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#include "Utils.h"
#include "Bank.h"
// some globals that don't really belong anywhere
int nks; // min abundance
uint32_t max_couv = 2147483646; // note: uint_abundance_t is 32 bits in SortingCount.cpp
struct timeval tim;
uint64_t nbkmers_solid = 0, b1_size = 0;
const char *solid_kmers_file = (char *)"solid_kmers_binary";
const char *false_positive_kmers_file = (char *)"false_positive_kmers";
const char *bloom_file = (char *)"bloom_data";
const char *assembly_file = (char *)"contigs.fa";
const char *branching_kmers_file = (char *)"branching_kmers"; // (only useful for multiple assemblies with same bloom&debloom structure (ie debugging))
const char *binary_read_file = (char *)"reads_binary";
const char *histo_file_name = (char *)"histo";
const char *breakpoints_file_name = (char *)"breakpoints";
const char *assoc_kmer_file = (char *)"paired_kmer";
// prefix-based output files naming
char prefix[1024];
char fileName[1024];
char *return_file_name(const char *suffix)
{
if (strlen(prefix)>0)
sprintf(fileName,"%s.%s",prefix,suffix);
else
sprintf(fileName,"%s",suffix);
return fileName;
}
int readlen;
#ifndef NO_BLOOM_UTILS
template <typename T, typename U>
// T can be Bloom, BloomCpt, BloomCpt3 or LinearCounter (just needs to support add(kmer_type) and possibly contains(kmer_type))
// U can be BloomCpt or BloomCpt3
void bloom_pass_reads(Bank *Sequences, T *bloom_to_insert, U *bloom_counter, char *stderr_message)
{
int64_t NbRead = 0;
int64_t NbInsertedKmers = 0;
Sequences->rewind_all();
char * rseq;
long i;
kmer_type kmer, graine, graine_revcomp;
while (Sequences->get_next_seq(&rseq,&readlen))
{
for (i=0; i<readlen-sizeKmer+1; i++)
{
kmer = extractKmerFromRead(rseq,i,&graine,&graine_revcomp);
if (bloom_counter != NULL)
{
// discard kmers which are not solid
if( ! bloom_counter->contains_n_occ(kmer,nks)) continue;
}
bloom_to_insert->add(kmer);
NbInsertedKmers++;
}
NbRead++;
if ((NbRead%10000)==0) fprintf (stderr,stderr_message,13,NbRead);
}
// fprintf (stderr,"\nInserted %lld %s kmers in the bloom structure.\n",(long long)NbInsertedKmers,"(redundant)");
}
template <typename T> // T can be Bloom, BloomCpt or BloomCpt3
void bloom_pass_reads_binary(T *bloom_to_insert, BloomCpt *bloom_counter, char *stderr_message)
{
//fprintf(stderr,"binary pass \n");
int64_t NbRead = 0;
int64_t NbInsertedKmers = 0;
kmer_type kmer;
// read solid kmers from disk
BinaryBank * SolidKmers = new BinaryBank(return_file_name(solid_kmers_file),sizeof(kmer),0);
while(SolidKmers->read_element(&kmer))
{
// printf("kmer %lld\n",kmer);
bloom_to_insert->add(kmer);
NbInsertedKmers++;
NbRead++;
if ((NbRead%10000)==0) fprintf (stderr,stderr_message,13,(long long)NbRead);
}
// fprintf (stderr,"\nInserted %lld %s kmers in the bloom structure.\n",(long long)NbInsertedKmers,"solid");
SolidKmers->close();
}
int estimated_BL1;
uint64_t estimated_BL1_freesize;
float NBITS_PER_KMER = 11 ; // number of bits per kmer that optimizes bloo1 size
// loading bloom from disk
template <typename T> //bloocpt or bloocpt3
Bloom *bloom_create_bloo1(T *bloom_counter, bool from_dump)
{
BinaryBank * SolidKmers ;
if(from_dump && nsolids) // from dump and known number of solid kmers
{
//nsolids is sotred in a config file
//number of solid kmers cannot be computed precisely from bloom file, imprecision of 0-7
