MODERN APPROACHES TO CHLAMYDIOSIS TREATMENT

     Chlamydia in the medical and popular literature usually relate to diseases transmitted through sexual contact. At the same time almost all reputable studies clearly show - chlamydia is currently the most common disease of the diseases transmitted primarily through sexual contact. A detect chlamydia every second of the surveyed women with inflammatory diseases of the urogenital area, with two thirds of women suffering from infertility, in 9 out of 10 women with recurrent miscarriages. In men, every second is caused by chlamydia urethritis. Not so long ago associated with the infection cause chronic diseases of the respiratory and digestive systems, the development of atherosclerosis and its complications, cancer and many other serious disorders. Is it all so seriously? And only a sexually transmitted infection chlamydia can be? How dangerous are its complications? And is it possible to cure this disease?
Numerous studies on the basis of which revealed the prevalence of chlamydia, held at the end of the last century. A first classification of microorganisms, which allowed to justify the treatment, set up in 80-th years (Table 1).

Table 1.
Classification of chlamydia, adopted by the International Association of Microbiological Societies (IAMS) in 1980


At present, it is safe to say that many of populyarizuemyh at the time of the facts and opinions (many of which are repeated unnecessarily and recklessly in the popular press of the present time) have undergone major changes. This was the result of studies and acquisition of new knowledge about chlamydia. Referred to changing attitudes affected the revision of the virulence of the infection, clinical course and prognosis of diseases caused by them, their epidemiology, etc. [1, 2]. And that is likely to be elaborate.
Modern classification of chlamydia adopted relatively recently - in 2000 - and includes four families and five genera of microorganisms. In each genus is represented by one to six species, differing in the structure of genes, which allowed to systematize the available clinical information to a qualitatively new level. In particular, as previously tipiruemyh isolated pathogens Chlamydia genus Chlamydophila, includes nizkovirulentnye types of microorganisms. Previously, a number of reasons (morphological complexity and technical nature), they are not separated. And if they were isolated, it is not on the grounds of virulence. For example, Ch. psittaci featured on Ch. trachomatis on the basis of resistance to sulfadiazine. Although not all of C. psittaci are resistant to it. A Ch. pneumoniae were classified according to its appearance under the electron microscope, although it was found that they differed in the different research groups [3, 4].

Table 2.
Classification of chlamydia, adopted at the IV European Congress of "Chlamydia-2000" in Helsinki

The essence of this separation is that only the original family of Chlamydia (in the overwhelming number of cases of Chlamydia trachomatis) can cause specific diseases in humans. A classified in the genus Chlamydophila only mimic the presence of chlamydia by their presence (or rather "hlamidiopodobnoy") infection.
Etiopathogenesis. Despite the fact that in this matter is not clear, from a practical point of view (the prevention and treatment), chlamydia is now the former problem can be considered solved. Some problems exist in the use of conventional methods for diagnosis of infection, as discussed more below. Similarly, because of the complexities of former and current topical diagnosis of chlamydia is unacceptable, some previously accumulated arrays melon.
All species of Chlamydia are obligate intracellular bacteria. Have two forms of life (elementary and reticular cells), and biphasic cycle of development, have a penchant for persistence. Elementary body is metabolically inactive, the extracellular form. Have a diameter of 250 - 500 nm and a rounded shape. In the body get chlamydia in the form of elementary bodies and, therefore, has called this form of infection. Reticular body is intracellular, metabolically active form. It has a larger diameter - from 300 to 1000 nm - and may have a different shape. These unique of its kind microbes occupy an intermediate position between viruses and bacteria, possessing some properties of both. The presence of the cell wall at the stage of the reticular cells does chlamydia like bacteria accessible to antibiotics. A difference in the morphology of various phases of development determines the difficulty of diagnosis. For example, some types of microorganisms of this group have a third form.
At present, the ability to cause specific diseases in humans has been proved only for the three types of chlamydia.
