In 1988, Italy has been carrying out a review of all published cases of NCC since the beginning of the century and 11 cases were reported, most of them in Sicily. [130] Figueredo de Coimbra warned of an increase in the incidence of NCC in Portugal by the presence of population from African colonies. [130] Stanol and collaborators propose a new prognostic classification of NCC (Español, 1986). Madrazo and Sandoval published a new classification of neurocysticercosis in 1989. [146] In the 1980s there was much emphasis on the immunological diagnosis of the NCC existed a large number of publications highlighting mainly studies on the enzyme linked immunosorbent assays [147-155] and immunoelectron In 1990, Medina et al. studied 100 consecutive Mexican patients with epilepsy that started after the age of 25 years. All patients underwent clinical evaluation, computed tomography, and electroencephalography; additionally, cerebro-spinal fluid analysis was performed in 82 of them.

**Figure 13.** Shows some of most relevant scientists in the field of cysticercosis who attended to the meeting in Atlanta, Georgia on February 24, 2008. From left to right: Phillip S Craig, Akira Ito, Ana Flisser, Peter Schantz and Lee Willigham III (Courtesy of Dr Flisser).

The volume of research and publications on cysticercosis, taeniosis and NCC increased gradually in this decade reaching a large number therefore it is not possible sum it up in the limited space we have but we should not conclude this historical review without prior highlight the meritorious work performed by some prominent researchers in this stage such as: Ana Flisser, Peter Schantz, Julio Sotelo, Hugo García, Oscar del Bruto, Akira Ito, Jefferson Proaño, Vedantam Rajshekhar, Ravindra Kumar Garg, Ted Nash, Carlton Evans, Osvaldo Takayanagui, Clinton White, Lee Willingham, Phillip S Craig, Gagandeep Singh, Sudesh Prabhakar, Hélène Carabine, Andrea Wrinkle, Arturo Carpio, Agnes Fleury, David Botero, Svetlana Agapejev, Gustavo Román, P Dorny, M Raghava, V Prabhakaran, Victor Tsang, Jorge Morales-Montor, Tetsuya Yanagida, Marshall Lightowlers, Elsa Sarti, Joachim Blocher, Yasuhito Sako, Minoru Nakao, Feng Zheng, Kazuhiro Nakaya, Jim Allan, Dolores Correa, Armando Gonzalez, Robert Gilman, García E, Márquez C, Fragoso G, Sciutto E, Cuetter A, Andrews R. Ferrer E., Cabrera Z., T Kelesidis, Tony Velasco, Cortés M, Alain Dessein, Pierre-Marie Preux PM, Jose A Serpa, Linda S Yancey, Pratibhasinghi Singhi, Dumas M, Carlos Larralde, Tapia G, Eon Keeseon, Carlos Larralde, Tammi Krecek, M Gupta, P Agarwal, GA Khwaja, P Sharma, R Shukla, D Singh, MV Padma, NK Misra, GK Ahuja, IK Phiri, Helena Ngowi, Sam Mukaratirwa, LM Michael, S Siziya, Humberto Foyaca, Emilia Noormahomed, Aline S de Aluja, Molina Alvarez, J Lasso, F Quet, M Guerchet, EB

Ngoungou, A Nicoletti, N Wadia, Jorge Burneo, Tonicarlo Velasco, Theodoros Kelesidis, P Huc, M Druet-Cabanac, M Strobel, Li T, Phil Craig, X Chen, D Qiu, J Qiu, MO Sato, T Wandra, H Bradshaw, L Li, N Nakao, J Waikagul, D Wattanakulanich, W Maipanich, K Nakaya, MO Sato, Y Sako, M Okamoto, K Swastika, C I Dewiyani, T Yanagida, K Nakaya, Hiroshi Yamasaki, James C. Allan, Marcello Otake Sato, Minoru Nakao, Yasuhito Sako, Kazuhiro Nakaya, Dongchuan Qiu, S Desai, A Cuetter, AE Gonzalez, S. Geerts, NC Kyvsgaard, DP McManus, ZS Pawlowski. However over this relationship there are still a significant number of researchers that do not appear in the listing.

Introduction to Cysticercosis and Its Historical Background 25

The 7th General Assembly on cysticercosis/taeniosis was held at the Faculty of Health Sciences, Walter Sisulu University, Mthatha, South Africa, from 20 – 22 July 2011. The meeting was organized by the Cysticercosis Working Group in Eastern and Southern Africa (CWGESA) in collaboration with the Faculty of Health Sciences of Walter Sisulu University. Veterinary, medical, agricultural and socioeconomic researchers met in Mthatha to discuss the Roadmap for Elimination of *Taenia solium* cysticercosis/taeniosis (TSCT) in Eastern and Southern Africa and to evaluate progress and make recommendations on intervention

**Figure 15.** Members of the CWGESA attended to this meeting. Seated from left to right: Lee Willigham

We have the advantage that the disease has been virtually isolated in the communities around Mthatha where there are also human and material resources needed to eradicate the cysticercosis if we receive the necessary support of governmental institutions and nongovernmental organizations committed to this goal. Eradicating the cysticercosis of the surrounding area of Mthatha is equivalent to delete the T solium in South Africa and this

Mexico is the country that more progress has been achieved in this regard and their doctors

For prevention and eradication of cysticercosis are needed peoples dedicated and committed to this cause, which should be headed only by honorable scientists stripped of

III, Samson Mukaratirwa, Humberto Foyaca, Helena Ngowi and Faustin Lekule.

could be the first country in the world to achieve it.

any trait of hubris, arrogance and self-sufficiency.

and researchers take pride that flag.

**4. Honor to honor deserves**

research for the elimination of the disease.

Most of these scientists have been grouped to work together on the same objective: *Taenia solium*. Some have more than 20 years of experience and more than 20 publications. Among the groups with better results are the groups in México, Perú, and the three groups from Ecuador (Quito, Cuenca, and Guayaquil). There are other groups from the Eastern and Southern Africa (CWGESA), Asia and Oceania, and the group of Europe of recent constitution (Willingham LA III et al., 2008).

The CWGESA was established in 2002 to promote communication, collaboration and coordination of integrated research and control activities to combat cysticercosis, a serious and sometimes fatal disease transmitted between pigs and people by the zoonotic tapeworm, Taenia solium.

**Figure 14.** Cysticercosis Working Group for Eastern and Southern Africa International Training Workshop organized by CWGESA and held on June 2004 Mthatha where delegates from 11 African countries participated.

The 7th General Assembly on cysticercosis/taeniosis was held at the Faculty of Health Sciences, Walter Sisulu University, Mthatha, South Africa, from 20 – 22 July 2011. The meeting was organized by the Cysticercosis Working Group in Eastern and Southern Africa (CWGESA) in collaboration with the Faculty of Health Sciences of Walter Sisulu University. Veterinary, medical, agricultural and socioeconomic researchers met in Mthatha to discuss the Roadmap for Elimination of *Taenia solium* cysticercosis/taeniosis (TSCT) in Eastern and Southern Africa and to evaluate progress and make recommendations on intervention research for the elimination of the disease.

**Figure 15.** Members of the CWGESA attended to this meeting. Seated from left to right: Lee Willigham III, Samson Mukaratirwa, Humberto Foyaca, Helena Ngowi and Faustin Lekule.

We have the advantage that the disease has been virtually isolated in the communities around Mthatha where there are also human and material resources needed to eradicate the cysticercosis if we receive the necessary support of governmental institutions and nongovernmental organizations committed to this goal. Eradicating the cysticercosis of the surrounding area of Mthatha is equivalent to delete the T solium in South Africa and this could be the first country in the world to achieve it.

Mexico is the country that more progress has been achieved in this regard and their doctors and researchers take pride that flag.

## **4. Honor to honor deserves**

24 Novel Aspects on Cysticercosis and Neurocysticercosis

constitution (Willingham LA III et al., 2008).

tapeworm, Taenia solium.

countries participated.

Ngoungou, A Nicoletti, N Wadia, Jorge Burneo, Tonicarlo Velasco, Theodoros Kelesidis, P Huc, M Druet-Cabanac, M Strobel, Li T, Phil Craig, X Chen, D Qiu, J Qiu, MO Sato, T Wandra, H Bradshaw, L Li, N Nakao, J Waikagul, D Wattanakulanich, W Maipanich, K Nakaya, MO Sato, Y Sako, M Okamoto, K Swastika, C I Dewiyani, T Yanagida, K Nakaya, Hiroshi Yamasaki, James C. Allan, Marcello Otake Sato, Minoru Nakao, Yasuhito Sako, Kazuhiro Nakaya, Dongchuan Qiu, S Desai, A Cuetter, AE Gonzalez, S. Geerts, NC Kyvsgaard, DP McManus, ZS Pawlowski. However over this relationship there are still a

Most of these scientists have been grouped to work together on the same objective: *Taenia solium*. Some have more than 20 years of experience and more than 20 publications. Among the groups with better results are the groups in México, Perú, and the three groups from Ecuador (Quito, Cuenca, and Guayaquil). There are other groups from the Eastern and Southern Africa (CWGESA), Asia and Oceania, and the group of Europe of recent

The CWGESA was established in 2002 to promote communication, collaboration and coordination of integrated research and control activities to combat cysticercosis, a serious and sometimes fatal disease transmitted between pigs and people by the zoonotic

**Figure 14.** Cysticercosis Working Group for Eastern and Southern Africa International Training Workshop organized by CWGESA and held on June 2004 Mthatha where delegates from 11 African

significant number of researchers that do not appear in the listing.

For prevention and eradication of cysticercosis are needed peoples dedicated and committed to this cause, which should be headed only by honorable scientists stripped of any trait of hubris, arrogance and self-sufficiency.

In our opinion, the person who has been most prominent in the research work related to the tapeworm *Taenia solium* is the Dr. Ana Flisser as shown below; she is a full time, upper rank, senior scientist at the National Autonomous University of Mexico, UNAM since 1972.

Introduction to Cysticercosis and Its Historical Background 27

PAHO Award for Administration 1999. For her work to transform the network of public health laboratories in her country to support priority health programs, including administration, teaching, and research. Director, National Institute for Epidemiology

128 original articles in international and Mexican refereed journals. 60 reviews or invited

62 book chapters (two chapters in this book are not included). 8 books as editor or coeditor.