estimated_BL1 = max( (int)ceilf(log2f(nsolids*NBITS_PER_KMER)), 1);
estimated_BL1_freesize = (uint64_t)(nsolids*NBITS_PER_KMER);
b1_size = (uint64_t) estimated_BL1_freesize; nbkmers_solid = nsolids ; // for correct printf in print_size_summary
printf("load bloom from dump, containing %lli solid kmers b1_size %lli\n",nsolids,b1_size);
}
else
{
// get true number of solid kmers, in order to precisely size the bloom filter
SolidKmers = new BinaryBank(return_file_name(solid_kmers_file),sizeof(kmer_type),0);
estimated_BL1 = max( (int)ceilf(log2f(SolidKmers->nb_elements()*NBITS_PER_KMER)), 1);
estimated_BL1_freesize = (uint64_t)(SolidKmers->nb_elements()*NBITS_PER_KMER);
//printf("nelem %lli nbits %g \n",(long long)(SolidKmers->nb_elements()),NBITS_PER_KMER);
}
//printf("Allocating %0.1f MB of memory for the main Bloom structure (%g bits/kmer)\n",(1LL<<estimated_BL1)/1024.0/1024.0/8.0,NBITS_PER_KMER);
if(estimated_BL1_freesize ==0) estimated_BL1_freesize =10;
//printf("freesize %lli estimated_BL1_freesize %0.1f MB of memory for the main Bloom structure (%g bits/kmer)\n",(long long)estimated_BL1_freesize,(estimated_BL1_freesize)/1024.0/1024.0/8.0,NBITS_PER_KMER);
Bloom *bloo1;
#if CUSTOMSIZE
bloo1 = new Bloom((uint64_t)estimated_BL1_freesize);
#else
bloo1 = new Bloom(estimated_BL1);
#endif
bloo1->set_number_of_hash_func((int)floorf(0.7*NBITS_PER_KMER));
if (from_dump)
bloo1->load(return_file_name(bloom_file)); // just load the dump
else
{
bloom_pass_reads_binary(bloo1, bloom_counter, (char*)"%cInsert solid Kmers in Bloom %lld"); // use the method reading SolidKmers binary file, was useful when varying Bloom size (!= dumped size)
//bloo1->dump(return_file_name(bloom_file)); // create bloom dump
SolidKmers->close();
}
return bloo1;
}
// wrapper for default behavior: don't load from dump
template <typename T> //bloocpt or bloocpt3
Bloom *bloom_create_bloo1(T *bloom_counter)
{
return bloom_create_bloo1(bloom_counter, false);
}
template Bloom *bloom_create_bloo1<BloomCpt>(BloomCpt *bloom_counter); // trick to avoid linker errors: http://www.parashift.com/c++-faq-lite/templates.html#faq-35.13
template Bloom *bloom_create_bloo1<BloomCpt>(BloomCpt *bloom_counter, bool from_dump);
// --------------------------------------------------------------------------------
// below this line: unused kmer counting code
FILE * F_kmercpt_read;
FILE * F_kmercpt_write;
//in last partition : create solid kmers file, and load solid kmers in bloo1 bloom
void end_kmer_count_partition(bool last_partition, Hash16 *hasht1)
{
int value;
int cptk=0;
int64_t nso=0;
/////////////////////////begin write files
rewind (F_kmercpt_read);
rewind (F_kmercpt_write);
#ifndef MINGW
ftruncate(fileno(F_kmercpt_write), 0); //erase previous file
#else // tempfix? fileno is not accepted by mingw
fclose(F_kmercpt_write);
F_kmercpt_write = fopen("kmer_count2","wb+");
#endif
BinaryBank * SolidKmers = NULL;
kmer_type graine;
if (last_partition)
SolidKmers = new BinaryBank(return_file_name(solid_kmers_file),sizeof(kmer_type),1);
while(fread(&graine, sizeof(graine),1, F_kmercpt_read)){
fread(&cptk, sizeof(cptk), 1, F_kmercpt_read);
hasht1->remove(graine,&value); // if graine is present, get value of graine and remove graine, else value=0
cptk += value;
fwrite(&graine, sizeof(graine), 1, F_kmercpt_write);
fwrite(&cptk, sizeof(cptk), 1, F_kmercpt_write);
if (last_partition && cptk >= nks)