1. Chlamydia trachomatis - found only in humans and causes a wide spectrum of disease (urogenital disease, conjunctivitis, some forms of arthritis, myocarditis, etc.). Although the frequency and nature of their different in nature. 18 known serotypes of this organism and that from this point of view it is convenient to consider the characteristics of the course of infection. For example, serotypes A, B, Ba, and C are the causative agents of trachoma (an infectious disease that can lead to blindness). Vectors are insects, and the main route of infection - rubbing eyes with dirty hands. At the same time, from an epidemiological point of view, some sort of danger is only serotype Ba (North America) and, to a much lesser extent, serotype A (Middle East and North Africa). Serotypes L1, L2 and L3 can cause a relatively rare Lymphogranuloma venerum (venereal limfogranulemu) and, as casuistry, haemorrhagic proctosigmoiditis.
And that serotypes D, E, F, G, H, I, J and K cause urogenital chlamydiosis, which is the main route of transmission is sexual. In addition, they are able to conjunctivitis (serotype D) and neonatal pneumonia (serotype E). Very rarely are the cause of paratrahomy adults. And from a medical point of view it is a disease of epidemic interest [2, 5, 6].
The other two representatives of the genus Chlamydia - Ch. suis and Ch. muridarum - do not cause infectious processes in humans, being highly specific for the disease, respectively, pigs and rodents of the family Muridae (mice and hamsters).
2. Chlamydophila pneumoniae - in spite of their belonging to the "hlamidiopodobnym" microorganisms capable of causing endemic (sporadic) cases of pneumonia-specific. Locally, the incidence may be 60-84 cases per 1000 population. Infection occurs mainly bronchitis and mild forms of pneumonia (with a tendency to chronicity of the process). Infection is airborne and air-dust paths in the presence of mandatory environmental conditions (warm, and dry air). The most vulnerable are those aged 20 to 49 years. At the same time found that 70-90% of infected process is latent, without severe symptoms. It is assumed that in rare cases can cause specific meningoencephalitis, arthritis, myocarditis, and Guillain-Barre syndrome. Currently, however, these data are reviewed [1, 7].
3. Chlamydophila psittaci - an absolute zoonotic disease - an infection transmitted to humans from animals. The potential to cause disease in birds and can then be transmitted to humans, causing quite formerly known disease - psittacosis. Modes of transmission is airborne and air-dust. In the United States each year not more than 200 cases, although experts believe the actual number is greater. Endemic among the parrots and pigeons, causing sporadic disease in other mammals (except man), as well as turtles. According Prukner-Radovcić E. , et al. [8], in the big cities infected pigeons reached 15.83% in the absence of antigen-positive individuals among wild birds. As a potential source of infection for humans is important for the elderly, children and immunocompromised patients, as well as poultry. Serovars C. psittaci C and D are occupational hazards for workers and for the slaughter of people in close contact with birds. Serovar E (also known as Cal-10, MP or MN) isolated from different poultry flocks around the world. Although he was connected with the epidemic psittacosis in people 20-30s of the last century, the concrete tank had not been identified [9, 10].
Psittacosis often begins with flu-like symptoms and becomes life-threatening in the development of chlamydial pneumonia. It is sometimes not quite right referred to as "atypical." Very rarely is believed to be possible specific arthritis, pyelonephritis, entsefalomiokardit. Although these data are now being actively monitored. Infected birds treated with tetracycline or special forms of chlortetracycline in the form of granules that are added to the feed for 45 days. In addition, you must disinfect the cage, dishes, and care items (such as Lysol solution).
Below mentioned microorganisms, previously positioned as a human infection, but is currently excluded from among them:
1. Chlamydophila abortus - cause disease only in animals and epidemichny of ruminants - causes abortion (fetal death) in horses, rabbits, guinea pigs, mice. Is widespread in cattle and is associated with clinical mastitis cows. Transmitted among mammals, and oral infections, but not detected in birds. Currently, there is a question of general vaccination of cattle since the disease brings significant economic losses [3, 11]
2. Chlamydophila pecorum - cause disease in animals, such as Ch. abortus (including the cow mastitis). Cows, horses, goats, pigs, and koalas. Different serotypes cause infertility, inflammation of the urinary tract, as well as abortion, conjunctivitis, encephalomyelitis, enteritis, pneumonia and polyarthritis [11, 12].
3. Chlamydophila felis - the cat causes rhinitis and conjunctivitis, which is also transmitted in the form of specific, but does not require treatment, conjunctivitis people. According to some reports, infection of cats with conjunctivitis is 12-30%, and among unvaccinated domestic cats antibodies were detected in 10%. Infection without symptoms is, according to the PCR, 2-3% [2, 13].