11 for doctor in science, 12 for master in science, 12 as member of advisory committee, 2 for

20 research projects from: DGAPA/UNAM, CONACYT, European Community,

Immunoparasitology since 2004, Faculty of Sciences, UNAM. Immunoparasitology in

483 papers in national and international congresses. 182 national and international invited lectures 44 graduate and bachelor courses. 52 lectures at different graduate courses and various memberships of Societies of Parasitology, Tropical Medicine and Immunology of Mexico, USA and Great Britain, as well as of the International Federations in these areas. (Source: World Federation of Parasitology). Again, we want to emphasize that not only fatten the results to the person, other qualities such as modesty, friendship, solidarity, honesty, and the desire to selflessly help young researchers are also qualities that make an example to follow that I wish to report to the new generations of scientists worldwide.

We would like to make it very clear that we have received from Dr. Flisser some photos to illustrate this chapter but that the decision to include this modest but deserved tribute to her maintained and hard work, is ours. Therefore she had no role in chapter design, information collection, decision to publish, or preparation of the manuscript. By her modesty, we know that she will not fully agree with this decision but we could not ignore her history. To pay a tribute to their achievements including her personal contribution to the reduction of the morbidity and mortality of the cysticercosis in Mexico and everything she has done by and

Diagnosis and Reference (INDRE), SSA, 1995-2000.

Cysticercosis and taeniosis due to *Taenia solium*

medical specialties, 15 at the bachelor level

IDRC/Ottawa, WHO/PAHO, Merck-Darmstadt/Merck-México.

graduate programs since 2005, Faculty of Medicine, UNAM.

**Research area**

**Publications**

**Advisor of thesis**

**Financial support**

**Recent teaching activities**

for the rest of humanity, is right.

**Academic activities**

3 books as author or coauthor

articles

The authors of this chapter believe that in order to be considered a researcher whose example should be followed by the younger generations are they must comply with certain requirements among which protrude the modesty, honesty, solidarity and a desire to help the younger generations of researchers to its best development unconditionally. In our opinion that example is Dr. Ana Flisser.

## **5. Recent awards and distinctions received:**

Dr. Ana Flisser won the University Award 2011 in Research in Natural Sciences.

Coordinator of the first MD/PhD Program in Mexico, UNAM, 2011.

First Vice-President of the World Federation of Parasitologists and President of the XIII International Congress of Parasitology to be held in Mexico City in August 2014

Award Ciudad Capital "Heberto Castillo Martínez" Research in Health and Environment, 2008 President, From Alaska to Chiapas: The First North American Parasitology Congress, Merida, Yucatan, June 21-25, 2007

Director of Research, Hospital General "Dr. Manuel Gea Gonzalez" SSA, 2001-2005.

**Figure 16.** Dr. Ana Flisser Department of Microbiology and Parasitology, Faculty of Medicine, National Autonomous University of Mexico.

PAHO Award for Administration 1999. For her work to transform the network of public health laboratories in her country to support priority health programs, including administration, teaching, and research. Director, National Institute for Epidemiology Diagnosis and Reference (INDRE), SSA, 1995-2000.

#### **Research area**

26 Novel Aspects on Cysticercosis and Neurocysticercosis

opinion that example is Dr. Ana Flisser.

Merida, Yucatan, June 21-25, 2007

Autonomous University of Mexico.

**5. Recent awards and distinctions received:**

In our opinion, the person who has been most prominent in the research work related to the tapeworm *Taenia solium* is the Dr. Ana Flisser as shown below; she is a full time, upper rank, senior scientist at the National Autonomous University of Mexico, UNAM since 1972.

The authors of this chapter believe that in order to be considered a researcher whose example should be followed by the younger generations are they must comply with certain requirements among which protrude the modesty, honesty, solidarity and a desire to help the younger generations of researchers to its best development unconditionally. In our

First Vice-President of the World Federation of Parasitologists and President of the XIII

Award Ciudad Capital "Heberto Castillo Martínez" Research in Health and Environment, 2008 President, From Alaska to Chiapas: The First North American Parasitology Congress,

**Figure 16.** Dr. Ana Flisser Department of Microbiology and Parasitology, Faculty of Medicine, National

Dr. Ana Flisser won the University Award 2011 in Research in Natural Sciences.

International Congress of Parasitology to be held in Mexico City in August 2014

Director of Research, Hospital General "Dr. Manuel Gea Gonzalez" SSA, 2001-2005.

Coordinator of the first MD/PhD Program in Mexico, UNAM, 2011.

Cysticercosis and taeniosis due to *Taenia solium*

#### **Publications**

128 original articles in international and Mexican refereed journals. 60 reviews or invited articles

62 book chapters (two chapters in this book are not included). 8 books as editor or coeditor. 3 books as author or coauthor

#### **Advisor of thesis**

11 for doctor in science, 12 for master in science, 12 as member of advisory committee, 2 for medical specialties, 15 at the bachelor level

#### **Financial support**

20 research projects from: DGAPA/UNAM, CONACYT, European Community, IDRC/Ottawa, WHO/PAHO, Merck-Darmstadt/Merck-México.

#### **Recent teaching activities**

Immunoparasitology since 2004, Faculty of Sciences, UNAM. Immunoparasitology in graduate programs since 2005, Faculty of Medicine, UNAM.

#### **Academic activities**

483 papers in national and international congresses. 182 national and international invited lectures 44 graduate and bachelor courses. 52 lectures at different graduate courses and various memberships of Societies of Parasitology, Tropical Medicine and Immunology of Mexico, USA and Great Britain, as well as of the International Federations in these areas. (Source: World Federation of Parasitology). Again, we want to emphasize that not only fatten the results to the person, other qualities such as modesty, friendship, solidarity, honesty, and the desire to selflessly help young researchers are also qualities that make an example to follow that I wish to report to the new generations of scientists worldwide.

We would like to make it very clear that we have received from Dr. Flisser some photos to illustrate this chapter but that the decision to include this modest but deserved tribute to her maintained and hard work, is ours. Therefore she had no role in chapter design, information collection, decision to publish, or preparation of the manuscript. By her modesty, we know that she will not fully agree with this decision but we could not ignore her history. To pay a tribute to their achievements including her personal contribution to the reduction of the morbidity and mortality of the cysticercosis in Mexico and everything she has done by and for the rest of humanity, is right.

Introduction to Cysticercosis and Its Historical Background 29

http://www.ispub.com/journal/the\_internet\_journal\_of\_neurology/volume\_12\_number \_2\_4/article/par asitic-zoonoses-of-the-brain-another-challenger.html. (Accessed 22

[2] Foyaca-Sibat H, editor. Treatment of Epilepsy Secondary to Neurocysticercosis, Novel

[3] Foyaca-Sibat H editor.Clinical Features of Epilepsy Secondary to Neurocysticercosis at

[4] Foyaca-Sibat H. editor. Epilepsy Secondary to Parasitic Zoonoses of the Brain, Novel

[5] Foyaca-Sibat H, Ibañez-Valdés L de F. Clinical trial of praziquantel and prednisone in rural patients with neurocysticercosis presenting recurrent epileptic attacks The Internet

http://www.ispub.com/journal/the\_internet\_journal\_of\_neurology/volume\_1\_number\_ 2\_39/article/cli nical\_trial\_of\_praziquantel\_and\_prednisone\_in\_rural\_patients\_with\_ neurocysticercosis\_presenting\_w ith\_recurrent\_epileptic\_attacks.html. (accessed 22

[6] Foyaca-Sibat H Ibañez-Valdés L de F. Intraventricular neurocysticercosis in HIV

[7] Bautista IR. Neurocisticercosis una realidad y sus limitantes en el diagnóstico. SIRIVS. Curso: Investigación II . Maestría en Salud Animal. Universidad Nacional Mayor de San

[8] Sotelo J. Eradication of cysticercosis is an attainable goal. BMJ 2003;326(7388):511-512. [9] Hotez PJ; Bottazi ME, Franco-Paredes C, Ault SK, Periago MR. The Neglected Tropical Diseases of Latin America and the Caribbean: A Review of Disease Burden and Distribution and a Roadmap for Control and Elimination. *PLoS Neglected Tropical*

[10] Román G, Sotelo J, Del Brutto O, Flisser A, Dumas M, Wadia N. et al. A proposal to declare neurocysticercosis an international reportable disease. *Bull World Health Organ*

[11] Shanley JD, Jordan MC: Clinical aspects of CNS cysticercosis. *Arch Inter Med*

[12] Loo L, Braude A: Cerebral cysticercosis in San Diego: a report of 23 cases and review of

[13] McComick G, Zec Chi-S, Heiden J. Cysticercosis Cerebri. Review of 127 cases. *Arch*

[14] McCormick GF. Cysticercosis: review of 230 patients. *Bull Clin Neurosci.* 1985;50:76–76.

[16] Uddin J, Gonzalez AE, Gilman RH, Thomas LH, Rodriguez S, Evans CA, Remick DG, Garcia HH, Friedland JS. Mechanisms regulating monocyte CXCL8 secretion in

[15] Carpio A. Neurocysticercosis: an update. *Lancet Infect Dis* 2002;2:751-62.

http://www.ispub.com/journal/the\_internet\_journal\_of\_neurology/volume\_2\_number\_ 1\_37/article/int raventricular\_neurocysticercosis\_in\_hiv\_positive\_patients.html. (ac es

January 2012).

January 2012).

sed 22 January 2012).

2000; 78: 399-406.

1980;140:1309- 1315.

*Neurol* 1982; 39:534-539.

Treatment of Epilepsy. Rikeja; InTech, 2011.

Journal of Neurology 2002;2):41-50.

the Insular Lobe, Novel Aspects on Epilepsy; InTech,2011.

patients. The Internet Journal of Neurology 2003;2(1):23-31.

Marcos Facultad de Medicina Veterinaria.2008

*Diseases* 2008; 2: e300 doi:10.1371/journal.pntd.0000300.

the literature. *Medicine* (Baltimore) 1982;61:341-350.

Aspects on Epilepsy, Humberto Foyaca-Sibat. Rikeja; InTech,2011.