// if last partition, also need to search for solid kmers in remaining of hasht1, so this is not enough:
{
SolidKmers->write_element(&graine);
nso++;
}
}
hasht1->dump(F_kmercpt_write); // dump remaining of hasht1
if (last_partition)
{
nso+=hasht1->getsolids(NULL,SolidKmers,nks); // get remaining solids of hasht1
fprintf(stderr,"nsolid kmers = %lli \n",(long long)nso);
SolidKmers->close();
#ifndef MINGW
ftruncate(fileno(F_kmercpt_read), 0); //erase previous file
#else // tempfix? fileno is not accepted by mingw
fclose(F_kmercpt_read);
F_kmercpt_read = fopen("kmer_count2","wb+");
#endif
}
}
template <typename T>
void exact_kmer_count(Bank *Sequences, T *bloom_counter, unsigned long max_memory)
{
FILE * count_file = fopen("kmer_count","wb+");
FILE * count_file_2 = fopen("kmer_count2","wb+");
FILE * F_tmp;
F_kmercpt_read = count_file ;
F_kmercpt_write = count_file_2;
Sequences->rewind_all();
unsigned int max_kmer_per_part = max_memory*1024LL*1024LL /sizeof(cell<kmer_type>);
int numpart = 0;
char * rseq;
long i;
int64_t NbRead = 0;
int64_t NbInserted = 0;
int64_t NbInserted_unique = 0;
kmer_type kmer, graine, graine_revcomp;
// that code makes hasht1 occupy full memory. should probably be reduced (but we're deprecating that procedure, right?)
int NBITS_HT = max( (int)ceilf(log2f((max_memory*1024L*1024L)/sizeof(cell_ptr_t))), 1);
Hash16 *hasht1 =new Hash16(NBITS_HT);
// partitioned exact kmer counting based on Bloom filter for solidity:
// the bloom filter enables membership test for a set S of supposedly solid kmers (contains false positives)
// read the (redundant) kmers from the reads, and load only those in S, in chunks, into a hash table
// at each pass, update a file containing the true count of non-redundant supposedly solid kmers (S)
// at the end, analyze the file to keep only those with true count >= solid
while (Sequences->get_next_seq(&rseq,&readlen))
{
for (i=0; i<readlen-sizeKmer+1; i++)
{
kmer = extractKmerFromRead(rseq,i,&graine,&graine_revcomp);
// discard kmers which are not solid
if( ! bloom_counter->contains_n_occ(kmer,nks)) continue;
//insert into hasht1
NbInserted_unique += hasht1->add(kmer);
NbInserted++;
if(hasht1->nb_elem >max_kmer_per_part) //end partition
{
fprintf(stderr,"End of Kmer count partition %lli / %i \n",(long long)(hasht1->nb_elem),max_kmer_per_part);
if(numpart==0)
hasht1->dump(F_kmercpt_write);
else
end_kmer_count_partition(false,hasht1);
//swap file pointers
F_tmp = F_kmercpt_read;
F_kmercpt_read = F_kmercpt_write;
F_kmercpt_write = F_tmp;
/////////end write files
//reset hash table
hasht1->empty_all();
numpart++;
} ///end partition
}
NbRead++;
if ((NbRead%10000)==0) fprintf (stderr,"%cLoop through reads for exact kmer count %lld",13,(long long)NbRead);
}
fprintf (stderr," \nTotal Inserted in hash (ie output of Bloom) unique %lli / %lli redundants \n",(long long)NbInserted_unique,(long long)NbInserted);
///////////////////////// last partition
end_kmer_count_partition(true,hasht1);
delete hasht1;
}
void bloom_count(Bank *Reads, unsigned long max_memory)
{
#define NBITS_BLOOMCPT 23 // 33 :4GB (4 bits/elem) // size of the bloom counter table to count kmers
fprintf(stderr,"nbits bloom counter: %i \n",NBITS_BLOOMCPT);
BloomCpt3 * bloocpt = new BloomCpt3(NBITS_BLOOMCPT);
BloomCpt3 * bloocpt2 = new BloomCpt3(NBITS_BLOOMCPT);
bloocpt->setSeed( 0x4909FEA3A68CC6A7LL);
bloocpt2->setSeed( 0x0CD5DA28467C5492LL);
