Reasearch of the balsam

Journal of Ethnopharmacology 101 (2005) 16–26
The Jerusalem Balsam: From the Franciscan Monastery in the
old city of Jerusalem to Martindale 33
A. Moussaieff a, E. Fride b, Z. Amarc, E. Levd, D. Steinberg e, R. Gallily f, R. Mechoulama,.
a Department of Medicinal Chemistry and Natural Products, School of Pharmacy, Hebrew University, Jerusalem 91120, Israel b Department of Behavioral Sciences, The College of Judea and Samaria, Ariel, Israel c Department of Land of Israel Studies, Bar-Ilan University, Ramat Gan, Israel
d Department of Eretz Israel Studies, University of Haifa, Israel e Institute of Dental Sciences, Faculty of Dentistry, Hebrew University-Hadassah, Jerusalem, Israel f Lautenberg Center for Immunology, Medical School, the Hebrew University, Jerusalem, Israel Received 4 January 2005; received in revised form 3 March 2005; accepted 5 March 2005 Available online 15 June 2005
The Jerusalem Balsam, a remedy based on an ethanolic extract of a herbal mixture, was formulated in 1719 in the pharmacy of the SaintSavior monastery in the old city of Jerusalem. Having gained fame, the Jerusalem Balsam was replicated and prepared in Europe.
One can still find variations of the formula in current pharmacopoeias (B.P., 1998. The Stationary Office, London, p. 1510; Sweetman, S.C., Blake, P.S., McGlashan, J.M., Parsons, A.V., 2002. Martindale: The Extra Pharmacopeia, 33rd ed. Pharmaceutical Press, London, p. 1101). We report here, five different formulas, all referred to as “The Jerusalem Balsam”. Three of those formulas were translated and two of these translations are presented in the text. A third one is available as Supplementary data online. As the formulas originate from different historical periods, the Jerusalem Balsam may be a good case study of the development of pharmaceutical formulations over a 250 years period.
One of the formulas, found in a manuscript form in the archive of the monastery, contains four plants: olibanum (Boswellia spp.), myrrh (Commiphora spp.), aloe (Aloe sp.) and mastic (Pistacia lentiscus L.). We conducted pharmacological assays on this four-plant formula. It showed anti-inflammatory, as well as anti-oxidative, and anti-septic properties. © 2005 Elsevier Ireland Ltd. All rights reserved. Keywords: Aloe sp.; Balsam; Boswellia spp.; Commiphora spp.; Myrrh; Olibanum; Mastic; St. Savior;
Traditional medicine
1. Introduction
1.1. Balsams

Balsams have been in medical use for centuries for the treatment of wounds, various skin conditions and other uses. Although the term “balsam” originally referred to a formulation of natural products with medicinal effects, the term has undergone a number of changes down the centuries. Now the
term is understood to indicate drug mixtures containing either Abbreviations: JB, Jerusalem Balsam; ROS, reactive oxygen species; OD, optical density; BHI, brain heart infusion . Corresponding author. Tel.: +972 2 6758634; fax: +972 2 6757076.
E-mail address: (R. Mechoulam).
natural or synthetic resins of varied composition. Balsams consist chiefly of resins, gums, essential and volatile oils, and aromatic acids (mainly cinnamic and benzoic) (Reynolds and Prasad, 1982).

1.2. St. Savior Monastery’s pharmacy Elzear Horn, a Franciscan monk of German origin, who
resided in Jerusalem between 1724 and 1744, has described at length the pharmacy of the Franciscans, as one of the finest in the entire Christian world (Horn, 1962), as did the Swedish botanist Frederick Hasselquist, who visited the institution in 1750 (Hasselquist, 1866; Schittny, 1993b).  The pharmacy ceased to function sometime during the mid- 0378-8741/$ – see front matter © 2005 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.jep.2005.03.028 A. Moussaieff et al. / Journal of Ethnopharmacology 101 (2005) 16–26 17
dle of the 20th century. The last documents we found in the monastery archives that pertain to the pharmacy are from 1932.