**Figure 17.** Dr. Ana Flisser: Always surrounded by talented young people who receive her teachings, experience and wisdom.

## **Author details**

Humberto Foyaca Sibat and Lourdes de Fátima Ibañez Valdés *Walter Sisulu University, Faculty of Health Sciences, Nelson Mandela Academic Hospital Division of Neurology., Mthatha,South Africa*

## **Acknowledgement**

Special thanks are due to the Cuban Ministry of Health and the Institute of Tropical Medicine "Pedro Kouri", and authorities of Nelson Mandela Academic Hospital, School of Medicine, Faculty of Health Sciences and Directorate: Research Development from Walter Sisulu University for their kind support. Finally, we wish to declare publicly our eternal and deepest gratitude to Lorna María Foyaca García, Fátima Susana Foyaca Ibañez and Thabo Humberto Foyaca Ibañez for their delight support. We like to thank to Dr Ana Flisser for her contribution to this chapter.

## **6. References**

[1] Foyaca-Sibat H, Ibañez-Valdés LdeF & J. Moré-Rodríguez : Parasitic Zoonoses Of The Brain: Another Challenger?. The Internet Journal of Neurology. 2010 Volume 12 Number 2.

http://www.ispub.com/journal/the\_internet\_journal\_of\_neurology/volume\_12\_number \_2\_4/article/par asitic-zoonoses-of-the-brain-another-challenger.html. (Accessed 22 January 2012).

[2] Foyaca-Sibat H, editor. Treatment of Epilepsy Secondary to Neurocysticercosis, Novel Treatment of Epilepsy. Rikeja; InTech, 2011.

28 Novel Aspects on Cysticercosis and Neurocysticercosis

experience and wisdom.

*of Neurology., Mthatha,South Africa*

**Acknowledgement**

contribution to this chapter.

**6. References**

Number 2.

**Author details**

**Figure 17.** Dr. Ana Flisser: Always surrounded by talented young people who receive her teachings,

*Walter Sisulu University, Faculty of Health Sciences, Nelson Mandela Academic Hospital Division*

Special thanks are due to the Cuban Ministry of Health and the Institute of Tropical Medicine "Pedro Kouri", and authorities of Nelson Mandela Academic Hospital, School of Medicine, Faculty of Health Sciences and Directorate: Research Development from Walter Sisulu University for their kind support. Finally, we wish to declare publicly our eternal and deepest gratitude to Lorna María Foyaca García, Fátima Susana Foyaca Ibañez and Thabo Humberto Foyaca Ibañez for their delight support. We like to thank to Dr Ana Flisser for her

[1] Foyaca-Sibat H, Ibañez-Valdés LdeF & J. Moré-Rodríguez : Parasitic Zoonoses Of The Brain: Another Challenger?. The Internet Journal of Neurology. 2010 Volume 12

Humberto Foyaca Sibat and Lourdes de Fátima Ibañez Valdés

http://www.ispub.com/journal/the\_internet\_journal\_of\_neurology/volume\_1\_number\_ 2\_39/article/cli nical\_trial\_of\_praziquantel\_and\_prednisone\_in\_rural\_patients\_with\_ neurocysticercosis\_presenting\_w ith\_recurrent\_epileptic\_attacks.html. (accessed 22 January 2012).

neurocysticercosis and the effect of antiparasitic therapy. J Immunol. 2010 Oct 1;185(7):4478-84. Epub 2010 Sep 8.

Introduction to Cysticercosis and Its Historical Background 31

[35] Sawhney IM, Singh G, Lekhra OP, Mathuriya SN, Parihar PS, Prabhakar S. Uncommon

[36] Seo HJ, Seo JH, Kim SW, Shin H. Isolated Spinal Neurocysticercosis : Unusual Ocular Presentation Mimicking Pseudotumor Cerebri. *J Korean Neurosurg Soc*. 2011 May; 49(5):

[37] Ahmad FU, Sharma BS. Treatment of intramedullary spinal cysticercosis: report of 2

[38] Amaral L, Maschietto M, Maschietto R, Cury R, Ferreira NF, Mendonça R, et al. Unusual manifestations of neurocysticercosis in MR imaging : analysis of 172 cases. *Arq*

[39] Amaya MA, Roa JL. Neurocisticercosis raquídea: Reporte de caso. *Rev Colomb Radiol.*

[40] Delobel P, Signate A, Guedj M, Couppie P, Gueye M, Smadja D, Pradinaud R. Unusual form of neurocysticercosis associated with HIV infection. *European Journal of*

[41] Dewan P, Shankar J, Sunil S, Gupta A Uncommon presentation of neurocysticercosis.

[42] Dickmann, G.H. Cisticercosis de la fosa craneana posterior. *Rev. Neurol.*1946;15:160-179. [43] Figueroa JJ, Davis LE, Magalhaes A. Extraparenchymal neurocysticercosis in

[44] Flisser A, Madrazo I, Plancarte A, Schantz P, Allan J, Craig P, Sarti E. Neurological symptoms in occult neurocysticercosis after single taeniacidal dose of praziquantel.

[45] Flisser A, Madrazo I. Evagination of Taenia solium in the fourth ventricle. N Engl J

[48] Ibañez-Valdés LdeF, Foyaca-Sibat H. Neurogenic heart secondary to insular stroke and neurocysticercosis. International Journal of Stroke (ISSN 1747-4930) 2006;1(1):68. [49] Lamejjati M, Aniba K, Haddi M. et al. Spinal Intramedullary arachnoid cyst in children.

[50] Leite CC, Jinkins JR, Escobar BE, et al. MR imaging of intramedullary and intradural-

[51] Musumdar D, Nadkarni T, Desai K, Dindorkar K, Goel A. Thoracic intramedullary

[52] Neal JH. An endoscopic approach to cysticercosis cysts of the posterior third ventricle.

http://hydra.fciencias.unam.mx:8080/xmlui/handle/123456789/62343?show=full [47] Foyaca-Sibat H, Ibañez-Valdés LdeF: "Generalized Cysticercosis With Cardiac Involvement". The Internet Journal of Neurology. ( ISSN: 1531-295X). 2007;7(2):6-11. http://www.ispub.com/journal/the\_internet\_journal\_of\_neurology/volume\_7\_number\_

2\_5/article/gen eralized\_cysticercosis\_with\_cardiac\_involvement.html

extra medullary spinal cysticercosis. *AJR* 1997;169:1713-1717.

cysticercosis. *Neurol Med Chir* (Tokyo)2002; 42 :575-579.

presentations of neurocysticercosis. *J Neurol Sci.* 1998;154(1):94-100.

cases and review of literature. *Surg Neurol*. 2007;67(1):74–77.

*South African Journal of Child Health*, 2011;5(2):12-13.

[46] Flisser A. Cisticercosis subcutáneo. *Ciencias,* UNAM 2011.

Albuquerque, New México. *J Neuroimaging.* 2011;21(1):38–43

296–298.

*Neuropsiquiatr.* 2003;61:533–541.

2011;22(1):3134-3137.

*Neurology*.2004; 11(1):55-58.

Lancet.1993;18:342-348.

Med. 1996 Sep 5;335(10):753-754

*Pediatr Neurosurg*. 2008;44(3):243–246.

*Neurosurgery* 1995;36(5):1040-1043.

[35] Sawhney IM, Singh G, Lekhra OP, Mathuriya SN, Parihar PS, Prabhakar S. Uncommon presentations of neurocysticercosis. *J Neurol Sci.* 1998;154(1):94-100.

30 Novel Aspects on Cysticercosis and Neurocysticercosis

1;185(7):4478-84. Epub 2010 Sep 8.

and management. *Ann Rev Med* 2000;51:187-206.

1988- 1998. *Presse Med* 1999;28(39):2141-2144.

update. *Medicine* (Baltimore). 2011;90(1):81-86.

services. *Trans R Soc Trop Med Hyg* 2008;102:228-232.

Internet Journal of Infectious Diseases 2003:2(2):15-23.

*Infect Dis* 1997;24:101-106.

25 January 2012).

Jun;17(6):1030-1036.

*Case Reports.* 2011;5:359.

*tropical* 2000;34:283–290.

Esp.184:15- 18, 1989.

2011;3(3):306– 308.

1986; 28:556-563.

neurocysticercosis and the effect of antiparasitic therapy. J Immunol. 2010 Oct

[17] White AC Jr. Neurocysticercosis: a major cause of neurological disease worldwide. *Clin*

[18] White AC Jr. Neurocysticercosis: updates on epidemiology, pathogenesis, diagnosis

[19] Foyaca-Sibat H, Ibañez-Valdés LdeF "Neurocysticercosis in HIV-positive patients" The

[20] Sarti E, Schantz PM, Plancharte A, Wilson M, Gutierrez R, Lopez AS, Roberts J, Flisser A. Prevalence and risk factor for *Taenia solium* taeniasis and cysticercosis in humans and

[22] Schantz PM, Moore AC, Muñoz JL, et al.: Neurocysticercosis in an Orthodox Jewish

[23] Rousseau MC, Guillotel B, Delmont J. Neurocysticercosis in the South-East of France

[24] García HH, Del Brutto OH; Cysticercosis Working Group in Peru. Neurocysticercosis:

[25] O'Neal S, Noh J, Wilkins P, Keene W, Lambert W, Anderson J, Compton Luman J, Townes J. Taenia solium Tapeworm Infection, Oregon, 2006-2009. *Emerg Infect Dis*. 2011

[26] Kelesidis T, Tsiodras S. Extraparenchymal neurocysticercosis in the United States. *J Med*

[27] Serpa JA, Graviss EA, Kass JS, White AC Jr. Neurocysticercosis in Houston, Texas: an

[28] Takayanagui O, Leite J. Neurocisticercose. *Revista da Sociedades Brasileira de Medicina*

[29] Pawlowski ZS. Control of neurocysticercosis by routine medical and veterinary

[30] García Albea E., Cisticercosis en España. Algunos datos Epidemiológicos. Rev Clin

[31] [31]Sánchez L, Abad L, Lozano E, Maldonado G. Neurocisticercosis intraventricular. Presentación de un caso localizado en el tercer ventriculo. *Radiologia*. 2002;44:309-313. [32] Sandeep GJ, Vipul CC, Bhakti SY, Himanshu VB. Imaging Findings of Disseminated Cysticercosis with Unusual Involvement of Spleen and Pancreas. *J Glob Infect Dis.*

[33] Sang-Wook K, Moon-Kyu K, Sae-Moon Oh, Se-Hyuck P. Racemose Cysticercosis in the

[34] Sarti E. La taeniasis y cisticercosis en México (revisión bibliográfica). *Salud Pública Mex*

Cerebellar Hemisphere. *J Korean Neurosurg Soc*. 2010; 48(1):59–61.

pigs in a village in Morelos, Mexico. *Am J Trop Med Hyg* 1992; 46:677-684.