// bloocpt->set_number_of_hash_func(4);
// bloocpt2->set_number_of_hash_func(6);
///////////////////////////////////first pass ; count kmers with Bloom cpt
bloom_pass_reads(Reads,bloocpt, (BloomCpt * ) NULL, (char*)"%cFirst pass %lld");
fprintf (stderr,"\n ____________ Second bloom counter _________\n");
bloom_pass_reads(Reads,bloocpt2, bloocpt, (char*)"%cSecond pass %lld");
STARTWALL(count);
fprintf(stderr,"\n------------------ second pass bloom counter \n\n");
delete bloocpt;
////////////////////////////////////// exact kmer count with hash table partitionning,
//also create solid kmers file and fills bloo1
exact_kmer_count(Reads,bloocpt2,max_memory);
//////////////////////////////////////
STOPWALL(count,"Counted kmers");
fprintf(stderr,"\n------------------ Counted kmers and kept those with abundance >=%i \n\n",nks);
////////////////////////////////////////////////////fin bloom insert
//delete bloocpt2;
}
void estimate_distinct_kmers(unsigned long genome_size, Bank *Reads)
{
int size_linearCounter = genome_size * 8; // alloc 8 bits * genome size for counting, i.e. ~ as much as the assembly Bloom size
LinearCounter *linearCounter = new LinearCounter(size_linearCounter);
bloom_pass_reads(Reads,linearCounter, (BloomCpt * ) NULL, (char*)"%cEstimating number of distinct kmers (%lld reads processed so far)");
long nb_distinct_kmers = linearCounter->count();
if (linearCounter->is_accurate())
printf("Estimated that %ld distinct kmers are in the reads\n",nb_distinct_kmers);
else
printf("Cannot estimate the number of distinct kmers. Allocate a larger counter\n");
delete linearCounter;
}
uint64_t extrapolate_distinct_kmers(Bank *Reads)
{
// start with 100MB RAM estimation and grow if necessary
return extrapolate_distinct_kmers_wrapped(100000000L, Reads);
}
uint64_t extrapolate_distinct_kmers_wrapped(unsigned long nbytes_memory, Bank *Reads)
{
unsigned long size_linearCounter = nbytes_memory * 8L; // alloc 8 bits * nbytes for counting
LinearCounter *linearCounter = new LinearCounter(size_linearCounter);
int stops = 100000;
// variant of bloom_pass_reads
int64_t NbRead = 0;
int64_t NbInsertedKmers = 0;
Reads->rewind_all();
char * rseq;
long i;
kmer_type kmer, graine, graine_revcomp;
long nb_distinct_kmers = 0;
long previous_nb_distinct_kmers = 0;
uint64_t estimated_nb_reads = Reads->estimate_nb_reads();
bool stop = false;
while (Reads->get_next_seq(&rseq,&readlen))
{
if (stop)
break;
for (i=0; i<readlen-sizeKmer+1; i++)
{
kmer = extractKmerFromRead(rseq,i,&graine,&graine_revcomp);
linearCounter->add(kmer);
NbInsertedKmers++;
if (NbInsertedKmers % stops == 0 && NbRead != 0)
{
previous_nb_distinct_kmers = nb_distinct_kmers;
nb_distinct_kmers = linearCounter->count()*estimated_nb_reads/NbRead;
//printf("estimated now: %ld\n",nb_distinct_kmers);
// the following condition will grossly over-estimate the number of distinct kmers
// I expect the correct result to be in the same order of magnitude
if (abs((int)(nb_distinct_kmers-previous_nb_distinct_kmers)) < previous_nb_distinct_kmers/20) // 5% error
stop = true;
if (!linearCounter->is_accurate())
stop = true;
}
}
NbRead++;
if ((NbRead%10000)==0) fprintf (stderr,(char*)"%cExtrapolating number of distinct kmers %lld",13,NbRead);
}
if (!linearCounter->is_accurate())
{
printf("Inaccurate estimation, restarting with %lu MB RAM\n",(2*nbytes_memory)/1024/1024);
delete linearCounter;
