in the control group (time point T 0 ). A 1-g aliquot was taken from each sample and transferred into a screw-capped test tube containing 9 mL of Stuart transport medium (Oxoid).

After homogenization, successive dilutions up to 10 -8 were prepared using the same eluent. Kanamycin-vancomycin blood agar, Bifidobacterium medium, and Lactobacillus selective medium (Probac) were the selective culture media used for microorganisms of the genera Bacteroides , Bifidobacterium , and Lactobacillus , respectively.

Once inoculated and identified, the plates were placed into GasPak anaerobic jars (Permution) and incubated at 37?C for 48 h in a low-oxygen and

high-carbon

dioxide

atmosphere generated by an Anaerobac system (Probac). After that, the plates were evaluated for bacterial growth and colony aspect.

Colony-forming units (CFUs) were counted for each plate, and the mean values for each type of microorganism were calculated. Microorganism concentration was expressed as log 10 CFU/g of feces. Observations related to colony morphology, Gram staining, and catalase testing were recorded for each plate. Mean values and standard deviations of log-transformed data and mean values at the original scale were calculated for Bacteroides, Bifidobacterium , and Lactobacillus counts [38]. Time points T 1 , T 2 , and T 3 (Group 1) were compared by using Friedman's nonparametric test with calculations of c 2 and p statistics. Comparisons of both

groups

at each time point were performed with the t -test for two independent samples (using log-transformed counts) with

calculations

of t and p statistics and/or Mann-Whitney's nonparametric method.

The comparisons were considered significant whenever p. The study was approved by the Research Ethics Committee of Hospital das Clinicas of the School of Medicine of UNESP in Botucatu.

The distribution of subjects by sex

was

homogeneous in both groups (50% males and 50% females). there was predominance of

the

41 to 50

year

(45%) and 21 to 30 year (30%) age ranges, whereas in Group 1 the 21 to 30 year (40%) and 41 to 50 year (30%) ranges predominated.



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