[21] Garg RK. Neurocysticercosis. *Postgrad Med J* 1998; 74(872):321-326.

community in New York City. *N England J Med* 1992;327:692-696.

updated concepts about an old disease. *Lancet Neurol*. 2005; 4:653-661.

http://www.ispub.com/ostia/index.php?xmlFilePath=journals/ijn/current.xml (accessed

	- [53] Park SY, Min HK, Jung HK, Kwan YS. Disseminated cysticercosis. *Journal of Korean Neurosurgical Society*. 2011;49(3):190-193.

Introduction to Cysticercosis and Its Historical Background 33

[72] Flisser A. Cysticercosis and Taenia solium. J Clin Neuroophthalmol. 1986 Sep;6(3):200 [73] Flisser A. Taeniasis and cysticercosis due to Taenia solium. Prog Clin Parasitol.

[74] Flisser A. Neurocysticercosis and epilepsy in developing countries. J Neurol Neurosurg

[75] Flisser A. Where are the tapeworms? Parasitol Int. 2006;55 Suppl:S117-20. Epub 2005

[76] Flisser A, Madrazo I, Delgado H. Cisticercosis humana. ED. *El Manual Moderno*. RSC-

[77] Chimelli L, Lovalho AF, Takayanagui OM. Neurocysticercosis: contribution of autopsies in the consolidated of mandatory notification in Ribeirao Preto-SP, *Brazil. Arq*

[78] Sanchez AL, Ljungstrom I, Medina MT. Diagnosis of human neurocysticercosis in

[80] Torres J. Recomendaciones para el tratamiento de la Neurocisticercosis. *Memorias 1as*

[81] Torres J. Avances en el tratamiento de la neurocysticercosis. *Memorias 2das jornadas de Medicina Tropical*, Caracas, 26 al 29 de julio de 1994.Torres M, Pérez C, Galdámez E, Gabor M, Miranda C, Cofré X et al. Teniosis: serie clínica en 35 pacientes. *Parasitol día*

[82] Vera A. Prevalencia de teniasis por Taenia solium y riesgo epidemiológico de cisticercosis en las comunidades del Valle del Río Temerla, Estados Carabobo y Yaracuy. Curso Medio de Salud Pública, Dpto. de Medicina Preventiva y Social VII, Escuela de Medicina "Witremundo Torrealba", Facultad de Ciencias de la Salud,

[83] Ferrer E., Cabrera Z., Cortés M M, Lares M, Harrison L, Parkhouse RME. Inmunodiagnóstico de cisticercosis basado en la detección de antígenos en comunidades rurales alrededor de Canoabo. Ed. Carabobo. *Act. Cient. Venezolana,*

[84] Ferrer E., Cortés M. M, Rojas G., Lares M., Fernández I., Harrison L., Parkhouse R.M.E. y Cabrera Z. Seroprevalencia y factores de riesgo de cisticercosis en dos comunidades rurales, Sanare y Río Tocuyo Edo. Lara 1998. *Act. Cient. Venezolana, AsoVAC*. 1999;50(2):

[85] Meza N., Rossi N., Sánchez N., Muñoz J., Hernández M. y Urdaneta H. Teniasis y cisticercosis humana en una población rural del Estado Táchira, Venezuela. *Act. Cient.*

[86] Carpio A, Hauser WA. Prognosis for seizure recurrence in patients with newly

[87] David J. Neurocysticercosis. "Dr. House Series" .Martes 22 de febrero del 2011.(última

endemic countries: clinical study in Honduras. *Parasitol Int* 1999;48(1):81-89.

*Jornadas Venezolanas de Medicina Tropical*, Caracas, 28 al 31 de julio de 1991.

http://www.medikamag.com/Page.asp?key=751&export=html

1994;4:77-116

Dec 6.

Psychiatry. 2001 May;70(5):707-712

048, ISO-90002:1994/NMX-CC-004:1998

*Neuropsiquiatr* 1998; 56(3B):577-584.

[79] MedikaMag.com. 2011 Rep. Dom.

Universidad de Carabobo.1994.

*Venezolana, AsoVAC.* 1999;50(2):265-267.

revisión en septiembre 22, 2011).

diagnosed neurocysticercosis. *Neurology* 2002;59:1730–1734.

*AsoVAC.* 1998;49(2): 297.

345.

2001;25:55-59.

*Transplant*. 2006;21 (8):2308-2310.

2003;116:637–638.

1982;9:401-407.

1996;38(3):207-216.

14(1):97-119.

*Medellin* 1989;2(II):75-82.

un caso. *Arch Neurocien* 2004;9(2):110-113.

autopsia.Patología (Mex) 1972; 10:27-40.

*Acta Med Colombiana* 1986;11:62-73.

Arq Neuropsiquiatr 1983; 41(1):50-63.

neurocysticercosis. *Arch Neurol* 1988;45:1130-1133.

VPH/83.49. *World Health Organization*, Geneva, 1983:1.207.

Medicina Bucaramenga Colombia; 1986, 14(1):51-60.

*Neurosurgical Society*. 2011;49(3):190-193.

intraventricular. *Rev Neurocir* 2007;9(4):107-113.

[53] Park SY, Min HK, Jung HK, Kwan YS. Disseminated cysticercosis. *Journal of Korean*

[54] Ramírez-Ceballos JA, Marquéz RMC.Tratamiento endoscópico de la neurocisticercosis

[55] Razdan S, Pandita KK, Pandita S. Reversible hemifacial spasm due to

[56] Ruangkanchanasetr P,Bangchuad T, Sithinamsuwan P, Benjasuratwong Y, Chuankrerkkul W, Ubolwatra S, Supaporn T. Hypothalamic neurocysticercosis presenting with polyuria: a first report of an unusual manifestation. *Nephrol. Dial.*

[57] Villanueva M, Alcocer JL, Martinez J, Horna A. Cisticercosis intramedular. Reporte de

[58] Wadia N, Desai S, Bhatt M: Disseminated cysticercosis. New observations, including CT scan findings and experience with treatment by praziquantel. *Brain* 1988, 111:597-614 [59] Xinou E, Lefkopoulous M, Gelagoti M, Drevelegas A, Diakou A, et al. Successful treatment of isolated cerebellar cysticercosis with albendazole. *Chin Med J* (Engl)

[60] Rabiela MT, Lombardo L, Flores F. Cisticercosis cerebral. Estudio de 68 casos de

[61] Lopez-Hernández A, Garaizar C. Childhood Cerebral Cysticercosis: clinical features and computed tomographic findings in 98 Mexican children. *Can J Neurol SC*

[63] Sotelo J, Penagos P, Escobedo F, Del Brutto OH: Short course of albendazole therapy for

[64] Agapejev S. Epidemiology of Neurocysticercosis in Brazil. *Rev Inst Med Trop Sao Paulo*

[65] Ramírez G, Pradilla G, Rodríguez M, González C. Cisticercosis. Estudio de 80 casos.

[66] Botero D. Cisticercosis en Colombia. Investigations realizadas hasta 1988. *An Acad Med*

[67] Takayanagui OM, Jardim E. Clinical aspects of nerocysticercosis: analysis of 500 cases.

[68] Gemmell M, Matyas Z, Pawloski Z, Soulsby EJL, Larralde C, Nelson G, Rosieky B. Guidelines for surveillance, prevention and control of taeniasis/cysticercosis.

[69] García HH, del Brutto OH. *Taenia solium* cysticercosis. *Infect Dis Clin North Am* 2000;

[70] Turkulov V, madle-Samardzija N, Canak G, Vukadinov J, Aleksic-Dordevic M. Clinical and diagnostic approaches to neurocysticercosis. *Med Pregl* 2001; 54(7-8):353-356. [71] Flisser A. Cisticercosis Humana: Epidemiología, diagnóstico e inmunología. Revista UIS

[62] Flisser A. Neurocysticercosis in Mexico. *Parasitol Today*1988; 4:131-137.

neurocysticercosis. *Ann Indian Acad Neurol*. 2009 Jul-Sep; 12(3): 191–192.

http://mentiraomedicina.blogspot.com/2011/02/neurocisticercosis.html

[88] Lombardo L, Matheus J.H., Cerebral Cysticercosis in México, Neurology 1961;11:825- 828.

Introduction to Cysticercosis and Its Historical Background 35

[106] Winkler AS, Kerschbaumsteiner K, Stelzhammer B, Meindl M, Kaaya J, Schmutzhard E. Prevalence, incidence, and clinical characteristics of epilepsy--a community-based

[107] Preux P-M and Druet-Cabanac M. Epidemiology and aetiology of epilepsy in sub-

[108] Hoberg EP, Alkire NL, de Queiroz A, and Jones A. Out of Africa: origins of the Taenia

[109] Heslip , S. 2001. Time-Space chart. ANP440. Hominid fossils. Spring Semester, 2001. http://www.msu.edu/~heslipst/contents/ANP440/index.htm (accessed 27 February

[110] Alles , D. 2001. Cenozoic. http://fire.biol.wwu.edu/trent/alles/Cenozoic.pdf (accessed

[111] Asfaw , B., White T, Lovejoy O, Latimer B, Simpson S, Suwa G. 1999. *Australopithecus garhi* : A New Species of Early Hominid from Ethiopia. SCIENCE 1999;284:629-635. [112] Cox, F.E.G. History of Human Parasitology. Clinical Microbiology Reviews. 2002; 15(4):595- 612. http://cmr.asm.org/content/15/4/595 (accessed 05 March 2012). [113] Discover magazine. http://blogs.discovermagazine.com/loom/2010/04/26/why-madag

[114] Ancient Hebrew Medicine. http://www.healthguidance.org/entry/6309/1/Ancient-

[116] Flisser A, Espinoza B, Tovar A, Plancarte A, Correa D. Host--parasite relationship in cysticercosis: an immunological study in different compartments of the host. Vet

http://www.greekmedicine.net/whos\_who/Hippocrates.html (accessed 27 February

http://en.wikipedia.org/wiki/Muhammad\_ibn\_Zakariya\_al-Razi (accessed 10 March

[118] Wikipedia- Pedanius Dioscorides. http://en.wikipedia.org/wiki/Dioscorides. (accessed

[119] Cordero M, Hidalgo A, Cisticercosis (C. cellulosae). 1999. Parasitología Veterinarian. Edit. Cordero del Campillo M, and Rojo V. Ed Mc Graw Hill-Interamericana, Madrid.