1.3. The Balsam of Jerusalem
The Balsam of Jerusalem was a widely known and important remedy for about two centuries in Europe and the Near East. It was formulated by Antonio Menzani di Cuna (1650–1729), a friar physician and pharmacist in the Saint Savior monastery in Jerusalem. “For 24 years Father Anthony
used his best exertions to find an effective balsam, and when after many experiments he emerged successfully, he made his discovery known in Milan in 1719” (Bagatti, 1946). It has been suggested, however, that the formula had earlier versions, for example, one mentioned in a 1608 medical treatise
by the physician Oswald Croll, which however does not contain Aloe, a component of the Menzani Formula (Schittny, 1992, 1993a). Eventually, Menzani published his formula in Milan in 1719 (Bagatti, 1946).
Having gained fame, the Jerusalem Balsam was replicated and prepared in Europe (Schittny, 1993a), but by the late 19th century itwas no longer a sought-after commodity. Bagatti (1946) writes about four different kinds of balsams formulated by Antonio Menzani di Cuna, all of them called
the “Jerusalem Balsam”. The Balsam was a kind of panacea, used in several differentways. Given topically, itwas used for healing ofwounds of all kinds, bruises, and all skin disorders.
Given systemically, it was used as a prophylactic in times of plagues, for treatment of stomach aches, worms, hemorrhoids, headaches, dizziness, ear and teeth problems, blood spitting, and cardiac disease (Hasselquist, cited in Bagatti, 1946).

1.4. The herbal constituents of the Balsam
The formula of the Balsam, found in the convent’s archive in manuscript form is a herbal mixture of four ingredients: “Olibanum”, “Myrrh”, “Mastic” and “Aloe”. Olibanum is the resin of Boswellia species (Burseraceae), native of Eastern Africa, where it is used as incense and for medical purposes. Its main medical uses are as a diuretic, for bilharzia, for stomach ache and for the treatment of
syphilis. In India, it is used for the treatment of rheumatism (Watt and Breyer-Brandwijk, 1962). Over the years, Boswellia species resin has been shown to be active in the treatment of inflammations (Singh and Atal, 1986), as well as in several diseases associated with inflammatory conditions
such as chronic collitis (Gupta et al., 2001). Previous reports indicate that the anti-inflammatory properties of Boswellia resin can be attributed to boswellic acid and its derivatives
(Reddy, 1989). These have been found in the resin of Boswellia species from Africa and from India (Buchele et al.,2003).
Myrrh is an oleo-resin, obtained from the stem of Commiphora molmol Engl. and other species of Commiphora (Burseraceae), which grow in north-east Africa and Arabia. It is used in incense and perfumes. It is chiefly employed in medicine in the form of mouth wash and has local antiseptic
qualities (Evans, 1996). It has controversial antiseptic and antibiotic properties and has been used as a vulnerary for wound healing (Newall et al., 1996). Another constituent of the Balsam is referred to as “aloe suc” (see Fig. 1). This can be interpreted in two different ways: either as Aloe “juice”, as succo is Italian for juice, or as Aloe succotrina Lam. (Aloe perryi Baker) (Asphodelaceae) – a species growing in the island of Succotrine. We decided to conduct our pharmacological experiments using Aloe vera
L. (Asphodelaceae), for several reasons: (A) the botanical identification of the plants in the text is somewhat vague and as Aloe vera L. is the species most widely investigated in the context of wound healing (a main medical use of the Balsam), it makes sense to examine it as a part of the formula.
It is also more readily available than Aloe succotrina. (B) Herbal materia medica mention abbreviations such as “Suc d’Aloes” referring to the juice of Aloe species (Steinmetz, 1954). Aloe vera L. provides two main herbal remedies of quite a different nature. “Aloes” is the solid residue obtained by evaporating the liquid, which drains from the leaves of various species of Aloe. Aloes is usually employed as purgative (Evans, 1996). The Aloe vera gel is obtained from the mucilage of the plant leaves and is rich in polysaccharides. A considerable amount of work has been done on the wound
healing properties of the Aloe vera gel, however, recent reviews find most published data controversial and unclear (Vogler and Ernst, 1999). Recently, an Aloe vera-derived drag-reducing polymer (DRP)was shown to prolong survival in rats with lethal hemorrhagic shock. DRP has visco-elastic properties, which may account for its putative wound healing action (Macias et al., 2004).
Mastic is the resin obtained from the stem and larger branches of Pistacia lentiscus L. (Anacardiaceae). It is a Mediterranean shrub that is used as an expectorant and stimulant
(Evans, 1996; Neuwinger, 2000).