return extrapolate_distinct_kmers_wrapped(2*nbytes_memory, Reads);
}
nb_distinct_kmers = linearCounter->count()*estimated_nb_reads/NbRead; // this is a very rough estimation
printf("Linear estimation: ~%ld M distinct %d-mers are in the reads\n",nb_distinct_kmers/1000000L,sizeKmer);
delete linearCounter;
return nb_distinct_kmers;
}
#endif
float needleman_wunch_mtg(string a, string b, int * nbmatch,int * nbmis,int * nbgaps, int deb )
{
float gap_score = -5;
float mismatch_score = -5;
float match_score = 10;
#define nw_score(x,y) ( (x == y) ? match_score : mismatch_score )
int n_a = a.length(), n_b = b.length();
float ** score = (float **) malloc (sizeof(float*) * (n_a+1));
for (int ii=0; ii<(n_a+1); ii++)
{
score [ii] = (float *) malloc (sizeof(float) * (n_b+1));
}
// float score[n_a+1][n_b+1]; //stack is too small
// float pointer[n_a+1][n_b+1];
for (int i = 0; i <= n_a; i++)
score[i][0] = gap_score * i;
for (int j = 0; j <= n_b; j++)
score[0][j] = gap_score * j;
// compute dp
for (int i = 1; i <= n_a; i++)
{
for (int j = 1; j <= n_b; j++)
{
float match = score[i - 1][j - 1] + nw_score(a[i-1],b[j-1]);
float del = score[i - 1][j] + gap_score;
float insert = score[i][j - 1] + gap_score;
score[i][j] = max( max(match, del), insert);
}
}
// traceback
int i=n_a, j=n_b;
float identity = 0;
int nb_mis = 0;
int nb_gaps = 0;
int nb_end_gaps = 0 ;
bool end_gap = true;
while (i > 0 && j > 0)
{
float score_current = score[i][j], score_diagonal = score[i-1][j-1], score_up = score[i][j-1], score_left = score[i-1][j];
if (score_current == score_diagonal + nw_score(a[i-1], b[j-1]))
{
//if(deb==1)printf("eat %i %i \n",i,j);
if (a[i-1]== b[j-1])
{
// if(deb==1)printf("match %c %c \n",a[i-1],b[j-1]);
identity++;
}
else
{
// if(deb==1)printf("mis %c %c \n",a[i-1],b[j-1]);
nb_mis++;
}
i -= 1;
j -= 1;
end_gap = false;
}
else
{
if (score_current == score_left + gap_score)
{
//if(deb==1)printf("add gap s1 at ij %i %i \n",i,j);
i -= 1;
}
else if (score_current == score_up + gap_score)
{
// if(deb==1)printf("add gap s2 at ij %i %i \n",i,j);
j -= 1;
}
if(!end_gap) //pour ne pas compter gap terminal
{
nb_gaps++;
}
}
}
//pour compter gaps au debut :
nb_gaps += i+j;
//if(deb==1)printf("add gaps i j %i %i \n",i,j);
identity /= max( n_a, n_b); // modif GR 27/09/2013 max of two sizes, otherwise free gaps
if(nbmatch!=NULL) *nbmatch = identity;
if(nbmis!=NULL) *nbmis = nb_mis;
if(nbgaps!=NULL) *nbgaps = nb_gaps;
for (int ii=0; ii<(n_a+1); ii++)
{
free (score [ii]);
}
free(score);
//printf("---nw----\n%s\n%s -> %.2f\n--------\n",a.c_str(),b.c_str(),identity);
return identity;
}
float needleman_wunch(string a, string b )
{
float gap_score = -5;
float mismatch_score = -5;
float match_score = 10;
#define nw_score(x,y) ( (x == y) ? match_score : mismatch_score )
int n_a = a.length(), n_b = b.length();
float ** score = (float **) malloc (sizeof(float*) * (n_a+1));
for (int ii=0; ii<(n_a+1); ii++)
{
score [ii] = (float *) malloc (sizeof(float) * (n_b+1));
}
// float score[n_a+1][n_b+1]; //stack is too small
// float pointer[n_a+1][n_b+1];
for (int i = 0; i <= n_a; i++)
score[i][0] = gap_score * i;
for (int j = 0; j <= n_b; j++)
score[0][j] = gap_score * j;
// compute dp
for (int i = 1; i <= n_a; i++)
{
for (int j = 1; j <= n_b; j++)
{
float match = score[i - 1][j - 1] + nw_score(a[i-1],b[j-1]);
float del = score[i - 1][j] + gap_score;