[120] Wadia, NH, Singh, G. "Taenia Solium: A Historical Note" Taenia Solium Cysticercosis:

http://www.greekmedicine.net/whos\_who/Galen.html. (accessed 13 March 2012). [122] Hort AF: Theophrastus (1916). ed. *Theophrastus: Enquiry into Plants*. 1, Book I-V. New

From Basic to Clinical Science CABI Publishing, 2002. 157-168.

York: Loeb Classical Library/G.P. Putnam's Sons. cited in:

[123] Greek Medicine-Avicenna. Available at the URL:

http://en.wikipedia.org/wiki/Theophrastus (accessed 15 March 2012).

door-to-door study in northern Tanzania. *Epilepsia.* 2009;50(10):2310-2313.

tapeworms in humans. *Proc. R. Soc*. Lond.B 2001;268:781–787.

as carstapeworms-matter-to-you. (accessed 20 March 2012).

HebrewMedicine.html (accessed 07 March 2012). [115] Greek Medicine. Hippocrates. Available at the URL:

Parasitol. 1986 Mar;20(1-3):95-102.

[117] Wikipedia- Muhammad ibn Zakariya al-Razi.

Saharan Africa. *Lancet Neurol* 2005; 4:21-31.

2012).

2012).

2012).

10 March 2012).

pages 493-495.

[121] Great Medicine-Galen.

28 February 2012).

[106] Winkler AS, Kerschbaumsteiner K, Stelzhammer B, Meindl M, Kaaya J, Schmutzhard E. Prevalence, incidence, and clinical characteristics of epilepsy--a community-based door-to-door study in northern Tanzania. *Epilepsia.* 2009;50(10):2310-2313.

34 Novel Aspects on Cysticercosis and Neurocysticercosis

*Neuropsiquiatr* 1990; 48(1):1-10.

Hospital. *Arch Neurol* 1988; 45:777-780.

828.

290.

336.

15(4)747-756.

2005;175(5):3273-3281.

http://mentiraomedicina.blogspot.com/2011/02/neurocisticercosis.html

childhood. *Trans R Soc Trop Med Hyg* 1987;81(3):371-373.

and inactive forms. *Arch Intern Med.* 1985;145:442–445.

comparative evaluation. *Arq Neuropsiquatr* 1997; 55(3A):349-356.

prognostic factor in 54 patients. *Rev Neurol* 1997; 25(147):1681-1484. [98] Del Brutto OH. Neurocysticercosis. *Rev Neurol* 1999; 29(5):4456-4466.

neurocysticercosis: analysis of 203 patients. *Neurology* 1992; 42:389-392

neurocysticercosis. *J Assoc Physicians India* 1999;47:476–479.

criteria for Neurocysticercosis. *Neurology* 2001; 57(2):177-183.

cysticercosis. *Ann Tropical Medical Parasitology* 1999;93*:* 247-258.

Peru. Taenia solium cysticercosis. *Lancet.* 2003; 362: 547-556.

region. *Assoc Physicians India* 2002;50:1398-1400.

[88] Lombardo L, Matheus J.H., Cerebral Cysticercosis in México, Neurology 1961;11:825-

[89] Takayanagui OM. Neurocysticercosis: I. Clinical and laboratory course of 151 cases. *Arq*

[90] Takayanagui OM, Leite JP. Neurocysticercosis. *Rev Soc Bras Med Trop* 2001;34(3):283-

[91] Kalra V, Paul VK, Marwah RK, Kichhar Gs, Bhargava S. Neurocysticercosis in

[92] Scharf D. Neurocysticercosis. Two hundred thirty-eight cases from a California

[93] Sotelo J, Guerrero V, Rubio F. Neurocysticercosis: a new classification based on active

[94] Villa AM, Monteverde DA, Rodríguez W, Boero A, Sica RE. Neurocysticercosis in a hospital of the city of Buenos Aires: study of 11 cases. *Arq Neuropsiquiatr* 1993;51(3):336-

[95] Singh G. Neurocystycercosis in South-Central America and the Indian subcontinent. A

[96] Singh G, Kaushal V, Ram S, Kaushal RK, Dhanuka AK, Khurana S. Cysticercus immunoblot assay in patients with single, small enhancing lesions and multilesional

[97] Del Brutto OH. Neurocysticercosis in children: clinical and radiological analysis and

[99] Del Bruto OH, Santibáñez R, Rubio CA, Aguirre R, Diez E, Alarcón TA. Epilepsy due to

[100] Del Bruto OH, Rajshekhar V, White AC Jr, Tsang VC, et al. Proposed diagnostic

[101] Sanchez, A.L.; Lindback, J.; Schantz, P.M.; Sone, M.; Sakai, H.; Medina, M.T. & Ljungstrom I. A population based case-control study of *Taenia solium* taeniosis and

[102] Varma A, Gaur KJ. The clinical spectrum of Neurocysticercosis in the Uttraranchal

[103] García HH, Evans CA, Nash TE, Takayanagui OM, White AC jret al., Current Consensus Guidelines for Treatment of Neurocysticercosis*. Clin Microbiol Rev* 2002;

[104] García HH, Gonzalez AE, Evans CA, Gilman RH; Cysticercosis Working Group in

[105] Uddin J, Garcia HH, Gilman RH, Gonzalez AE, Friedland JS. Monocyte-astrocyte networks and the regulation of chemokine secretion in neurocysticercosis. *J Immunol.*

http://www.greekmedicine.net/whos\_who/Galen.html. (accessed 13 March 2012).

http://www.greekmedicine.net/whos\_who/Avicenna.html accessed 15 March 2012).

Introduction to Cysticercosis and Its Historical Background 37

[145] Flisser A, Pérez-Montfort R, Larralde C. The immunology of human and animal

[146] Flisser A, Woodhouse E, Larralde C. Human cysticercosis: antigens, antibodies and

[147] Flisser A, Laclette JP, Larralde C, Vogt M, Willms K. Integral study of cysticercosis. A

[148] Flisser A. Immunology of human cysticercosis. Bol Estud MED Biol. 1983 Jul-Oct;32(7-

[149] Pérez Sánchez E., Zúñiga G, Sarmiento J, Hernández C. Correlaciones Preliminares entre Clínica, Escanografía e Inmunología. Rev. UIS-Medicina.Buaramanga (Colombia);

[150] Vaz JA, Ferreira WA. Immunodiagnóstico da neurocisticircose: Teste inmunoenzimático com antigenos quimicamente ligados a suportes para pesquisa de anticorpos em soro e líquido cefalorraquiano. Rev. Inst. Med.Trop.Sao Paulo; 1988,30

[151] Gonzales Armando E., Cama Vialiano., Hilman Robert H., et al. Prevalence and comparisons of serologic assays, necropsy and tongue examination for the diagnosis of

[152] Foyaca-Sibat H, Cowan LD, Carabin H, Serrano-Ocaña G, , Krecek RC, Willingham A. "Accuracy of serological exam for the diagnosis of neurocysticercosis in outpatients with epilepsy, Eastern Cape Province, South Africa" PLOS Negleted

[153] Tsang, Víctor C., Brand Joy ., Boyer Ann E., An Enzime Linked inmunoelectrotransfer Blot Assay and Glycoprotein Antigens for Diagnosing HumanCysticercosis (Taenia

[154] Tsang VCW, Brand AJ, Boyer AE. An Enzyme-linked immunoelectrotransfer blot assay by glycoprotein antigens for diagnosing human cysticercosis (*Taenia solium*). *J*

[155] Tsang C.W., and Brand A. Joy ., A rapid inmunoblot Assay (Western Blot), to Detected

[156] Tsang C W Víctor., Pilcher Joy A., Zhou Wei., et al Efficacy of the inmunoblot assay for cuysticercosis in pigs and in pigs and mpodiulated expression of distinct IgM, IgG activities to taennia solium antigens in exparimental infections. Veterinary Inmonology

[157] Wilson M., Bryan Ralph T., Fried Janet A., et al., Clinical evaluation of the cysticercosis Enzyme-Linked Inmunoelectrotransfer Blot in Patients with neurocysticercosis. The

[159] Flisser A, Gyorkos TW. Contribution of immunodiagnostic tests to epidemiological/intervention studies of cysticercosis/taeniosis in Mexico. Parasite

Taenia Solium (cysticercosis), Journal of Inmuno Assay; 1989, 10(2-3-:237.

[158] Flisser A. Overview of immunology. Arch Med Res. 1997 Spring;28(1):145, 147

porcine cysticercosis in Perú. Am. Jour. Trop Med Hyg; 1990, 43(2):194-199.

Soliu). Journal H of Infectous Diseases; 1989, 159 (1): 50-59.

summary of a scientific event.Bol Chil Parasitol. 1981 Jul-Aug;36(3-4):63-65.

cysticercosis: a review. Bull World Health Organ. 1979;57(5):839-856

non-responders. Clin Exp Immunol. 1980 Jan;39(1):27-37

8 Suppl 1):143-76

1986,14(1):115- 127.

Trop.Dis.Dec.2009;3(3):1-7.

*Infect Dis*.1989;159: 50– 59.

and Inmunopathology; 1991, 29:69-66.

Immunol. 2007 Dec;29(12):637-649.

Journal of infectous Diseases; 1991, 164:1007-1009.

(1)1-10.

http://en.wikipedia.org/wiki/Rudolf\_Leuckart (accessed 29 March 2012).