2. Materials and methods
2.1. Manuscripts
Original manuscripts were located and copied in the archive and library of the St. Savior’s convent in Jerusalem. The archive documents were translated. We used conversion tables, used in the convent’s pharmacy to translate old pharmacological signs (Fig. 1).

2.2. Plants materials
Herbal constituents of the Balsam were purchased from Pamir (Tel-Aviv, Israel) and the Balsam was formulated by extracting the herbal mixture in absolute ethanol, following 18 A. Moussaieff et al. / Journal of Ethnopharmacology 101 (2005) 16–26 Fig. 1. Formula 1 of the Jerusalem Balsam and a conversion table of pharmaceutical signs located in and photocopied from the archives of St. Saviors convent in Jerusalem. formula 1 (found in the archive). For wound healing experiments, two  variations were tested, following the manuscript formula: (A) Boswellia carterii Birdw. resin (18 g), Pistacia lentiscus L. resin (12 g), Aloe vera gel (9 g) and Commiphora molmol Engl. resin (3 g) was extracted in 300 ml ethanol for three days. (B) The herbal mixture was as in (A), but Aloe
vera gelwas replaced by Aloe vera juice. Formula Bwas used for the anti-inflammatory, anti-bacterial and anti-oxidative experiments. The dry weight of 1ml of formula B was 92.5 mg.

Pic 1- Formula of the Jerusalem Balsam and a conversion table of pharmaceutical signs located in and photocopied from the archives of St. Saviors convent

in Jerusalem.

2.3. Anti-in.ammatory model In order to examine the anti-inflammatory effect of the Balsam, we used an external ear mouse model of inflammation (Hanus et al., 1999). Sabra white mice (five mice in each

group) were injected intraperitoneally (i.p.) with formulation B (see above) 1 h prior to arachidonic acid administration on the ears (5 mg in 5 _l of ethanol).

The control ear was administered 5 _l of ethanol. After arachidonic acid administration,

the inflamed ears were examined every 15 min for A. Moussaieff et al. / Journal of Ethnopharmacology 101 (2005) 16–26 19 thickness (using a thickness gauge, [Mititoyo, Japan]) and redness (using a rating scale from 0–2), for 90 min.

2.4. Anti-bacterial assays For the anti-bacterial assay, we could not use Broth agar to check the microbial suspension for its optical density, as the proteins in it aggregate in the presence of the Balsam. Therefore we used an inhibition zone technique on Mueller–Hinton

agar plates. Paper discs, 1.4 cm diameter, were placed on the agar plates. Before placing the discs, we applied 20_l of the Balsam solution formulation B (see above) and 20 _l of chlorhexidine 0.2% solution on the test discs and 20 _l of ethanol on the control group. Microbes were homogenously

dispersed on the agar. The plates were then incubated at 37 .C for 48 h. After 48 h, we checked the agar for presence of colonies.

We used a live-dead bacterial viability kit (Molecular Probes, Oregon, USA) to confirm the above agar-plates tests:

50_l of a microbe suspension (O.D. = 1.5) was dripped on glass slides. 100 _l of the Balsam (formula B) was applied to 20 ml brain heart infusion (BHI) medium. Microbes were stained according to manufacturer’s protocol and confocal scanning laser microscope was used to view the slides.