4. Chlamydophila caviae - guinea pigs cause inflammation of the eye and conjunctival sac. It is specific to them and non-invasive for other animals. It is also possible infection of the genital tract with the development of clinical pigs, similar to human infection Ch. trachomatis. The microorganism affects only the mucosal epithelium and is not invasive. Despite extensive research 2003-2006, failed to show C. caviae as a pathogen for humans [14].
Epidemiology. In general, the role of pathogen Chlamydia trachomatis is a ubiquitous Ch. trachomatis. Extensive studies show that only young people who are infected with chlamydia in the world no less than 30%. At the same time in the reproductive age of women in different regions from 30 to 60% and men - at least 50%. The incidence of chlamydia in Russia does not have an epidemic level, some, such as gonorrhea, but is 2.5 times lower incidence of trichomoniasis - 99-101 case per 10,000 inhabitants. In Ukraine, it is more than 50 cases per 100 000 population. In the country, according to some estimates, roughly 4-8% of infected population [15, 16].
In the UK, chlamydia is the most common of all sexually transmitted infections. 4-4.5 thousand are diagnosed each year of diseases with a tendency to increase their number and frequency of complicated forms (about 2.6%). Most often the disease is detected in the groups of men and women 20-24 years - 16-19 years [2, 7] (Fig. 1).


Fig. 1. The incidence of chlamydia in index DALY (a weighted measure of death, illness and disability) in different regions of the world (per 100 thousand population)

In the United States become ill each year 3-4 million people with chlamydia with a tendency to increase. Ch. trachomatis is the cause of half of NGU in men. It is detected in 3-5% of healthy men who were hospitalized in ordinary hospitals and up to 15-20% of men admitted to the STI clinic. Do gay men Ch. trachomatis in the urethra is found in 4-5% of cases, and in the rectum - 4-7%. The material, taken from the cervix, Chlamydia were detected in 5% of women without any symptoms of the disease and at least 20% of women receiving treatment at STI clinics. The highest infection was observed among the population aged 17-25 years, where Ch. trachomatis detected 10 times more common than gonorrhea. According to various estimates, about 5% of U.S. adults are infected with chlamydia. In this infection is the greatest factors belonging to the following groups: 1) people under 25 years old, 2) the citizens, and 3) African-Americans and 4) people with low socio-economic status. In 50-70% of children born to mothers ifitsirovannymi, get an infection from birth. In a number of specific cases are reported of the eyes, rectum, vagina and the back of the neck. And 30-40% of these children develop complications of chlamydia in the form of conjunctivitis and pneumonia [3, 13].
Established ways of transmission of chlamydia are:
1. Sexual contact (vaginal or anal sex, much less - oral). Characteristic of Ch. trachomatis.
2. Airborne (characteristic under special conditions, for Ch. Pneumoniae, as well as Ch. Psittaci).
Contact-household, in which the infection is transferred with contaminated hands, objects, etc. currently denied. Therefore, the cause of contamination can not be toilet seats, swimming pools, sauna, common utensils and towels. Understandable exception to this rule is trachoma, which is characteristic for some African countries.
The probability of infection through unprotected sexual contact (vaginal, anal) with patients with chlamydia is about 50%. At the same time, we know that in monogamous sexual relationships of about 3-5% of cases of chlamydia occur in only one partner. In the presence of mixed infections are more likely to gonorrhea or trichomoniasis disease with a decrease in the likelihood of chlamydia. One study showed that women, patients with gonorrhea, infected their sexual partners of men in 81% of cases, while suffering from chlamydial cervicitis passed the infection in only 28% of cases. And if the women had both infections, the transmission of gonorrhea occurred more frequently (77%) than that of chlamydia (28%). The incubation period of chlamydia is from 1 to 3 weeks.
Symptoms of chlamydiosis are very diverse, as relates to persistent chlamydia infections, ie, flowing long and asymptomatic. In the latter case, this concerns 25-50% of affected men and 67-80% of women. In other cases, as a rule, the manifestations of the disease is minimal. This may be a discomfort in the abdomen, a violation of urination (itching or pain, frequent urge), whitish or clear discharge from the urethra. Women can change the nature of menstrual bleeding (their shortening and change of color) and intermenstrual appear. The appearance of scanty discharge from the urethra and vagina are usually short-lived and observed after 7-30 days after infection. It was asymptomatic chlamydia causes a significant risk of specific complications. Although it is worth noting that they are much more pronounced for women [2, 7, 15].