[145] Flisser A, Pérez-Montfort R, Larralde C. The immunology of human and animal cysticercosis: a review. Bull World Health Organ. 1979;57(5):839-856

36 Novel Aspects on Cysticercosis and Neurocysticercosis

*Neurol Sci.* 2010 Sep;37(5):557-561.

[132] Giovanni Battista Morgagni.

(accessed 25 March 2012).

[136] Friedrich Albert von Zenker.

15:160-179, 1946.

(Mex). 1975;6(1):1-12.

1993.

March 2012).[138] Rudolf Leuckart

*Ecuatoriana de Ciencias Neurológicas.* 1978;1:13-25.

complement fixation test. Neurology 1956;6:725-737.

15 March 2012).

Quito, 1991.

2012). [137]

http://www.greekmedicine.net/whos\_who/Avicenna.html accessed 15 March 2012). [124] Umami Medicine. http://www.greekmedicine.net/whos\_who/Avicenna.html (accessed

[126] Del Brutto O, Sotelo J. Neurocysticercosis Update. *Rev. Inf. Dis*.1988;10(6):1075-1087 [127] Olive J.L., Angulo, R.P. introduction and general aspects Panel discussion.

[128] McLachlan RS. Julius Caesar's late onset epilepsy: a case of historic proportions. *Can J*

[130] García E, Cruz, Historia de la Cisticercosis. *Teniasis y Cisticercos en el Ecuador*, ed. MSP,

[131] Mora R, et al., "Neurocisticercosis" .En infecciones del sistema nervioso central. *Ed.*

http://en.wikipedia.org/wiki/Giovanni\_Battista\_Morgagni (accessed 25 March 2012). [133] Abraham Gottlob Werner. http://en.wikipedia.org/wiki/Abraham\_Gottlob\_Werner

[134] René Laennec. http://en.wikipedia.org/wiki/Ren%C3%A9\_Laennec (accessed 25 March

[135] Rudolf Virchow. Wikipedia. http://creationwiki.org/Rudolf\_Virchow (accessed 28

http://en.wikipedia.org/wiki/Friedrich\_Albert\_von\_Zenker(accessed 29 March 2012). [137] Lasso J. Contribución a la historia de la cisticercosis cerebral.Vol XXXI, 1994 http://escuela.med.puc.cl/publ/cuadernos/1994/pub\_02\_94.html. (accessed 30 March 2012). [138] Alarcón T. Cisticercosis cerebral en el Ecuador, revisión de 32 casos estudiados. *Revista*

[140] Dickmann, G.H. Cisticercosis de la fosa craneana posterior. *Rev. Neurol* (B. Aires),

[141] Nieto D. Cysticercosis of the nervous system. Diagnosis by mean of spinal fluid

[143] Proaño J, Proaño J.V, Molina Alvarez, Lasso J. Guía de Intervención: normas técnicas, métodos y procedimientos para el programa de control y vigilancia epidemiológica del complejo teniasis/cisticercosis. Editorial Ministerio de Salud Pública, Quito-Ecuador

[144] Flisser A, Tarrab R, Willms K, Larralde C. Immunoelectrophoresis and double immunodiffusion in the diagnosis of human cerebral cysticercosis. Arch Invest Med

http://en.wikipedia.org/wiki/Rudolf\_Leuckart (accessed 29 March 2012).

[139] Carvajal J. La cisticercosis humana en México (simposio),1988; 124 (5,6), 1988.

[142] Obrador S. Cysticercosis cerebri. Acta Neurochirurgica. 1962; 10(3): 320-364.

[125] Macias V, Maqueo M. Cisticercosis cerebral. *Rev Invest Clin.* 1958;4:443-454.

Cysticercosis of the nervous system. J. Neurosurg 19:632-34, 1962.

[129] Flisser A. Neurocysticercosis in Mexico. *Parasitol Today*1988; 4:131-137.

*Fondo Educativo Interamericano.* Págs.267-283, Bogotá 1978.

	- [160] Medina MT, Rosas E, Rubio-Donnadieu F, Sotelo J. Neurocysticercosis as the main cause of late-onset epilepsy in Mexico. Arch Intern Med. 1990 Feb;150(2):325-7.

**Chapter 2**

© 2013 Morales-Montor et al., licensee InTech. This is an open access chapter distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

© 2013 Morales-Montor et al., licensee InTech. This is a paper distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use,

distribution, and reproduction in any medium, provided the original work is properly cited.

**Neuroimmunoendocrine Interactions in Murine**

**Cysticercosis: From the Lab Bench Work to Its**

**Possible Applications in Controlling Porcine**

**Cysticercosis and Human Neurocysticercosis**

The immune and neuroendocrine systems are interconnected by a network in which hormones, antigens, receptors, cytokines, antibodies and neuropeptides modulate immune response in connection with neuroendocrine changes while maintaining homeostasis (Besedovsky & del Rey, 1996). Two of the main components of this network are the hypothalamic-pituitary-adrenocortical (HPA) and the hypothalamic-pituitary-gonadal axes

Interactions between the immune system and both the HPA and HPG axes are characterized by their activation and starting of the stress response, which, in turn, has immunomodulating activities that are important in preventing excessive immune responses (Chikanza & Grossman, 2000; Morales-Montor *et al.*, 2004b). Furthermore, the important functions of both axes have been shown for adaptation and maintenance of homeostasis during critical illness and viral, bacterial, parasitic and autoimmune diseases (Besedovsky & del Rey, 1996). An important aspect of cell communication that has emerged as a result of studying neuro-endocrine-immune interactions is the redundancy of the use of some chemical messengers to communicate among them. As an example, neurotrophins are chemical messengers first identified and characterized in the nervous system. Members of this protein family are also expressed and secreted by immune and endocrine cells, and

Karen Nava-Castro, Romel Hernández-Bello, Saé Muñiz-Hernández, Lorena López-Griego,

Additional information is available at the end of the chapter

(HPG) (Besedovsky & del Rey, 2000; Rivier & Rivest, 1993).

have immunological and endocrinological functions (Haddad *et al.*, 2002).

http://dx.doi.org/10.5772/50700

**1. Introduction**

Jhonatan A. Hernández-Valdés, Valeria López-Salazar, Nelly Tiempos-Guzmán and Jorge Morales-Montor


**Author details**

Vicente Enrique Machaca Arceda1 \*†, Miguel Angel Machaca Arceda2† and Pamela Claudia Laguna Laura2†

1 Universidad La Salle, Arequipa, Perú

2 Universidad Nacional de San Agust'in, Arequipa, Perú

\*Address all correspondence to<EMAIL_ADDRESS>
† These authors contributed equally.

© 2021 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

*COVID-19 Pandemic: Analysis and Statistics of Confirmed Cases DOI: http://dx.doi.org/10.5772/intechopen.98891*


*3.2. PHA Production by P. resinovorans Using Various Types of Carbon*

Microbial metabolism for the accumulation of PHA occurs in two different ways: (1) short-chain carbon sources are usually utilized by biosynthesis for accumulation of PHA by the phaABC pathway, through acetyl-CoA to acetoacetyl-CoA by phaA (thiolase), while phaB (oxidoreductase) and phaC (polymerase) are mainly involved in intracellular processes; and (2) long-chain carbon sources, such as fatty acids or alkanes, are utilized through the β-oxidation pathway, and various acyl-CoA transferases convert to intermetabolites, including the mcl-PHA precursor [17]. *P. resinovorans* has both metabolisms for PHA accumulation; therefore, there is a need to evaluate the PHA production of different carbon sources. *P. resinovorans* was cultured using 2% of glucose, 20 (*v*/*v*)% of each alkane (n-octane, n-decane, or n-dodecane), and 20 (*v*/*v*)% of mixed alkane to compare PHA production and the monomer composition in accordance with the carbon

source (Table 1). With glucose as the carbon source, 1.56 g/L of PHA was produced that contained 1.47% 3-hydroxyhexanoate (3HHx), 10.60% 3-hydroxyoctanoate (3HO), and 87% 3-hydroxyoctanoate (3HD). Among the carbon sources with 20% of a single alkane (n-octane/n-decane/n-docane), the best cell growth and PHA production were found using n-octane as the sole carbon source. This produced a CDW of 1.74 g/L, PHA of 0.37 g/L, and PHA content of 22.4% that contained 4.59% of 3HHx, 86.05% of 3HO, and 9.36% of 3HD. When the mixed alkane was used, 0.69 g/L of PHA was produced that contained 11.77% of 3HHx, 74.39% of 3HO, and 13.84% of 3HD. Although cell growth was not the highest, the mixed alkane showed an approximately 50% increase in PHA production compared with using n-octane.

**Table 1.** Cell growth, PHA production, and monomer composition by *P. resinovorans* in various carbon sources.

3HHx: 3-hydroxyhexanoate, 3HO: 3-hydroxyoctanoate, 3HD: 3-hydroxydecanoate.

#### *3.3. Mixture Analysis of Alkanes (n-octane, n-decane, and n-dodecane) as a Carbon Source for mcl-PHA Production*

To investigate the effect of the three alkanes on cell growth, PHA production, and PHA content, *P. resinovorans* was grown as 10 culture compositions based on the mixture analysis model, as described in "Materials and methods" (Table 2). For each culture, we measured the cell growth, amount of PHA, and PHA content. The results of the statistical analysis were shown by contour plots and the predicted highest cell growth, PHA titer, and PHA content using *P. resinovorans* were determined (Figure 3).

**Table 2.** Monomer composition of 10 conditions by mixture analysis.

3HHx: 3-hydroxyhexanoate, 3HO: 3-hydroxyoctanoate, 3HD: 3-hydroxydecanoate.

Contour plots for cell growth predicted the best ratio to be when n-octane, n-decane, and n-dodecane are 10.36%, 5.48%, and 4.5%, respectively, which produced CDW of 5.28 g/L (Figure 3A). In the case of 9.15% of n-octane, 6.44% of n-decane, and 4.29% of n-dodecane, the PHA titer was 2.39 g/L of PHA (Figure 3B). In addition, the PHA content calculated by the amount of PHA contained in the cell was predicted to be a maximum of 52.33% content (*w*/*v*) when n-octane, n-decane, and n-dodecane are 8.63%, 6.86%, and 4.50%, respectively (Figure 3C). The optimal alkane composition for total PHA production predicted by the model is a combination of n-octane, n-decane, and n-dodecane, rather than pure alkane or a combination of two different alkanes [16,22,23]. The reason why a

combination of alkane compounds in the medium is beneficial is not currently clarified, but is estimated to be due to the better activities of alkane hydroxylase of *P. resinovorans* when the mixed alkane exists [24,25].