Bacteria, Staphylococcus aureus and Streptococcus mutans ATCC 27315, were used as skin and oral bacteria models, respectively. They were obtained from the Department of Oral Biology, Faculty of Dental Medicine, the Hebrew University, Jerusalem.

2.5. Anti-oxidative assay In order to examine the anti-oxidative effect, mouse macrophage cells, RAW 264.7 cells (5ª105 cells) were suspended in 500 _l of Hanks balanced cell solution (without phenol red). Various doses of the Balsam formulation

B in 20_l of ethanolic solution were added and immediately thereafter, 30 _l of zymozan and 10 _l of Luminol were added to the suspension. Berthold Biolumat LB 9500 T spectrophotometer was used to determine levels of reactive oxygen species (ROS) at 25 .C (Avron and Gallily, 1995).

As control, 20 _l of ethanol was added to 1ml of buffer. Cell vitality was examined by erythrosine B staining (0.4%).

2.6. Wound-healing experiments To examine the effect of the Jerusalem Balsam on wound healing we used two models: as a model for a cut alone (healing by first intention) we used a model based on the bursting pressure of abdominal wall defects (Seror et al., 2003). As a model for wound healing that involves the contraction of a wound (healing by second intention) we used a burn model (Eldad et al., 1998).

2.6.1. Animals

For the anti-inflammatory assay, female Sabra mice (2 months old) were used. For the surgical wound-healing model, we used Sabra rats, weighing 200±30 g. We used guinea pigs, weighing 900±60 g for the burn wound healing model. The experiments on animals were performed according

to standards determined by the Committee on Ethics in Animal Research of the Hebrew University of Jerusalem. All animals were purchased from Harlan, Jerusalem.

2.6.2. Statistical analysis

Data from the ear inflammation assay were analyzed using two-way analyses of variance for repeated measures (time). At all time points, the groups were compared using Newman–Keuls post-hoc tests.

3. Results

3.1. The different formulas of the Jerusalem Balsam We found five different formulas with the name

“Jerusalem Balsam”. They are:

3.1.1 A formula found in a manuscript in the monastery archive (Fig. 1). The manuscript is dated 1778 (see also Schittny, 1993a).

3.1.2 A formula found in the city archive of Venice dated

early 18th century (Schittny, 1992; Schittny, 1993a)

(Fig. 2).

3.1.3 A formula found in a pharmaceutical book in the monastery library, dated 1904 (Cesaris, 1904) (Fig. 3).

3.1.4 A formula taken to Germany by a pilgrim friar and modified by a local pharmacist in 1860 (Schittny, 1992; Schittny, 1993a) (Fig. 4).

3.1.5 Formulas that are still a part of official pharmacopoeias (B.P., 1998; Sweetman et al., 2002). One can find these formulas designated as “Compound Benzoin Tincture”, which seems to be a synonym for the “Jerusalem Balsam”. Indeed Ansel et al. (1995) give the following background information for “Compound Benzoin Tincture”: “The tincture originated in the fifteenth or sixteenth century and through the years probably has acquired more synonyms than any other official preparation.

A few of these are indicated as follows. Synonyms: Friar’s Balsam; Turlington’s Drops; Persian

Balsam; Swedish Balsam; Jerusalem Balsam; Wade’s Drops; Turlington’s Balsam of Life.” We present here, for the first time to our knowledge, translated texts of formulas one and three that were found in the monastery itself. In spite of the vast differences between the formulas they carry the same title. These differences are discussed in the following section.

3.2. Formula 1

The text is in 18th century Italian and makes use of the pharmaceutical language and signs known at the time. The language differs from present-day Italian, and the pharmaceu- 20 A. Moussaieff et al. / Journal of Ethnopharmacology 101 (2005) 16–26 Fig. 2. The Venice formula of the Jerusalem Balsam, dated early 18th century (Schittny, 1993a,b Schitny, 1993). Fig. 3. The formula of the Jerusalem Balsam as recorded in a pharmaceutical book (Cesaris, 1904), found in the library of St. Saviors convent. Fig. 4. A formula of the Jerusalem Balsam used in Germany as reported by Schitny (1993). See text. A. Moussaieff et al. / Journal of Ethnopharmacology 101 (2005) 16–26 21 tical signs are rendered according to a decoding table from the monastery

Pic 2- The Venice formula of the Jerusalem Balsam, dated early 18th century (Schittny, 1993a,b Schitny, 1993).