float insert = score[i][j - 1] + gap_score;
score[i][j] = max( max(match, del), insert);
}
}
// traceback
int i=n_a, j=n_b;
float identity = 0;
int nb_mis = 0;
int nb_gaps = 0;
int nb_end_gaps = 0 ;
bool end_gap = true;
while (i > 0 && j > 0)
{
float score_current = score[i][j], score_diagonal = score[i-1][j-1], score_up = score[i][j-1], score_left = score[i-1][j];
if (score_current == score_diagonal + nw_score(a[i-1], b[j-1]))
{
if (a[i-1]== b[j-1])
identity++;
else
nb_mis++;
i -= 1;
j -= 1;
end_gap = false;
}
else
{
if (score_current == score_left + gap_score)
i -= 1;
else if (score_current == score_up + gap_score)
j -= 1;
}
}
identity /= max( n_a, n_b); // modif GR 27/09/2013 max of two sizes, otherwise free gaps
for (int ii=0; ii<(n_a+1); ii++)
{
free (score [ii]);
}
free(score);
//printf("---nw----\n%s\n%s -> %.2f\n--------\n",a.c_str(),b.c_str(),identity);
return identity;
}
void Progress::init(uint64_t ntasks, const char * msg)
{
gettimeofday(×tamp, NULL);
heure_debut = timestamp.tv_sec +(timestamp.tv_usec/1000000.0);
fprintf(stderr,"| %-*s |\n",98,msg);
todo= ntasks;
done = 0;
partial =0;
for (int ii=0; ii<16;ii++) partial_threaded[ii]=0;
for (int ii=0; ii<16;ii++) done_threaded[ii]=0;
subdiv= 100;
steps = (double)todo / (double)subdiv;
if(!timer_mode)
{
fprintf(stderr,"[");fflush(stderr);
}
}
void Progress::finish()
{
set(todo);
if(timer_mode)
fprintf(stderr,"\n");
else
fprintf(stderr,"]\n");
fflush(stderr);
todo= 0;
done = 0;
partial =0;
}
void Progress::finish_threaded()// called by only one of the threads
{
done = 0;
double rem = 0;
for (int ii=0; ii<16;ii++) done += (done_threaded[ii] );
for (int ii=0; ii<16;ii++) partial += (partial_threaded[ii] );
finish();
}
void Progress::inc(uint64_t ntasks_done, int tid)
{
partial_threaded[tid] += ntasks_done;
done_threaded[tid] += ntasks_done;
while(partial_threaded[tid] >= steps)
{
if(timer_mode)
{
struct timeval timet;
double now;
gettimeofday(&timet, NULL);
now = timet.tv_sec +(timet.tv_usec/1000000.0);
uint64_t total_done = 0;
for (int ii=0; ii<16;ii++) total_done += (done_threaded[ii] );
double elapsed = now - heure_debut;
double speed = total_done / elapsed;
double rem = (todo-total_done) / speed;
if(total_done > todo) rem =0;
int min_e = (int)(elapsed / 60) ;
elapsed -= min_e*60;
int min_r = (int)(rem / 60) ;
rem -= min_r*60;
fprintf(stderr,"%c%-5.3g %% elapsed: %6i min %-4.0f sec estimated remaining: %6i min %-4.0f sec ",13,100*(double)total_done/todo,min_e,elapsed,min_r,rem);
}
else
{
fprintf(stderr,"-");fflush(stderr);
}
partial_threaded[tid] -= steps;
}
}
void Progress::inc(uint64_t ntasks_done)
{
done += ntasks_done;
partial += ntasks_done;
while(partial >= steps)
{
if(timer_mode)
{
gettimeofday(×tamp, NULL);
heure_actuelle = timestamp.tv_sec +(timestamp.tv_usec/1000000.0);
double elapsed = heure_actuelle - heure_debut;
double speed = done / elapsed;
double rem = (todo-done) / speed;
if(done>todo) rem=0;
int min_e = (int)(elapsed / 60) ;
elapsed -= min_e*60;
int min_r = (int)(rem / 60) ;
rem -= min_r*60;
fprintf(stderr,"%c%-5.3g %% elapsed: %6i min %-4.0f sec estimated remaining: %6i min %-4.0f sec ",13,100*(double)done/todo,min_e,elapsed,min_r,rem);
}
else
{
fprintf(stderr,"-");fflush(stderr);
}
partial -= steps;
}
}
void Progress::set(uint64_t ntasks_done)
{
if(ntasks_done > done)
inc(ntasks_done-done);
}
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