**Figure 3.** Mixture contour plots of mixed alkane composition for PHA production. The optimal composition ratio between n-octane, n-decane, and n-dodecane was determined as 20% (*v*/*v*) as a total concentration. (**A**): CDW (g/L), (**B**): PHA (g/L), and (**C**): PHA content (%) of different mixed alkane compositions.

#### *3.4. Jar-Scale Fermentation for mcl-PHA Production in Media-Optimized Conditions*

Carbon, nitrogen, and phosphorus are essential nutrients for growth, and their limitation can trigger mcl-PHA production in micro-organisms. However, the specific molecular mechanisms that drive this synthesis in *Pseudomonas* species under unfavorable growth conditions remain poorly understood. Therefore, it is necessary to determine the optimal concentration of nitrogen and phosphorus, because mcl-PHA production is related to their concentration when alkanes are used as carbon sources. To determine the optimal conditions, nitrogen concentrations were observed from 0% to 1.0% and phosphorus concentrations from 0% to 3.0% (Figure 4). When the concentration of nitrogen in the culture medium increased from 0 to 0.05%, PHA production tended to increase accordingly. The maximized PHA produced in 0.05% nitrogen was 1.31 g/L, but in the medium where the nitrogen concentration was more than 0.1%, PHA production decreased (Figure 4A). When the concentration of phosphorus in the medium increased from 0 to 1%, the production of PHA increased as well (Figure 4B). The maximum PHA production was 1.14 g/L at a concentration of 1% phosphorus. However, PHA production tended to decrease when the concentration of phosphorus was more than 2%. These results suggest that the optimal phosphorus concentration in the medium is 1.0%.

Using the optimized alkane combination and nitrogen and phosphorus concentration, *P. resinovorans* was cultured in a 7 L jar fermenter (3 L of working volume). The CDW, amount of PHA, and PHA content reached 3.5 g/L, 2.1 g/L, and 60%, respectively (Figure 5). Previous reports focused on *P. oleovorans* for the production of mcl-PHA with n-octane, and these show production of up to 16.8 g/L of PHA by fed-batch culture, while n-decane and n-dodecane were not considered because of low PHA production (Table 3). Compared with other *Pseudomonas* species, it was proven that use of *P. resinovorans* is more effective for production of mcl-PHA when mixed alkane is used as a carbon source.

**Table 3.** mcl-PHA production using various alkanes by Pseudomonas species.

CDW: final cell dry weight, PHA formed: final concentration of PHA, 3HB: 3-hydroxybutyrate, 3HHx: 3-hydroxyhexanoate, 3HO: 3-hydroxyoctanoate, 3HN: 3-hydroxynonanoate, 3HD: 3-hydroxydecanoate, 3HUD: 3-hydroxyundecanoate, 3HDD: 3-hydroxydodecanoate and 3HTD: 3-hydroxytetradecanoate.

#### *3.5. Physical Properties of Produced mcl-PHA by P. resinovorans*

The thermal properties of the produced mcl-PHA were determined by means of DSC (Table 4 and Supplementary Figure S2). The Tg of the samples ranged from −43.44 to −34.29, which can be regarded as values typical for this type of PHA. The sample also showed a Tm in the range of 39.62 with a ΔHm of 13.2 J/g. In addition, these values are typical for mcl-PHAs and indicate classical rubber- to latex-like characteristics [26]. The Mw, Mn, and

polydispersity (Mw/Mn) were similar for the copolyesters, despite the differences in their monomer compositions. The Mn and Mw were 267,649 and 630,526 Da, respectively.

**Table 4.** Physical properties of mcl-PHA from *P. resinovorans* using mixed alkane.

In addition, Z average (Mz) and Z + 1 average (Mz + 1) molar mass were determined as 1,110,977 and 1,729,603 Da, respectively. The polydispersity index (PDI) was calculated (PDI = Mw/Mn) as 2.36, more or less similar to that of the PHA value of mcl-PHAs biosynthesized by other known *Pseudomonas* species.

**Figure 4.** Effects of nitrogen and phosphorus limitation on PHA production. (**A**): CDW, PHA titer, and PHA content (%) in range of 0 to 1% nitrogen concentration. (**B**): CDW, PHA titer, and PHA content (%) in range of 0 to 3% phosphate concentration.

**Figure 5.** PHA production by *P. resinovorans* in optimized condition. Batch culture was performed in 7 L scale jar fermenter (3 L as working volume) with optimized culture medium.

#### **4. Conclusions**

Recent studies have shown that the use of renewable carbon sources can produce a significant reduction in actual production costs. A very interesting research topic would be for industrially produced waste oil or oil produced by pyrolysis of waste plastics to be used as a fermentation substrate for production of PHA, which is a promising bioplastic. Therefore, it is important to find PHA-producing micro-organisms and establish culture conditions in accordance with the carbon composition of waste oil or oil from waste plastics.

In this study, the ability of *P. resinovorans* to produce PHA from alkanes (n-octane, n-decane, and n-dodecane), especially mixed alkane, was identified, and a model was built to improve mcl-PHA production. Compared with other *Pseudomonas* species, *P. resinovorans* was much more effective in utilizing mixed alkane, and it can be an attractive microbial host for production of PHA. Although the regulation of PHA production based on alkane utilization has not been discovered, the results of this study are meaningful for use as basic data for the application of various renewable carbon resources containing alkanes.

**Supplementary Materials:** The following supporting information can be downloaded at: https: //www.mdpi.com/article/10.3390/polym14132624/s1, Figure S1: Pyrolysis oil of waste-plastic and its composition, Figure S2: DSC analysis of purified PHA produced by *P. resinovorans* using mixed alkane.

**Author Contributions:** J.-M.J.: Writing—Original Draft, Investigation, Data Curation. S.-J.P.: Methodology, Investigation, Data Curation. Y.-S.S.: Data Curation. Y.-H.Y.: Writing—Review and Editing, Methodology. J.-J.Y.: Conceptualization, Supervision, Project administration, Funding acquisition. All authors have read and agreed to the published version of the manuscript.

**Funding:** This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korean government (Ministry of Science and ICT or MSIT; NRF-2020R1A2C2102381) and the Korea Institute of Industrial Technology (KITECH) through Development of eco-friendly production system technology for a total periodic resource cycle (21-0140).

**Institutional Review Board Statement:** Not applicable.

**Informed Consent Statement:** Not applicable.

**Conflicts of Interest:** The authors declare no conflict of interest.


**3.2 Characteristics of SW-GW interactions at the regional scale**

The interaction of SW-GW is related to low topographical pathways varying from 10 to 100 km or more at a regional scale. The recharge or discharge

**Figure 6.** *River water and groundwater interactions at the regional scale.*

trends of groundwater, regional topographic, hydrological conditions are mainly characterized by the regional groundwater flow system at a regional scale. The hydrological processes of the large watershed such as precipitation, surface run-off, infiltration, evapotranspiration, base flow, streamflow, and channel conditions are described in **Figure 6**. All hydrological processes can cover on the regional scale of a large watershed and a small stream reach conditions can cover local scales. Slow hydrological response to recharging areas will result in an insignificant seasonal impact on the regional GW flow system. The regional flow system is developed with long-distance SW flows and low charging levels on higher topographical slopes. It explains why recharging and discharge levels fluctuate more at the local level than at the regional level. The recharge or discharge rate investigation of the regional flow system can be analyzed using environmental tracer isotopes and hydro-chemical characteristics (major ions and heavy metals) [6].


**Meet the editors**

Cândida Lucas is a Portuguese, born at Porto in 1958. She has a PhD degree in Biology/Microbiology from the New University of Lisbon/Gulbenkian Institute of Science (1988), following a degree in Biology (University of Lisbon [1982]). She is a full professor of the Biology Department from the University of Minho, Portugal, since 2015. Then, since 1990, she contributed to create

postgraduation courses and programmes on Molecular Biology/Genetics. She directed the Centre of Molecular and Environmental Biology Research (CBMA) (2006–2013) and is presently a codirector of the Institute of Science and Innovation for Bio-Sustainability (IB-S) from the University of Minho (http://ib-s.uminho.pt/). Her scientific interests covered diverse aspects of yeast biology, from plasma membrane transporters and osmotolerance to the extracellular matrix and differentiation (h-index 18).

Célia Pais has a PhD degree in Biology/Microbiology from the University of Minho, Portugal (1990), and a postgraduate degree in Plant Pathology from the Imperial College, London, UK, following a degree in Biology at the University of Coimbra, Portugal (1978). She is an associate professor with habilitation at the Biology Department, University of Minho, where she was the

vice-president of the School of Sciences (2002–2006) and the head of the Biology Department (2012–2014). She participated in the creation of the PhD Programme in Environmental and Molecular Biology that she coordinated from 2008 to 2012. She develops her scientific activity at the Centre of Molecular and Environmental Biology Research (CBMA), focusing on the study of yeast diversity and the development of new molecular tools for diagnosis and genotyping.

## Contents

#### Chapter 7 **HMGB Proteins from Yeast to Human. Gene Regulation, DNA Repair and Beyond 139**

Vizoso-Vázquez Ángel, Barreiro-Alonso Aida, Rico-Díaz Agustín, Lamas-Maceiras Mónica, Rodríguez-Belmonte Esther, Becerra Manuel, González-Siso María Isabel and Cerdán María Esperanza

#### **Section 3 Yeast Versus Plant - Unknown Territory 167**

Chapter 8 **Endophytic Yeast and Hosts: A Mutualistic Association Friendly to the Environment 169**

Esperanza del Pilar Infante Luna


**3. Discussion**

Data presented in Table 2 clearly indicate that there are major di fferences in the composition of the essential oils of three studied cultivars. All three grades of resin sample gave the highest value for α-pinene, similar findings reported by already mentioned authors [11] and a comparable range for α-pinene was noted in di fferent fractions of the essential oil of *B. sacra* [13]. The percentage of monoterpenes and sesquiterpenes reported for the Grade 1 essential oil of Najdi showed similar values as reported for *Boswellia serrata* [14]. However, there is a major di fference in the values recorded for α-pinene in this study, which was 79.59%, while only 5.3% was reported for *B. serrata* [15].