Pic 3- The formula of the Jerusalem Balsam as recorded in a pharmaceutical book (Cesaris, 1904), found in the library of St. Saviors convent.

Pic 4- A formula of the Jerusalem Balsam used in Germany as reported by Schitny (1993). See text.

3.2.1. Translation

(For clarity we have paraphrased the translation or made comments on the text; our comments and wording are in square brackets). Homogeneous Balsam of Jerusalem [the name implies that

more formulas were known to the author as “the Jerusalem Balsam”]. Alcohol distilled from wine – 6 pounds, incense [Boswellia spp.] – 6 ounces, mastic [Pistacia lentiscus L.] – 4 ounces, Aloe [The original “aloe suc” could be rendered in two ways: (A) Aloe vera juice – succo in Italian;

(B) Aloe succotrina – a species growing in the island of Succotrine. See also Introduction] – 3 ounces, Myrrh [Commiphorra spp.] – 1 ounce. Then, put it [the Balsam] in a glass in the light of sun, inside

a “Bain-Marie” [a hot water bath] for a few days, until the “digestion” is good [until adequate maceration of the botanical drugs] and when it is well digested, keep the precipitate with the solvent.

It is of value in healing wounds of all sorts, cuts, wounds caused by arquebus [a kind of a gun], gunshot injuries, as long as they are not lethal, if applied swiftly. It heals fistulas and is excellent for all skin diseases; and against the biting of poisonous animals and rabies [in those cases],

it should be swallowed. in a soup [it is used for the healing of] hemorrhoids, the ears, the teeth, sciatic , “resipola” [Erysipela – a streptococcal infection of the skin], for insect bites. For [use during] the time of the plagues, or when infection is suspected, drink morning and evening 10 drops in soup [and it] will protect from becoming infected. For stomach aches, aches on the sides [above the waist] the gas expeller [common language for carminatives], [the Balsam] kills worms, [is used for] headaches, vertigo etc.

3.3. Formula 2

We have translated the plant list and the accompanying text. However, due to space limitations, we do not present the translation of this formula here. The translation of this formula is available as Supplementary data online. The first part of the text gives details of the different constituents

of the Balsam. Forty ingredients are listed under the names used for them in the pharmacy of St. Savior monastery in the beginning of the 18th century, the time when the manuscript was written (there is no record of the exact year). All but two of the ingredients are of herbal origin. We have

tentatively identified the different ingredients of the formula. However, the exact identification of the species is problematic in some cases for the following reasons: a. The botanical identification of the plants at the time of writing of the formula is not certain, and the present day botanical nomenclature system cannot be directly applied. b. The different herbal products were bought in markets and from sellers, whose botanical expertise is uncertain, hence, related species of various plants may have been employed. Different suppliers provided the pharmacy with herbs from all over theworld and most likely sold different plant species under the same name. c. As the plant materials were bought in the form of leaves, fruits, resins, etc., it was probably difficult for the pharmacists,

as knowledgeable as they were, to always perform a valid botanical identification. In some cases, one common name referred to several plant species whose differentiation is impossible today.

3.4. Formula 3

3.4.1. Balsamo di Gerusalemme Translation. Beaten Juniperus fruit 15 g, myrrh, aloe, gentiana, rabarbaro [Rheum spp.], agarico [Agaric spp. – a mushroom. Most common in medical use is the A. muscaria],

saffron. Of each [of these six herbs] 5 g, China Calissaja 20 g, alcohol 35% 1500 g, anise water 400 g, common water 5250 g, sugar 5250 g. Macerate the drug for 8 days in alcohol, add the sugar dissolved in water and filter. This formula is apparently for internal use. Although this is not explicit from the text, it is obvious from its ingredients – the sugar and the anise. It is probably a kind of syrup and

not a Balsam for topical use, unlike the previous formulas.