Proctitis is more typical for homosexuals and is manifested in different severity of pain in the rectum and false desires (tenesmus), but can occur without subjective symptoms. And the most frequent clinical manifestation of infection in women is cervicitis, which is observed in 30-60% of cases in combination with gonorrhea. Independently, he normally bessimptomen. Scarce leucorrhoea observed in only 37% of women. Most often there is a slight itching or discomfort in the genital area.
A very specific symptoms determines the articular syndrome with chlamydia in both men (mostly) and women. Historically, it is called Reiter's syndrome or oculo-urethro-synovial syndrome. Previously, it was linked to Shigella, Yersinia and Salmonella, but now clearly understood as the result of chlamydia. Clinically, Reiter's syndrome is a combination of symptoms of urethritis (all patients), joints (in 90-95% of patients) and eyes (conjunctivitis in 30-40%). Symptoms do not always appear simultaneously, but it begins with urethritis, and then joined by other symptoms. Most - 1-2 weeks after the start of urethritis, and occasionally a few months later. When articular pain syndrome with predominant involvement of multiple major joints (usually 4-5). Involved, rather than simultaneously, one after another. The most commonly affected knee (70%), followed by ankle (50-60%), small joints of the feet (40%), shoulder (20%), wrist (15%), hip (15%), elbow (n = 7 % of patients). There is Unsharp pain in the joints, the skin over it becomes hot to the touch, sometimes congested. Maybe there is swelling of the joints, very rarely - effusion. Many patients develop pain after physical exertion, often accompanied by myositis, muscle atrophy. He suffers from a general state of health of patients. In 20% of patients with articular syndrome temperatures can reach 38-40 ° C.
Possible damage to the skin and mucous membranes. The most typical are clinic balanitis, changes in the oral mucosa with erosions and desquamation of the plots, and keratoderma. It begins with a rash of red spots on the palms, soles, or across the skin surface. The spots turn into blisters, then pimples conical horn, or thick, crusted plaques. Often the affected nails (thickening, brittleness nail plate).
For Ch. pneumoniae, as mentioned above, in 10-30% of infection is characterized by the development of mild forms of pneumonia. At the same time all the possible development of joint syndrome and, less frequently, other lesions. But they all proceed more smoothly compared to Ch. trachomatis.
Complications. For men, the risk of specific complications is much lower, but increases with repeated infection with chlamydia. Previously it was thought that chlamydia in men often leads to prostatitis, which is now not recognized. Similarly, question the possibility of infertility in the background scarring of the vas deferens [6, 17].
Epididymitis. Chlamydial infection is prevalent in much the role of the causes of inflammation of the epididymis in young men. Only after 35 years, as evidenced by the literature, begin to dominate the bacterial agents. For such a combination of epididymitis necessarily with urethritis. There is swelling, tenderness and pain in the scrotum (usually on one side only), often with fever. The pain is so severe that the patient does not walk, but may be weak, whining, and not cutting nature.
Endometritis and salpingitis develops in the absence of adequate treatment for women and are the result of the further spread of chlamydia from the cervix. It is now established the prevalence of chlamydial infection is, and not as previously thought, the bacterial microflora. In certain cases, the infection spreads to the surface of the liver with the development of perigepatita (about 5% of women with salpingitis and endometritis). Develops pain in the upper right quadrant of the abdomen, which is accompanied by nausea, vomiting and fever. As a result of chronic inflammation of the fallopian tubes, infertility can develop. According to various estimates, its incidence is 10-40% of untreated cases of chlamydia in women. In addition, the background of specific inflammation (salpingitis) increases the risk - in descending order - an ectopic pregnancy, chronic pelvic pain, pelvic peritonitis, miscarriages and premature births. About 5% of the women of this population have a form of liver disease - perihepatitis - which is considered as a separate form of chlamydia, and as a complication [4, 18]
Diagnosis. Each of the methods of diagnosis of chlamydia has a number of advantages and disadvantages, have varying degrees of sensitivity depending on the duration of the disease and the localization process. For this reason, use at least two ways from different groups.
Methods for direct detection
Microscopy of stained preparations. Stained cytological smears by the method of Romanovsky - Giemsa.