The increased time of extraction through hydrodistillation in three cultivars has resulted in a reduction of monoterpenes, while at the same time the content in % of sesquiterpenes is increased. Moreover, on the basis of data collected, it can be stated that the same species of plants behave di fferently in terms of the yield of active components of essential oils under di fferent environments. The Najdi sample showed a higher percentage for monoterpenes in all three grades compared to the other two frankincense samples.

Concerning the antimicrobial activity, it was interesting to note that both *S. aureus* and *P. aeruginosa* tested strains were susceptible to most of the essential oils obtained from the three different cultivars. As there are not many antimicrobials concurrently active against the above-mentioned pathogens, these results are relevant because there is a pressing demand for effective antimicrobials towards these microorganisms present on the WHO/OMS priority bacteria list. Therefore, tested essential oils could be a source for new anti-staphylococcal and anti-pseudomonal molecules. Grade 2 of Sahli showed a MIC at 0.264 mg/mL for *P. acnes*, an important pathogen of dermatologic relevance. The activity could be explained by a higher concentration of sesquiterpenes in the extracts obtained by subsequent hydrodistillation of 4 h.

A previous article on four essential oils from different *Boswellia* species showed a significant activity against *Candida tropicalis* and *C. albicans* [16]. In our study we found that all tested samples showed an interesting antifungal activity against *C. albicans* and *M. furfur* with MIC values ranging from 27.28 to 0.240 mg/mL. A high percentage of α-pinene in all Grade 1 essential oils (and in general in monoterpenes) could explain the activity against tested fungal pathogens. Our findings could enable the use of frankincense oil blends in many fields, such as starting products for dermatological bacterial and fungal infections treatments.

In conclusion, GC-MS analysis of *B. sacra* essential oil indicates α-pinene as its primary component. α-Pinene has been found to be one of the integral components of many other antimicrobial essential oils, and thus can be inferred as one of the main players in the antimicrobial activity of *B. sacra* essential oils. Indeed, many of the monoterpenoids putatively identified in our study have been previously reported to have a potent broad antimicrobial spectrum activity. Moreover, the occurrence of drug resistance towards an essential oil in a microorganism is very rare because of its multicomponent nature that necessitates modification of numerous targets [17].

#### **4. Materials and Methods**

## *4.1. Sample Collection*

The different *B. sacra* gum resins were purchased from local market, collected over a period from March to May 2016, and compared with authenticated samples preserved in the Herbarium Center of Nizwa University, Oman, under Voucher number UC29.

The sample named Najdi was obtained from a plateau behind the Dhofar Mountains, where a rapid decline in rainfall and moisture was noted in this region. At the same time, temperature variances rise and desert weather changes the climate of the plain, including the southern slopes. Though annual rainfall in the Jebel region ranges from 500–750 mm, precipitation in the Nejd is recorded only in traces. The sample named Sahli was obtained from valleys (Shabi) of the Dhofar region. The normal annual precipitation is around 110 mm but can be from 70 to 360 mm. July to August is usually the rainy period. The sample Houjri is from Jebel Samhan of the Dhofar region. The Jebel hill range forms a distinct agro-climatic region. Thick fogs are detained back through ranges from inner desert areas. Precipitation is predominantly high and it can be from 600 to 700 mm, the highest in the country, which supports an enduring flora cover.

#### *4.2. Extraction of Essential Oils*

The essential oils from different cultivars of *B. sacra* were obtained through a hydrodistillation method using Clevenger-type apparatus. A total of 250 g of finely grounded frankincense oleogum resin was added to 2500 mL distilled water in a 5000 mL bottom flask placed in a heating mantle. Hydrodistillation was performed under atmospheric pressure at boiling temperature (about 100 ◦C) using a closed steam jacket. Following two hours of hydrodistillation, a Grade 1 essential oil of each oleogum resin was collected, whereas Grade 2 and Grade 3 essential oils were collected after 4 and 6 h of successive distillation of the same resin sample. Therefore, a total of nine gum resin essential oils

were obtained. The resulted essential oils were collected, dried with anhydrous sodium sulfate, and kept in a dark glass bottle at 4◦ C until analyzed by gas chromatography–mass spectrometry (GC-MS).

#### *4.3. GC*/*MS Analysis*

GC/MS analysis of obtained essential oils was carried out by using a Thermo Scientific DSQ II single quadrupole system of an EI (electron ionization) type, employed in full scan. The temperatures of the injector and ion source were 265 ◦C and 260 ◦C, respectively. The capillary column was a ZB-WAX (30 m x 0.25 mm i.e., film breadth 0.25 μm, (Phenomenex, Italy)). The temperature of the oven was set and the column temperature started at 50 ◦C, was raised at 3 ◦C/min to 250 ◦C, and after that a second gradient was used to 300 ◦C at 40 ◦C per minute, which was then seized for 3 min under isothermal conditions. The flow rate of helium, the carrier gas, was 1 mL/min. A sample of 1μL was inoculated with a split ratio of 1:100. The temperature of the ion source was 260 ◦C, the temperature of the MSn transfer line was 265 ◦C and the temperature of the injector was 270 ◦C. Ionization voltage was 70 eV and the mass range scanned was 35–550 *m*/*z*. Each oil component was identified on the basis of its retention index, relative to C8-C36 n-alkanes, from MS library searches using the NIST and Wiley GC-MS databases, and by comparison with mass spectral data in the literature.

The percent composition of the essential oil was computed by a standardization process from the GC peak areas, supposing identical mass response factor for all components. Triplicate analyses were prepared for each oil sample.

#### *4.4. Antimicrobial Activity, Minimum Inhibitory Concentrations (MICs) Determination*

The MICs of essential oils for seven reference bacterial strains (two strains of *Staphylococcus aureus* (ATCC 25923; ATCC 6538), two reference strains of *Pseudomonas aeruginosa* (ATCC 15442; ATCC 9027) and *Staphylococcus epidermidis* ATCC 12228, *Staphylococcus hominis* ATCC 27844, and *Propionibacterium acnes* ATCC 11827) were defined by means of an already described technique for a dilution antimicrobial susceptibility test for bacteria that grow aerobically (CLSI), with few amendments [18]. Concisely, 0.1 mL of essential oil was mixed with 0.1mL of culture medium (Mueller Hinton broth, Sigma-Aldrich, Milan, Italy) present in a well of a sterile 96-wells plate, and a 1:2 dilution series with broth medium was performed. A growth control and a negative sterile well control was included for each plate. A bacterial suspension (10 μL) from a suspension in NaCl 0.9% of isolated clusters, preferred from a 18 to 24 h agar plate and attuned to get a turbidity equivalent to a 0.5 McFarland standard, was added into the wells and the final test concentration of bacteria was approximately 5 × 106 CFU/mL bacterial. The 96-well plates were incubated at 37 ◦C for 24 h and MICs were read by a microplate spectrophotometer (GloMax®-Multi Detection System, Promega Italia s.r.l, Milan, Italy) as the lowest concentration of the sample that fully suppresses bacterial growth in the microdilution wells and whose optical density (OD) at 570 nm is comparable to OD values of negative control wells (only medium, without inoculum).

Antifungal activity against *Candida albicans* (ATCC 10231) and *Malassezia furfur* (ATCC 14521) were evaluated in terms of MICs by using a similar micro-method but by employing Sabouraud broth (Sigma-Aldrich, Milan, Italy) or Sabouraud broth with added 2% olive oil in the case of *M. furfur*, as growth media, and with an incubation time of 48 h.


**7. Conclusions**

Different hypotheses were given to elucidate the physiological substrates for the ABCC6 transporter and its pathological involvement in PXE. However, mechanistic details for *ABCC6* mutations leading to ectopic mineralization were yet to be resolved [6,47,48]. In our studies, we not only investigated the structural components of ABCC6 transporter

but also lighten the mechanisms, which might be involved in ectopic mineralization. In our experiments, knockdown of *ABCC6* or its inhibition by probenecid decreased the extracellular ATP concentration and altered the expression of *NT5E* and *TNAP*. By these findings, we can propose that the lack of ABCC6 transport activity could lead to a promineralizing state through alteration of extracellular purine metabolites, which ultimately affect TNAP enzymatic activity.

Moreover, in both knockdown and probenecid-treated HepG2 cells, the expression of lamin was found decreased, which suggests that reduced ABCC6 transport activity also leads to cell senescence in PXE and can be beneficial to prevent cancer progression. Interestingly, in both the knockdown and probenecid-treated HepG2 cells, we found a decrease in migration rate, which is restored after ATP administration.

Collectively, these findings suggest that the ABCC6 transporter is not only required to maintain some important circulatory factors like PPi, but is also required to maintain proper functioning of other factors, which are involved in the conversion of extracellular nucleotides, and to feed purine pool to maintain homeostasis between Pi and PPi ratio. Moreover, our studies strengthen the idea that the ABCC6 transporter is not only involved in PXE pathophysiology, but can also be considered a target for anti-cancer therapy.

**Author Contributions:** Conceptualization, F.B. and A.O.; data curation, A.O.; writing—original draft preparation, V.A. and P.K.; writing—review and editing, P.K. and M.A.C.M.; supervision, F.B.; funding acquisition, F.B. and A.O. All authors have read and agreed to the published version of the manuscript.

**Funding:** This research was funded by INBIOMED PROJECT (MIUR, ARS01\_01081).

**Conflicts of Interest:** The authors declare no conflict of interest.


**5. Conclusion**

As been discussed during this chapter, there are multiple environmental and occupation factors that are well known to cause asthma and worsening of asthma symptoms. Many triggers been recognized that can be divided into allergic and non allergic which is important at the moment of symptoms interrogation. A methodology approach and personalized approach need to be done for early and proper recognition of this factors. Educational programs on avoidance and recognition needs to be provided to the general population as well, education regarding common symptoms

when dealing with this exposure which will eventually lead for better patient respiratory control and quality of life.