3.5. Formula 4

It is a variation of the original formula and is reported by Schittny (1992). It actually represents two different formulas. The first was probably brought to Germany from the monastery by a monk named Augustin Staude. According to Schittny, Staude gave the formula he brought from the St.

Savior convent to a pharmacist in his home town Glatz, who revised the formula in 1860 (Schittny, 1992). The second formulawas a revised version of the first in order to make it more suitable for internal use (Schittny, 1992).

3.6. Formula 5

This variation of the Jerusalem Balsam formula appears under the name “Compound Benzoin Tincture”, and has several synonyms. Two of these are “Friar’s Balsam” and “Jerusalem Balsam” (Ansel et al., 1995) (see above). It thus appears that variations of the Jerusalem Balsam are

still in use.

3.6.1. Pharmacological results We conducted several pharmaceutical model experiments with formula 1 (as described above). This formulawas chosen not only because it was described in an undoubtedly authentic document, but also because of its relative simplicity. It is a tincture (ethanol extract) of four herbal ingredients. The formula exhibits anti-inflammatory action (in in vitro and in

22 A. Moussaieff et al. / Journal of Ethnopharmacology 101 (2005) 16–26 Fig. 5. (a) and (b) The anti-inflammatory effect of the Jerusalem Balsam (JB) on ear thickness (a) and ear redness (b). Redness was scaled from 0 to 2. 0 Represents no redness compared to the non-inflamed ear and 2 represents a red ear. The scores are compared with those of indomethacin. p < 0.05, one-way Mann–Whithey.

vivo models), anti-oxidative action and anti-bacterial action (Figs. 5–8). To test the Balsam’s effectiveness onwound healing we used a surgical wound healing model on healthy rats and on diabetic rats, representing healing by first intention (Seror et al., 2003) and a thermal wound healing model on guinea pigs, representing healing by second intention (Eldad et al., 1998).We found no significant effect onwound healing, despite the Balsam’s wide use for this purpose. A. Moussaieff et al. / Journal of Ethnopharmacology 101 (2005) 16–26 23 Fig. 6. The antiseptic effect of the Jerusalem Balsam on Staphilococcus aureus as seen on paper discs put on agar plates. The disc treated with the Balsam has almost no colonies on it comparable to the disc treated with chlorhexidine, as opposed to the disc treated with ethanol alone. (1) 20 _l balsam, (2) 20 _l ethanol, (3) 20 _l chlorhexidine 0.2%.

Pic 5- (a)and (b) The anti-inflammatory effect of the Jerusalem Balsam (JB) on ear thickness (a) and ear redness (b). Redness was scaled from 0 to 2. 0

Represents no redness compared to the non-inflamed ear and 2 represents a red ear. The scores are compared with those of indomethacin. p < 0.05, one-way


Pic 6- The antiseptic effect of the Jerusalem Balsam on Staphilococcus aureus as seen on paper discs put on agar plates. The disc treated with the Balsam

has almost no colonies on it comparable to the disc treated with chlorhexidine, as opposed to the disc treated with ethanol alone. (1) 20 l balsam, (2) 20 l

ethanol, (3) 20 l chlorhexidine 0.2%.

3.7. In.amed ear model results One group of mice was administered the Balsam, after concentrating it down to 1/4 of its volume in an evaporator. It was administered at a dose of 8 _l (0.74 mg of dry

extract)/10 g mouse weight, in a vehicle consisting of 1:1:18 ethanol extract, Cremophor and saline, respectively.Asecond group served as control andwas administered the vehicle.We compared the swollen ear to the non-swollen ear in each of the mice and compared the differences between the ears in the different groups. We present the results of the inflamed ear model in Fig. 5a and b. We observed a significant inhibitory effect of the Balsam on arachidonic acid-induced swelling of the external mouse ear – both in thickness and in redness, indicating the Balsam’s anti-inflammatory properties.

