II. MICROSCOPIC STUDIES OF THE ANCIENT BlOOD OF LANCIANO

A minute Blood fragment has been included in paraffin according to Lenzi's technique.
At the Emallume-Eosine staining, no cellular element can be detected, with the exception of a yellow-dark-green granular material with foreign bodies of vegetable origin.

III. MICROCHEMICAL STUDIES ON BLOOD

Teichmann's reaction modified by Bertrand with hydrochlorate hematine and Takayama's reaction with hemo-chromogen, have been carried out on the ancient Blood in Lanciano with negative results, together with human Blood samples normally dried, which gave positive results.
Oxidase research (Stone and Burke's tests) gave highly positive results on the test sample and on control human normally dried Blood.

IV. THIN LAYER CHROMATOGRAPHY STUDY OF HAEMOGLOBIN FROM THE ANCIENT BLOOD

Haemoglobin and alkaline-haematic standards prepared according to Dacie's technique, were run on silica gel, in parallel with eluted liquid of the Blood in Lanciano, transformed also in alkaline-haematin according to the already mentioned technique, being this the best suited preparation for chromatographic goals.
The liquid used in the running reaction was methanol-acetic acid-water, in the ratio 90:3:7. After 90 minutes, the sample (ancient Blood in Lanciano) had migrated similarly to the standards thus showing the same Rf (0,88). [Fig.19]

The test was highly reproducible as shown by the results of triplicate experiments and of different running conditions.

CONCLUSIONS (II. III. IV.)

The negativity of Teichmann-Bertrand's test and Takayama's test does not exclude the presence of Blood, as these tests can become negative due to sample denaturation.
A positive oxidase test, generally indicating the presence of Blood, can occur also with organs contaminated by vegetable extracts or metal traces.
Paper thin layer chromatography (Franchini) is a validated test to identify the presence of Blood even in badly preserved samples which are not positive for haemoglobin.
This study confirms the real hematological nature of the ancient Blood in Lanciano.

V. IMMUNOLOGICAL DEFINITION OF THE SPECIES TO WHICH THE ANCIENT BLOOD AND FLESH IN LANCIANO DO BELONG

The elution fluid of Blood and Flesh in Lanciano has been employed in the zonal precipitation - reaction by Uhlenhuth, a recognized technique in modern laboratory methodology (Gradwohls).
Antihuman protein antiserum from Behringwerke has been used for testing samples and controls.[Fig.20]

The zonal precipitation reaction has given a positive result in 5 minutes in the test-tube n.1 (Blood) n.2 (Flesh) and n.3 (human serum), but it was negative in the test-tube n.4 (elution liquid of Blood and rabbit serum), n.5 (elution liquid of Flesh and rabbit serum) n.6 (ox serum and human antiprotein serum), n.7 (isotonic saline and antihuman protein serum).

CONCLUSION (V.)

The results of the zonal precipitation test according to Uhlenhuth, based on appropriate controls, confirm that the Eucharistic Miracle Blood and Flesh in Lanciano belong to the human species.

VI. DETERMINATION OF BLOOD GROUP IN THE ANCIENT BLOOD AND FLESH IN LANCIANO.

The absorption-elution reaction proposed by Siracusa and standardized by Fiori and coll. has been used to define the Blood group (ABO) in the elution fluid of the ancient Blood and Flesh in Lanciano.
The method has been widely accepted as fully valid to define the blood group in those cases which do not allow to perform classical tests, such as tissues, biological fluids, dried Blood spots.
It has been found that Blood and Flesh in Lanciano have both A and B agglutinogens; therefore, they belong to AB blood group. [Fig.21]

VII. ELECTROPHORETIC ANALYSIS OF THE PROTEINS OF THE ANCIENT BLOOD IN LANCIANO.

After dialysis in Colover's microcells against carbowax, the elution fluid of the ancient Blood, concentrated five times, gives an electrophoretic tracing in cellulose-acetate (Veronal buffer, ph 8.6; power: 0.5 ampère for 30 minutes, Ponceau S stain, Cromoscan photometer recording. [Fig.22]

The percentage composition of proteins in the liquid tested is similar to that known for human normal Blood serum (Henry).
The protein content is as follows: albumin 61.93 % (normal values 52-68%). Alfa 1 globulin 2.38% (normal values 2.4-5.3%), Alfa 2 globulin 7.14% (normal values 6,6-13,5%), Beta globulin in 7,14% (normal values 8.5-14.5%), Gamma globulin 21.42% (normal values 10.7-21%).
Globulin-albumin ratio is of 1.62 the normal value being 1.13-1.73.

VIII. DETERMINATION OF MINERAL CONTENT (CALCIUM, CHLORIDE, PHOSPHORUS, MAGNESIUM, POTASSIUM, SODIUM) IN THE ANCIENT BLOOD IN LANCIANO

Table 1 shows the levels of minerals (calcium, phosphorous, magnesium, sodium) found in dried samples of normal human Blood compared with those of the Ancient Blood in Lanciano.
The levels have been evaluated in comparison with those present in whole Blood (in: Tables Scientiphiques Geigy, VIth edition pag 582-590, 1960).

Table 1.
Mineral quantitative values (calcium, chloride, phosphorus, magnesium, potassium, sodium) in the Ancient Blood in Lanciano and in 10 human Blood normally dried samples.

                      Ca     Cl       P      Mg      K       Na
SAMPLE               mg%   mEq\1     mg%   mEq\1   mEq\1   mEq\1
----------------- ------ ------- ------- ------- ------- -------
Blood in Lanciano 114,29    4,25    1,99    0,96    5,76   46,44
Blood n.  1         4,42   31,8     8,42    1,57   12,80   55,04
Blood n.  2         3,96   30,75    8,37    1,52   12,16   48,16
Blood n.  3         3,73   31,50    8,75    1,15   14,08   48,16
Blood n.  4         4,66   43,20    8,10    1,39    5,12   116,9
Blood n.  5         3,73   37,70    9,72    1,39    4,80   99,76
Blood n.  6         3,26   31,76    8,42    1,54    3,84   79,12
Blood n.  7         4,89   35,2     9,07    1,82    4,00   65,36
Blood n.  8         3,96   36,00   10,00    1,66    4,16   68,80
Blood n.  9         3,82   34,40    9,55    1,34    4,00   79,12
Blood n. 10         3,35   32,80    9,47    1,64    3,84   73,96
----------------- ------ ------- ------- ------- ------- -------
Arithmetical average of 10 samples
                    3,97   34,51    8,98    1,50    6,88   73,43
----------------- ------ ------- ------- ------- ------- -------
Whole Blood
normal values       9-11  77-90,6  28-48   1,7-3,4   43     85
----------------- ------ ------- ------- ------- ------- -------

From Table 1 the following conclusions can be drawn:

a) During the slow drying process of 10 human normal whole Blood samples, in open glass tubes, kept at 50deg.C, at 37deg.C or at room temperature during three months, a loss in phosphorus, calcium, chloride, potassium, and, to a lesser extent, in magnesium and sodium can be recorded;

b) The Eucharistic Miracle Blood in Lanciano showed markedly decreased levels of chloride, phosphorus, magnesium, potassium and sodium as compared to human blood samples, normally dried. On the other hand, calcium levels were highly increased in the Blood from Lanciano (mg. 114, 29%).


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