The culture diagnosis. Isolation of the pathogen to the cell culture medium McCoy or HeLa. The advantage is its high sensitivity and specificity, and lack of - the high cost due to the requirements for material and technical equipment of laboratories and staff.
Methods for DNA diagnostics. Have high sensitivity and specificity. Require the taking of material for analysis directly from the source localization of the pathogen.
A method of direct immunofluorescence (DFA) allows to identify chlamydial inclusions by the characteristic yellow-green glow. A significant drawback is the subjectivity of evaluation, which is largely dependent on the skills lab, and low sensitivity for small amounts of antigens of Chlamydia. At the present time is rarely used.
Methods of indirect detection
Immunosorbent assay (ELISA). A widely used method, although the accuracy did not exceed 50-70%. Based on the detection of antibodies produced in response to the introduction of the pathogen. In the presence of a combination of antibodies of classes A, M and G gives an indication of advanced disease. For example, the IgG titers indicate the presence of chlamydia, and IgA titers indicates infection within the previous 2 weeks.
The method of polymerase chain reaction (PCR). It is also widely used technique, the accuracy of which reaches 90-95%. For several reasons the method is strongly recommended to combine with ELISA.
Using these methods due to their rather difficult reproducibility. Methodological violations in conjunction with a false-positive reactions of some diagnostic kits manufacturers, according to some reports, cause up to 30% of the overdiagnosis of the disease. What makes the prescribed treatment is not justified. And that is why there is often recurrence of diagnostic recommendations in two different laboratories [15, 16].
In addition to these subjective reasons rather inaccurate diagnosis of chlamydia, there are more substantial. For, despite the high accuracy of the PCR, it does not reach 100%, which are required to exclude infection. In this context, low titres of antibodies (1:20 and below) is recommended to repeat the study after 1 month. At the same time, while maintaining a negative PCR and monotone low titers of antibodies in the blood according to the ELISA is possible to ascertain the absence of infection. And to avoid unnecessary treatment with a positive ELISA result is absolutely necessary to confirm the PCR-scrapings. In this case a negative PCR test does not exclude the presence of chlamydia, which requires monitoring (more frequent and repeated) is according to the EIA. Among other reasons, some errors should also mention increasing titer of IgG to a recent infection, especially those newly transferred, against the effect of immunosuppressive viruses. Similar effects occur after antibiotic treatment, which also have immunosuppressive properties. However, in the latter case, such an increase in titer is not beyond the laboratory error (rate fluctuations).
Do not forget to check other sexually transmitted diseases, since the frequency of a combination of chlamydia with gonorrhea or trichomoniasis is very high.
Treatment for chlamydiosis is quite simple and in most cases does not cause problems. The basis of treatment is antibiotics - macrolides, tetracyclines, or - although, contrary to popular opinion, can be used, and representatives of other groups. For 95% of patients a course of antibiotics will be sufficient for a complete cure. If there are complications, as a rule only in women who may need treatment, supplementing antibiotics (immunotherapy, restorative drugs, physical therapy, etc.), but this therapy is situational. The definition of "sensitivity" of chlamydia to antibiotics is not required because they are absolutely sensitive to tetracyclines and macrolides.
Tetracyclines (doxycycline) and macrolides (azithromycin) is officially considered drugs of choice for treatment of chlamydia. For an alternative means include erythromycin (erythromycin and erythromycin ethyl succinate), and some fluoroquinolones (Table 3). But be aware that some drugs from the last-mentioned group, such as ciprofloxacin, are not sufficiently effective [7, 18].
Table 3 identifies some of the most commonly used in the present treatment regimens.
Table 3.
Used antibiotic regimens


Against chlamydiosis on background of chronic respiratory disease (COPD, asthma, etc.) are recommended regimen of antibiotics for at least 14 days. Shorter courses of treatment, as experience shows, these conditions do not provide adequate sanitation, and do not protect against recurrent chlamydiosis.
In pregnancy, macrolides, and used as an exception, allowed the use of penicillin (amoxicillin). At the same time features of the effect of the latter on the life cycle of chlamydia, with their inherent incidental activation of dendritic cells and the secondary immune response, causes the growth rate of the articular and Reiter syndrome. In addition, they are not efficient enough. That is why the use of penicillins involves the need to re-culture results in 3-4 weeks. As an alternative, you can use in pregnancy clindamycin, which provides a high cure rate (99%), but this strategy is much more expensive therapy [17, 19].