3.8. Antiseptic results No bacterial colonies were formed on the agar beneath discs on which the Balsam was applied, as opposed to the agar beneath the control discs, where bacterial colonies grew

(Fig. 6). We assume that the Balsam solution has a local antiseptic activity. This was confirmed by glass slides treated with the Balsam as described above (Fig. 7). Biofilm growth on glass slides treated with the Balsam, demonstrate less bacterial colonies as opposed to the control slides (treated with

ethanol). The number of colonies on the Balsam-treated slides was similar to that in a positive control using chlorhexidine. This antiseptic action of the balsam, together with its anti-inflammatory properties apparently made it a valuable remedy for the healing of wounds, as well as for treating

infectious diseases.

3.9. Anti-oxidative effect

The Balsam exhibited a marked anti-oxidative effect in a RAW cell model in three experiments (Fig. 8). The antioxidative effect was shown to be dose dependent. By administrating 2% extract to the plates, we obtained an almost absolute inhibition of reactive oxygen species (ROS) formation, as opposed to the control (2% ethanol). Under the same conditions, addition of cannabidiol (CBD), a known anti-oxidative agent (Hampson et al., 1998; Malfait et al., 2000) and Following 24 h of incubation of

CBD with the cells, ROS generation was inhibited by a maximum of 84%, using a nontoxic amount of 8 _l of CBD. This result indicates a robust anti-oxidative effect of the Balsam. The wound healing models we used showed no difference between treated animals and non-treated animals (data not

shown). 24 A. Moussaieff et al. / Journal of Ethnopharmacology 101 (2005) 16–26

Fig. 7. Confocal microscope pictures of Streptococcus mutans on glass slides. (a) 100 _l chlorhexidine 0.2%, (b) 100 _l Balsam, (c) 100 _l ethanol. The slide treated with the Balsam is similar to the slide treated with chlorhexidine and shows almost no bacteria on it, as opposed to the control slide, treated with ethanol. Dead bacteria appear red and live ones appear green. Fig. 8. RAWmonoocyte cells were stimulated with Zymozan in presence of the Jerusalem Balsam. As control, we used stimulated cells in the presence of ethanol (vehicle). Production of ROSwas measured by luminometer. ROS productionwas inhibited up to 98.5%, as seen after 10 min. This is superior to

some compounds known to inhibitROS production, for example cannabidiol – a known anti-oxidative agent (Hampson et al., 1998; Malfait et al., 2000) that was tested under the same conditions. One representative experiment of three is shown.

Pic 7- Confocal microscope pictures of Streptococcus mutans on glass slides. (a) 100 l chlorhexidine 0.2%, (b) 100 l Balsam, (c) 100 l ethanol. The slide
treated with the Balsam is similar to the slide treated with chlorhexidine and shows almost no bacteria on it, as opposed to the control slide, treated with ethanol.
Dead bacteria appear red and live ones appear green

4. Discussion
The reports written at the time and the numerous variations of the Jerusalem Balsam provide evidence for its popularity and importance. However, we suggest that there are also other explanations for this variety. Thus, formulas 1 and 2 were presumably formulated within a short time period and both originate in the monastery.Yet, formula 2 consists of 40 ingredients,
while formula 1 consists of only four. It would not be far fetched to attribute this difference to a socio-economic consideration, namely that the four-ingredient formula was obviously cheaper to prepare and could be administered to indigent patients, while the more elaborate formulation was
for patients who could afford to pay for it. Another reason for the variety of formulas with the same designation could be the brand name “Jerusalem Balsam”. The formula had apparently
become successful in Europe and the Near East and the name of Jerusalem – a holy city to Christians, Moslems and Jews – presumably served as a marketing tool.

The Jerusalem Balsam was an important pharmaceutical formulation inWestern medicine for more than two centuries (Hasselquist, 1866; Bagatti, 1946; Horn, 1962). However, A. Moussaieff et al. / Journal of Ethnopharmacology 101 (2005) 16–26 25 time had an effect on the nature of the formula, both on its ingredients and its administration. During the 18th century, treating wounds was of utmost importance and the Balsam was extensively used for those purposes. However, by the
late 19th century and the beginning of the 20th century this was no longer the case. This could be attributed to the new knowledge of microbial infections of wounds and the use of
detergents and antiseptics, which partly satisfied the need for topical wound healing drugs, formerly filled by the Balsam. It is also possible that the Balsam was not found to be very effective. Its later use was presumably mostly internal for conditions such as stomach-aches. Why would such a famous remedy evolve in the old city of Jerusalem? The city was not a modern scientific center at the time (nor was there such a center in the whole of the Levant). The pharmacy of St. Savior’s monastery was one of the finest in Europe and the Near East for almost two hundred years (Hasselquist, 1866; Bagatti, 1946; Horn, 1962). It is unlikely that the immense cost, in funds and manpower, of supporting this kind of medical facility would have been spent without any incentive. We assume that the St. Savior’s pharmacy was seen as a foothold for the Catholic Church and Catholic countries, enabling them to strengthen their influence in Jerusalem. We conducted experiments on several different pharmacological models, using an extract of the four-constituent herbal formula (formula 1). It showed significant anti-inflammatory action on a mouse ear inflammation model. It also had marked
antiseptic and anti-oxidative effects. In view of the antiinflammatory and antiseptic activities of the Balsam, we are nowexamining these activities in each of the individual plants and are in the process of identifying active constituent(s).

In the models used by us, the Balsam did not showany significant effect onwound healing.We assume that the wide use of the Jerusalem Balsam in wound healing was (at least partially) due to its anti-inflammatory and antiseptic activities. Contamination is a major factor inhibitingwound healing and inflammation is an essential process during the first steps of wound healing, but can also impair proper wound healing if over-expressed (Cohen et al., 1994; Adzick, 1997; Cortan et al., 1999). In view of the recent work by Macias et al. (2004), which indicates wound-healing properties of Aloe gel, it is
possible that the Balsam does have healing properties (not observed in our models), using either Aloe vera juice or gel. It is also possible that Aloe vera L. is not the species used by the monks in the original Jerusalem Balsam formula (see above). The use of the Balsam as a panacea could be explained by its anti-inflammatory together with the anti-oxidative effect, and antiseptic effects. Altogether, it was likely to promote health in people suffering from various ailments.

5. Conclusion
To the best of our knowledge, this is the first time five different formulas of the Jerusalem Balsam are presented. This work demonstrates the importance of historical texts in bioprospecting medical herbs. This approach could be modified and improved by using new high-throughput ways
as recently described by Buenz et al. (2004). Medicinal plant mixtures, as used in the old pharmacy of St. Savior are common in almost all medical traditions. Chinese herbal medicine is a prime example of such a tradition. However, the problems posed by mixtures of herbs, each comprising a composite of
many compounds, deters many research ers in the field. Is the activity of the mixture due to a single chemical entity or to synergism of compounds from different plant species? Or is it an “entourage effect” (Ben-Shabat et al., 1998), in which non-active constituents enhance the activity of an active constituent? In some cases, however, it has been possible to identify active chemical entities from a mixture of herbal constituents. For a recent example, see Nobuhiro et al. (2004). We feel that this line of research should not be neglected, since herbal mixtures have been central in herbal medicine
in the past, and still are, in many parts of the world. The Jerusalem Balsam is a prime example of such a mixture. As mentioned above, the Jerusalem Balsam may be not only of pharmaceutical interest but may also represent an example of the use of medicine as a tool to widen Western cultural (and
hence political) sphere of influence in the Ottoman Empire  and particularly in Jerusalem.

The Jerusalem Balsam, and the medicinal plants from which it is produced, are still of pharmaceutical interest and might be a source of yet undiscovered compounds with potential medical use. Of the different herbs constituting the Balsam, two, namely Boswellia spp. and Commiphora spp.,
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