A single dose. A number of randomized trials confirm the high efficacy of azithromycin after a single oral dose of 1 g (treated in 97-100% of cases), a significant reduction in the incidence of side effects from the gastrointestinal tract and the absence of adverse effects on the fetus. Also recommended during pregnancy [15, 20].
With concomitant gonorrhea is recommended to further single dose fluoroquinolone (ciprofloxacin 500 mg, 500 mg of levofloxacin or gatifloxacin 400 mg), if other treatment is antibiotics. The combination of the infection with chlamydia commonly observed in approximately 50% of cases. There are data that show an increase in the frequency of transmission of gonococcal infection of the partner against chlamydia. This requires in most cases, a more thorough preventive re-infection [5, 16, 21].
In the accompanying mycoplasmosis correction schemes mentioned above is not required because they provide a cure both infections simultaneously.
The prognosis for cure for chlamydiosis favorable. As a rule, only long-term untreated chlamydial infection over time leads to the emergence of various types of complications caused by the spread of germs to other organs and tissues. And much more likely in those women.
Prevention of chlamydiosis and its complications. The same as for other sexually transmitted diseases. It should be clearly aware that the infection is no way to contact-household (with the usual kiss, taking baths, a towel, utensils, toilet seats, etc.). Available in the popular literature of the nature of the warnings are sometimes too much to prevent unsanitary conditions.
In most cases, within the next two months after the recovery should proceed with the surrender of strokes (once a month to check for recurrence of chlamydia appears to be sufficient). Titers of antibodies to chlamydia decreased by ELISA for 6-12 months.
Recommendations for prevention of primary infection.
1.      Use only latex condoms during sexual intercourse. Most of the implemented during outside the pharmacy distribution network ("no latex") are not a barrier to chlamydia.
2.      Avoid sexual contact with partners "high risk". These are men and women 20-25 years, persons, leading promiscuous and people of lower socioeconomic status.
Recommendations for prevention of secondary infection
3.      Report the infection you have found all their sexual partners. They also need to be diagnosed for further treatment (if necessary). This way you tell your re-infection.
4.      Regular sexual partners - after examination - must be treated simultaneously.
5.      Inspect again if symptoms of chlamydia and continued there after some time. The presence of the disease in the past does not give immunity, ie, does not protect you from re-infection.
6.      Be sure to completely finish the entire course of antibiotics that is prescribed for you even though you feel fine and have no symptoms.
Further research and some scientific data. With regard to potency, promotions and pathogenetic effects of chlamydia on a number of diseases rather large-scale studies have been underway for more than 30 years. In this respect, particular interest is Ch. pneumoniae, disease manifestations in humans is mediated by a variety of conditions. Not so long ago have been completed in-depth study of the relationship between chlamydial infection and the development of atherosclerosis and associated coronary heart disease. Despite some data, where it was shown that infection with Ch. pneumoniae may be a factor in the development atherosclerotic plaques, it was recognized that significant differences between groups of patients were detected. But research in the prevention of complications of atherosclerosis (myocardial infarction, stroke) with antibiotics still in progress [5, 12].
There is evidence on the impact of Ch. pneumoniae in the formation of arterial aneurysms. Suffice common in the age groups 65 years and older, as well as atherosclerotic narrowing of blood vessels, they are often the cause of various complications. Back in 1996 Ong et al. [22] reported the detection of microorganisms in 11 out of 25 investigated abdominal aneurysms. Since that time held more than 12 special studies, which also confirmed certain laws, including those in the experiment [18, 23].
Ch. pneumoniae with increased frequency is detected in the cerebrospinal fluid of patients with multiple sclerosis. In this treatment tetracycline antibiotics has given some encouraging results. Although those who do not yet allow clear to talk about the role of chlamydia in the etiopathogenesis of this disease [4, 6].
A significant number of studies conducted to determine the relationship of infection Ch. pneumoniae with lung cancer. In a meta-analysis found a slight increase in incidence in the background serologically confirmed the presence of infection. But while the need for more extensive study of this issue. Similar, very interesting data were obtained in respect of diabetes mellitus type 2 and some forms of obesity. However, all these studies are so far beyond the practical applications [14, 21, 24].

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